Planta (2016) 243:847–887

DOI 10.1007/s00425-015-2452-8

REVIEW

Sandalwood: basic biology, tissue culture, and genetic

transformation
Jaime A. Teixeira da Silva1 • Mafatlal M. Kher2 • Deepak Soner2 •

Tony Page3 • Xinhua Zhang4 • M. Nataraj2 • Guohua Ma4

Received: 27 July 2015 / Accepted: 16 December 2015 / Published online: 8 January 2016
Ó Springer-Verlag Berlin Heidelberg 2016

Abstract mechanisms underlying the success or failure of traditional

Main conclusion Sustainable resource preservation of propagation, including a synopsis of the process of hemi-
Santalum species that yield commercially important parasitism in S. album, and of the suitability of host plants
forest products is needed. This review provides an to sustain the growth of seedlings and plants under forestry
understanding of their basic biology, propagation, production. For the mass production of economically
hemi-parasitic nature, reproductive biology, and important metabolites, and to improve uniformity of
biotechnology. essential oils, the use of clonal material of similar genetic
background for cultivation is important. This review
Many species of the genus Santalum (Santalaceae) have
summarizes traditional methods of sandalwood production
been exploited unremittingly for centuries, resulting in the
with complementary and more advanced in vitro tech-
extinction of one and the threatened status of three other
nologies to provide a basis for researchers, conservationists
species. This reduction in biodiversity of sandalwood has
and industry to implement sustainable programs of
resulted from the commercial exploitation of its oil-rich
research and development for this revered genus.
fragrant heartwood. In a bid to conserve the remaining
germplasm, biotechnology provides a feasible, and effec-
Keywords In vitro  Micropropagation  Santalum
tive, means of propagating members of this genus. This
album  Santalum spicatum  Somatic embryogenesis 
review provides a detailed understanding of the biological
Tissue culture

Electronic supplementary material The online version of this

article (doi:10.1007/s00425-015-2452-8) contains supplementary
material, which is available to authorized users.

& Jaime A. Teixeira da Silva 1

P. O. Box 7, Miki-cho Post Office, 3011-2, Ikenobe,
[email protected] Kagawa-ken 761-0799, Japan
& Mafatlal M. Kher 2
B.R. Doshi School of Biosciences, Sardar Patel University,
[email protected] Sardar Patel Maidan, Vadtal Rd., P.O. Box 39,
Vallabh Vidyanagar, Gujarat 388120, India
& Deepak Soner
3
[email protected] Forests and People Research Centre, University of Sunshine
Coast, Maroochydore DC, Locked Bag 4, Sunshine Coast,
& Tony Page
QLD 4558, Australia
[email protected]
4
Key Laboratory of Plant Resources Conservation and
& Xinhua Zhang
Sustainable Utilization, South China Botanical Garden, the
[email protected]
Chinese Academy of Sciences, Guangzhou 510650, China
& M. Nataraj
[email protected]
& Guohua Ma
[email protected]

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848 Planta (2016) 243:847–887

The historical, cultural, medicinal, and economic reviews (Rai and Sarma 1990; Ganeshaiah et al. 2007;
importance of sandalwood as a basis Arun Kumar et al. 2012; Rashkow 2014). Indian sandal-
for conservation wood was extensively exploited in the Pacific throughout
the first half of the 19th century although initial evidence of
Sandalwood trees of the genus Santalum belong to the sandalwood trade originated much earlier, with the begin-
Santalaceae. This family is composed of 29 genera with ning of Buddhism into China from India (Ritter 1836;
approximately 400 species, 19 of which are specific to the Thomson et al. 2005a). This occurred in the first century
Santalum genus (Fox 2000; Harbaugh 2007; Harbaugh and AD typified by smoldering sandalwood incense in temples.
Baldwin 2007; Harbaugh et al. 2010; Butaud 2015; Trade then extended to the Pacific when Americans and
Table 1). Encyclopedia Britannica online (2013) lists 36 Australians began to trade with China, leading to the dis-
genera. Harbaugh and Baldwin (2007) placed those num- covery of sandalwood in the Pacific, including Hawaii, and
bers as ‘‘15 extant species, approximately 14 varieties, and Australia (Thomson et al. 2005a). Commercial exploitation
one recently extinct species, distributed throughout India, of sandalwood has, however, resulted in the acute degra-
Australia, and the Pacific Islands.’’ Nageswara Rao et al. dation of natural populations of many species, including
(2011) listed 16 species and several varieties. According to those in India (Rashkow 2014), Indonesia (Ora 2012),
The Plant List (2015), currently only 12 species names are Papua New Guinea (Gunn et al. 2002), and Vanuatu
accepted, while 41 remain unresolved (Supplementary (Gillieson et al. 2008).
Table 1). These discrepancies suggest that some attention Heartwood does not exist in young trees of S. album and
to the taxonomy of the Santalum genus is required. only mature trees (30–50 years old) produce the heartwood
The economically most prominent species include rich in fragrant essential oil (Burdock and Carabin 2008;
Indian sandalwood or East Indian sandalwood (Santalum Zhang et al. 2012b). The concentration of essential oil
album L.), and Australian sandalwood [S. spicatum (R.Br.) within the heartwood of mature sandalwood varies between
A.DC.]. Indian sandalwood has various levels of impor- trees, ranging from 0.5 to 5 % in S. album (Sindhu
tance in cosmetic, perfumery, and aromatherapy industries, Veerendra and Anantha Padmanabha 1996), 0.05–8 % in S.
in religion, as well as in traditional medicine (reviewed in austrocaledonicum (Page et al. 2010b), and 0.1–8.2 % in S.
Dhanya et al. 2010; Arun Kumar et al. 2012; Heena Kausar lanceolatum (Page et al. 2007). Phytochemical analyses of
et al. 2014). Indian sandalwood was used for carving the heartwood of several sandalwood species (S. album, S.
wooden idols, the manufacture of richly carved boxes, spicatum, S. austrocaledonicum, and S. insulare) reveal
work tables, and cabinets (Chada 1972), and for burning in that more than 230 compounds, mainly terpenoids, have
certain Hindu and Buddhist rituals or to carve deities and been identified so far (reviewed by Baldovini et al. 2011).
temples (Kushalapa 1998). The wood paste was also used The Flavor and Extract Manufacturers’ Association, the
as an ointment to dissipate heat (Ral 1990). Sandalwood United States Food and Drug Administration, as well as the
essential oil, which has a rich tradition of uses spanning Council of Europe have approved sandalwood essential oil
more than 4000 years as mentioned in Sanskrit texts, is an for use in food-based products (Burdock and Carabin
important ingredient of cosmetic produces, herbal medi- 2008). The essential oils of sandalwood, including biore-
cine, and perfumes (Ritter 1836; Burdock and Carabin actors, will be reviewed separately (Teixeira da Silva et al.
2008). The ancient Egyptians imported the wood and used unpublished review).
it in medicine, embalming and ritual burning to venerate Excessive exploitation of natural stands and the lack of
the gods (Arun Kumar et al. 2012). Historically, sandal- initiatives to establish, until fairly recently, artificial stands
wood has a rich tradition of trade with the East, dating as has led to a decrease in natural stocks and thus an increase
far back as the 5th century BC (Edwards 1951) when its in market prices (Gillieson et al. 2008; Arun Kumar et al.
aromatic heartwood and oils were already recognized as 2012; Subasinghe 2013). Anantha Padmanabha (2000,
prized commodities. Sandalwood trade in India was started 2014) described a decline in sandalwood production in
as early as the 13th century by Indian rulers trying to India over several decades: 4000 t (1950) ? 2000
monopolize Indian sandalwood resources to ensure eco- t (1990) ? *1000 t (1999). More recent figures regarding
nomic strength for power and warfare, the classic case sandalwood supply in India are difficult to determine, since
being the mighty Vijaya Nagara Empire (13–16 century much of the traded wood comes from illegally harvested
AC) of the Deccan region (Ganeshaiah et al. 2007). Real- sources, and estimates suggest around 1000 tonnes of
izing the value of sandalwood, Tippu Sultan, the King of legally traded wood (AAG 2006; McKinnell 2011). In
Mysore (India), declared the sandalwood tree as a royal Indonesia, the sandalwood trade in East Nusa Tenggara
tree in 1772 (Buchanan 1884; Adkoli 1977). More histor- Province decreased rapidly during the 1990s contributing
ical details on Indian sandalwood are available in other almost 50 % of total regional revenue for the early 1990s to

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Planta (2016) 243:847–887 849

Table 1 Santalum species and distribution: modified from Harbaugh and Baldwin (2007), Harbaugh et al. (2010) and Butaud (2015)
Species number Species Variety Distribution

1 S. album L Australia, Indonesia, India

2 S. austrocaledonicum Viell. austrocaledonicum New Caledonia, Vanuatu
minutum N.Hallé New Caledonia
pilosulum N.Halle New Caledonia
glabrum Hürl New Caledonia
3 S. boninense (Nakai) Tuyama Bonin Islands
4 S. lanceolatum R.Br. Australia
5 S. macgregorii F.Muell. Papua New Guinea
6 S. obtusifolium R.Br. Australia
7 S. yasi Seem. Fiji, Tonga
8 S. freycinetianum Gaudich. freycinetianum Hawaiian Islands (O‘ahu, Moloka‘i)
lanaiense Rock Hawaiian Islands (Lana‘i, Maui)
pyrularium (A.Gray) Stemmerm. Hawaiian Islands (Kaua‘i)
9 S. haleakalae Hillebr Hawaiian Islands (Maui)
10 S. ellipticum Gaudich. ellipticum Hawaiian Islands
littorale (Rock) Skottsb Hawaiian Islands (O‘ahu)
11 S. paniculatum Hook. & Arn. paniculatum Hawaiian Islands (Hawai‘i)
pilgeri (Rock) Stemmerm Hawaiian Islands (Hawai‘i)
12 S. fernandezianum F.Phil. Juan Fernandez Islandsa
13 S. insulare Bertero ex A.DC. insulare Society Islands (Tahiti)
alticola Fosberg & Sachet Society Islands (Tahiti)
deckeri Fosberg & Sachet Marquesas Islands
hendersonense (F.Br.) Fosberg & Sachet Pitcairn Islands
marchionense (Skottsb.) Skottsb. Marquesas Islands
margaretae (F.Br.) Skottsb. Austral Islands (Rapa)
mitiaro Sykes Cook Islands (Mitiaro)
raiateense (J.Moore) Fosberg & Sachet Society Islands (Raiatea, Mo‘orea)
raivavense F.Br. Austral Islands (Raivavae)
14 S. acuminatum (R.Br.) A.DC. Australia
15 S. murrayanum (T.Mitch.) C.A.Gardner Australia
16 S. spicatum (R.Br.) A.DC. Australia
17 S. leptocladum Gand. Australia
18 S. involutum H. St. John Hawaiian Islands (Kaua‘i)
a
Extinct species

13 % in the late 1990s, and total production continued to using Traditionaloven.com (2015)]; (AAG 2006; McKin-
decline from 7465 tonnes in the 9 years 1987–1997 to just nell 2011). The average auction price for the heartwood of
2178 tonnes in the 6 years 2001–2007 (Ora 2012). wild Indian sandalwood rose from $9,400/tonne in 1990
According to the Tropical Forestry Services, essential (Ral 1990) to approximately $150,000/tonne in July 2014
oil is extracted from the heartwood of Indian sandalwood (on small volumes based on an auction held in Tamilnadu,
by distillation and trades for over $5000/kg on the inter- India), which indicates a significant annual compounded
national market (TFS 2015a). The annual supply of wild- growth rate (TFS 2015b). A 12 % annual increase in the
harvested Australian sandalwood species approaches 2000 minimum price paid to landowners for S. austrocale-
tonnes of S. spicatum (McKinnell 2011), and while a donicum in Vanuatu has also been recorded, from an
500 m3 license exists for S. lanceolatum (DPI&F 2004; equivalent of US$4/kg in 2000 (Mele 2001; Berry 2005) to
Timber-Queensland 2012), others suggests that actual US$20/kg in 2015 (Tosul 2015). The price of S. spicatum
annual yields fluctuate between 120 and 400 tonnes varies considerably between product grades, and in
[equivalent to between 49.86 and 166.21 m3; conversion 2007–2008, export prices ranged from USD 3000 to

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850 Planta (2016) 243:847–887

10,000/tonne (McKinnell 2011), which has increased to FPC) pers. comm. 2015). Planted sandalwood in Vanautu
USD 8000 to 17,000/tonne in 2014–2015 (Zauba.com (S. austrocaledonicum) comprises largely small-scale
2015). The price of Indian sandalwood is ten times higher woodlots with an estimated area of approximately 550 ha
than that of Australian sandalwood while the heartwood of planted by 2006 with planting continuing to date (Page
Indian sandalwood yields more essential oil with a higher et al. 2010a).
proportion of a- and b-santalols—important constituents of This review aims to explore the basic biology and
sandalwood oil fragrance—than other sandalwood species propagation of the genus Santalum, how in vitro tissue
(Baldovini et al. 2011; FAO 2015; TFS 2015a). This pos- culture has been used to produce clonal plant material, and
itive economic perspective makes Indian sandalwood an what promises this technology holds in aiding the conser-
attractive option for commercial growers aiming at inter- vation and mass propagation of threatened Santalum
national perfumery, cosmetic, and pharmaceutical markets germplasm.
(Christian 2015).
Indian sandalwood tree is a hemi-parasitic plant that
requires a host for sustained growth (discussed in more Basic biology and propagation
detail in the section ‘‘Hemi-parasitism and host plant
dependence’’), making cultivation practices highly spe- Basic biology, flowering control, reproductive
cialized. These factors, together with its strong demand in mechanisms, and breeding
Asia and the Middle East for the global fragrance industry,
have led to the mass deforestation of Indian sandalwood Sexual reproduction
from natural habitats. Indian sandalwood was first classi-
fied as vulnerable by the International Union for Conser- The onset of reproductive maturity in several sandalwood
vation of Nature (IUCN) in 1998 (IUCN 2015a). The species (S. album, S. austrocaledonicum, S. macgregorii, S.
Convention on International Trade in Endangered Species spicatum, and S. yasi) occurs between 2 and 5 years (Jiko
(CITES) also considers closely related S. austrocale- 1993; Barrett and Fox 1995; Doran and Brophy 2005).
donicum, S. yasi, and S. insulare as endangered (CITES Sexual reproduction in sandalwood may be considered to
2013). Four Santalum species, namely S. album (vulnerable be opportunistic with several species flowering across most
due to factors like fire, grazing, exploitation of wood and months of the year, with two (sometimes three) peak
smuggling; IUCN 2015a), S. fernandezianum (extinct due periods following favorable rainfall events/periods. In S.
to cutting for the aromatic wood; IUCN 2015b), S. mac- austrocaledonicum (Doran and Brophy 2005), S. macgre-
gregorii (endangered due to overexploitation of the scented gorii (Bosimbi 2005), and S. yasi (Bulai and Nataniela
wood for incense; IUCN 2015c), and S. haleakalae (vul- 2005), a low level of reproduction may be found across
nerable; IUCN 2015d), are listed in the IUCN Red Data most months of the year. The periods of peak fruit pro-
List. The use of traditional technologies such as seed ger- duction occur in both wet and dry seasons with the most
mination, as well as the use of applied biotechnologies, prominent of these two peaks occurring in the wet months
such as tissue culture, would allow production to be stan- for S. album (in India and Timor) (Suriamihardja and
dardized, and perhaps even reverse species decline through Suriamihardja 1993) and S. yasi (in Fiji) (Bulai and
the production of cultivated stands. Nataniela 2005) and the dry months for S. austrocale-
In southern China, great efforts are being made to donicum (in Vanuatu) (Daruhi 1993; Doran and Brophy
increase production (Zhang et al. 2007) while plantations 2005). Reproduction in S. spicatum (Applegate et al. 1990;
have expanded in India, China, Indonesia, and Australia Loneragan 1990) and S. lanceolatum (Applegate and
over the past 20 years (Dhanya et al. 2010; Lu 2011). McKinnell 1993) is highly dependent upon rainfall events,
Indian sandalwood (S. album) plantings in Australia are and the timing and location of seed crops are highly vari-
currently approaching 11,000 ha with two main producers able. The period of fruit development in tropical sandal-
Tropical Forestry Services (TFS 2015c) having just over wood species typically occurs over a 2- to 3-month period
9000 hectares and Santanol who acquired Elders Forestry (Corrigan et al. 2005), whereas it can take up to 6 months
estate in 2013 (Jackman 2013) comprising approximately for the fruit to mature in S. spicatum (Barrett and Fox
1800 ha (Werren 2011). In South China, plantation of S. 1995).
album on a large scale began in 2013 increasing rapidly The general breeding system of Santalum species may
to 5000 ha, planted mainly in mountain areas (Guohua be described as facultatively allogamous (incompletely
Ma, unpublished data). According to Western Australian outbreeding), with variation between families and indi-
Forest Products Commision (FPC), the 2015 Australian viduals at the level of self-incompatibility (Ma et al. 2006;
sandalwood (S. spicatum) plantings in Australia are Muir et al. 2007; Tamla et al. 2011) and with no capacity
approaching 20,000 ha (Erasmus (General Manager of for apomixis or parthenocarpy (Ma et al. 2006; Tamla et al.

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Planta (2016) 243:847–887 851

2011). The preferential outcrossing nature of the breeding Self-incompatibility

system and the capacity for self-fertilization is advanta-
geous, providing the genus a capacity to colonize new The sandalwood species S. album and S. spicatum have
islands. been recorded to have self-incompatibility mechanisms
Across all sandalwood species studied, only a low pro- operating at both pre- and post-fertilization (Rugkhla et al.
portion of controlled cross-pollinated flowers successfully 1997). In S. lanceolatum, Warburton (2000) found evi-
develop into mature seed. The proportions range from dence of self-incompatibility or pistil dysfunction and
1.3 % in S. spicatum (Rugkhla et al. 1997), 7.5 % in S. Tamla et al. (2011) identified possible self-incompatibility.
lanceolatum (Tamla et al. 2011), and 9.4–14 % in S. album Given that S. album, S. lanceolatum, and S. spicatum
(Rugkhla et al. 1997; Kulkarni and Muniyamma 1998; Ma occupy distinct phylogenetic clades (Harbaugh and Bald-
et al. 2006). Percent seed set in S. album following open- win 2007), it is possible that self-incompatibility is wide-
pollination was 2–5.2 % (Sindhu Veerendra and Anantha spread within the genus.
Padmanabha 1996; Ma et al. 2006), which was lower than Genetic variation in the expression of self-incompati-
that reported for controlled cross-pollination (9.4–14 %). bility in sandalwood is likely, with 20 % of S. lanceolatum
The low percentage seed set in sandalwood can therefore demonstrating ‘self’ fertility (Tamla et al. 2011), Muir
be influenced by the effectiveness of pollen vectors to et al. (2007) finding a high level of inbreeding within one
influence successful cross-pollination. family of S. spicatum, and Ma et al. (2006) reporting that
Sandalwood species are largely pollinated by a diverse 24 % of self-pollinated flowers set seed in S. album. This
range of insect vectors (Sindhu Veerendra and Anantha flexibility in breeding strategy would be of advantage in
Padmanabha 1996; Kulkarni and Muniyamma 1998; continental Australian species dispersing and colonizing
Tassin 2005) including bees (Apidae, Xylocopidae, many islands in south-east Asia and Pacific (Harbaugh and
Anthorphoridae), ants (Formicidae), wasps (Eumeninae, Baldwin 2007).
Vespidae and Sphecidae), flies (Sarcophagidae, Cal-
liphoridae, Muscidae and Syrphidae), and moths and Interspecific incompatibility/hybridization
butterflies (Danaidae, Pieridae and Papilionidae) (Jyothi
et al. 1991; Bhaskar 1992; Baskorowati 2011; Ratnan- The development of viable hybrid progeny has been
ingrum and Indrioko 2014). Ratnaningrum and Indrioko reported for crosses between S. album with each of S.
(2014) found that bees were the most frequent flower austrocaledonicum (Tamla et al. 2011), S. lanceolatum
visitors in S. album, followed by ants, butterflies, moths, (Tamla et al. 2011), and S. yasi (Bulai and Nataniela 2005;
wasps, and flies. Insect visitors to S. album flowers were Doran et al. 2005) despite total geographic isolation and
found to be most prevalent in the morning (Baskorowati substantial morphological variation between them.
2011) in the following sequence: moths (06:00–08:00), A phylogenetic study of the genus Santalum using
bees (07:00–10:00), wasps (08:00–10:00), and butterflies nuclear ribosomal and chloroplast DNA sequences
(09:00–12:00), while ants and flies were observed tending revealed that the earliest genetic divergence was of S.
flowers throughout the day (Ratnaningrum and Indrioko acuminatum and S. spicatum splitting from all remaining
2014). lineages (Harbaugh and Baldwin 2007). This early diver-
This aspect of sandalwood reproductive biology is an gence of S. spicatum reflects a relatively distant genetic
important consideration for the conservation of wild stands, relationship with S. album and incongruity between their
since the distribution structure and density of reproduc- mating systems (Rugkhla et al. 1997). This incongruity can
tively mature trees is likely to influence cross-pollination, be observed as an incompatibility mechanism between
seed set, and potential seedling recruitment. pollen and style, and possibly within the developing zygote
While effective cross-pollination is an important factor (Rugkhla et al. 1997).
influencing seed set in sandalwood, substantial abscission The phylogenetic clade containing the species S. aus-
of immature fruit (75–80 % fruit drop) can be found in S. trocaledonicum and S. lanceolatum (sensu stricto) was
album and S. spicatum following controlled outcross found to diverge from the clade containing S. album and S.
pollination (Rugkhla et al. 1997). Given that these authors yasi between 6.3 and 9.5 million years ago (Harbaugh and
found 10–40 % of cross-pollinated flowers had evidence Baldwin 2007). Given that no clear reproductive barriers
of successful fertilization, it is possible that the abscission exist between all four of these species (Tamla et al. 2011),
of immature fruit can be caused by maternal resource it is possible that no further divergence of these species’
limitation when pollination is not a limiting factor. San- breeding system has occurred during that time. Given that
talum species can therefore be described as mass-flower- these two clades comprise 12 of the 15 extant Santalum
ing with typically less than 10 % developing into viable species (Harbaugh and Baldwin 2007), it may be possible
seed. that hybridization between species is the norm rather than

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the exception in this genus. This feature of the breeding karyotypes belonged to the 2B type of Stebbins’ karyotypic
system can facilitate the introgression of traits between symmetry (Stebbins 1971), suggesting that S. album was a
species and the development of hybrids. Introduction of primitive taxon. Somatic cells of S. yasi and the sponta-
exotic sandalwood species within the natural range of a neous F1 hybrid, S. album 9 S. yasi, were diploid, with
compatible species will likely result in uncontrolled gene 2n = 20 (Zhang et al. unpublished data). These results
flow between them and modify the genetic structure and allow species and this hybrid in Santalum to be exploited
diversity of the local species. and utilized in future plant breeding programs.

Karyological studies in sandalwood Propagation

Karyological studies are a prerequisite for the creation of Seed propagation

innovative new varieties in plant genetics and breeding.
Polyploidy is widely acknowledged as one form of Seeds are important organs of higher plants since they
breeding in plants. Previous studies showed that the contain the zygotic embryo, a vital part of the reproductive
somatic cells of Santalum were diploid, with 2n = 20, life cycle that is renewed when seeds are dispersed fol-
including S. album (Rao 1942b), S. ellipticum, S. lowed by seed germination. The seeds of many sandalwood
freycinetianum, and S. paniculatum (Carr 1978). Forty species are naturally dispersed by bird-based endozoochory
chromosomes were observed in many haustorium cells of (Batabyal et al. 2014). In contrast, the seeds of S. spicatum
S. album (Srimathi and Sreenivasaya 1962). Harbaugh are dispersed by a small marsupial known as a woylie
(2008) reported four ploidy levels in Santalum ranging (Murphy et al. 2005). Seed can be induced to germinate
from diploid (n = 10) to octoploid (n = 40) based on flow artificially under controlled environmental conditions, but
cytometric analysis of DNA content of 16 Santalum species this depends on the stage of fruit development and even on
and 5 varieties across Pacific islands (S. album, S. yasi, S. the source and size of seed (Manonmani and Vanangamudi
spicatum, S. paniculatum, S. obtusifolium, S. murrayanum, 2002), usually after breaking dormancy (Jayawardena et al.
S. macgregorii, S. leptocladum, S. lanceolatum, S. halea- 2015). Manonmani and Vanangamudi (2001) noted that
kalae, S. insulare var. marchionense, S. insulare var. ra- black fruits (most mature) showed a higher germination
iateense, S. freycinetianum var. freycinetianum, S. percentage than brown or red fruits (73.2, 68.2, and
freycinetianum var. lanaiense, S. freycinetianum var. 29.0 %, respectively). However, Hirano (1990) showed
pyrularium, S. ellipticum var. ellipticum, S. ellipticum var. that simple removal of the testa, and a dip of naked seed in
littorale, S. boninense, S. austrocaledonicum, and S. DithaneTM M45 (a fungicide) for 5 min, could result in
acuminatum) (Harbaugh 2008), the majority of which were 67 % germination for Indonesian S. album, 26 days to
diploid and tetraploid, with 2n = 20 and 2n = 40, germination and seed viability of 324 days (assessed by the
respectively. The results from this study suggest that two total number of days between first and last seedling). These
allopolyploid events between distantly related species and values, also in Hirano’s study, were 77 %, 75 and 824 days
four putatively autopolyploid events occurred in Santalum, for S. paniculatum, and 38 %, 155 and 387 days for S.
indicating that the Santalum island colonists have a ten- haleakalae; these species originate from Hawai’i. Hirano
dency to be polyploid. Polyploids in Santalum are likely to (1990) used a substrate consisting of vermiculite, perlite,
be better suited than diploids for long-distance dispersal and peat plus osmocote (18-6-12) (2:2:1), noting that
and successful establishment on oceanic islands, since chelated iron (Sequestrene 138 Fe) was essential for seed
polyploidy may increase the likelihood of their establish- germination and seedling growth. S. album seeds germi-
ment and long-term survival across the Pacific islands by nated more rapidly when the testa was removed, and seeds
decreasing inbreeding depression (Harbaugh 2008). were soaked in 0.05 % gibberellic acid (GA3) for 12–16 h
A detailed karyomorphological study on S. album (Nagaveni and Srimathi 1981; Das and Tah 2013). A
revealed that the interphase nucleus was a simple chro- similar treatment was suggested for S. macgregorii seed
mocenter type while the prophase chromosomes were of germination by Gunn et al. (2002). Gamage et al. (2010)
the interstitial type (Zhang et al. 2010). A mixoploid of reported that the germination efficacy of stored S. album
2n = 2x = 20 and 2n = 4x = 40 was found in the shoot- seeds decreases over time, reaching 0 % after 28th weeks,
tip meristem cells of some S. album individuals, of which suggesting that seeds should be sown once shed rather than
about 5 % was found to be tetraploid (4n = 40). The using stored seeds and that germination trials should be
karyotypic formulae analyzed showed that centromeres started early (Gamage et al. 2010). Seed germination of S.
were predominantly in a median position with a few sub- spicatum in natural conditions is very poor, and only
median centromeres, i.e., 2n = 20 = 18 m ? 2sm and 1–5 % seeds germinate (Loneragan 1990). Liu et al. (2009)
2n = 40 = 32 m (2SAT) ? 8sm, respectively. The found that a substrate mixture of burnt soil, peat, and

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coconut dust (1:1:1, w/w), including 2 % calcium super- a source of scions improved graft success (60 %) more than
phosphate, resulted in 98 % survival (as well as greater scions obtained directly from field-grown trees. The suc-
height and biomass than other substrates) of S. album cess rate of in vitro grafting was also affected by scion size
seedlings when Kuhnia rosmarinifolia Vent. was used as and rootstock age. Under suitable conditions, scions and
the primary host plant and after treating surface-sterilized hypocotyls united to form complete plants with 2–4 leaves
(0.1 % mercuric chloride for 5–6 min and three washes in within 6–8 weeks (Sanjaya et al. 2006a). Grafting of S.
distilled water) seeds with 1 mg/L GA3 for 24 h. Lu et al. austrocaledonicum has also been demonstrated using a top-
(2013) soaked S. album seeds in 0.1 % GA3 for 12 h using wedge graft with actively growing semi-hardwood stems
the protocol of Radomiljac (1998) then surface-sterilized (Tate et al. 2006). These authors suggested that the success
them with 3 % NaOCl for 5 min, washed them with dis- of grafting was dependent upon the skill of the propagator
tilled water, and then germinated them on sterilized sand with the percentage of successfully grafted unions varying
soaked with distilled water at 28–30 °C for 3–4 weeks. between 60 and 90 %.
When germination conditions are suitable and in associa- Thus, techniques for the establishment of a seedling
tion with an appropriate host, this method can be useful for stock are well established, although mechanization of the
the large-scale seed germination of S. album (Ral 1990), as process is less explored. To address this gap, St. Jack et al.
summarized in Fig. 1 (specific for Karnataka, India). (2013) devised a seed metering device that would optimize
Athelstone et al. (1994) assessed the effects of plant growth the mechanization of seed planting and germination.
regulators (ethrel, silver thiosulfate, 6-benzyladenine,
kinetin, GA3, and GA4) on S. accuminatum seed germi- Vegetative propagation
nation. They found that cracking seed and treatment with
580 lM GA3 (duration of treatment not reported) resulted Various authors have reported on successful vegetative
in 90 % seed germination within 43 days. Jayawardena propagation of S. album by cuttings. Uniyal et al. (1985)
et al. (2015) found that scarification and 500 mg/L GA3 demonstrated that S. album can be propagated by root
could increase germination to 100 % from near-zero levels cuttings with 60 % success when they used Seradix B2Ò at
in non-scarified control S. album seeds. Chauvin and Ehr- the time of planting during the first week of April under
hart (1998) observed differences in germination rates of Indian conditions. S. lanceolatum can be propagated by
two provenances of S. austrocaledonicum var. austro- root suckers (Warburton et al. 2000). Havea (2012)
caledonicum: 55 % germination in the ‘‘Ile des Pins’’ reported that S. yasi, S. album, and their hybrids can be
provenance after 2 years and 0 % germination in the vegetatively propagated using apical cuttings from younger
‘‘Mare’’ provenance with the exception of 19 % germina- seedlings (up to 12 months of age). They recommended
tion in one lot after 15 months. Nasi (1995) had noted that treating the cuttings with indole-3-butyric acid and a-
25 % germination of S. austrocaledonicum var. pilosulum naphthaleneacetic acid (1.0 mg/L each) and using a sub-
was possible without pre-treatment, but only after 1 year; strate composed of sand:peatmoss (30:70, w/v).
however, removing the endocarp, nicking the seeds, not Batabyal et al. (2014) proposed that S. album can be
storing seeds, and soaking seeds in 100 mg/L GA3 all propagated using 15-cm-long stem cuttings from 3- to
increased the germination percentage at 28 °C. Natural 4-year-old trees by treatment with 1.5 mg/L indole-3-acetic
stands of S. album growing in East Nusa Tenggara Pro- acid (IAA) and 1.5 mg/L GA3, resulting in most leaves per
vince in Indonesia showed regeneration percentages rang- branch, although rooting percentages among the treatments
ing widely from 4.85 to 48.4 % (Wawo 2008). were not reported. These authors were also able to increase
In vivo or in vitro germinated seedlings can be used as the number of branches per cutting by applying 1.5 mg/L
ideal rootstock for grafting (Sanjaya and Rai 2003). San- IBA and kinetin. Rao and Srimathi (1977) attempted air
jaya et al. (2006a) managed to attain a 50 % in vivo layering to clonally propagate S. album, in which branches
micrografting rate of success in S. album when 4- to 5-cm- of 2 cm thick from different seasons’ growth were used,
long scions, collected from a candidate plus tree (CPT) of observing that March–April (rainy season) were most
50–60 years of age, were grafted onto 90-day-old nursery- favorable for air layering under Indian conditions. Collins
grown rootstock. Sanjaya et al. (2006a) reported that scion et al. (2000) found substantial variation in the percentage
size, rootstock age, and scion collection season are vital of stem cuttings with adventitious roots between experi-
factors that affect graft success. Grafted plants were kept ments, genotypes and species. Differences between
under greenhouse conditions for 6–8 weeks to allow for experiments were recorded for S. album (9.5 and 0 %
graft union. In vitro micrografts were accomplished by rooting), S. austrocaledonicum (63.5 and 20.2 %), and S.
placing 1- to 2-cm-long scions collected from nodal cut- yasi (46.1 and 10.1 %). Differences were observed
tings (obtained from CPT) onto the hypocotyl of 45-day- between 15 S. austrocaledonicum genotypes with root
old in vitro rootstocks. The use of in vitro-grown shoots as induction ranging from 25 to 88.9 %. It has been proposed

123
854 Planta (2016) 243:847–887

Host plants are Can be planted at a

indicated in Table 2 spacing of 3 m2 along Sandalwood tree
with host

Ready for transplanting to the field Seeds available (April-May and Sept-Oct)

GA3-treated seeds

Sown in beds (sunken or raised 250

Host pruned periodically g/m2 bed of sand + red earth (can
apply fungicide + nematicide)

Seedlings suffering from

damping-off due to fungus
Germination starts after 30 days; 4-6 Surface sterilization
and nematicide infection can
Seedling reaches leaves form in about 60-70 days
be sprayed with fungicide
and nematicide about 30-45 cm in
height in 6-7 months
Transplant to poly bags of 15 x
30 cm with host seeds sown in
the bag to provide primary host
In vitro germinated seedlings
on MS medium or any nutrient
medium can serve as root
These plants can be used stock. Seedlings must be >1.5
as rootstock for grafting; month old.
minimum age: > 3 months

Fig. 1 Cultivation of sandalwood from seeds (modified from Ral 1990)

that S. yasi and S. austrocaledonicum are more amenable to many species of the Fabaceae, Mimosaceae, Casuari-
stem cutting propagation than S. album, S. lanceolatum, naceae, Meliaceae, Myrtaceae, Apocynaceae, and Rham-
and S. macgregorii (Collins et al. 2000; Thomson et al. naceae are proven good hosts. However, not all species in
2005b). The differences observed in the series of experi- these families are good hosts. For example, in the Faba-
ments conducted by Collins et al. (2000) indicate that it is ceae, many species, including Butea monosperma and
difficult to generalize the optimum conditions required for Tamarindus indica, are poor hosts (Table 2).
successful stem cutting propagation across the Santalum Annapurna et al. (2004) noticed that the container size
genus. for potting influenced the success of pot growth, with
600 ml pots being an ideal size for S. album seedlings,
Hemi-parasitism and host plant dependence producing plantlets 20 cm high within 6 months when in
the presence of pigeon pea [Cajanus cajan (L.) Millsp.] as
First noted by Scott (1871), and further confirmed by the host plant. These authors also recommended a potting
Brandis (1903), Santalum species are hemi-parasitic, mixture of sand, soil, compost, burnt rice husk, and char-
whereby they derive part of their water and nutrient coal (5:3:10:1:1). Despite these optimized conditions, the
requirements from a host plant. Under cultivation, sandal- authors noted that root development was quite poor, indi-
wood species require hosts during all stages of production cating that one possible reason might be the use of an
including the pot stages, in the first years of establishment inappropriate host. Radomiljac (1998) had previously used
and at maturity (Ehrhart and Fox 1995; Fox et al. 1996; Alternanthera nana R.Br. and Sesbania formosa (F.
Radomiljac et al. 1998a, 1999a; Annapurna et al. 2006), Muell.) N. Burb. as the primary host in pot experiments
although the choice of potting medium can also be an that used 1500 ml pots in a substrate consisting of sand,
influencing factor (Annapurna et al. 2005). Sandalwood peat, and perlite (3:2:2), successfully increasing the sur-
can potentially be grown across a range of environments vival, height, and diameter of S. album plants; in that study,
provided locally adapted host species can be identified for Atalaya hemiglauca (F.Muell.) F.Muell. ex Benth, Acacia
each stage of cultivation (Table 2). Among these families, hemignosta F.Muell, and Crotalaria retusa L. were not

123
 Table 2 Host plants (suitable and unsuitable) for sandalwood (chronological listing)
Species Country of Suitable hosts Family Unsuitable hosts Family References
test

S. album India Azadirachta indica A.Juss Meliaceae Butea monosperma (Lam.) Taub. Fabaceae Parthasarathi et al.
Cassia fistula L. Fabaceae Dodonea viscosa Jacq. Sapindaceae (1974)
C. siamea (Lam.) lrwin et Barneby Fabaceae Gmelina arborea Roxb. Lamiaceae
Dalbergia latifolia Roxb. Fabaceae Melia azedarach L. Meliaceae
Planta (2016) 243:847–887

Ficus bengalensis L. Moraceae Tamarindus indica L. Fabaceae

Grevillea robusta A.Cunn. ex R.Br. Proteaceae
Pithecellobium dulce (Roxb.) Benth. Fabaceae
Pongamia pinnata (L.) Panigrahi Fabaceae
Syzygium cumini (L.) Skeels. Myrtaceae
Wrightia tinctoria (Roxb.) R.Br. Apocynaceae
Zizyphus mauritiana Lam. Rhamnaceae
S. spicatum Australia Acacia acuminata Benth. Mimosaceae NR Struthers et al. (1986)
S. album India Acacia nilotica (L.) P.J.H.Hurter & Mabb. Fabaceae NR Anantha padmanabha
Bauhinia biloba L. Fabaceae et al. (1988)
Cassia siamea (Lam.) Lrwin et Barneby Fabaceae
Casuarina equisetifolia L. Casuarinaceae
Dalbergia sissoo Roxb. Fabaceae
Melia dubia Cav. Meliaceae
Pongamia pinnata (L.) Panigrahi Fabaceae
Terminalia arjuna (Roxb.) Wight & Arn. Combretaceae
T. alata Heyne ex Roth Combretaceae
Wrightia tinctoria (Roxb.) R.Br. Apocynaceaea
S. acuminatum Australia Acacia rostellifera Benth. Mimosaceae Melaleuca viminea Lindl. Myrtaceae Tennakoon et al. (1997)
A. pulchella R. Br. Mimosaceae
Allocasuarina campestris (Diels) L. Casuarinaceae
Johnson
855

123
 Table 2 continued
856

Species Country of Suitable hosts Family Unsuitable hosts Family References

test

123
S. acuminatum Australia Acacia rostellifera Benth. Mimosaceae NR Tennakoon et al. (1997b)
A. pulchella R. Br. Mimosaceae
Allocasuarina campestris (Diels) L. Casuarinaceae
Johnson
Alyogyne hakeaefolia (Giord.) Alef. Malvaceae
Anthocercis viscosa R. Br. Solanaceae
Baeckea tetragona Benth. Myrtaceae
Cryptandra mutila Reisseck. Rhamnaceae
Dodonaea aptera Miq. Sapindaceae
Exocarpos sparteus R. Br. Santalaceae
Leucopogon ovalifolius Sonder. Epacridaceae
Lysinema ciliatum R. Br. Epacridaceae
Melaleuca viminea Lindley Myrtaceae
Spyridium cordatum (Turca.) Benth. Rhamnaceae
S. spicatum Australia Acacia acuminata Benth. Fabaceae Allocasuarina huegeliana (Miq.) L.A.S. Casuarinaceae Brand et al. (1998, 2000,
Johnson 2003); Brand (2009)
Eucalyptus loxophleba subsp. loxophleba Myrtaceae
S. album Australia Acacia trachycarpa E. Pritzal Mimosaceae Eucalyptus camaldulensis Dehnh. Myrtaceae Radomiljac et al. (1998b,
A. ampliceps Maslin. Mimosaceae 1999a,b,c)
Sesbania formosa (F. Muell) N. Burb. Papilionaceae
S. acuminatum Australia Melia azedarach L. Meliaceae NR Loveys et al. (2001a)
Myoporum parvifolium R.Br. Scrophulariaceae
S. acuminatum Australia Acacia pycnantha Benth. Mimosaceae Eucalyptus fasciculosa F. Mueller Myrtaceae Loveys et al. (2001b)
Atriplex vesicaria Benth. Amaranthaceae Eucalyptus camaldulensis Dehnh. Myrtaceae
Heterodendrum oleifolium Desf. Sapindaceae
Pittosporum phylliraeoides DC. Pittosporaceae
Rhagodia spinescens var. deltaphylla Amaranthaceae
S. acuminatum Australia Acacia cyclops A Cunn. Ex G. Don Mimosaceae NR Loveys et al. (2002)
Atriplex nummularia Lindl. Amaranthaceae
Myoporum parvifolium R. Br. Scrophulariaceae
Templetonia retusa (Vent.) R. Br. Fabaceae
S. acuminatum Australia Acacia acuminata Benth. Mimosaceae NR Woodall and Robinson
(2002)
Planta (2016) 243:847–887
 Table 2 continued
Species Country of Suitable hosts Family Unsuitable hosts Family References
test

S. acuminatum Australia Acacia acuminata Benth. Mimosaceae Acacia cyclops A.Cunn. ex G. Don Mimosaceae Woodall and Robinson
A. asepala Maslin Mimosaceae A. jibberdingensis Maiden & Blakely Mimosaceae (2003)
A. assimilis S.Moore Mimosaceae Alyxia buxifolia R.Br. Apocynaceae
A. cochlearis (Labill.) H.L.Wendl. Mimosaceae Astroloma prostratum R.Br. Epacridaceae
Planta (2016) 243:847–887

A. declinata R.S. Cowan & Maslin Mimosaceae A. epacridis (DC.) Druce Epacridaceae
A. erinacea Benth. Mimosaceae Dichopogon fimbriatus (R.Br.) J.F.Macbr. Anthericaceae
A. glaucoptera Benth. Mimosaceae Dryandra armata R.Br. Proteaceae
A. lasiocalyx C.R.P.Andrews Mimosaceae Enchylaena tomentosa R.Br. Chenopodiaceae
A. microbotrya Benth. Mimosaceae Eucalyptus perangusta Brooker Myrtaceae
A. patagiata R.S. Cowan & Maslin Mimosaceae E. salmonophloia F.Muell. Myrtaceae
A. pulviniformis Maiden & Blakely Mimosaceae E. tetragona (R.Br.) F.Muell Myrtaceae
A. saligna H. Wendl. Mimosaceae Grevillea nudiflora Meisn. Proteaceae
A. sulcata R.Br. Mimosaceae Hybanthus floribundus (Lindl.) F.Muell. violaceae
Allocasuarina campestris (Diels) Casuarinaceae Melaleuca pentagona Labill. Myrtaceae
L.A.S.Johnson
A. huegeliana (Miq.) L.A.S.Johnson Casuarinaceae Nemcia hookeri (Meisn.) Crisp Papilionaceae
A. humilis (Otto & F.Dietr.) Casuarinaceae Opercularia spermaocea Rubiaceae
L.A.S.Johnson
A. microstachya (Miq.) L.A.S.Johnson Casuarinaceae Oxylobium microphyllum Benth. Papilionaceae
A. thuyoides (Miq.) L.A.S.Johnson Casuarinaceae Petrophile fastigiata R.Br. Proteaceae
Banksia attenuate R.Br. Proteaceae P. seminuda Lindl. Proteaceae
Dampiera lavandulacea Lindl. Goodeniaceae Rhagodia baccata (Labill.) Moq. Henopodiaceae
Daviesia incrassate Sm. Papilionaceae Spyridium majoranifolium (Fenzl) Rye Rhamnaceae
D. teretifolia Benth. Papilionaceae Synaphea petiolaris R.Br. Proteaceae
Dodonaea humifusa Miq. Sapindaceae
D. ptarmicaefolia Turcz. Sapindaceae
Gastrolobium parviflorum (Benth.) Crisp Papilionaceae
Hakea lissocarpha R.Br. Proteaceae
H. nitida R.Br. Proteaceae
H. preissii Meisn. Proteaceae
H. trifurcata (Sm.) R. Br. Proteaceae
Halgania cyanea Lindl. Boraginaceae
Hibbertia rupicola (S.Moore) Dilleniaceae
C.A.Gardner
Jacksonia furcellata (Bonpl.) DC. Papilionaceae
Labichea lanceolata Benth. Caesalpiniaceae
857

123
 Table 2 continued
858

Species Country of Suitable hosts Family Unsuitable hosts Family References

test

123
Lasiopetalum rosmarinifolium (Turcz.) Sterculiaceae
Benth.
Olearia axillaris (DC.) Benth. Asteraceae
O. muelleri Benth. Asteraceae
Senna artemisioides subsp. filifolia Randell Caesalpiniaceae
Templetonia retusa (Vent.) R. Br. Papilionaceae
Thomasia angustifolia Steud. Sterculiaceae
S. album India Azadirachta indica A. Juss Meliaceae Artocarpus integrifolia L. Moraceae Nagaveni and
Cajanus cajan (L.) Millsp. Papilionaceae Acacia auriculiformis A. Cunn. Ex Benth. Fabaceae Vijayalakshmi (2003)
Casuarina equisetifolia L. Casuarinaceae Swietenia mahogani L. Meliaceae
Eucalyptus camaldulensis Dehnh. Myrtaceae
Pongamia pinnata L. Fabaceae
Tectona grandis L. Verbenaceae
Wrightia tinctoria (Roxb.)R.Br. Apocynaceae
S. album China Acacia auriculaeformis A. Cunn. Fabaceae Artocarpus heterophyllus (Wall.) R. N. Park. Moraceae Li (2003)
Acacia confusa Merr. Fabaceae Camellia odorata Abel. Theaceae
Albizia lebbek (L.) Benth. Fabaceae Mangifera indica Linn Anacardiaceae
Caesalpinia pulcherrima L. Caesalpiniaceae Ormosia fordiana Oliv. Papilionaceae
Casuarina junghuhniana Miq. Casuarinaceae Pterocarpus echinatus Pers. Fabaceae
Cajanus cajan (L.) Mill sp. Fabaceae Syzygium cumini (L.) Skeels Myrtaceae
Kuhnia rosmarnifolia Vent. Asteraceae
Lantana camara L. Verbenaceae
S. album China Gardenia jasminoides Ellis Rubiaceae NR Ma et al. (2005)
Hibiscus rosa-sinensis L. Malvaceae
Phyllanthus reticulatus Poir. Euphorbiaceae
S. album China Acacia confusa Merr. Fabaceae Bischofia polycarpa (H. Lévl.) Airy Shaw Euphorbiaceae Lu (2011); Lu et al.
Dalbergia odorifera T. Chen Fabaceae Dracontomelon duperreranum Pierre Anacardiaceae (2013, 2014)
S. album China Kuhnia rosmarnifolia Vent. Asteraceae NR Zhang et al. (2012a)
Acacia confusa Merr. Fabaceae Bauhinia blakeana Dunn Leguminosae Chen et al. (2014)
Calliandra haematocephala Hassk. Mimosaceae Delonix regia (Hook.) Raf. Leguminosae
Caesalpinia sappan L. Fabaceae Leucaena leucocephala Lam. Mimosaceae
Erythrina corallodendron L. Papilionaceae Ormosia pinnata (Lour.) Merr. Papilionaceae
Planta (2016) 243:847–887
Planta (2016) 243:847–887 859

effective as pot hosts. Annapurna et al. (2006) indicated

that Mimosa pudica L. was the most effective host among

Rocha et al. (2014, 2015)

seven hosts tested, claiming that C. cajan has three dis-

Ouyang et al. (2015)

advantages: (1) rapid growth thus competing with sandal-
wood for light and nutrients; (2) susceptibility to fungi and
insect pests; and (3) need for intensive management (fre-
References

quent pruning). In contrast, Acacia acuminata was an

effective host for S. spicatum (Brand et al. 1998, 2000,
2003; Brand 2009; Table 1).
Thymelaeaceae

In South China, sandalwood plants of different ages of

development (seedlings to adult plants) need different
hosts. In the seedling (pot) stage, a good host plant Kuhnia
Family

rosmarinifolia Vent. (Asteraceae) is usually used as the

seedling host (Zhang et al. 2012a; Chen et al. 2014; Yang
et al. 2014). This species is a good host plant since it is
propagated by cuttings, it roots easily, and no seeds are
produced, thus avoiding its dispersal. Even after seedlings
are transferred to the field, K. rosmarinifolia serves as a
Aquilaria sinensis (Lour.) Gilg.

good host for 1–2 years until the parasitized host dies. At
this stage, some shrubs (Tephrosia candida Roxb. DC,
Calliandra haematocephala, Cajanus cajan Millsp) and
small trees (Caesalpinia sappan Linn, Cerasus yedoensis
Matsum.) need to be cultivated near sandalwood trees.
Unsuitable hosts

Among possible hosts, species of the Fabaceae are com-

monly used possibly since they have nitrogen-fixing Rhi-
zobium species in their nodules which improve the soil
NR

environment (Li 2003; Chen et al. 2014). However, some

legume species are poor hosts such as Delonix regia (Boj.
Caprifoliaceae
Casuarinaceae

ex Hook.) Raf., Bauhinia blakeana Dunn and Cassia

surattensis F.Muell. (Chen et al. 2014). Most genera of the
Fabaceae
Family

Rutaceae are good hosts, including Citrus, Clausena, and

Murraya.
Lu et al. (2013) found that nodulation of the host was an
important factor determining the transfer of nitrogen from
the host to S. album, increasing the amount of N transferred
by about 6-fold compared to control (non-nodulated)
treatments. Seedlings were planted 10 cm apart from the
host (Dalbergia odorifera T. Chen) in a substrate of ver-
Casuarina equisetifolia L.

Lonicera japonica Thunb.

miculite and perlite (2:1, v/v, pH 6.8). Nodulation of the

Dalbergia sissoo Roxb.

host was enhanced by Bradyrhizobium elkanii DG, which

was originally isolated from the active nodules of Ptero-
Suitable hosts

carpus macrocarpus Kurz. This provides a simple, but

important and often overlooked, management strategy for
the field production of S. album. Biomass production of
shoots, roots, and haustoria of S. album were greater when
NR not reported in the study

grown with N2-fixing hosts than non-N2-fixing hosts (Lu

et al. 2014; Table 1). The rhizospheric pH value of the host
Country of

affects the growth of S. album. Optimal pH in the rhizo-

Table 2 continued

China

spheric soil of almost all fine hosts ranged from 5.0 to 6.0
India
test

(Mei et al. 2011). Generally, the advantage of N2-fixing

species over non-N2-fixing species is evidenced by the
S. album
S. album
Species

literature (Table 2). The number and size of haustoria,

height, and base diameter of S. album showed significant

123
860 Planta (2016) 243:847–887

differences among different N2-fixing hosts (Chen et al. the hemi-parasitism by haustoria that form between San-
2014). talum species and the host root. Sahai and Shivanna (1984)
A recent comparative study showed that a suitable host observed that the addition of 30 lg/ml xenognosin, the
among leguminous and non-leguminous trees could pro- active fraction of tragacanth gum (an exudate of a legume,
mote the growth of S. album. Ouyang et al. (2015) found Astragalus sp.), could effectively induce haustoria in
that net photosynthetic rate, stomatal conductance, tran- 3-month-old S. album seedlings in the absence of the host.
spiration rate, and plant height in S. album in China were Tennakoon et al. (1997a) found that S. acuminatum
similar in two suitable hosts, D. sissoo, a leguminous tree, derived nitrogen primarily from woody N2 fixers (legumes
and Lonicera japonica, a non-leguminous vine. It was and Allocasuarina), although each host transported a
suggested that black-stained material (BSM) in the inter- characteristic set of organic nitrogenous solutes, but little
action of haustoria penetrating into hosts might play an or no nitrate. They concluded that only a limited direct flow
important role in establishing the sandalwood host con- of amino compounds took place between xylem streams of
nection. In the good hosts, L. japonica and D. sissoo, the hosts and S. acuminatum (Tennakoon et al. 1997b). There
connection between the finger parenchymal cells of the were close resemblances between S. album and legume
haustoria and the host root cell was tight and was related to hosts (S. formosa, A. trachycarpa, A. ampliceps) in con-
the presence of a prominent BSM while BSM in the poor centration and composition of xylem sap amino acids, and
host Aquilaria sinensis was thin. The origin of BSM and its in the amino acid spectra of the corresponding parasite
functions remain unclear. Thus, the metabolism underlying endophytic tissue, and low N levels in xylem sap of
the host preference of sandalwood needs further E. camaldulensis and dissimilarities between its amino acid
investigation. composition and that of partnered S. album (Radomiljac
The dry matter gains of S. album grown at 33 weeks et al. 1998b). The analyses of net C and N gains of S.
were significantly improved when S. album was separately album and the C: N ratios of xylem solutes of S. album
partnered with the three legume hosts, Sesbania formosa, between the parasite-host associations showed that the
Acacia trachycarpa, and A. ampliceps compared with the heterotrophic gains of C from xylem of the three legume
hemi-parasite grown with Eucalyptus camaldulensis or hosts was highest in S. album partnered with A. ampliceps
without a host (Radomiljac et al. 1999b). They proposed S. (57.9 % of total carbon) and lowest in S. album and S.
formosa to be best host, followed by A. ampliceps, A. formosa (34.6 %) over a 9-week period (Radomiljac et al.
trachycarpa, and E. camaldulensis (Radomiljac et al. 1999b). This is similar to the study of Tennakoon et al.
1999b). Based on assays of leaf, stem, bark, and root tissue (1997b) in which S. acuminatum probably would gain more
of S. album and its hosts, net increases in mineral contents than one-third of its C requirement for dry matter pro-
of S. album over the first 9 weeks were obtained when duction heterotrophically from the xylem of its hosts.
attached to a beneficial host. In particular, some elements Foliar N concentrations of S. album were significantly
such as Ca, K, P, and Na were greatest when associated greater than corresponding hosts and higher when on the
with hosts richest in corresponding elements. S. album N2-fixing hosts than on E. camaldulensis, or without a host
foliage became more abundant in Na and in some cases (Radomiljac et al. 1999c). When S. austrocaledonicum,
also in P and N compared with associated hosts. By con- distributed in New Caledonia, was associated with A.
trast, net losses or only small gains of P, K, Ca over the spirorbis, psydrax odorata, Diospyros pustulata, or Cleis-
study interval in S. album grown alone or associated with tanthus stipitatus, an analysis of foliar mineral composition
E. camaldulensis indicated poor nutrient uptake ability showed that nitrogen levels were particularly high when A.
through its own root system. A previous study on mineral spirorbis served as the host (Veillon and Jaffré 1995).
nutrition of S. spicatum with its preferred host, Acacia These results indicate that Santalum directly gained xylem
acuminata, showed high levels of K and Na, a high K/Ca N and C from N2-fixing legumes hosts and that the parasite
ratio, and low levels of Zn in the parasite compared to the obtained little N and C from the xylem sap of non-bene-
host plant (Struthers et al. 1986). In this interaction, host ficial hosts such as E. camaldulensis.
seedlings are planted in the first year while untreated seeds In addition, xylem sap of hosts contained variable
of S. spicatum are sown directly in the second or third year, amounts of sucrose, glucose, and fructose, whereas that of
although Woodall and Robinson (2002) found that simul- matching parasites was dominated by fructose in S. album
taneous sowing of both S. spicatum and A. acuminata (Radomiljac et al. 1998b). Similarly, the levels of sucrose,
improved germination, survival, and growth of the former. fructose, glucose, malate, and citrate were high in all saps,
Woodall and Robinson (2003) noted that S. spicatum par- and fructose was especially prominent in S. acuminatum
asitized 68 hosts in remnant populations in Western Aus- (Tennakoon et al. 1997b). Using 14C, glucose was found to
tralia (Table 2). These studies provide some evidence that move from the host Myoporum parvifolium to S. acumi-
a beneficial host can supply nutrients as a direct result of natum (Loveys et al. 2001a). Dissimilarities were also

123
Planta (2016) 243:847–887 861

evident in the proportional amounts of xylem-borne Dynamics of the haustoria

organic acids and sugar and organic acid composition of
leaves between hosts and S. album (Radomiljac et al. Root haustoria are structures of parasitic plants that form
1998b). Chloride, sulfate and phosphate were found in the physical connections with the roots of other plants and
xylem of S. acuminatum (Tennakoon et al. 1997b). This provide a conduit between parasite and host (Irving and
implied that substantial metabolic patterns of incoming Cameron 2009). These haustoria are important organs for
xylem solutes were variable between Santalum species. transporting water and nutrients from host to parasite. Due
Tennakoon et al. (1997b) found that 40–80 % of proline to the hemi-parasitic nature of sandalwood, like other
was from the infiltration of xylem of haustoria-bearing root members of the Santalales, their cultivation is more com-
segments of a major host (A. rostellifera) and nitrate plex compared with other woody trees. Many studies have
reductase activity was induced in haustoria following host focused on haustorial anatomy and development and par-
xylem feeding of nitrate in S. acuminatum. That study asite physiology to better understand the mechanisms of
concluded that haustoria act as a major site for the syn- hemi-parasitism in sandalwood.
thesis and export of proline and might therefore play an Morphological and anatomical characters of haustorium
important role in osmotic adjustment of the parasite and its have been widely investigated in Santalum (Barber 1906,
related acquisition of water from hosts in Santalum. An 1907; Rao 1942a; Tennakoon et al. 1997a, b; Tennakoon
interesting finding was the transfer of an insecticidal and Cameron 2006; Zhang et al. 2012a; Yang et al. 2014).
compound with a molecular weight of 705.4, from the host Haustoria have an inverted conical shape and their apex is
Melia azedarach to S. acuminatum, which increased the horizontally round before contacting host root. Otherwise,
mortality of apple moths fed on the fruit of S. acuminatum if haustoria can encounter a suitable host, its horizontal
(Loveys et al. 2001a). face will attach to the host root and closely clasp it to
In S. album, photosynthetic rate and water-use efficiency penetrate and establish a vascular connection with that of
were generally higher than in corresponding hosts while the host root, and finally form a mature haustorium. The
transpiration rates were not noticeably different between general anatomy of Santalum haustorium was similar in
parasite-host associations (Ouyang et al. 2015). However, these studies even when sandalwood partnered with dif-
diurnal profiles of gas exchange and leaf water potential of ferent hosts. Research has showed that Santalum is a
hosts and parasites showed closely coordinated diurnal xylem-feeding parasite relying on the interfacial par-
stomatal responses of the parasite water relations to its enchyma of finger cells of haustoria to actively transport
host, thus suggesting that transpiration rates of the parasite water and solutes from the host root (Tennakoon and
generating leaf water potential gradients led to continuous Cameron 2006).
uptake of water and nutrients from the host (Radomiljac Ouyang et al. (2015) studied the differences between the
et al. 1999c). Transpiration and photosynthetic rates of S. suitability of hosts for S. album. Well-formed and fre-
acuminatum were consistently lower than those of its quently replaced haustoria formed in a good host Dalber-
principal hosts, but water-use efficiencies were very similar gia sissoo that likely enhanced metabolism and nutrient
to that of one host, A. rostellifera (Tennakoon et al. 1997a). transport. Ultrastructural observations showed differences
A comparative study on water relations and gas exchange in the interfacial region and internal structure between
characteristics of S. acuminatum and its hosts at three finger parenchymal cells and host root cells, vascular tis-
examined sites, i.e., Middleback, Aldinga and Adelaide, in sue, the reaction of host root cells, and penetration modes.
central Southern Australia showed that S. acuminatum had Recently, a detailed study of haustorial development
a higher osmotic potential than the hosts at Middleback and showed obvious morphogenic differences between haus-
Aldinga sites during summer and winter, with a stable wa- torium formation and lateral root initiation: while the for-
ter potential difference of 1.7 MPa between them over a mer develops laterally from the root cortex of S. album, in
measurement period of 24 h, in which mannitol, Na?, K? the latter, the lateral root originates from the pericycle of
and Cl- dominated in the osmotic potential of the parasite the mother root (Zhang et al. unpublished data). A large
(Loveys et al. 2001b). Moreover, stomatal conductance and number of starch and proteinaceous compounds are present
assimilation of S. acuminatum were also lower those of the throughout haustorial development, especially in the stage
hosts at both sites during summer and winter measure- in which haustoria invade the host root. Many lysosomes
ments. However, transpiration for S. acuminatum was dif- can be observed and large-scale digestion of host cells
ferent between the two sites investigated. The occurs at the interface between the parasite and the host
instantaneous water-use efficiency [(0.13–2.2 lmol (CO2) (Zhang et al. 2012a). These results suggest that the haus-
mmol-1 (H2O)] of S. acuminatum was higher than that of torium is an active metabolic organ in sandalwood and that
the host at Middleback, expressed as lower transpirational mechanical force and enzymatic activity may be needed for
water loss. the haustoria to be able to penetrate the host tissues. Yang

123
 Table 3 Explant source, size, and surface sterilization procedures for preparation of tissue culture studies of sandalwood (chronological listing)
862

Species Explant source Explant type, size and density; culture vessel Surface sterilization and preparation References

123
S. album Mature fruit. Age of mother plant NR Seeds, ZEs, endosperms. Exact size of explant Fruits ? CW 10 min Rangaswamy and Rao
NR. Culture tube and dimensions NR. 15 ml (1963); Rao (1965);
media/tube. Explant density NR Rao and Rangaswamy
(1971)
S. album Stem segments from 4-week-old in vitro Stem segments (size NR). 5 mm long NR Rao and Bapat (1978);
seedlings. Age of mother plant NR hypocotyls segments from seedlings for plant Bapat and Rao (1979);
regeneration and callus culture. Culture vessels Shekhawat et al.
NR (tubes from photos). Explant density NR (2008, 2010)
S. album Shoot segments and shoot tips of [20-year-old 5–6 mm diameter shoots cut into 5–6 mm long Shoot segments and shoot tips ? DW ? 0.1 % Lakshmi Sita et al.
trees explants (shoot segments). Shoot HgCl2 10 min ? SDW (multiple washes) (1979, 1980b);
tips = 5 mm. Culture vessels NR (text tubes Lakshmi Sita and
and Petri dishes from photos). Explant density Sobha Rani (1983);
NR Lakshmi Sita (1986)
S. album Green seeds (6-8 mm). Age of mother plant NR Endosperm from seeds. Other conditions, as for NR Lakshmi Sita et al.
Lakshmi Sita et al. 1979 (1980a); Lakshmi Sita
(1986)
S. album NR Achlorophyllous shoots derived from callus NR Bhaskar and Rao (1983)
S. album Juvenile shoots from mature tree, green fruits, The cut ends of shoot segments were sealed with Shoot segments, green fruit and mature Rao and Raghava Ram
mature seeds molten wax, Explant size, explant density, seeds ? detergent soap (conc. and duration (1983)
culture vessel type and dimensions NR (test NR) ? tap water (duration NR) ? 0.1 %
tubes and Petri dishes assumed from photos) HgCl2 (w/v) 5-8 min ? SDW (multiple
washes)
S. album Seeds ? in vitro seedlings. Age of mother plant 5 mm hypocotyl segments from 4-week-old HgCl2 (conc. NR) 5 min ? SDW (6X) ? Bapat and Rao (1984)
NR in vitro seedlings (shoot bud initiation). sown on BM
Hypocotyl segments with excised regenerated
shoot buds (for SE). Culture vessel NR (tubes
from photos). Explant density NR
S. album Juvenile shoots from 30-year-old trees. Cut 1 cm long nodal segments; 1.5 cm long Shoots ? detergent water ? RTW ? Rao et al. (1984)
ends immediately sealed by molten wax. internode segments. Culture vessel and explant 0.1 % HgCl2 8 min ? 6X SW
density NR
S. album 4-week-old in vitro seedling. Juvenile shoots Protoplasts from 5 mm long hypocotyls Protocol for seedling and stems NR. Leaves: Bapat et al. (1985)
from 30-year-old tree; first 3–4 young leaves segments of seedlings, callus culture of stem 0.1 % HgCl2 ? SDW (multiple washes)
from apex. Age of mother plant NR and leaves. Culture vessel NR (tubes and Petri
dishes from photos). Explant density NR
S. album Shoot segments of 20-year-old trees Shoot segments ? callus ? suspension NR Ozias-Akins et al.
culture ? protoplast isolation ? protoplast (1985)
culture ? SEs. Explant size, culture vessel
and explant density NR
S. album Shoot segments of 20-year-old trees. Protoplast from 4 to 5-days-old suspension NR Rao and Ozias-Akins
Embryogenic cell suspension cultures from culture. Subculture in 100 ml Erlenmeyer flask (1985)
3-year-old callus cultures containing 6–8 ml suspension/14 ml medium
(cell suspension culture). 2 ml/6 cm Petri dish.
1–5 9 105 protoplasts/ml (protoplast culture)
Planta (2016) 243:847–887
 Table 3 continued
Species Explant source Explant type, size and density; culture vessel Surface sterilization and preparation References

S. album Young shoots from 20-year-old mother plant Internode segments (5 mm long). Density and HgCl2 (conc. NR) 7–8 min ? SDW (multiple Bapat and Rao (1988)
immediately sealed at both cut ends culture vessel NR. SEs embedded and washes)
suspended at 10 beads/flask (100 ml)
S. album Stem internodes of young shoots from 20-year- Details not available from original protocol Details not available from original protocol Bapat and Rao (1992a);
old tree (Bapat and Rao 1989) (Bapat and Rao 1989) Bapat et al. (1996)
Planta (2016) 243:847–887

S. album 4-week-old fruits. Age of mother plant NR Endosperm. Exact size of explants, explant NR Sankara Rao et al.
density and culture vessels NR (1996); Shiri and Rao
(1998)
S. album Tender and thicker stem shoots. Age of mother Shoot tips, nodal explants, and leaves. Test tube RTW ? few drops detergent ? 1-2 DW wash Muralidharan (1997)
plant NR (borosilicate glass, 150 mm 9 25 mm or (for all explants) ? 0.05 % HgCl2 5 min for
100 mm 9 25 mm). Explant density NR tender stem; 0.1 % HgCl2 5 min for thicker
stem and leaves; 0.5–1.0 % HgCl2 5–10 min
for shoot tip and nodal explants. Rinses NR
S. album Seeds. Age of mother plant NR Zygotic embryo from 10 days in vitro NR Rai and McComb
germinated seeds (1997)
S. album Nodes and hypocotyls from in vitro-germinated Explant size, density NR. Culture vessel NR NR Das et al. (1998)
‘Elite seedlings. Age of mother plant NR
Kerala’
S. album Seeds and nodes from mature, superior tree. To compare with field-derived explants, in vitro- Seeds without pericarp ? single nodal Rugkhla and Jones
614, S. Seeds harvested from fruit with a green to derived leaves and nodal segments were also segments: commercial soap (1 %) (1998)
spicatum reddish pericarp used. Exact size of explants and age of mother 5–10 min ? 1 % NaOCl 15 min ? 5X SDW
S107 plants NR. Explant density and culture vessels
NR
S. album Stem of 30-year-old tree. Stem and hypocotyl segments: size NR NR Bapat and Rao (1999)
Age of seedlings NR
S. album Seeds ? in vitro seedlings. Seedling hypocotyls (10–15 mm long). Density NR Das et al. (1999)
Age of mother plant NR and culture vessel NR for embryogenic callus
production. Embryogenic callus used for
bioreactor (liquid media) and test tube (solid
media) trials
S. album Seeds ? in vitro seedlings. Seedling hypocotyls (size NR). Density and NR Das et al. (2001)
Age of mother plant NR culture vessel NR for embryogenic callus
production. Embryogenic callus used for liquid
media trials in 250-ml Erlenmeyer flasks
S. album Endosperm. Age of mother plant NR Endosperm callus. Culture vessel and explant NR Anil and Rao (2000)
density NR
S. album Endosperm from 4-week-old fruit Endosperm ? callus ? callus proliferation and NR Anil et al. (2000)
SE induction ? PEM ? SE maturation.
Culture vessel and explant density NR in
suspension culture. Torpedo and cotyledonary
stage SE at 21 days used
863

123
 Table 3 continued
864

Species Explant source Explant type, size and density; culture vessel Surface sterilization and preparation References

123
S. album 20-year-old candidate plus tree Nodal segments (1.5–2.0 cm, trimmed to RTW 30 min ? soap ? 4 % NaOCl Radhakrishnan et al.
1–1.5 cm after disinfection) 4 min ? 0.1 % HgCl2 10 min ? SDW (2001)
2–3 min
S. album Seeds ? in vitro seedlings. Age of mother plant Hypocotyls and stem nodes (size NR). Culture NR Ilah et al. (2002)
NR vessel NR
S. album Mature seeds from 50 to 60-year-old elite tree Zygotic embryos. 200-ml glass jars with 35 ml Seeds: deionized water overnight ? 2 % Rai and McComb
medium/jar for induction of somatic NaOCl ? 0.005 % Tween-20 (2002)
embryogenesis. 25 9 100 mm test tubes with 10-15 min ? 3X SDW. Seeds air-dried in
15 ml medium/test tube for somatic embryo laminar flow bench for 10 min prior to
conversion to plantlets excision of zygotic embryos
S. album Seeds. Age of mother plant NR GA3 treated seeds. Culture vessel and density Seeds ? 0.15 % HgCl2for 8 min ? SDW Sanjaya and Rai (2003)
NR
S. album Leaves from 3 to 4-week-old in vitro seedlings Intact leaves or half leaves (sections cut Seeds: 0.1 % HgCl2 4 min ? 3X SDW Mujib (2005)
perpendicular to the mid-vein) 5–15 mm long;
adaxial and abaxial surfaces of the laminae
tested separately
S. album In vitro shoots derived from in vitro subcultured Shoot tip size and age of donor plant NR. Jam NR Primawati (2006)
shoots. Age of mother plant NR bottles (13 cm high, 7 cm diameter)
S. album Nursery seeds (for rootstocks; 2006a) and Nodal shoot segments (2.5–3.5 cm long) with Decoated seeds for in vitro rootstocks (2006a): Sanjaya et al. (2006a, b)
50–60-year-old tree (high-yielding oil: dormant axillary buds, harvested in Nov-Jan. 0.05 % GA3 overnight ? 0.075 % HgCl2
4–5 %; 2006a, 2006b) for scions 25 9 100 mm test tubes (one seed/tube; 25 ml 6-7 min ? 6-8X SDW. Nodal segments from
medium/tube) 50-60-y-old tree for scions (2006a, 2006b):
70 % EtOH 30-40 s ? 0.075 % HgCl2
6–7 min ? 6-8X SDW
S. album Mature stem segments from elite tree (age NR) 1 cm long stem segments (nodes or internodes?). 0.1 % HgCl2 5 min ? 2-3X SDW Shekhawat et al. (2008)
100 ml Erlenmeyer flasks: 200 mg of friable
embryogenic callus tissue in 20 ml of liquid
medium
S. album Young shoots from elite mature tree (age NR) 1 cm long stem segments, 80 9 150-mm Young shoots ? RTW 30 min ? 0.1 % HgCl2 Ma et al. (2008)
(diameter 9 height) culture jars, explant 10 min ? SDW 3X
density NR
S. album Top shoots, young leaves, and seeds. Age of Top shoots, leaves of 0.5 cm2, seeds Top shoots and leaves ? 0.1 % HgCl2 Mo et al. (2008)
mother plant NR 10 min ? SDW 5X
Seeds ? RTW ? peel pericarps ? 0.1 %
HgCl2 10 min ? SDW 5-6X
S. album Fruits collected in May from tree (age NR) Seed. Viability tested with 2,4,5-triphenyl Fruits soaked overnight in water ? DW Nikam and Barmukh
tetrazolium chloride (TTC) before use. 1 or 2 3X ? soap solution 30 min ? 0.1 % HgCl2 (2009)
seeds/culture tube 3 min ? SDW 3X. Seeds ? 0.1 % HgCl2
3 min ? SDW 3X ? 4 mM GA3
12 h ? SDW 3X
Planta (2016) 243:847–887
 Table 3 continued
Species Explant source Explant type, size and density; culture vessel Surface sterilization and preparation References

S. album Immature seeds and mature seeds, 10-year-old ZEs Seeds ? peel pericarps ? 70 % EtOH Mo et al. (2010)
tree 1 min ? 5-10 % NaClO 5–20 min ? 0.1 %
HgCl2 5-15 min ? SDW 4-5X
S. album Seeds of 8-year-old plants ? in vitro seedlings Internodes (8–10 mm) from 2-months-old Seeds: RTW 20 min ? * 2 % Tween-20 Janarthanam and
seedlings. 25 9 150 mm culture tubes. 1 min ? SDW ? 0.1 % HgCl2 5 min ? 4X Sumathi (2011);
Planta (2016) 243:847–887

Explant density NR SDW. Pericarp removed Janarthanam et al.

(2012)
S. album Potted greenhouse plants (age NR) Nodes, internodes, juvenile leaves and shoot tips Seeds: Samantray and Upadhyaya (2010) Revathy and Arumugam
protocol. 4 explants: RTW ? 70 % (2011)
EtOH ? 0.1 % HgCl2 5 min ? 3-5X SDW
S. album, 10–15 cm side branches of 10–12 year-old trees Juvenile nodal segments (5–7 cm long) ? cut Leaf nodes ? tap H2O ? bleach (4.2 % Baiculacula (2012)
S. yasi, collected in Jan and May–June into 1 cm nodal segments 0.5 cm above and NaClO) time NR ? tap
R1 0.5 cm below the nodes ? leaf nodes. Culture H2O ? SDW ? 70 % EtOH
hybrids in McCartey bottles or polycarbonate tubes. 1 min ? 100 mg/l citric acid ? 4.2 %
Explant density NR NaClO ? 2–3 drops Tween time NR ? SDW
a few times
S. album Leaves from 3 to 4-week-old in vitro and ex Leaf disks (squares of 5–8 mm). Petri dishes Seeds: 0.1 % HgCl2 4 min ? 3X SDW. Ex Bele et al. (2012)
vitro seedlings vitro leaves (1st and 2nd from apex): collected
in DW ? RTW 30 min ? DDW ? 2 %
Tween-20 20 min ? 1 % Bavistin 10 min on
shaker at 30 rpm ? 70 % EtOH
1 min ? 0.2 % HgCl2 10 min (in 100 psi
vacuum) ? 4-5X SDW
S. album Hypocotyl segments from 5-week-old seedlings Hypocotyls and crown section (3–10 mm long), Seeds: 2.5 % NaOCl 2 h ? 3 washes in SW Crovadore et al. (2012)
cotyledon and leaf pieces (10–16 mm2 surface (20, 10, 5 min each) ? testa removed for
area), root segments (3–5 mm long) from 5 in vitro SG
and 10 DAG. Test tubes and Petri dishes
S. album Leaves from mature trees (age NR) 10 cm2 trimmed to 4 cm2. Final sterilized 0.1 % Tween-20 5 min ? 4–5 washes in Singh et al. (2013)
explant = 10-15 mm. One explant per test DW ? 1 g/l Bavistin, 0.2 g/l cefotaxime, and
tube (150 9 25 mm) 0.2 g/l kanamycin 10 min ? 3X
SDW ? 0.1 % HgCl2 3–4 min ? 6-8X SDW
S. album Nodes from young branches (age NR) 2-3 cm nodes According to Singh et al. (2013) Singh et al. (2015)
865

123
866 Planta (2016) 243:847–887

BM basal medium (in mg/l: KINO3 (1900), NH4NO3 (1650) CaC12.2H2O (440), MgSO47H2O (370), KH2PO4 (170), MnSO44H2O (25), H3BO3 (10), ZnSO47H2O (13.9), Na2EDTA (18.6),
myo-inositol (100), nicotinic acid (5), folic acid (5), glycine (2), pyridoxine–HCl (0.5), thiamine-HCI (0.5), biotin (0.05), sucrose (20 g/l), Difco bacto agar (6 g/l) modified WB according to

chloride, NaOCl sodium hypochlorite, NR not reported in the study, PEM pro-embryogenic masses, rpm revolutions per minute, RTW running tap water, SDW sterilized (by autoclaving)
Rangaswamy (1961)), CW chlorine water, DAG day(s) after germination, DW distilled water, DDW double distilled water, EtOH ethyl alcohol (ethanol), GA3 gibberellic acid, HgCl2 mercury
et al. (2014) found that the inter-collapsed layers, an
important structure within haustoria, might be involved in
Peeris and Senarath cell inclusion and energy concentration at the inner
meristematic region and these cell inclusions and energy
References

were recycled to affect penetration, to reinforce the phys-

(2015)

ical connection between the haustorium and host root, and

to supply space for haustorial development in S. album.
Haustoria can form within 30 days after germination
15 min ? 70 % EtOH 10 min ? 3X SDW

without the need for induction by haustoria-inducing fac-

Soap water 5 min ? RTW 30 min ? 2 g/l

tors (HIFs) (Barrett and Fox 1997). The molecular mech-

Captan TM 30 min ? 15 % CloroxTM

anisms involved in haustorial development of S. album

Surface sterilization and preparation

were explored based on global transcriptome analysis by de

novo assembly (Zhang et al. 2015). A substantial number
of differentially expressed genes (DEGs) were involved in
cell wall metabolism and protein metabolism, as well as
mitochondrial electron transport functions during hausto-
rial development. This is in agreement with anatomical
observations of haustoria development in which prolific
protein synthesis actively occurred and cell walls in the
host root tissue were degraded and cell membranes disin-
tegrated in the interface of haustoria and the host root
(Zhang et al. 2012a). Radomiljac et al. (1999b) noticed that
(green pericarp still attached to mother plant)

the number of haustoria formed by Santalum on roots of

Explant type, size and density; culture vessel

2 cm nodes and mature and immature seeds

different hosts was poorly correlated with host quality. A

previous study indicated that endogenous levels of IAA,
CK, GA3, and ABA were higher in haustoria than in
seedling roots, suggesting that phytohormones play an
important role in haustorial development (Zhang et al.
2012a, 2015). The authors found that auxin signal trans-
duction might be essential for haustorial initiation and that
GAs play an important role throughout haustorial devel-
opmental processes (Zhang et al. 2015). This result also
provides vital clues for artificially increasing the number of
haustoria to improve growth by exogenous application of
plant growth regulators in an in vitro environment. Genes
encoding nodulin-like proteins might be important candi-
plants was treated with fungicide (Tilt 10 ml/l)
Nodes from 2-year-old plant potted along with

(composition of Albert solution or Reference

once in a week and Topsin 5 g/l and Thiram
1.4 g/l spreyed alternatively to mother plant.

date genes for haustorial morphogenesis in S. album based

Desmodium and Alternanthera sp. Mother

on nodulation genes (ENODs) and related proteins found in

Albert solution 50 ml/l applied weekly

is NR). Explants collected in morning

distilled water, SW sterile water, ZE zygotic embryo

legume nodule formation (Legocki and Verma 1979; Kalo

et al. 2005; Smit et al. 2005; Zhang et al. 2015). The elu-
cidation of this interesting question in the future probably
could explain one cause for haustorial formation without of
the presence of HIFs in the root hemi-parasite, S. album
(Zhang et al. 2015).
Explant source

Applied biotechnologies
Table 3 continued

In vitro culture, tissue culture,

and micropropagation
S. album
Species

This section provides an overview of the principal trends

and key observations of the literature as described in

123
 Table 4 In vitro conditions for tissue culture studies of sandalwood (chronological listing)
Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

S. album mWM ? 20 % CM ? 400 mg/l CH (SGM). PP NR. Diffused light (LI NR). SG produced seedlings in 5 weeks. In seed Rangaswamy and Rao
mWM ? 2 mg/l 2,4-D ? 5 mg/l Kin ? 0.25 % 25 ± 2 °C. 50–60 % RH culture, 30–40 % of seeds initially started to (1963); Rao (1965)
YE (seed culture). pH 5.8. 4 % sucrose. Difco germinate but did not convert to seedlings.
bacto agar 0.8 % 60–70 % of cultures showed no sign of
Planta (2016) 243:847–887

germination. Callus formed from endosperm

after 3–4 weeks: subculture produced friable
callus in 4–6 weeks. Excised ZEs unresponsive
S. album mWM ? CH or CM or CH ? CM (SGM and PP NR. Diffuse light (400–600 lux). SG on BM ? CH or CM or both. ZEs and Rao and Rangaswamy
subculture for cotyledon-derived callus). 25 ± 2 °C. RH NR endosperm did not respond. Plantlets obtained (1971)
mBM ? 2,4-D or Kin or YE alone, or in from embryoids formed from ZE-derived callus.
combination (ZE and endosperm culture). Germinated seedlings did not develop haustoria.
mBM ? CH ? CM (excised ZE). CH 400 mg/ Undifferentiated callus from cotyledons
l; CM 20 %; YE 0.25 %. pH 5.8. Carbon source produced embryoids then plantlets. Excised ZE
NR. 0.8 % Difco bacto agar cultured for 3–4 weeks formed normal seedlings
then plantlets after 2 weeks
S. album BM or BM ? 1 mg/l [IAA, IBA, NAA, NOA, PP NR. Fluorescent light. 1000 lux. Only hypocotyl explants responded well on BM Rao and Bapat (1978)
Kin, BA, Zea, SD 8339, or AdS], 0-2 mg/l 25 ± 2 °C. RH 55–60 % fortified with cytokinins. Shoot segments did not
(NAA ? Kin) or 0–2 mg/l (NAA ? BA) (SIM). respond. Callus prolific on 2,4-D ? BA media.
BM ? 1 mg/l 2,4-D or pCPA or 1 mg/l (2,4- 2 mg/l BA produced 15–20 buds/explant.
D ? Kin) or 1 mg/l (2,4-D ? BA) (CIM). Complete plantlets regenerated in 4–8 weeks.
BM ? 0.5 mg/l NAA ? 0.5–5 mg/l IBA (RIM) Rooting obtained only in few cultures (data NR).
Acclimatization NP
S. album Liquid BM (same as Rao and Bapat PP NR. Fluorescent light. 1000 lux. After 4 weeks of callus subculture, globular SEs Bapat and Rao (1979)
1978) ? NAA or IAA (1 mg/l), 2,4-D (1 mg/l), 25 ± 2 °C. RH 55–60 % converted to plantlets and planted in soil (30 %
BA (1 mg/l), GA**** (1 mg/l), CM (10 %), on BM ? 1 mg/l IAA). Plantlets acclimatized in
NAA (1 mg/l ? BA (1 mg/1), or CM soil after 8 weeks (watered with Hoagland’s
(10 %) ? CH (400 mg/1) (SEMM) (liquid solution; other acclimatization conditions NR)
culture in gyrator shaker at 120 rpm
continuously for 4 w). Solidified BM ? BA (1-
12 mg/l), 2,4-D (1 mg/l), 2,4-D (1 mg/l) ? CM
(10 %), 2,4-D (1 mg/l) ? CH (400 mg/1), or
2,4-D (1 mg/l) ? YE (400 mg/l) (SEIM).
BM ? IAA (1 mg/1), NAA (1 mg//1) or BA
(1 mg/1) (SEMM)
S. album MS ? 1 mg/l 2,4-D ? 0.2–0.5 mg/l Kin (CIM). 12-h PP. CWFT. 400 lux. 26 ± 1 °C Callus formed from shoot tips and shoot segments. Lakshmi Sita et al.
MS ? 1 mg/l GA3 (SEIM). MS or MS ? 1 mg/ Embryoids formed within 4–5 weeks from (1979)e; Laxmi Sita and
l GA3 ? 1-2 mg/l IAA ? 0.1 mg/l 2,4-D or callus. Organogenesis not quantified. Sobha Rani (1983);
MS ? 0.5 mg/l BA ? 1 mg/l NAA (SEMM). Acclimatization NP. Callus used to isolate Lakshmi Sita (1986)
WB or WB ? 0.5 mg/l IAA (RIM). pH 6.0. 2 % protoplasts in 1983 study
sucrose. Gelling agent NR
867

123
 Table 4 continued
868

Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

123
S. album MS ? 1-2 mg/l 2,4-D or MS ? 0.5–2 mg/l As for Lakshmi Sita et al. 1979 Embryoid formation in 4–6 weeks. Organogenesis Lakshmi Sita et al.
BA ? 1 mg/l NAA (CIM). MS ? 1 mg/l 2,4-D not quantified. Acclimatization NP (1980a)e, b; Lakshmi
(callus subculture). MS ? 1–2 mg/l GA3 Sita (1986)
(SEIM). MS ? 0.3 mg/l BA ? 1 mg/l
IAA ? 0.3 mg/l Kin ? 1 mg/l GA3 (SEMM).
WB ? 0.5 mg/l IAA (RIM). Shake cultures
using callus in 250-ml flasks at 70 rpm and pH
5.2 (1980b). Other conditions as for Lakshmi
Sita et al. (1979), or unclear
S. album 1–3 mg/l NAA (RIM). Other conditions NR NR In vitro-derived shoots cultured in pots with host Bhaskar and Rao (1983)
Cassia siamea, leading to plantlet formation
S. album MS ? 1 mg/l 2,4-D alone or in combination with Continuous light. 1000 lux. 25 ± 2 °C. Within 3 weeks, callus formed on surface of Rao and Raghava Ram
0.2 mg/l Kin (CIM). MS ? 1 mg/l 55 % RH. At SE germination stage, endosperm. During 6 weeks, callus proliferated (1983)
IAA ? 1 mg/l BA (SEIM). MS ? 1 mg/l cultures kept in dark for 72 h then well. SE formation observed within 4 weeks
IAA ? 0.5 mg/l IBA ? 0.5 mg/l GA3 (SE transferred to continuous light for 4–6 from callus obtained on CIM. Plantlets with 2–3
germination). pH, 5.8. 2 % sucrose (CIM) or weeks pairs of leaves with roots developed within 4
5 % (SEIM, SE germination). 0.6 % agar weeks and transferred to vermiculite then
acclimatized in earthen pots with soil
S. album mBM ? 1 mg/l NAA or 1 or 2 mg/l BA (SIM). PP NR. CWFT. 1000 lux. 25 ± 2 °C. RH Normal plantlets produced (time period NR). Bapat and Rao (1984)e
mBM ? (0.5 mg/l NAA ? 5 mg/l IBA) or 55–60 % Acclimatized for 3 w in Hoagland’s nutrient
1 mg/l BA (RIM). mBM ? 0.5 mg/l or 1 mg/l solution (Hoagland and Arnon 1938). 10 %
IAA (SEIM). mBM ? 0.4 % YE ? 400 mg/l survived in field without host plant. 90 % of the
CH ? 400 mg/l CA (SEMM). mBM ? 1 mg/l basal region of hypcotyl explants differentiated
IAA or NAA (PCM) into buds. Insignificant effect of seedling age.
Rapid response on liquid medium than on agar
medium. 100 % response on hypocotyl explant
orientated with root end in direct contact with
medium. 20 cm long plantlets obtained on RIM.
4 % sucrose ? 1650 mg/l NH4NO3 best for SE
induction
S. album MS ? 0.5 or 1 mg/l BA (SIM). MS ? 1 mg/l 2,4- Initial 72 h darkness for root induction. Shoot buds obtained from nodes did not respond Rao et al. (1984)e
D or MS ? 1 mg/l 2,4-D ? 0.2 mg/l Kin All other conditions as in Bapat and Rao to rooting. Shoots obtained from SEs of
(CIM). MS ? 1 mg/l IAA ? 1 mg/l BA (1984) internode-derived callus resulted in successful
(SEIM). 2 % sucrose. 0.7 % agar. pH 5.6. plantlet conversion and 10 % survival in field
MS ? 1 mg/l IAA ? 0.5 mg/l IBA ? 0.5 mg/l conditions
GA3 (SEMM). 5 % sucrose. 0.6 % agar
Planta (2016) 243:847–887
 Table 4 continued
Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

S. album MS ? 1 mg/l 2,4-D or BA (CIM for hypocotyl). As for Bapat and Rao (1984) Effect of explant source on protoplast isolation Bapat et al. (1985)e
MS ? 1 mg/l 2.4-D or 1 mg/l 2,4-D ? 0.5 mg/l efficiency. Maximum number of SEs induced on
Kin (CIM for stem). MS ? 1 mg/l 2,4-D (PCM stem-derived callus cultured on ‘ MS ? 1 mg/l
for hypocotyl callus). MS ? 1 mg/l IAA. Plantlet conversion from stem callus-
Planta (2016) 243:847–887

IAA ? 1 mg/l BA (SEIM from stem callus). derived SEs. No SE induction from leaf- and
Modified V47 medium ? 1 mg/l 2,4- hypocotyl-derived callus
D ? 1 mg/l NAA ? 1 mg/l BA (protoplast
proliferation medium for stem callus).
MS ? 1 mg/l IAA or MS ? 1 mg/l
IAA ? 1 mg/l BA, or ‘ MS ? 1 mg/l IAA or
‘ MS ? 10 % CM ? 500 mg/l CH (SEIM
from stem callus). SEMM NR. Medium for leaf-
derived protoplast culture NR
S. album CIM NR ? suspension culture (composition NR SEs obtained from protoplast derived culture. Ozias-Akins et al. (1985)e
NR) ? subculture (0.5 or 1 mg/l 2,4- Organogenesis not NR but complete plants
D) ? liquid or gelled media (0.1–2 mg/l BA) claimed to have formed from SEs
(details NR) ? protoplast isolation ? V47
medium (750 mOs/kg H2O, adjusted using
mannitol) (PCM) ? MS medium (IBA ? BA
conc. NR). Gelling agent, pH, carbon source and
other conditions NR
S. album MS ? 2,4-D (0.5–2.5 mg/l) ? 1 mg/l folic acid PP NR. Continuous diffused light. LI NR. Club and heart shaped stages of SE obtained from Rao and Ozias-Akins
(liquid SEIM). 3.4 % sucrose (CSC). Liquid MS 26 °C. Gyrotary shaker at 150 rpm (for different callus cell lines on media with BA and (1985)
or MS ? agar either with Zea (0.2, 0.35 mg/l), suspension culture) 2,4-D. Secondary SEs obtained from cell
BA (0.1, 0.5, 1.0, 2.0 mg/l) or BA (0.1, 0.5, 1.0, suspension culture on MS containing 0.1 and
2.0 mg/l) ? 0.01 mg/l 2,4-D (SEM). MS or ‘ 1 mg/l BA. SE successfully converted to
MS, alone or MS or ‘ MS ? BA (0.1 or 1 mg/ plantlets. SEs from protoplast did not form roots
1) or 1 mg/1 IAA or 1 mg/1 GA3 ? 0.2 mg/l
BA (SEMM). MS ? agar or filter-paper bridge
on mWBM ? 0.5 mg/l IAA or combination of
0.1 mg/l IAA ? 0.5 mg/l IBA ? 0.5 mg/l GA3
(SGM). Protoplast culture: V47 medium (750
mOs/kg H2O). pH 5.8 (PCM). MS ? 1 mg/l
IAA ? 1 mg/l BA or 400 mg/l casamino acid
(SEIM from protoplasts). MS ? 1 mg/l BA
(SEMM from protoplasts). WM ? 0.5 mg/l
IAA (plantlet conversion from callus-derived
SE). ‘ MS (only major elements?) or
WM ? 1 mg/l IAA or 0.5 mg/l IBA or 1/3 mg/l
GA3 ? 4 or 5 % sucrose ? 0.6 % agar
(germination for SE from protoplasts).
Subculture cell suspension cultured every 4–5
days
869

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sandalwooda acclimatization and variation

123
S. album MS ? 1 mg/l 2,4-D (CIM). MS ? 0.3 mg/l IAA Suspension of beads under continuous Embryogenic cells encapsulated in 3 % alginate Bapat and Rao (1988)e
or 0.5 mg/l GA (SEIM, as a suspension culture). light. 950 lux. 25 ± 2 °C bead showed plantlet conversion after 16 weeks.
MS ? 0.5 mg/l BA ? 0.5 mg/l IAA (secondary However, low synseed germination (10 %)
SEs). Gelling agent and pH NR. 5 % sucrose for
SE germination
S. album MS ? 87.6 lM sucrose ? 4.52 lM 2,4-D (CIM). Suspension of beads under continuous SEs isolated and desiccated for 10, 20 or 30 days. Bapat and Rao (1992a);
MS ? IAA 2.85 lM ? BA light. 1000 lux. 25 ± 2 °C. 80 rpm One lot of desiccated SEs encapsulated in 3 % Bapat et al. (1996)e
2.22 lM ? 87.6 lM sucrose (SEM). sodium alginate gel (control = non- (using protocol
MS ? 87.6 lM sucrose or WM ? 58.4 lM encapsulated SEs). Both encapsulated and non- originally from Bapat
sucrose (SEIM) for 4 w. WM ? 58.4 lM encapsulated SEs showed revived growth after and Rao (1989)
sucrose ? 18.92 lM ABA (plantlet conversion) rehydration on WM and developed into plants.
2. Synthetic seed: Encapsulation of SE according The desiccation tolerance and regeneration of
to Bapat and Rao (1988) plants was dependent on the pre-treatment given
to SEs. In Bapat et al. 1996, the addition of a
cyanobacterial extract (Plectonema boryanum
strain UTX594) promoted callus formation in
the absence of PGRs
S. album MS ? 1 mg/l 2,4-D ? 1 mg/l BA or 2 mg/l 2,4- NR 4 stages of somatic embryogenesis claimed, with Sankara Rao et al. (1996)e
D ? 0.5 mg/l Kin (CIM). MS ? 1 mg/l 2,4- distinct biochemical profiles
D ? 3 % sucrose (SEIM). PGR-free MS ? 2 %
mannitol (SE differentiation). PGR-free ‘MS
with or without mannitol (SE maturation). 0.6 %
agar
S. album MS ? 0.5 lM BA ? 0.5 lM Kin (SIM). 16-h PP. CWFT. 18 lE m-2 s-1. Callus produced from intermodes on CIM. 4 shoot Muralidharan (1997)
MS ? 0.5-4 lM 2,4-D (CIM). MS ? 0.5–2 lM 25 ± 2 °C buds/nodal explant on SIM. Shoot tips failed to
IBA (RIM). pH 5.7. 2 % sucrose. 0.5 % agar. respond. Plant regeneration was not possible
Subcultured every 4 weeks
S. album MS ? 20 % CM ? CH or 1 lg/l Kin (SGM; 16-h PP. Light source NR. 40 lmol m-2 91.6 % of explants induced SEs, forming 14.23 Rai and McComb 1997e,
1997). MS ? 2 mg/l BA (1997) or 4.5 lM TDZ s-1. 25 ± 1 °C SEs/explant after 8 w. 91.6 % of primary SEs (2002)e
(2002) (SEIM). PGR-free MS (SEMM). formed secondary SEs, forming 20.5 SEs/
‘MS ? 0.5 mg/l IAA (1997) or 2.8 lM GA3 primary SE after 8 weeks. 81.2 % of SEs
(2002) (SE conversion). pH 5.8. 3 % sucrose. germinated after 8 w. From the same treatment,
0.8 % agar (SEIM, SEMM) or 0.15 % Phytagel 72.3 % of plants survived in the field after initial
(SE conversion) acclimatization in sand ? soil ? FYM (2:1:1)
and 1-2 seeds of red gram (Cajanus cajan) as the
primary pot host. Most details not explained in
1997 paper
S. album MS minerals ? B5 vitamins ? 0.5 mg/l BA PP NR. CWFLT. 1500 lux. 26 ± 2 °C. 25–60 shoots/SEs from hypocotyls, and only 3–5 Das et al. (1998)e
‘Elite (SIM). MS minerals ? B5 vitamins ? 0.5 mg/l 60–70 % RH from nodes
Kerala’ BA ? 0.5 mg/l IAA (SEIM). pH 6.0. 4 %
sucrose
Planta (2016) 243:847–887
 Table 4 continued
Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

S. album MS ? 5 lM BA (SIM). MS ? 5 lM SEIM: PP and light source NR. S. album nodal segments: 100 % explants formed Rugkhla and Jones (1998)
614, S. BA ? 2 lM GA3 (for leaf enlargement). 2 lmol m-2 s-1. SEMM: 16-h PP. green SEs, 72 % formed white SEs (2 lM
spicatum MS ? 1-2 lM TDZ (SEIM). MS ? 6 lM Light source NR. 50 lmol m-2 s-1. TDZ), 64 % formed friable embryogenic tissue
S107 IAA ? 1 lM Kin (SEMM). MS ? 6 lM GA3 25 ± 1 °C (2 lM TDZ). S. album seed: 0 % explants
Planta (2016) 243:847–887

(SE germination) ? 3 lM GA3 (SE elongation) formed green SEs, 96 % formed white SEs and
(S. album) and with 250 mg/l CH ? 5 % CW friable embryogenic tissue (2 lM TDZ). S.
(S. spicatum). 6-w subculture in SEIM, then 3-w spicatum nodal segments: 20 % of explants
subculture during SE maintenance. pH 5.8. 2 % formed primary and secondary SEs. Rooting
sucrose. 0.2 % gelrite was problematic but overall plantlet growth was
improved on liquid medium. Acclimatization
NP
S. album MS ? 1 mg/l 2,4-D ? 1 mg/l BA (CIM). 14-h PP. CWFT. 5 (callus, SEs) ? 200 Pricked torpedo–cotyledonary stage SEs Shiri and Rao (1998)e
MS ? 1 mg/l 2,4-D for 3 w (SEIM). PGR-free (plantlets) lE m-2 s-1. 26 ± 2 °C (0.75–1.2 cm) from a Sankara Rao et al. (1996)-
MS ? 2 % mannitol (SE differentiation). PGR- derived line. Plantlets ardened to the greenhouse
free ‘MS with or without simply by growing on paper bridges overlaying
mannitol ? WB ? 0.5 mg/l IAA liquid WB. SEs genetically transformed with
(SE ? plantlets). 3 % sucrose. Agar conc. NR Agrobacterium tumefaciens harboring
LBA4404/pKIWI105 (see text for details).
Success of regeneration and genetic
transformation not quantified
S. album MS ? 1 mg/l IAA (SEIM). No other SEIM 16-h PP. CWFT. 24 lmol m-2 s-1. 3000 seedlings derived from bioreactor in 6 w (vs Das et al. (1999)e
conditions described. Initial pH (bioreactor) 5.8. 24 ± 2 °C. 60–70 % RH 800 in 12 w on solid medium): 59.3 % vs
2000 ml in a 3.5-L airlift bioreactor (25 g 88.9 % abnormality and 31.1 vs 7.7 SEs/10 ml
embryogenic callus/l). MS ? 1 mg/l in former vs latter, respectively
BA ? 0.5 mg/l ABA (bioreactor). Air-flow = 1
vvm. 3 % sucrose
S. album MS ? 1 mg/l BA ? 1 mg/l 2,4-D (CIM). PP and light source NR. Diffused light of Embryogenic callus consisted of two types of Anil and Rao (2000)
MS ? 1 mg/l 2,4-D (SEIM). Liquid MS (PGR- 5 lEm-2 S-2. 26 °C ± 2 °C cells: (a) more cytoplasm that formed clumps;
free) ? 2 % mannitol (SEMM). Orbital shaker (b) less cytoplasm and elongated shape. Torpedo
at 100 rpm. 3 % sucrose. 3.9 mM CaCl2. and bipolar stages of SE collected after 21 days
Gelling agent and pH NR of culture from first type of cells
S. album MS ? 4.52 mM 2,4-D (CIM; 2-w subcultures). Light conditions NR. 25 ± 2 °C 57.35 % optimization of SE production. SEs Das et al. (2001)e
MS ? 2.85 mM IAA ? 3.99 mM BA (SEIM 1; produced in 14 days
subcultures NR). WPM ? 2.85 mM
IAA ? 3.99 mM BA ? 40 mM
glutamine ? 0.33 M mannitol ? 11.6 mM
nitrate ? 7.89 mM ammonium ? 1.31 mg/l
ABA (SEIM 2; subcultures NR). 50 ml medium
in 250-ml Erlenmeyer flasks (12 g embryogenic
callus/l; 75 rpm). 3 % sucrose (CIM, SEIM 1) or
4 % (SEIM 2). pH NR
871

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sandalwooda acclimatization and variation

123
S. album MS ? 4 mg/l BA (CIM, SIM). MS ? 3 mg/l IBA 16-h PP. Light source NR. 1000 lux. Callus induced after 3 weeks. The inclusion of a Radhakrishnan et al.
(RIM). pH 5.7. 2 % sucrose. 0.9 % agar 25 ± 2 °C seed extract of Cajanus cajan (red gram) in RIM (2001)
did not improve rooting
S. album BM NR ? 2.26 lM 2,4-D or 2.68 lM pCPA PP and light source NR. 24 lmol m-2 2-w-old initial callus induced on CIM transferred Ilah et al. (2002)e
(CIM). BM NR ? 2.70 lM NAA ? 2.22 lM s-1. 24 ± 2 °C. 70–80 % RH to SEIM. Liquid medium superior to solid
BA (SEIM). WPM (PGR-free) (SEMM). medium (depending on stage of SE). 53–68 %
5.71 lM IAA (RIM). pH, carbon source, gelling of cultures showed abnormalities (aggregation
agent NR of pro-embryos and SEs, growth arrest,
browning of SEs, SEs with no or poorly
developed roots, roots with undeveloped shoot).
83 % rooting
S. album PGR-free MS (seed germination). 16-h PP. Light source NR. 40 lmol m-2 45-d-old seedlings used as rootstock in Sanjaya and Rai (2003)
MS ? 11.12 lM BA (SIM). 3 % sucrose. 0.6 % s-1. 25 ± 2 °C micrografting. The larger the scion used, the
agar more successful the micrograft: 1-2 cm scions
resulted in 60 % graft success
S. album MS ? 0.44 and 2.22 lM BA (SIM). pH 5.8 (MS) 16-h PP. CWFT. LI NR. 25 ± 2 °C Direct shoot formation from leaf explants Mujib (2005)
or 5.2 (WPM). WPM ? 5.71 lM IAA (RIM). (13–20/explant) within 20–25 days. No auxins
3 % sucrose. 0.8 % agar could induce shoots. Approx. 4-fold higher
shoot bud production in liquid medium than in
solid medium. Epiphyllous shoots produced
when explants placed in horizontal position, but
not when in the vertical position, but shoots
from these buds developed slowly
S. album MS ? 1.5 mg/l BA ? 0.2 mg/l Kin ? 100 mg/l NR. 1.4 shoots/shoot tip. No rooting or acclimatization Primawati (2006)
glutamine (shoot tip development medium). 3 % experiments. 17 % browning and contamination
sucrose
S. album PGR-free MS (SGM for rootstocks). 12-h PP. CWFT. 60 lmol m-2 s-1. Seeds were germinated in vivo and in vitro to Sanjaya et al. (2006a)
MS ? 0.53 lM NAA ? 11.09 lM BA (SIM). 28 ± 1 °C assess micrografting, and 45-days-old seedlings
MS (liquid) ? 2 % sucrose on paper bridges served as rootstocks. In vitro shoots (0.5–2.0 cm
(post-grafting). Subculture every 4 w. pH 6.0. long) derived from third subculture were used as
3 % sucrose. 0.6 % agar scions. 60 % of grafts were successful. 8-week-
old grafted plantlets were successfully
acclimatized (% success NR) in sterilized
soilrite (60 lmol m-2 s-1; 25 ± 1 °C)
S. album MS ? 0.53 lM NAA ? 11.09 lM BA (SIM). 12-h PP. CWFT. 60 lmol m-2 s-1. 4.92 shoots/sterilized nodal shoot segment in Sanjaya et al. (2006b)
SIM ? 283.93 lM AA ? 118.1 lM 28 ± 1 °C. 60–65 % RH initial culture and 4.63 shoots/in vitro nodal
CA ? 104.04 lM cysteine ? 342.24 lM shoot segment during shoot multiplication.
glutamine ? 10 % CM (SMM). MS ? 2 % 41.7 % of shoots could root in vitro. 50 % of
sucrose after 48-h pulse in 98.4 lM IBA. shoots could root in soilrite after treatment with
Subculture every 4 w. pH 6.0. 3 % sucrose. 1230 lM IBA. 100 % survival of field-grown
0.6 % agar plantlets
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Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

S. album MS ? 5.0 lM BA ? 0.5 lM NAA (adventitious 14-h PP. CWFT. 40 lmol m-2 s-1. 5.7 shoots/shoot explant after 1 month of culture. Ma et al. (2008)
shoot propagation) ? monthly subculture. 25 ± 2 °C Some adventitious roots within 1–3 months.
MS ? 250 lM IBA (RIM). pH 5.6. 3 % 8 % of shoots formed roots. 95 % survival of
sucrose. 0.6 % agar acclimatized plantlets in sand: peat soil: organic
Planta (2016) 243:847–887

matter (1:1:1) after 2 months of acclimatization

in greenhouse and watered each day.
Greenhouse growth conditions NR
S. album MS ? 1 mg/l BA, MS ? 1 mg/l 2,4-D, or 16-h PP. CWFT. 1500 lux. 25 ± 1 °C On MS ? TDZ for SE induction: 91.2 % Mo et al. (2008)
MS ? 0.5 mg/l TDZ (CIM). Subcultured twice induction. SE germination = 100 % on
for globular SE induction. MS ? TDZ, MS ? GA3 ? 4 % sucrose, but no roots formed
MS ? GA3, or ‘ MS ? GA3 (SE germination).
pH NR. 4 % sucrose. Gelling agent NR
S. album MS ? 1 mg/l 2,4-D ? 0.2 % Phytagel (SEIM). 16-h PP. Light source NR. 45 lmol m-2 Embryogenic suspension cultures genetically Shekhawat et al. (2008,
4-8-w-old callus transferred to mMS (liquid) at s-1. 25 ± 2 °C transformed with Agrobacterium tumefaciens 2010)
100 rpm (suspension cultures). MS ? 1 mg/l harbouring pCAMBIA 1301 (2008) or
BA ? 1 mg/l IAA ? 5 mg/l hyg (SEMM). pD35SHER (2010) (see text for details). Shoots
‘MS ? 1 mg/l BA ? 0.1 mg/l GA3 (SIM). formed in 10–15 % of transformed tissues, with
WB ? 0.1 % AC (plantlet development). pH 100 % conversion to plantlets
5.7. 3 % sucrose
S. album MS ? 4 lM BA (SGM). pH 5.8. 3 % sucrose. 16-h PP. CWFT. 55 lmol m-2 s-1. 80.7 % of seeds germinated with GA3 (pre)- Nikam and Barmukh
0.65 % agar 25 ± 2 °C treatment vs 46 % in control. Polyembryony (2009)
observed in 2–3 % of seeds. Plantlets could be
maintained for about 2 years in the absence of a
host after acclimatization of 1-months-old
seedlings in soil
S. album MS ? 1 mg/l 6-BA, MS ? 1 mg/l 2,4-D or Darkness for SGM. Transfer to 16-h PP Mature seeds could germinate on MS ? GA3 after Mo et al. (2010)
MS ? GA3 (SGM). pH NR, carbohydrate, after germination. CWFT. 40 lmol m-2 7–10 days culture and seedling induction was
gelling agent NR s-1. 25 ± 1 °C 82.5 % after 45 days of culture
S. album MS ? 1 mg/l BA ? 0.5 mg/l GA3 (SGM; 2011, 16-h PP. CWFT. 40 lmol m-2 s-1. 72 % SGM. 41.0 shoots/explant after 45 days Janarthanam and Sumathi
2012). MS ? 1 mg/l 2iP (SIM, SMM; 2011). 25 ± 1 °C. 55–60 % RH (2011) or 52.3 shoots/explant (2012). 4.2 roots/ (2011); Janarthanam
MS ? 1 mg/l TDZ (SIM; 2012). CIM ? 10 % shoot after 6 weeks (2011) or 6 roots/shoot after et al. (2012)
CM (SEM; 2011, 2012). ‘MS ? 0.5 mg/l 40 days (2012). 70 % survival of acclimatized
IBA ? 0.25 mg/l NAA (RIM; 2011). plantlets in red soil ? vermiculite ? FYM
‘MS ? 0.5 mg/l IAA (RIM; 2012). pH 5.6. (1:1:1) at 60–70 % RH (other conditions NR)
3 % sucrose. 0.9 % agar and watered with dilute MS (2011, 2012)
873

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123
S. album MS ? 4.44 lM BA ? 2.69 lM NAA (SGM). 16-h PP. CWFT. 2000 lux. 25 ± 2 °C 80 % SG. 60 % of internodes showed an Revathy and Arumugam
MS ? 13.5 lM 2,4-D (SEIM). MS ? 2.22 lM embryogenic response on SEIM within 4–6 (2011)e
BA ? 1.44 lM GA3 (SEM). ‘MS ? 2.46 lM weeks. 65 % shoot elongation after 25 days.
IBA (RIM). pH 5.6. 3 % sucrose. 0.9 % agar 60 % of shoots rooted in RIM. 60 % survival of
acclimatized plantlets in sterile
soil ? sand ? vermiculite (1:1:1)
S. album, S. PGR-free MS (SIM). MS ? 2 lM 16-h PP. CWFT. 30 lmol m-2 s-1. 25 °C Shoot formation in 8 m. 13.5 % survival after Baiculacula (2012)
yasi, R1 BA ? 0.02 lM NAA (RIM). pH 5.7. 2 % shoot formation for S. yasi (results for S. album
hybrids sucrose. 0.7 % agar and R1 hybrids NR). 19.1 % survival after root
formation for S. album (results for S. yasi and
R1 hybrids NR)
S. album MS ? 1 mg/l 2,4-D ? 0.5 mg/l TDZ (direct and Darkness for 1 w. 16-h (CIM, SIM) or Direct somatic embryogenesis (11.44 %), indirect Bele et al. (2012)e
indirect SEIM). MS ? 2 mg/l 2,4-D ? 0.5 mg/l 12-h (plantlets) PP. CWFT. LI and somatic embryogenesis (54.23 %): 160.08
TDZ (indirect SIM). MS ? 2 mg/l 2,4- source NR. 25 ± 2 °C SEs/explant; indirect organogenesis (20.38 %);
D ? 0.5 mg/l NAA (direct SIM; plantlet direct organogenesis (9.48 %); regeneration of
regeneration via direct organogenesis). plantlets via direct organogenesis (36.69 %);
MS ? 2 mg/l TDZ ? 1.0 mg/l GA3 (plantlet plant regeneration via somatic embryogenesis
regeneration via somatic embryogenesis). (163.63 %) or 141.25 % via indirect
MS ? 1 mg/l TDZ ? 0.5 mg/l GA3 ? 0.5 mg/l organogenesis. Roots could not be induced from
NAA (plantlet regeneration via indirect any medium. Acclimatized plantlets in
organogenesis). pH 5.8. 3 % sucrose. 0.75 % sand ? soil ? FYM (sterilized mixture; 1:1:1),
agar but survival not quantified. Protocol very
complex and values [100 % difficult to
interpret
S. album B5 ? 3 % sucrose. 0.78 % Bacto agar (SGM). 16-h PP. CWFT ? purple photosynthetic 96 % SG in vitro. 1 % infection. 2 % Crovadore et al. (2012)
MS ? 0.5 lM 2,4-D ? 10 lM Kin (CIM). lamps. 6000 lux. 27 °C polyembryony (discarded). No in vitro rooting
B5 ? 0.5 lM 2,4-D ? 10 lM Kin (CMM). possible from callus-derived shoots. Seedlings
MS ? 2.5 lM Kin (SIM from green callus). pH planted with grafted 2-year-old Citrus as pot
5.7. 3 % sucrose. 0.78 % Bacto agar host. GC–MS used to analyze the composition
of callus following extraction with pentane
S. album WPM ? 990 mg/l K2SO4 ? 100 mg/l myo- 16-h PP. CWFT. 36 lmol m-2 s-1. 2013: 100 % of explants formed callus; 24.6 shoot Singh et al. (2013, 2015)
inositol ? 0.4 mg/l TDZ (CIM). 25 ± 2 °C. 40–60 % RH buds/callus; 20.7 shoots/explant; 91.7 % of
WPM ? 2.5 mg/l BA ? 0.4 mg/l NAA (SIM). shoots rooted. 2015: 100 % of explants formed
WPM ? 1.5 mg/l IBA (RIM). pH 5.8. 3 % callus; 16 shoot buds/callus; 82.37 % of shoots
sucrose. 0.8 % agar rooted. [90 % of plantlets acclimatized in
sterile soil and coco-peat (1:1) (2013) or 85 %
of plantlets acclimatized in sterile soil, sand and
coco-peat (1:1:1) (2015) survived after 4–5
weeks
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Studied Culture medium, PGRs, additives, subculturesb Culture conditionsc Experimental outcome, maximum productivity, References
sandalwooda acclimatization and variation

S. album MS ? 2.5 mg/l 2,4-D ? 3 mg/l Kn (CIM). 25 ± 1 °C, LI ? source NR Dark for Callus induction observed with 8 weeks and SEs Peeris and Senarath
MS ? 0.5 mg/l BA ? 1 mg/l IAA ? 0.5 mg/l callus and SE induction. PP = 16 h for within 2 weeks. About 58 % of SEs germinated (2015)
Kn (SEIM). MS ? 2 mg/l GA3 (SGM). SE germination and plant development within 2 weeks. About 76 % of plantlets
MS ? 0.4 mg/l BA ? 0.2 mg/l IAA (plantlet produced with healthy shoots and roots.
Planta (2016) 243:847–887

development medium). pH 5.8. 3 % sucrose. Acclimatization NR

0.78 % jelly moss
2,4-D, 2,4-dichlorophenoxyacetic acid, 2iP N6-(2-isopentenyl) adenine, AA ascorbic acid, ABA abscisic acid, AdS adenine sulfate, B5 medium or Gamborg medium (Gamborg et al. 1968), BA
N6-benzyladenine (BA is used throughout even though BAP (6-benzylamino purine) may have been used in the original, according to Teixeira da Silva, 2012a, BM basal medium (in mg/l:
KINO3 (1900), NH4NO3 (1650) CaC122H2O (440), MgSO47H2O (370), KH2PO4 (170), MnSO44H2O (25), H3BO3 (10), ZnSO47H2O (13.9), Na2EDTA (18.6), myo-inositol (100), nicotinic
acid (5), folic acid (5), glycine (2), pyridoxine–HCl (0.5), thiamine-HCI (0.5), biotin (0.05), sucrose (20 g/l), Difco bacto agar (6 g/l) modified WB according to Rangaswamy (1961)); CA, citric
acid; CH, casein hydrolysate; CIM callus induction medium, CM coconut milk, CMM callus multiplication medium, CSC cell suspension culture, CWFT white fluorescent tubes, FYM farmyard
manure, GA3 gibberellic acid, GC–MS gas chromatography-mass spectrometry, hyg hygromycin, IAA indole-3-acetic acid, IBA indole-3-butyric acid, Kin kinetin (6-furfurylaminopurine), LI
light intensity, mBM modified basal medium (BM), MES 2-(N-morpholino)ethane sulfonic acid, MS Murashige and Skoog (1962) medium, mMS modified MS, mWM modified White’s basal
medium (White 1963), NAA a-naphthaleneacetic acid, NOA napthoxy acetic acid; NP not performed, NR not reported in the study, PCM protoplast culture medium, pCPA para chlorophe-
noxyacetic acid, PGR plant growth regulator, PP photoperiod, RH relative humidity, RIM root induction medium, rpm revolutions per minute, SD 8339 6-benzyl-9-tetrahydropyran adenine, SE
somatic embryo, SEIM somatic embryo induction medium, SEMM somatic embryo multiplication medium, SEM shoot elongation medium, SGM seed germination medium, SIM shoot induction
medium, SMM shoot multiplication medium, SW-1 sandalwood suspension culture-1, SW-2 sandalwood suspension culture-2, TDZ thidiazuron (N-phenyl-N0 -1, 2,3-thiadiazol-5-ylurea), TPM
tissue proliferation medium, V47 medium (3 mM MES ? 0.5 mg/l 2,4-D ? 0.5 mg/l BA, Binding 1974), vvm volume of air per unit medium per unit time, WB White’s basal medium (White
1963), WPM woody plant medium (Lloyd and McCown 1980), YE yeast extract, Zea zeatin
b
Even though calli was used in the original, the term callus has been used here based on recommendation of Teixeira da Silva 2012b
c
The original light intensity reported in each study has been represented since the conversion of lux to lmol m-2 s-1 is different for different illumination (main ones represented): for
fluorescent lamps, 1 lmol m-2 s-1 = 80 lux; the sun, 1 lmol m-2 s-1 = 55.6 lux; high-voltage sodium lamp, 1 lmol m-2 s-1 = 71.4 lux (Thimijan and Heins 1983)
d
Most likely GA3
e
Claims of somatic embryogenesis without sufficient proof (cytological, histological, genetic), i.e., only photos of macromorphologyaGenera: S. = Santalum
875

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876 Planta (2016) 243:847–887

greater detail in Tables 3 and 4. The most recent reviews the source of disinfected floral tissues for either in vitro
on sandalwood tissue culture and related biotechnologies breeding experiments or more specialized in vitro tech-
were published by Rao and Bapat (1992), (1993), (1995), niques such as anther or ovary culture. The latter two
Bapat and Rao (1992a), and Bapat (1993) techniques have not yet been applied to any Santalum
species. The establishment of an in vitro propagation pro-
Perspectives and culture establishment tocol traditionally requires several stages: culture initiation
(including the appropriate choice of explant, surface ster-
Most in vitro studies have been conducted on S. album with ilization, culture conditions and medium composition),
only one report on S. spicatum (Tables 3, 4). In vitro tissue multiplication, rooting (in vitro and ex vitro), acclimati-
culture and micropropagation are popular methods for the zation, and field establishment. These are outlined next for
large-scale propagation and improvement of existing plant Santalum spp.
genotypes, serving as the basal method for genetic trans-
formation experiments, often cutting the time to obtain Choice of explant and surface sterilization
novel germplasm through conventional breeding. By pro-
viding a sterile culture environment, in vitro tissue culture The choice of an explant depends primarily on the desired
also allows for developmental events to be studied, more so objective (e.g., shoot tips or nodal explants for the pro-
when thin cell layers are used (Teixeira da Silva and duction of true-to-type clonal plants) or on the availability
Dobránszki 2013), and for the production of in vitro of healthy (disease-free) material. The age and physiolog-
flowers (Teixeira da Silva et al. 2014), which can serve as ical status of the mother plant need to be considered, such

Fig. 2 In vitro adventitious

shoot propagation and root
formation in Santalum album.
a Adventitious shoots were
induced on MS medium
supplemented with 5.0 lM BA
and 0.5 lM NAA.
b Adventitious shoots were
propagated and subcultured on
MS medium supplemented with
5.0 lM BA and 5.0 lM IBA.
c Root formation on rooting
medium supplemented with
250 lM IBA. Unpublished
figure/photos (Guohua Ma)

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Planta (2016) 243:847–887 877

as actively growing vs dormant shoots, which often (Table 3). Even though the choice of sterilant, its concen-
depends on seasonal availability. For the micropropagation tration, treatment time and rinses are all essential aspects of
of sandalwood, various explants have been used. Details a surface sterilization protocol (Teixeira da Silva et al.
about explant source, the type of explant used, and their 2015), such basic information is frequently missing from
size are described in Table 3. In general, the micropropa- many protocols, as is a description of the efficiency of the
gation of any plant species employs explants with a pre- sterilization protocol and the quantification of infection
determined meristem such as a node or a shoot tip, either (Table 3).
from mature trees or from in vitro or ex vitro germinated
seedlings; other explants such as internodes, stem seg- Culture conditions
ments, or leaves of leaf disks are generally utilized to
induce adventitious shoots or somatic embryos. In sandal- Specific and tested culture conditions are required to ini-
wood, the explants used encompassed nodes and shoot tips tiate a plant tissue culture because photoperiod, light
from mature trees or seedlings (Rao et al. 1984; Muralid- intensity, light source, and temperature play important
haran 1997; Das et al. 1998; Rugkhla and Jones 1998; roles in the successful establishment of an in vitro culture.
Radhakrishnan et al. 2001; Ilah et al. 2002; Primawati The most commonly used conditions in the tissue culture of
2006; Sanjaya et al. 2006a, b; Revathy and Arumugam S. album were 16-h photoperiod and temperatures between
2011; Peeris and Senarath 2015; Singh et al. 2015), zygotic 24 and 28 °C (Table 4), although some protocols used a
embryos and seed endosperm (Rangaswamy and Rao 1963; 12-h photoperiod (Lakshmi Sita et al. 1979, 1980a, b;
Rao 1965; Rao and Rangaswamy 1971; Rao and Raghava Sanjaya et al. 2006a, b) or very rarely a 14-h photoperiod
Ram 1983; Sankara Rao et al. 1996; Rai and McComb (Shiri and Rao 1998). Rao et al. (1984) used a 72-h dark
1997; Shiri and Rao 1998; Anil et al. 2000; Anil and Rao treatment for root induction. Optimal (recommended)
2000; Rai and McComb 2002; Mo et al. 2008), mature and in vitro regeneration protocols for S. album would be Singh
immature seeds as well as seedling-derived explants such et al. (2013, 2015) for callus induction, Rugkhla and Jones
as internodes, hypocotyls, leaves, and stem segments (Rao (1998) for somatic embryogenesis, and Sanjaya et al.
and Bapat 1978; Bapat and Rao 1979, 1984; Lakshmi Sita (2006a, b) for direct shoot induction from leaf explants.
et al. 1980a; Bapat et al. 1985; Das et al. 1999, 2001;
Sanjaya and Rai 2003; Mujib 2005; Mo et al. 2008, 2010; Medium composition
Shekhawat et al. 2008; Nikam and Barmukh 2009; She-
khawat et al. 2010; Revathy and Arumugam 2011; Janar- Medium composition includes the type of basal medium
thanam and Sumathi 2011; Bele et al. 2012; Crovadore used or the composition and concentration of macro- and
et al. 2012; Janarthanam et al. 2012), and explants such as micronutrients, carbon source, gelling agent (type and
leaves, stem segments, and internodes from mature trees concentration) or liquid culture, plant growth regulators
(Lakshmi Sita et al. 1979; 1980b; Lakshmi Sita and Shobha (PGRs), and additives. Mostly Murashige and Skoog
Rani 1983; Rao and Raghava Ram 1983; Ozias-Akins et al. (1962) (MS) basal medium was used for S. album culture
1985; Rao and Ozias-Akins 1985; Lakshmi Sita 1986; initiation, multiplication and rooting (Table 4). Only one
Bapat and Rao 1988, 1992b, 1999; Bapat et al. 1996; Ma study (Singh et al. 2013) used woody plant medium
et al. 2008; Shekhawat et al. 2008; Singh et al. 2013). In (WPM) (Lloyd and McCown 1980). Singh et al. (2013)
rare cases (Bhaskar and Rao 1983), in vitro-raised shoots used WPM with 990 mg/L K2SO4 to induce callus from
derived from callus were used explants although details leaves of S. album while WPM with PGRs was used to
about callus induction were not reported. induce shoots and roots (Table 4). Crovadore et al. (2012)
Surface sterilization, an essential step for the initiation used B5 medium (Gamborg et al. 1968) for in vitro seed
of an aseptic culture, varies depending on the explant type germination, while MS medium was used for callus cul-
and source (Table 3). In sandalwood, surface-sterilized ture, shoot induction, and rooting. In general, 3 % sucrose
fruits can be used for aseptic seed germination and to was used as the preferred carbon source, but 4–5 % sucrose
isolate endosperm (Rao and Bapat 1978, 1979; Bapat et al. was also reported in a few cases (Table 4). In most studies,
1985; Das et al. 1999; Ilah et al. 2002; Mujib 2005; Cro- semisolid medium was used and gelled with agar (Table 4),
vadore et al. 2012). Once an aseptic seedling has germi- although Laxmi Sita et al. (1980a, b) used liquid culture
nated, it can be used as a source of explants that does not medium for S. album somatic embryogenesis. Adventitious
require further surface sterilization (Rangaswamy and Rao S. album shoots can be induced on MS medium supple-
1963; Rao 1965; Rao and Rangaswamy 1971, 1999; Anil mented with 5.0 lM BA and 0.5 lM NAA, subcultured on
et al. 2000; Anil and Rao 2000). For surface sterilization, MS medium supplemented with 5.0 lM BA and 5.0 lM
HgCl2 in concentrations ranging from 0.05 to 0.5 % (w/v) IBA, and rooted in the presence of 250 lM IBA (Fig. 2).
are most frequently used, irrespective of the explant Optimal basal medium for S. album would be WPM

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878 Planta (2016) 243:847–887

Fig. 3 Callus suspension,

induced somatic embryogenesis,
and plant regeneration in
Santalum album. a Callus
suspension in a flask in MS
liquid medium supplemented
with 4.5 lM 2,4-D.
b Suspension cells transformed
into embryogenic callus on solid
MS medium supplemented with
5.0 lM TDZ and 0.5 lM IBA
after 1 month. c Somatic
embryos induced on MS
induction medium
supplemented with 5.0 lM TDZ
and 0.5 lM IBA. d Somatic
embryos germinated on 1/4 MS
medium supplemented with
0.3 % activated charcoal and
free of sucrose. e Plantlets
regenerated from germinated
somatic embryos on 1/4 MS
medium supplemented with
0.3 % activated charcoal and
free of sucrose. Unpublished
figure/photos (Guohua Ma)

supplemented with 0.4 mg/l TDZ for callus induction 1980a, b; Rao and Raghava Ram 1983; Rao et al. 1984;
(Singh et al. 2013, 2015), or MS supplemented with 5 lM Bapat et al. 1985; Ozias-Akins et al. 1985; Rao and
BA for the induction of somatic embryos (Rugkhla and Ozias-Akins 1985; Lakshmi Sita 1986; Bapat and Rao
Jones 1998) or 0.53 lM NAA and 11.09 lM BA for direct 1988, 1992; Bapat et al. 1990, 1996; Sankara Rao et al.
shoot induction from leaf-derived explants (Sanjaya et al. 1996; Rai and McComb 1997; Das et al. 1998, 1999;
2006a, b). Shiri and Rao 1998; Rugkhla and Jones 1998; Anil and
Rao 2000; Das et al. 2001; Ilah et al. 2002; Rai and
Somatic embryogenesis McComb 2002; Mo et al. 2008; Shekhawat et al. 2008,
2010; Revathy and Arumugam 2011; Bele et al. 2012;
Somatic embryogenesis, an important in vitro plant Singh et al. 2013, 2015; Peeris and Senarath 2015).
regeneration pathway, is utilized for large-scale propaga- Details about culture conditions and media composition to
tion and has various applications in cryoconservation, induce somatic embryogenesis are provided in Table 4.
synthetic seed production, and as a source of protoplasts. Despite these prolific reports, mainly for S. album, many
A total of 33 in vitro Santalum propagation studies have have not been supported by very strong, or convincing,
claimed somatic embryogenesis (Rao and Rangaswamy proof, such as histology, cytology, flow cytometry, or
1971; Bapat and Rao 1979; Lakshmi Sita et al. 1979, molecular markers.

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Anil and Rao (2000) claimed that a Ca2?-mediated and 0.55 M sorbitol. That study appears to have been a pre-
signaling pathway may be involved in somatic embryoge- amble for the more comprehensively reported study by Rao
nesis after callus induced in the presence of 2,4-D took up and Ozias-Akins (1985) in which the authors reported the
four-fold levels of labeled Ca2? resulting in the formation same ideal enzyme cocktail for protoplast isolation from
of pro-embryogenic cell masses. Spherical organelles in the embryogenic cell suspension cultures derived from shoot
endosperm are oil bodies in which calcium-dependent segments of a 20-year-old S.album tree.
protein kinase (CDPK) (first purified in somatic embryos
by Anil and Rao (2001), and termed swCDPKs) is Genetic transformation
expressed during seed development (Anil et al. 2003). As a
result of the action of CDPK, the existence of these oil There are only three reports on the genetic transformation
bodies and the Ca2?-mediated signaling pathway, as much of sandalwood, all three targeting S. album and using
as 30 % of the dry weight of the endosperm was made up Agrobacterium tumefaciens-mediated gene transfer. Shiri
of oil (Anil et al. 2003). Earlier studies by this group (Anil and Rao (1998) used LBA4404/pKIWI105 to introduce
et al. 2000) indicated that CDPK could not be detected in gusA and nptII genes into somatic embryos, confirming
somatic embryos, or in the soluble proteins of shoots and integration using a range of biochemical assays and
flowers, but only in zygotic embryos, seedlings, and molecular tools [GUS histochemical staining, NPTII
endosperm. S. album callus suspension that develops in MS expression assay, PCR, Southern blot, dot blot (equivalent
liquid medium supplemented with 4.5 lM 2,4-D can be to Northern blot)]. Disarmed A. tumefaciens strain EHA105
induced to form embryogenic callus on solid MS medium (Hood et al. 1993) harboring the pCAMBIA 1301 binary
supplemented with 5.0 lM TDZ and 0.5 lM IBA, while vector carrying the gusA gene driven by single CaMV 35S
somatic embryos form on MS medium supplemented with promoter and an hptII gene driven by a double-enhanced
5.0 lM TDZ and 0.5 lM IBA (Fig. 3). 35S promoter was used to transform embryogenic S. album
callus (Shekhawat et al. 2008). Gene integration was con-
Protoplasts firmed by PCR, RT-PCR and Southern blot analysis, and
GUS activity was shown to be stable over 7 months of
There are only four studies available in which protoplast culture. In an ensuing study, Shekhawat et al. (2010)
culture was used for the regeneration of plants. Laxmi Sita transformed embryogenic cell suspension cultures—also
and Sobha Rani (1983) isolated protoplasts from young with EHA105—with a hepatitis B small surface antigen
leaves of mature S. album trees. In their protocol, leaves (HBsAg) gene, encoding for a pharmaceutically important
were incubated in 2 % cellulase R10, 0.5 % macerozyme, recombinant protein. Using electroporation and selection
0.5 % pectinase, and 1 % hemicellulase in cell protoplast on kanamycin-supplemented medium, the authors con-
washing (CPW) salts with 10 % mannitol at pH 5.6, firmed transgene expression using RT-PCR, Western blot
incubated for 16 h at 25 °C with shaking. Protoplasts could analysis, and ELISA. Supplementing medium with 30 mM
be isolated by using only 1 % cellulase R10 and 0.5 % trehalose almost doubled the level of HBsAg expressed.
pectinase in V47 medium (Binding 1974). Leaves and Genetic transformation is thus weakly explored in sandal-
callus yielded 2 9 105 and 3 9 106 protoplasts/ml, wood species. However, the introduction of biotic (dis-
respectively. Protoplasts formed cellular colonies in eases, pests, viruses) and abiotic (drought, cold, frost,
6 weeks after culture in liquid MS medium containing salinity) resistance through transgenic means would widen
2 mg/l 2,4-D, 0.2 mg/L BA and 10 % mannitol. The the possible niches where sandalwood could be cultivated,
regeneration of organs or plantlets was not reported. Also taking advantage of usually hostile conditions to harvest
in S. album, Bapat et al. (1985) used 1 % cellulase R10, sandalwood products.
0.5 % macerozyme, and 0.5 M sorbitol or mannitol to
isolate protoplasts from stem callus, 2 % cellulase R10,
1 % pectinase, 1 % hemicellulase, 0.9 % CaCl22H2O, and Conclusions and future perspectives
0.55 M sorbitol for hypocotyl callus and 2 % cellulase
R10, 1 % macerozyme, 1 % hemicellulase and 0.8 M Sandalwood from several Santalum species is a commer-
sorbitol for leaf mesophyll, all in V47 medium. The highest cially important forest product that has been traded for
yield was from stem callus (8.73 9 106), and the authors many centuries. International trade has been based on the
claimed the production of somatic embryos and the sub- harvest of wild stands, resulting in excessive exploitation
sequent development into plantlets. Ozias-Akins et al. and severe reductions in abundance and biodiversity across
(1985) also claimed to derive somatic embryos from callus most species. This has led to sustained price rises for
induced from protoplasts derived from shoot segments sandalwood products and increasing interest in sandalwood
using 1 % cellulase, 1 % macerozyme, 0.5 % driselase, cultivation, both as an industrial crop and on a smaller

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880 Planta (2016) 243:847–887

scale within agroforestry systems. Successful cultivation of Guangdong Province Science and Technology Program (number:
sandalwood will depend on a clear understanding of its 2015B020231008). The authors thank the assistance of Dr. Budi
Winarto (IOCRI, Indonesia) with assistance in the interpretation of
basic biology and propagation, details of which have been Indonesian literature and Dr. Robert Nasi (Director, Center for
examined in depth in this review. The hemi-parasitic nature International Forestry Research (CIFOR), Indonesia) for providing
of sandalwood necessitates the planting of host plants to difficult-to-access literature. TP recognizes the generous contributions
support its growth. While this makes cultivation of san- of the Australian Center for International Agricultural Research
(ACIAR). The authors are thankful to the anonymous reviewers for
dalwood more complex relative to other forestry species, constructive comments and suggestions.
many suitable species have been recorded in the literature
to support sandalwood growth during the three main stages Compliance with ethical standards
of its rotation (pot, intermediate, and final hosts). Sandal-
Conflict of interest The authors declare no conflicts of interest.
wood can potentially be grown across a range of environ-
ments provided locally adapted suitable host species are
used from the nursery phase until harvest. It is evident that
Santalum spp. preferentially form haustorial connections References
with N2-fixing legumes (Table 2) and source significant N
and C from these hosts. The breeding system can be AAG (Australian Agribusiness Group) (2006) Market overview—the
Australian sandalwood industry. Australian Agribusiness Group,
described as incompletely outbreeding, with variation in
Melbourne. http://sandalwood.org.au/wp-content/uploads/2012/
the level of self-compatibility recorded between families 06/AAG-2007-Market-Overview.pdf (last accessed 14 Decem-
and individuals. The capacity for self-fertilization within ber, 2015)
the genus is likely to have contributed to its capacity to Adkoli NS (1977) Sandalwood in Karnataka. Retrospect and prospect.
In: Proceedings of the All India Sandal Seminar. Bangalore,
colonize new territory/islands after long-distance seed
India, pp 86–94
dispersal by birds. No clear reproductive barriers exist Anantha Padmanabha HS (2000) Sandalwood and its marketing trend.
between four commercial tropical species, and it may be My For 36:147–152
possible that the capacity for hybridization between species Anantha Padmanabha HS (2014) Indian sandalwood market trend and
production. In: Ramakantha V (ed) International seminar on
is widespread in this genus. This feature of the breeding
sandalwood: current trends and future prospects. institute of
system can facilitate introgression of traits between species wood science & technology: Indian Council of Forestry
and development of hybrids. Introduction of exotic san- Research & Eduction, Bangalore, India, p 74 (Abstract)
dalwood species within the natural range of a compatible Anantha Padmanabha HS, Nagaveni SH, Rai SN (1988) Influence of
host plants on the growth of sandal. My For 24:154–160
species will likely result in uncontrolled gene flow between
Anil VS, Rao KS (2000) Calcium-mediated signaling during sandal-
them and modify the genetic structure and diversity of local wood somatic embryogenesis. Role for exogenous calcium as
species. The judicious application of biotechnology and second messenger. Plant Physiol 123:1301–1312. doi:10.1104/
cultivation may provide a viable alternative to extractive pp.123.4.1301
Anil VS, Rao KS (2001) Purification and characterization of a Ca2?-
exploitation, leading to survival and proliferation of
dependent protein kinase from sandalwood (Santalum album L.):
important members of the Santalum genus. This review evidence for Ca2?-induced conformational changes. Phytochem-
highlights the successes and failures within the fields of istry 58:203–212. doi:10.1016/S0031-9422(01)00231-X
plant tissue culture, as a means of mass propagating Anil VS, Harmon AC, Rao KS (2000) Spatio-temporal accumulation
and activity of calcium-dependent protein kinases during
valuable germplasm. The ability to induce somatic
embryogenesis, seed development, and germination in sandal-
embryos would allow synthetic seeds to be produced, wood. Plant Physiol 122:1035–1044. doi:10.1104/pp.122.4.1035
aiding further germplasm storage, as cryopreservation Anil VS, Harmon AC, Rao KS (2003) Temporal association of Ca2?-
units, and standardization (Sharma et al. 2013). Transgenic dependent protein kinase with oil bodies during seed develop-
ment in Santalum album L.: its biochemical characterization and
strategies, and genomic and transcriptomic analyses would
significance. Plant Cell Physiol 44:367–376. doi:10.1093/pcp/
allow for genes encoding for more rapid growth or for pcg046
resistance to pests and diseases, such as spike disease, or to Annapurna D, Rathore TS, Joshi G (2004) Effect of container type
abiotic stresses, to be inserted, thus producing more robust and size on the growth and quality of seedlings of Indian
sandalwood (Santalum album L.). Aust For 67:82–87. doi:10.
and faster-growing trees.
1080/00049158.2004.10676211
Author contribution statement All authors contributed Annapurna D, Rathore TS, Joshi G (2005) Refinement of potting
medium ingredients for production of high quality seedlings of
equally to all parts of the development and revision of this sandalwood (Santalum album L.). Aust For 68:44–49. doi:10.
review. 1080/00049158.2005.10676225
Annapurna D, Rathore TS, Joshi G (2006) Modern nursery practices
Acknowledgments XZ and GM are sincerely thankful for funding in the production of quality seedlings of Indian sandalwood
supported by the National Natural Science Foundation of China (grant (Santalum album L.)—stage of host requirement and screening
numbers 31470685, 31270720, and 31100498), Natural Science of primary host species. J Sustain For 22:33–55. doi:10.1300/
Foundation of Guangdong Province (S2012010009025), and the J091v22n03_03

123
Planta (2016) 243:847–887 881

Applegate GB, McKinnell FH (1993) The management and conser- Barber CA (1907) Studies of root parasitism. The haustorium of
vation status of Santalum species occurring in Australia. In: Santalum album L. 2. The mature haustorium. Mem Dept Agric
McKinnell FH (ed) Sandalwood in the Pacific region. Proceed- India Bot Ser l, Part 2. pp 1–58
ings of a symposium held on 2 June 1991 at XVII Pacific Barrett DR, Fox JED (1995) Geographical distribution of Santalaceae
Science Congress, Honolulu, Hawaii, ACIAR Proceedings No. and botanical characteristics of species in the genus Santalum.
49. Canberra, Australia, pp 5–12 Sandalwood seed nursery and plantation technology. In: Gjerum
Applegate GB, Chamberlain J, Feigelson J, Hamilton L, McKinnell L, Fox JED, Ehrhart Y (eds) Proceedings of a Regional
FH, Neil PE, Rai SN, Statham PC, Stemmermann L (1990) Workshop for Pacific Island Countries, Noumea, New Caledo-
Sandalwood in the Pacific: a state-of-knowledge synthesis and nia, 1–11 August 1994. FAO, Suva, Fiji. pp 2–23
summary from the April 1990 symposium. In: Hamilton L, Barrett D, Fox JED (1997) Santalum album: kernel composition,
Conrad CE (eds) Proceedings of the Symposium on Sandalwood morphological and nutrient characteristics of pre-parasitic
in the Pacific, April 9–11. General Technical Report PSW-122. seedlings under various nutrient regimes. Ann Bot 79:59–66.
Pacific South Research Station, Forest Service, USDA, Hono- doi:10.1006/anbo.1996.0303
lulu, Hawaii, pp 1–11 Baskorowati L (2011) Flowering intensity and flower visitors of
Arun Kumar AN, Joshi G, Mohan Ram HY (2012) Sandalwood: history, Santalum album L. at ex situ conservation plot, Watusipat,
uses, present status and the future. Curr Sci 103:1408–1416 Gunung Kidul, Yogyakarta. J For Res 8:130–143
Athelstone MR, Loveys B, Jusaitis M (1994) Stimulation of Batabyal S, Dalal T, Tah J (2014) Responses of some phyto-hormones
germination of quandong (Santalum acuminatum) and other for vegetative propagation of an ancient precious wood plant:
Australian native plant seeds. Aust J Bot 42:565–574. doi:10. Santalum album L. Biosci Discov 5:170–174
1071/BT9940565 Bele D, Tripathi MK, Tiwari G, Baghel BS, Tiwari S (2012)
Baiculacula S (2012) Micropropagation of sandalwood. In: Thomson Microcloning of sandalwood (Santalum album Linn.) from
L, Padolina C, Sami R, Prasad V, Doran J (eds). Regional cultured leaf discs. J Agric Technol 8:571–583
workshop on Sandalwood Resource Development Research and Berry A (2005) Sandalwood development in Vanuatu. Paper
Trade in the Pacific and Asian Region. European Union, presented at the Regional workshop on Sandalwood Research,
Secretariat of the Pacific Community, James Cook University Development and Extension in the Pacific Islands and Asia
and the Australian Centre for International Agricultural (7–11 October, 2002), Noumea
Research. Port Vila, Vanuatu. pp 111–116 Bhaskar V (1992) Pollination biology and fertilization in Santalum
Baldovini N, Delasalle C, Joulain D (2011) Phytochemistry of the album L. (Santalaceae). Flora 187:73–78
heartwood from fragrant Santalum species: a review. Flavour Bhaskar V, Rao NS (1983) In situ development of callus shoots in
Fragr J 26:7–26. doi:10.1002/ffj.2025 sandal (Santalum album L.). Indian For 109:45–46
Bapat VA (1993) Studies on synthetic seeds of sandalwood (Santalum Binding H (1974) Regeneration von haploiden und diploiden pflanzen aus
album L) and mulberry (Morus indica L). In: Redenbaugh K (ed) protoplasten von Petunia hybrida L. Zeit Planzenfisiol 74:327–356
Synseeds. Applications of synthetic seeds to crop improvement. Bosimbi D (2005) Sandalwood (Santalum macgregorii) in Papua New
CRC Press Inc., Boca Raton, pp 381–407 Guinea. In: Proceedings of a Regional Workshop on Sandalwood
Bapat VA, Rao PS (1979) Somatic embryogenesis and plantlet Research, Development and Extension in the Pacific Islands and
formation in tissue cultures of sandalwood (Santalum album L.). Asia Noumea, New Caledonia, 7–11 October 2002
Ann Bot 44:629–630 Brand JE (2009) Effect of different Acacia acuminata variants as
Bapat VA, Rao PS (1984) Regulatory factors for in vitro multipli- hosts on performance of sandalwood (Santalum spicatum) in the
cation of sandalwood tree (Santalum album Linn.) I. Shoot bud northern and eastern wheatbelt, Western Australia. Aust For
regeneration and somatic embryogenesis in hypocotyl cultures. 72:149–156. doi:10.1080/00049158.2009.10676297
Proc Indian Acad Sci Plant Sci 93:19–27. doi:10.1007/ Brand JE, Ryan PC, Williams MR (1998) Establishment and growth
BF03053003 of sandalwood (Santalum spicatum) in south-western Australia:
Bapat VA, Rao PS (1988) Sandalwood plantlets from ‘‘synthetic the Northampton pilot trial. Aust For 62:33–37
seeds’’. Plant Cell Rep 7:434–436. doi:10.1007/BF00269531 Brand JE, Crombie DS, Mitchell MD (2000) Establishment and
Bapat VA, Rao PS (1989) In vitro strategies for sandalwood growth of sandalwood (Santalum spicatum) in south-western
propagation. In: Dhawan V (ed) Applications of biotechnology Australia: the influence of host species. Aust For 63:60–65.
in forestry and horticulture. Plenum Press, New York, doi:10.1080/00049158.2000.10674814
pp 145–156. doi:10.1007/978-1-4684-1321-2_11 Brand JE, Robinson N, Archibald RD (2003) Establishment and
Bapat VA, Rao PS (1992a) Biotechnological approaches for sandal- growth of sandalwood (Santalum spicatum) in south-western
wood (Santalum album L.) micropropagation. Indian For Australia: Acacia host trials. Aust For 66:294–299. doi:10.1080/
118:48–54 00049158.1999.10674760
Bapat VA, Rao PS (1992b) Plantlet regeneration from encapsulated Brandis D (1903) Treatment of the sandal tree. Indian For 29:3–6
and non-encapsulated desiccated somatic embryos of a forest Buchanan F (1884) A journey through the countries of Mysore,
tree: sandalwood (Santalum album L.). J Plant Biochem Canara and Malabar. East India Company, London
Biotechnol 1:109–113. doi:10.1007/BF03262907 Bulai P, Nataniela V (2005) Research, development, and extension of
Bapat VA, Rao PS (1999) Somatic embryogenesis in Santalum album sandalwood in Fiji—a new beginning. In: Thomson L, Bulai S,
L. Sandalwood Res Newsl 8:4 Sovea L (eds) Regional workshop on sandalwood research,
Bapat VA, Gill R, Rao PS (1985) Regeneration of somatic embryos development and extension in the Pacific Islands and Asia,
and plantlets from stem callus protoplasts of sandalwood tree Noumea, New Caledonia, 7–11 October 20. pp 83–91
(Santalum album L.). Curr Sci 54:978–982 Burdock GA, Carabin IG (2008) Safety assessment of sandalwood oil
Bapat VA, Iyer RK, Rao PS (1996) Effect of cyanobacterial extract (Santalum album L.). Food Chem Toxicol 46:421–432. doi:10.
on somatic embryogenesis in tissue cultures of sandalwood 1016/j.fct.2007.09.092
(Santalum album). J Med Aromat Plant Sci 18:10–14 Butaud J-F (2015) Reinstatement of the Loyalty Islands sandalwood,
Barber CA (1906) Studies of root parasitism. The haustorium of Santalum austrocaledonicum var. glabrum (Santalaceae), in
Santalum album L. 1. Early stages up to penetration. Mem Dept New Caledonia. PhytoKeys 56:111–126. doi:10.3897/phytokeys.
Agric India Bot Ser l, Part 1. pp 1–130 56.5924

123
882 Planta (2016) 243:847–887

Carr GD (1978) Chromosome numbers of Hawaiian flowering plants Dhanya B, Viswanath S, Purushothman S (2010) Sandal (Santalum
and the significance of cytology in selected taxa. Am J Bot album L.) conservation in southern India: a review of policies
65:236–242. doi:10.2307/2442457 and their impacts. J Trop Agric 48:1–10
Chada YR (ed) (1972) Santalum album. Wealth of India, Vol. 9. Doran JC, Brophy JJ (2005) Sandalwood—a global perspective. In:
Publication and Information Directorate, CSIR, New Delhi, Thomson L, Bulai S, Sovea L (eds) Regional workshop on
India, pp 208–222 sandalwood research, development and extension in the Pacific
Chauvin JP, Ehrhart Y (1998) Germination of two provenances of Islands and Asia, Noumea, New Caledonia, 7–11 October 2002.
Santalum austrocaledonicum var. austrocaledonicum. In: Secretariat of the Pacific, Suva, pp 29–49
Radomiljac AM, Ananthapadmanabho HS, Welbourn RM, Rao Doran J, Thomson L, Brophy J, Goldsack B, Bulai P, Faka’osi T,
KS (eds) Sandal and its Products. Proceedings of an international Mokoia T (2005) Variation in heartwood oil composition of
seminar held on 18–19 December 1997 organised by the Institute young sandalwood trees in the south Pacific (Santalum yasi, S.
of Wood Science and Technology (ICFRE) and Karnataka State album and F1 hybrids in Fiji, and S. yasi in Tonga and Niue).
Forest Department, Bangalore, India. Australian Centre for Sandalwood Res Newslett 20:3–7
International Agricultural Research, Canberra, Australia, DPI&F (Department of Primary Industries and Fisheries) (2004) The
pp 113–116 Queensland forest industry: An overview of the commercial
Chen R, Zhang XH, Ma GH (2014) Studies on parasitic relationship growing, management and processing of forest products in
between Santalum album L. and leguminous plants. J Trop Queensland. Queensland Department of Primary Industries and
Subtrop Bot 22:53–60 (in Chinese with English abstract) Fisheries, Brisbane. http://trove.nla.gov.au/version/13964164
Christian K (2015) Sandalwood the good oil for US funds. In: http:// (last accessed 14 December, 2015)
www.news.com.au/finance/business/sandalwood-the-good-oil- Edwards W (1951) The fragrant wood of Hawaii. Illus. by Jerry
for-us-funds/story-e6frfkur-1227179781909 (last accessed 14 Chong. Advertiser Weekly. Honolulu, Sept. 4
December, 2015) Ehrhart Y, Fox JED (1995) State of knowledge regarding
CITES (Convention on International Trade in Endangered Species) cultivation of sandalwood. In: Gjerum L, Fox JED, Ehrhart
(2013) Proceedings of the 16th meeting of the Conference of the Y (ed) Sandalwood seed nursery and plantation technology.
Parties Bangkok (Thailand), 3–14 March 2013, 10 pp. http:// Proceedings of a Regional Workshop for Pacific Island
www.environment.gov.au/biodiversity/wildlife-trade/cites (last Countries held at Nouméa, New Caledonia, 1–11 August
accessed 14 December, 2015) 1994. FAO, Suva, Fiji. RAS/92/361. Field Document N°8,
Collins S, Walker S, Haines R (2000) SPRIG vegetative propagation pp 275–291
completion report. Queensland Forestry Research Institute, Encyclopaedia britannica (online) (2013) Santalaceae. http://global.
Queensland britannica.com/EBchecked/topic/523114/Santalaceae (last
Corrigan H, Naupa S, Likiafu R, Tungon J, Sau B, Viji I, Sam C, accessed 14 December, 2015)
Kalamor L, Gerrand A, Mele L, Walker S, Collins S, Thomson L FAO (Food and Agriculture Organization of the United Nations).
(2005) A strategy for conserving, managing and better utilizing 2015. Sandalwood oil. Flavours fragances plant Orig. Chapter 6
the genetic resources of Santalum austrocaledonicum (sandal- Sandalwood Oil. http://www.fao.org/docrep/v5350e/V5350e00.
wood) in Vanuatu. In: Proceedings of the regional workshop on htm (last accessed 14 December, 2015)
sandalwood research, development and extension in the Pacific Fox JE (2000) Sandalwood: the royal tree. Biologist (London)
Islands and Asia (7–11 October 2002), Noumea, New Caledonia 47:31–34
Crovadore J, Schalk M, Lefort F (2012) Selection and mass Fox JED, Doronila AI, Barrett DR, Surata IK (1996) Desmanthus
production of Santalum album L. calli for induction of virgatus (L.) Willd. An efficient intermediate host for the
sesquiterpenes. Biotechnol Biotechnol Equip 26:2870–2874. parasitic species Santalum album L. in Timor, Indonesia.
doi:10.5504/bbeq.2012.0028 J Sustain For 3:13–23. doi:10.1300/J091v03n04_02
Daruhi G (1993) Sandelwud Bilong Vanuatu—a bright future. In: Gamage YMM, Subasinghe S. MCUP, Hettiarachi DS (2010)
Sandalwood in the Pacific Region. Proceedings of symposium, 2 Changes of seed germination rate with storage time of
June 1991 at the XVII Pacific Science Congress, Honolulu, Santalum album L. (Indian sandalwood) seeds. In: Proceedings
Hawaii, pp 26–29 of the 15th International Forestry and Environment Sympo-
Das SC, Tah J (2013) Effect of GA3 on seed germination of sandal sium, 26-27 November 2010. Published by Department of
(Santalum album L.). Int J Curr Sci 8:79–84 Forestry and Environmental Science. University of Sri
Das S, Das S, Mujib A, Pal S, Dey S (1998) Influences of carbon Jayewardenepura, Sri Lanka, pp 279–281
sources and pH on rapid mass propagation of Santalum album Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of
through somatic embryogenesis: application in biotechnology suspension cultures of soybean root cells. Exp Cell Res
and forestry. In: Radomiljac AM, Ananthapadmanabho HS, 50:151–158. doi:10.1016/0014-4827(68)90403-5
Welbourn RM, Rao KS (eds) Sandal and its Products. Proceed- Ganeshaiah KN, Uma Shaanker R, Vasudeva R (2007) Bio-resources
ings of an international seminar held on 18-19 December 1997 and empire building: What favoured the growth of Vijayanagara
organised by the Institute of Wood Science and Technology Empire? Curr Sci 93:140–146
(ICFRE) and Karnataka State Forest Department, Bangalore, Gillieson D, Page T, Silverman J (2008) An inventory of wild
lndia. Australian Centre for International Agricultural Research, sandalwood stocks in Vanuatu. Australian Centre for Interna-
Canberra, Australia, pp 66–68 tional Agricultural Research, Canberra. http://aciar.gov.au/pub
Das S, Pal S, Mujib A, Sahoo SS, Dey S, Ponde NR, Das Gupta S lication/fr2008-08 (last accessed 14 December, 2015)
(1999) A novel process for rapid mass propagation of Santalum Gunn BG, Bewang IF, Bunn Y (2002) A strategy for conserving,
album L. in liquid media and bioreactor. Acta Hortic managing the genetic resources of Santalum macgregorii (PNG
502:281–288. doi:10.17660/ActaHortic.1999.502.45 sandalwood) in Papua New Guinea. Report prepared for PNG
Das S, Ray S, Dey S, Das Gupta S (2001) Optimisation of sucrose, Forest Authority as part of ACIARs Domestication of Papua
inorganic nitrogen and abscisic acid levels for Santalum album New Guinea’s Indigenous Forest Species Project (ACIAR FST/
L. somatic embryo production in suspension culture. Process 1998/115). CSIRO Forestry and Forest Products, Canberra,
Biochem 37:51–56. doi:10.1016/S0032-9592(01)00168-6 pp 1–43

123
Planta (2016) 243:847–887 883

Harbaugh DT (2007) A taxonomic revision of Australian northern Jiko LR (1993) Status and current interest in sandalwood in Fiji. In:
sandalwood (Santalum lanceolatum, Santalaceae). Aust Syst Bot McKinnell FH (ed) Sandalwood in the Pacific Region. Proceed-
20:409–416. doi:10.1071/SB07009 ings of Symposium, 2 June 1991 at the XVII Pacific Science
Harbaugh DT (2008) Polyploid and hybrid origins of Pacific island Congress, Honolulu, Hawaii, ACIAR Proceedings No. 49.
sandalwoods (Santalum, Santalaceae) inferred from low-copy Australian Centre for International Agricultural Research, Can-
nuclear and flow cytometry data. Int J Plant Sci 169:677–685. berra, Australia, pp 13–18
doi:10.1086/533610 Jyothi PV, Atluri JB, Subba Reddi C (1991) Pollination ecology of
Harbaugh DT, Baldwin BG (2007) Phylogeny and biogeography of Santalum album (Santalaceae). Trop Ecol 32:98–104
the sandalwoods (Santalum, Santalaceae): repeated dispersals Kalo P, Gleason C, Edwards A, Marsh J, Mitra RM, Hirsch S, Jakab J,
throughout the Pacific. Am J Bot 94:1028–1040. doi:10.3732/ Sims S, Long SR, Rogers J, Kiss GB, Downie JA, Oldroyd GE
ajb.94.6.1028 (2005) Nodulation signaling in legumes requires NSP2, a
Harbaugh DT, Oppenheimer HL, Wood KR, Wagner WL (2010) member of the GRAS family of transcriptional regulators.
Taxonomic revision of the endangered Hawaiian red-flowered Science 308:1786–1789. doi:10.1126/science.1110951
sandalwoods (Santalum) and discovery of an ancient hybrid Kulkarni HD, Muniyamma M (1998) Floral biology and breeding
species. Syst Bot 35:827–838. doi:10.1600/036364410X539899 systems in sandal, Santalum album L. In: Radomiljac AM,
Havea (2012) The vegetative propagation of sandalwood species Ananthapadmanabho HS, Welbourn RM, Rao KS (eds)
Santalum yasi, S. album and F1 hybrid. In: Thomson L, Padolina Sandal and its Products. Proceedings of an international
C, Sami R, Prasad V, Doran J (eds). Regional workshop on seminar held on 18–19 December 1997 organised by the
sandalwood resource development research and trade in the Institute of Wood Science and Technology (ICFRE) and
Pacific and Asian Region. European Union, Secretariat of the Karnataka State Forest Department, Bangalore, lndia. Aus-
Pacific Community, James Cook University and the Australian tralian Centre for International Agricultural Research, Can-
Centre for International Agricultural Research. Port Vila, berra, Australia, pp 135–146
Vanuatu, 2012. pp 109–110 Kushalapa KA (1998) Trade liberalisation in sandalwood. In:
Heena Kausar S, Jahan N, Ahmed K, Aslam M, Ahmed P, Ahmed S Radomiljac AM, Ananthapadmanabho HS, Welbourn RM, Rao
(2014) Unani perspective and recent studies of sandal safed KS (eds) Sandal and its Products. Proceedings of an international
(Santalum album Linn.): a review. World J Pharm Pharm Sci seminar held on 18–19 December 1997 organised by the Institute
3:2133–2145 of Wood Science and Technology (ICFRE) and Karnataka State
Hirano RT (1990) Propagation of Santalum, sandalwood tree. USDA Forest Department, Bangalore, lndia. Australian Centre for
Forest Service General Technical Report PSW-122. pp 43–45 International Agricultural Research, Canberra, Australia,
Hoagland DR, Arnon DI (1938) The water-culture method for pp 24–26
growing plants without soil. Circular (California Agricultural Lakshmi Sita G (1986) Sandalwood (Santalum album L.). In: Bajaj
Experiment Station). p 347 YPS (ed) Biotechnology in agriculture and forestry, vol 1.,
Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Trees, Springer, Berlin, pp 363–374
Agrobacterium helper plasmids for gene transfer to plants. Lakshmi Sita G, Shobha Rani B (1983) Preliminary studies on
Transgenic Res 2:208–218. doi:10.1007/BF01977351 isolation and culture of protoplast from sandalwood (Santalum
Ilah A, Abdin MZ, Mujib A (2002) Somatic embryo irregularities in album). In: Potrykus I, Harms CT, Hinnen A, Hütter R, King PJ,
in vitro cloning of sandal (Santalum album L.). Sandalwood Res Shillito RD (eds) EXS 45: Experientia Supplementum: Proto-
Newslett 15:2–3 plast, Vol. 45. Birkhäuser Basel, Basel, pp 4–5. doi: 10.1007/
Irving LJ, Cameron DD (2009) You are what you eat: interactions 978-3-0348-6556-2
between root parasitic plants and their hosts. Adv Bot Res Lakshmi Sita G, Raghava Ram NV, Vaidyanathan CS (1979)
50:87–138. doi:10.1016/S0065-2296(08)00803-3 Differentiation of embryoids and plantlets from shoot callus of
IUCN (2015b) Santalum fernandezianum. IUCN Red List of Threat- sandalwood. Plant Sci Lett 15:265–270. doi:10.1016/0304-
ened SpeciesTM version 2015-4. http://www.iucnredlist.org/ 4211(79)90118-4
details/30406/0 (last accessed 14 December, 2015) Lakshmi Sita G, Raghava Ram NV, Vaidyanathan CS (1980a)
IUCN (2015c) Santalum macgregorii. IUCN Red List of Threatened Triploid plants from endosperm cultures of sandalwood by
SpeciesTM version 2015-4. http://www.iucnredlist.org/details/ experimental embryogenesis. Plant Sci Lett 20:63–69. doi:10.
38390/0 (last accessed 14 December, 2015) 1016/0304-4211(80)90070-X
IUCN (2015d) Santalum haleakalae. IUCN Red List of Threatened Lakshmi Sita G, Shobha J, Vaidyanathan CS (1980b) Regeneration of
SpeciesTM version 2015-4. http://www.iucnredlist.org/details/ whole plants by embryogenesis from cell suspension cultures of
30783/0 (last accessed 14 December, 2015) sandalwood. Curr Sci 49:196–198
IUCN (International Union for Conservation of Nature) (2015a) Legocki RP, Verma DPS (1979) A nodule-specific plant protein
Santalum album (sandalwood). IUCN Red List of Threatened (Nodulin-35) from soybean. Science 205:190–193. doi:10.1126/
SpeciesTM version 2015-4. http://www.iucnredlist.org/details/ science.205.4402.190
31852/0 (last accessed 14 December, 2015) Li YL (2003) Study on the introduction of sandalwood. Science Press,
Jackman M (CEO, Elders) (2013) Completion of sale of sandalwood Beijing, China, p 154 (in Chinese)
assets. Company statement, 28 March, 2013. p 1 Liu XJ, Xu DP, Xie ZS, Zhang NN (2009) Effects of different culture
Janarthanam B, Sumathi E (2011) High frequency shoot regeneration media on the growth of Indian sandalwood (Santalum album L.)
from internodal explants of Santalum album L. Int J Bot seedlings in Zhanjiang, Guangdong, southern China. For Stud
7:249–254. doi:10.3923/ijb.2011.249.254 China 11:132–138. doi:10.1007/s11632-009-0023-4
Janarthanam B, Dhamotharan R, Sumathi E (2012) Thidiazuron Lloyd G, McCown B (1980) Commercially-feasible micropropaga-
(TDZ)-induced plant regeneration from internodal explants of tion of mountain laurel, Kalmia latifolia, by use of shoot-tip
Santalum album L. J Biosci Res 3:145–153 culture. Int Plant Propag Soc Proc 30:421–427
Jayawardena MMDM, Jayasuriya KMGG, Walck JL (2015) Confir- Loneragan OW (1990) Historical review of sandalwood (Santalum
mation of morphophysiological dormancy in sandalwood (San- spicatum) research in Western Australia. Research Bulletin—
talum album, Santalaceae) seeds. J Natn Sci Found Sri Lanka Department of Conservation and Land Management, Western
43:209–215. doi:10.4038/jnsfsr.v43i3.7949 Australia

123
884 Planta (2016) 243:847–887

Loveys BR, Loveys BR, Tyerman SD (2001a) Water relations and gas Murphy MT, Garkaklis MJ, Hardy GESJ (2005) Seed caching by
exchange of the root hemiparasite Santalum acuminatum woylies Bettongia penicillata can increase sandalwood Santalum
(Quandong). Aust J Bot 49:479–486. doi:10.1071/BT00080 spicatum regeneration in Western Australia. Austral Ecol
Loveys BR, Tyerman SD, Loveys BR (2001b) Transfer of photosyn- 30:747–755. doi:10.1111/j.1442-9993.2005.01515.x
thate and naturally occurring insecticidal compounds from host Nagaveni HC, Srimathi RA (1981) Studies on germination of sandal
plants to the root hemiparasite Santalum acuminatum (Santa- (Santalum album L.) pre-treatment of sandal seeds. Indian For
laceae). Aust J Bot 49:9–16. doi:10.1071/BT99080 107:348–354
Loveys BR, Tyerman SD, Loveys BR (2002) Effect of different host Nagaveni HC, Vijayalakshmi G (2003) Growth performance of sandal
plants on the growth of the root hemiparasite Santalum (Santalum album L.) with different host species. Sandalwood
acuminatum (Quandong). Aust J Exp Agric 42:97–102. doi:10. Res Newsl 18:1–4
1071/EA01093 Nageswara Rao M, Soneji JR, Sudarshana P (2011) Santalum. In:
Lu JK (2011) The parasitism between hemiparasite Santalum album Kole C (ed) Wild crop relatives: genomic and breeding
and its hosts. PhD thesis, Chinese Academy of Forestry, Beijing, resources. Springer, Berlin, pp 131–144. doi:10.1007/978-3-
China (in Chinese with English abstract) 642-21250-5
Lu JK, Kang LH, Sprent JI, Xu DP, He XH (2013) Two-way transfer Nasi RP (1995) Germination and seed dormancy in Santalum
of nitrogen between Dalbergia odorifera and its hemiparasite austrocaledonicum: a synopsis. In: Gjerum L, Fox JED, Ehrhart
Santalum album is enhanced when the host is effectively Y (ed) Sandalwood seed nursery and plantation technology.
nodulated and fixing nitrogen. Tree Physiol 33:464–474. Proceedings of a Regional Workshop for Pacific Island Countries
doi:10.1093/treephys/tpt024 held at Nouméa, New Caledonia, 1-11 August 1994. FAO, Suva,
Lu JK, Xu DP, Kang LH, He XH (2014) Host-species-dependent Fiji. RAS/92/361. Field Document N°8, pp 59–74
physiological characteristics of hemiparasite Santalum album in Nikam TD, Barmukh RB (2009) GA3 enhances in vitro seed
association with N2-fixing and non-N2-fixing hosts native to germination in Santalum album. Seed Sci Technol 37:276–280.
southern China. Tree Physiol 34:1006–1017. doi:10.1093/ doi:10.15258/sst.2009.37.2.02
treephys/tpu073 Ora YANR (2012) An integrated management and conservation
Ma GH, He YM, Zhang JF, Chen FL (2005) Studies on semi-parasitic strategy for sandalwood (Santalum album L.) in West Timor,
sandalwood seedlings. J Trop Subtrop Bot 13:233–238 (in Indonesia. Paper presented at the Regional workshop on
Chinese with English abstract) Sandalwood Resource Development, Research and Trade in
Ma GH, Bunn E, Zhang JF, Wu GJ (2006) Evidence of dichogamy in the Pacific and Asian Region, Port Vila, Vanuatu, 22–25
Santalum album L. J Integr Plant Biol 48:300–306. doi:10.1111/ November 2010, Thomson L, Padolina C, Sami R, Prasad V,
j.1744-7909.2006.00201.x Doran J (eds), pp 65–89
Ma GH, Hu YJ, Xu QS (2008) Tissue culture and rapid propagation of Ouyang Y, Zhang XH, Chen YL, Teixeira da Silva JA, Ma GH (2015)
Santalum album L. Plant Physiol Commun 44:296 (in Chinese) Growth, photosynthesis and haustorial development of semipar-
Manonmani V, Vanangamudi K (2001) Influence of degree of fruit asitic Santalum album L. penetrating into roots of three hosts: a
maturity on seed germination, seedling vigour and storability of comparative study. Trees—structure function. doi: 10.1007/
sandal (Santalum album). J Trop For Sci 13:415–422 s00468-015-1303-3
Manonmani V, Vanangamudi K (2002) Effect of seed source and size Ozias-Akins P, Rao PS, Schneider O (1985) Plant regeneration from
on seed germination and seedling vigour of sandal (Santalum embryogenic suspension-derived protoplasts of sandalwood (San-
album). J Trop For Sci 14:150–155 talum album). In: Henke RR, Hughes KW, Constantin MJ,
McKinnell FH (2011) WA sandalwood industry development plan Hollaender A, Wilson C (eds) Tissue Culture in Forestry and
2008–2020. Australian Sandalwood Network, Forest Products Agriculture, Basic Life Sciences Vol. 32. Springer US, Boston,
Commission, Western Australia, pp 1–48 MA, pp 338–339 (poster abstract). doi: 10.1007/978-1-4899-0378-5
Mei QW, Zhang XH, Ma GH (2011) Influence of rhizospheric pH Page T, Southwell I, Russell M, Leakey R (2007) Evaluation of
value of host on growth of Indian sandalwood and preference to heartwood and oil characters in seven populations of Santalum
host. J Trop Subtrop Bot 19:565–570. doi:10.3969/j.issn.1005- lanceolatum from Cape York. In: Thomson L, Bulai S,
3395.2011.06.014 Wilikibau B (eds) Regional workshop on sandalwood research,
Mele L (2001) Media release: sandalwood trading season 2001. development and extension in the Pacific Islands and Asia, Nadi,
Vanuatu Department of Forests, Port Vila Fiji, 28 Nov – 1 Dec 2005. Secretariat of the Pacific Community
Mo XL, Zeng QQ, Qiu WF, Chen YZ (2008) Study on somatic (SPC), Australian Agency for International Development Assis-
embryogenesis from sandalwood and plantlet regeneration. Food tance (AusAID), German Agency for Technical Cooperation
Drug 10:35–37 (in Chinese with English abstract) (GTZ). pp 131–136
Mo XL, Zeng QQ, Chen YW, Chen YZ, Zhen Y (2010) Zygotic Page T, Potrawiak A, Berry A, Tate H, Tungton J, Tabi M (2010a)
embryo culture in vitro and rapid propagation of Santalum Production of sandalwood (Santalum austrocaledonicum) for
album. Subtrop Plant Sci 39:32–34. doi:10.3969/j.issn.1009- improved smallholder incomes in Vanuatu. For Trees Liveli-
7791.2010.02.009 (in Chinese with English abstract) hoods 19:299–316. doi:10.1080/14728028.2010.9752673
Muir K, Byrne M, Barbour E, Cox MC, Fox JED (2007) High levels Page T, Southwell I, Russell M, Leakey R (2010b) Geographic and
of outcrossing in a family trial of Western Australian sandal- phenotypic variation in heartwood and essential-oil characters in
wood (Santalum spicatum). Silvae Genet 56:222–230 natural populations of Santalum austrocaledonicum in Vanuatu.
Mujib A (2005) In vitro regeneration of sandal (Santalum album L.) Chem Biodivers 7:1990–2006. doi:10.1002/cbdv.200900382
from leaves. Turk J Botany 29:63–67 Parthasarathi K, Gupta SK, Rao PS (1974) Differential response in the
Muralidharan EM (1997) Micropropagation of teak, rosewood and cation exchange capacity of the host plants on parasitization on
sandal. Kerala Forest Research Institute (KFRI) Research Report sandal (Santalum album). Curr Sci 43:20
119. KFRI, Peechi,Thrissur, India, pp 1–20 Tennakoon KU, Pate JS, R SG (1997b) Haustorium-related uptake
Murashige T, Skoog F (1962) A revised medium for rapid growth and and metabolism of host xylem solutes by the root hemiparasitic
bioassays with tobacco tissue cultures. Physiol Plant shrub Santalum acuminatum (R. Br.) A. DC. (Santalaceae). Ann
15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x Bot 80:257–264. doi: 10.1006/anbo.1997.0433

123
Planta (2016) 243:847–887 885

Peeris M, Senarath W (2015) In vitro propagation of Santalum album Rao PS, Bapat VA (1993) Micropropagation of sandalwood (San-
L. J Natl Sci Found Sri Lanka 43:265–272. doi:10.4038/jnsfsr. talum album L.) and mulberry (Morus indica L.). In: Ahuja MR
v43i3.7954 (ed) Micropropagation of woody plants, forestry science, Vol.
Primawati E (2006) Perbanyakan cendana (Santalum album Linn.) 44. Kluwer Academic Publishers, Dordrecht, Netherland,
secara kultur in vitro dengan pemberian zat pengatur tumbuh pp 317–345. doi:10.1007/978-94-015-8116-5_19
sitokinin (BAP dan kinetin). PhD thesis, Institut Pertanian Rao PS, Bapat VA (1995) Somatic embryogenesis in sandalwood
Bogor, Indonesia. Departemen Konservasi Sumberdayahutan (Santalum album L.). In: Jain SM, Gupta PK, Newton RJ (eds)
Dan Ekowisatafakultas Kehutanan, pp 1–54 (in Indonesian) Somatic embryogenesis in woody plants, Vol. 2. Kluwer
Radhakrishnan S, Vanangamudi K, Parthiban KT (2001) Callogenesis Academic Publishers, Dordrecht, Netherland, pp 153–170.
and organogenesis in sandal (Santalum album). J Trop For Sci doi:10.1007/978-94-011-0491-3
13:391–393 Rao PS, Ozias-Akins P (1985) Plant regeneration through somatic
Radomiljac AM (1998) The influence of pot host species, seedling embryogenesis in protoplast cultures of sandalwood (Santalum
age and supplementary nursery nutrition on Santalum album album L.). Protoplasma 124:80–86. doi:10.1007/BF01279726
Linn. (Indian sandalwood) plantation establishment within the Rao PS, Raghava Ram NV (1983) Propagation of sandalwood (Santalum
Ord River Irrigation Area, Western Australia. For Ecol Manage album Linn) using tissue and organ culture technique. In: Sen SK,
102:193–201. doi:10.1016/S0378-1127(97)00158-8 Giles KL (eds) Plant Cell Culture in Crop Improvement, Basic Life
Radomiljac AM, McComb JA, Pate JS, Tennakoon KU (1998a) Sciences, Vol. 22. Plenum Press (Now Springer), New York,
Xylem transfer of organic solutes in Santalum album L. (Indian pp 119–124. doi:10.1007/978-1-4684-4379-0
sandalwood) in association with legume and non-legume hosts. Rao PS, Rangaswamy NS (1971) Morphogenic studies in tissue
Ann Bot 82:675–682. doi:10.1006/anbo.1998.0741 cultures of the parasitic Santalum album L. Biol Plant
Radomiljac AM, McComb JA, Shea SR (1998b) Field establishment 13:200–206. doi:10.1007/BF02933637
of Santalum album L.: the effect of the time of introduction of a Rao PS, Srimathi RA (1977) Vegetative propagation of sandal,
pot host (Alternanthera nana R. Br.). For Ecol Manage Santalum album L. Curr Sci 46:276–277
111:107–118 Rao PS, Bapat VA, Mhatre M (1984) Regulatory factors for in vitro
Radomiljac AM, McComb JA, McGrath JF (1999a) Intermediate host multiplication of sandalwood tree (Santalum album Linn.). II.
influences on the root hemi-parasite Santalum album L. biomass Plant regeneration in nodal and internodal stem explants and
partitioning. For Ecol Manage 113:143–153. doi:10.1016/S0378- occurrence of somaclonal variation in tissue culture raised
1127(98)00421-6 plants. Proc Indian Natl Acad Sci B 50:196–202
Radomiljac AM, McComb JA, Pate JS (1999b) Heterotrophic carbon Rashkow ED (2014) Perfumed the axe that laid it low: the
gain and mineral nutrition of the root hemi-parasite Santalum endangerment of sandalwood in southern India. Indian Econ
album L. in pot culture with different hosts. Aust For Soc Hist Rev 51:41–70. doi:10.1177/0019464613515553
62:128–138. doi:10.1080/00049158.1999.10674774 Ratnaningrum YWN, Indrioko S (2014) Variation on genotypes and
Radomiljac AM, McComb JA, Pate JS (1999c) Gas exchange and flowering characters affecting pollination mechanisms of san-
water relations of the root hemi-parasite Santalum album L. in dalwood (Santalum album Linn., Santalaceae) planted on ex situ
association with legume and non-legume hosts. Ann Bot gene conservation in Yogyakarta, Indonesia. Eurasian J For Res
83:215–224. doi:10.1006/anbo.1998.0815 17:19–34
Rai R, McComb J (1997) Direct somatic embryogenesis from mature Revathy E, Arumugam S (2011) Somatic embryogenesis and plantlets
embryos of sandalwood. Sandalwood Res Newsl 6:2–3 regeneration from seedling explants of Santalum album L. Int J
Rai VRV, McComb J (2002) Direct somatic embryogenesis from Curr Res 33:237–241
mature embryos of sandalwood. Plant Cell Tissue Organ Cult Ritter (1836) Das Sandelholz (Santalum album Linn. Dschandana im
69:65–70. doi:10.1023/A:1015037920529 Sanskr., Zandal im Arab., Dschandan im Hindi und Mongolis-
Rai SN, Sarma CR (1990) Depleting sandalwood production and chen) in Malabar und seine Verbreitungssphäre. Arch Pharm
rising prices. Indian For 116:348–354 56:293–305 (in German). doi: 10.1002/ardp.18360560313
Ral SN (1990) Status and cultivation of sandalwood in India. In: Rocha D, Ashokan PK, Santhoshkumar AV, Anoop EV, Sureshkumar
Hamilton L, Conrad CE (ed) Proceedings of the Symposum on P (2014) Influence of host plant on the physiological attributes of
Sandalwood in the Pacific, April 9–11. General Technical Report field-grown sandal tree (Santalum album). J Trop For Sci
PSW-122. Honolulu, Hawaii, pp 66–71 26:166–172
Rangaswamy NS (1961) Experimental studies on female reproductive Rocha D, Ashokan PK, Santhoshkumar AV, Anoop EV, Sureshkumar
structures of Citrus microcarpa BUNGE. Phytomorphology P (2015) Anatomy and functional status of haustoria in field
11:109–127 grown sandalwood tree (Santalum album L.). For Res 04:148.
Rangaswamy NS, Rao PS (1963) Experimental studies on Santalum doi:10.4172/2168-9776.1000148
album L.—establishment of tissue culture of endosperm. Phy- Rugkhla A, Jones MGK (1998) Somatic embryogenesis and plantlet
tomorphology 14:450–454 formation in Santalum album and S. spicatum. J Exp Bot
Rao LN (1942a) Parasitism in the Santalaceae. Ann Bot 6:131–150 49:563–571. doi:10.1093/jxb/49.320.563
Rao LN (1942b) Studies in the Santalaceae. Ann Bot 6:151–175 Rugkhla A, McComb JA, Jones MGK (1997) Intra- and inter-specific
Rao PS (1965) In vitro induction of embryonal proliferation in pollination of Santalum spicatum and S. album. Aust J Bot
Santalum album L. Phytomorphology 15:175–179 45:1083–1095. doi:10.1071/BT96079
Rao PS, Bapat VA (1978) Vegetative propagation of sandalwood Sahai A, Shivanna K (1984) Seed germination, seedling growth and
plants through tissue culture. Can J Bot 56:1153–1156. doi:10. haustorial induction in Santalum album, a semi-root parasite.
1139/b78-129 Proc Indian Acad Sci 93:571–580. doi:10.1007/BF03053220
Rao PS, Bapat VA (1992) Micropropagation of sandalwood (San- Samantray S, Upadhyaya C (2010) Methodological studies and
talum album L.). In: Bajaj YPS (ed) High-tech and microprop- research on micropropagation of Chandan (Santalum album L.):
agation II. Biotechnology in agriculture and forestry, Vol. 18. an endangered plant. Int J Sci Technol 1:10–18
Springer-Verlag, Berlin, Heidelberg, pp 193–210. doi:10.1007/ Sanjaya HSA, Rai VR (2003) In vitro and in vivo micrografting of
978-3-642-76422-6 Santalum album shoot tips. J Trop For Sci 15:234–236

123
886 Planta (2016) 243:847–887

Sanjaya Muthan B, Rathore TS, Rai VR (2006a) Factors influencing Tassin J (2005) Status of research on Santalum austrocaledonicum in
in vivo and in vitro micrografting of sandalwood (Santalum New Caledonia current knowledge and future prospects. In:
album L.): an endangered tree species. J For Res 11:147–151. Thomson L, Bulai S, Sovea L (eds) Regional workshop on
doi:10.1007/s10310-005-0208-1 Sandalwood Research, Development and Extension in the
Sanjaya Muthan B, Rathore TS, Rai VR (2006b) Micropropagation of Pacific Islands and Asia. Secretariat of the Pacific Community:
an endangered Indian sandalwood (Santalum album L.). J For Noumea, New Caledonia, Suva, Fiji, pp 135–141
Res 11:203–209. doi:10.1007/s10310-006-0207-x Tate H, Sethy M, Tungon J (2006) Grafting of sandalwood in
Sankara Rao K, Chrungoo NK, Sinha A (1996) Characterization of Vanuatu. Sandalwood Res Newslett 21:7
somatic embryogenesis in sandalwood (Santalum album L.). Teixeira da Silva JA (2012a) Is BA (6-benzyladenine) BAP (6-
In Vitro Cell Dev Biol Plant 32:123–128. doi:10.1007/ benzylaminopurine)? Asian Aust J Plant Sci Biotechnol 6
BF02822754 (special issue 1):121–124
Scott J (1871) Root parasitism by sandalwood. J Agric Hortic Soc Teixeira da Silva JA (2012b) Callus, calluses or calli: multiple
India 2:287 plurals? Asian Aust J Plant Sci Biotechnol 6 (special issue
Sharma S, Shahzad A, Teixeira da Silva JA (2013) Synseed 1):125–126
technology—a complete synthesis. Biotechnol Adv Teixeira da Silva JA, Dobránszki J (2013) Plant thin cell layers: a
31:186–207. doi:10.1016/j.biotechadv.2012.09.007 40-year celebration. J Plant Growth Regul 32:922–943. doi:10.
Shekhawat UKS, Ganapathi TR, Srinivas L, Bapat VA, Rathore TS 1007/s00344-013-9336-6
(2008) Agrobacterium-mediated genetic transformation of Teixeira da Silva JA, Kerbauy GB, Zeng S, Chen Z, Duan J (2014)
embryogenic cell suspension cultures of Santalum album L. In vitro flowering of orchids. Crit Rev Biotechnol 34:56–76.
Plant Cell Tissue Organ Cult 92:261–271. doi:10.1007/s11240- doi:10.3109/07388551.2013.807219
007-9330-4 Teixeira da Silva JA, Winarto B, Dobránszki J, Zeng S (2015)
Shekhawat UKS, Ganapathi TR, Srinivas L (2010) Expression of Disinfection procedures for in vitro propagation of Anthurium.
hepatitis B small surface antigen in Santalum album embryo- Folia Hortic 27:3–14. doi:10.1515/fhort-2015-0009
genic cell suspension cultures. Biol Plant 54:720–724. doi:10. Tennakoon KU, Cameron DD (2006) The anatomy of Santalum
1007/s10535-010-0128-6 album (sandalwood) haustoria. Can J Bot 84:1608–1616. doi:10.
Shiri V, Rao KSS (1998) Introduction and expression of marker genes 1139/b06-118
in sandalwood (Santalum album L.) following Agrobacterium- Tennakoon KU, Pate JS, Arthur D (1997) Ecophysiological aspects of
mediated transformation. Plant Sci 131:53–63. doi:10.1016/ the woody root hemiparasite Santalum acuminatum (R. Br.) A.
S0168-9452(97)00232-X DC and its common hosts in South Western Australia. Ann Bot
Sindhu Veerendra HC, Anantha Padmanabha HS (1996) The breeding 80:245–256. doi:10.1006/anbo.1997.0432
system in sandal (Santalum album L.). Silvae Genet 45:188–190 The Plant List (2015) Santalaceae. http://www.theplantlist.org/1.1/
Singh CK, Raj SR, Patil VR, Jaiswal PS, Subhash N (2013) Plant browse/A/Santalaceae/Santalum/ (last accessed 14 December,
regeneration from leaf explants of mature sandalwood (Santalum 2015)
album L.) trees under in vitro conditions. In Vitro Cell Dev Biol Thimijan RW, Heins RD (1983) Photometric, radiometric, and
Plant 49:1–7. doi:10.1007/s11627-013-9495-y quantum light units of measure: a review of procedures for
Singh CK, Raj SR, Jaiswal PS, Patil VR, Punwar BS, Chavda JC, interconversion. HortScience 18:818–822
Subhash N (2015) Effect of plant growth regulators on in vitro Thomson L, Bulai S, Wilikibau B (eds) (2005a) Proceedings of the
plant regeneration of sandalwood (Santalum album L.) via regional workshop on Sandalwood research, development and
organogenesis. Agrofor Syst In press. doi: 10.1007/s10457-015- extension in the Pacific islands from 28 November to 1
9853-3 December 2005. Nadi, Fiji, pp 1–146
Smit P, Raedts J, Portyanko V, Debellé F, Gough C, Bisseling T, Thomson L, Wainiqolo I, Smith A, Manu V, Havea M, Robson K
Geurts R (2005) NSP1 of the GRAS protein family is essential (2005b) Sandalwood work in SPRIG (South Pacific Regional
for rhizobial Nod factor-induced transcription. Science Initiative on Forest Genetic Resources). Paper presented at the
308:1789–1791. doi:10.1126/science.1111025 Regional workshop on Sandalwood Research, Development and
Srimathi RA, Sreenivasaya M (1962) Occurrence of endopolyploidy Extension in the Pacific Islands and Asia, Noumea, New
in the haustorium of Santalum album Linn. Curr Sci 31:69–70 Caledonia, 7–11 October 2002
St. Jack D, Hesterman DC, Guzzomi AL (2013) Precision metering of Timber-Queensland (2012) Queensland forest and timber industry
Santalum spicatum (Australian sandalwood) seeds. Biosyst Eng situation analysis. Queensland Government, Brisbane. https://
115:171–183. doi:10.1016/j.biosystemseng.2013.03.004 www.daf.qld.gov.au/__data/assets/pdf_file/0006/56490/qld-for
Stebbins GL (1971) Chromosomal evolution in higher plants. Edward est-timber-industry-situation-analysis.pdf (last accessed 14
Arnold Ltd., London December, 2015)
Struthers R, Lamont BB, Fox JED, Wijesuriya S, Crossland T (1986) Tosul D (2015) Sandalwood Harvesting Season Order 2015. Depart-
Mineral nutrition of sandalwood (Santalum spicatum). J Exp Bot ment of Agriculture, Livestock, Forest, Fisheries and Biosecu-
37:1274–1284. doi:10.1093/jxb/37.9.1274 rity, Republic of Vanuatu
Subasinghe SMCUP (2013) Sandalwood research: a global perspec- Traditionaloven.com (2015) http://www.traditionaloven.com/build
tive. J Trop For Environ 3:1–8 ing/masonry/concrete/convert-tonne-metric-t-concrete-to-cubic-
Suriamihardja H, Suriamihardja S (1993) Sandalwood in Nusa metre-m3-concrete.html (last accessed 14 December, 2015)
Tenggara Timur. In: McKinnell FH (ed) Sandalwood in the Tropical Forestry Services (TFS) (2015a) Why Indian Sandalwood?
Pacific Region. Proceedings of symposium, 2 June 1991 at the TFS Sandalwood Project 2015, Indian Sandalwood. Product
XVII Pacific Science Congress. ACIAR, Honolulu, Hawaii, Disclosure Statement. Tropical Forestry Services Ltd, 169
pp 39–43 Broadway, Nedlands WA 6009, Australia, pp. 14–15
Tamla HT, Cornelius JP, Page T (2011) Reproductive biology of Tropical Forestry Services (TFS) (2015b) The Sandalwood Market—
three commercially valuable Santalum species: development of Supply and Demand. TFS Sandalwood Project 2015, Indian
flowers and inflorescences, breeding systems, and interspecific Sandalwood. Product Disclosure Statement. Tropical Forestry
crossability. Euphytica 184:323–333. doi:10.1007/s10681-011- Services Ltd, 169 Broadway, Nedlands WA 6009, Australia,
0530-y pp. 16–17

123
Planta (2016) 243:847–887 887

Tropical Forestry Services (TFS) (2015c) TFS Sandalwood Project Woodall GS, Robinson CJ (2003) Natural diversity of Santalum
2015, Indian Sandalwood. Product Disclosure Statement. Trop- spicatum host species in south-coast river systems and their
ical Forestry Services Ltd, 169 Broadway, Nedlands WA 6009, incorporation into profitable and biodiverse revegetation. Aust J
Australia Bot 51:741–753. doi:10.1071/BT02118
Uniyal DP, Thapliyal RC, Rawat MS (1985) Vegetative propagation Yang XY, Zhang XH, Teixeira da Silva JA, Liang KM, Deng RF, Ma
of sandal by root cuttings. Indian For 111:145–148 GH (2014) Ontogenesis of the collapsed layer during haustorium
Veillon JM, Jaffré T (1995) Sandalwood (Santalum austrocale- development in the root hemi-parasite Santalum album Linn.
donicum Vieillard) in New Caledonia: taxonomy, distribution, Plant Biol 16:282–290. doi:10.1111/plb.12026
ecology. In: Gjerum L, Fox JED. Ehrhart Y (ed) Sandalwood Zauba.com (2015) Detailed import data of Santalum spicatum. https://
Seed Nursery and Plantation Technology. Proceedings of a www.zauba.com/import-santalum-spicatum-hs-code.html (last
Regional Workshop for Pacific Island Countries held at Nouméa, accessed 14 December, 2015)
New Caledonia, 1–11 August 1994. FAO, Suva, Fiji.RAS/92/ Zhang NN, Xu DP, Wang WW, Yao QD, Deng HD, Peng WX (2007)
361. Field Document volume 8, pp. 25–36 Cultivation technology and development strategy of sandalwood
Warburton CL, James EA, Fripp YJ, Trueman SJ, Wallace HM (Santalum album L.). Pract For Technol 6:13–15 (In Chinese)
(2000) Clonality and sexual reproductive failure in remnant Zhang XH, Teixeira da Silva JA, Ma GH (2010) Karyotype analysis
populations of Santalum lanceolatum (Santalaceae). Biol Con- of Santalum album L. Caryologia 63:142–148. doi:10.1080/
serv 96:45–54. doi:10.1016/S0006-3207(00)00049-5 00087114.2010.10589719
Wawo AH (2008) Study on seed germination and seedling growth Zhang XH, Teixeira da Silva JA, Duan J, Deng R, Xu X, Ma GH
models of sandalwood (Santalum album L.) of several mother (2012a) Endogenous hormone levels and anatomical characters
seed trees in Belu Regency. East NusaTenggara. Biodiversitas of haustoria in Santalum album L. seedlings before and after
9:117–122. doi:10.13057/biodiv/d090209 (in Indonesian with attachment to the host. J Plant Physiol 169:859–866. doi:10.
English abstract) 1016/j.jplph.2012.02.010
Werren M (2011) Some suggestions for plantation forestry in Tropical Zhang XH, Teixeira da Silva JA, Jia YX, Yan J, Ma GH (2012b) Essential
Queensland. Paper presented at the Northern Australia Forestry oils composition from roots of Santalum album L. J Essent Oil Bear
Stakeholder Forum. Cairns, Australia Plants 15:1–6. doi:10.1080/0972060X.2012.10644011
White PR (1963) The cultivation of animal and plant cells, 2nd edn. Zhang XH, Berkowitz O, Teixeira da Silva JA, Zhang M, Ma GH,
The Ronald Press, New York, p 228 Whelan J, Duan J (2015) RNA-Seq analysis identifies key genes
Woodall GS, Robinson CJ (2002) Same day plantation establishment associated with haustorial development in the root hemiparasite
of the root hemiparasite sandalwood (Santalum spicatum (R Br) Santalum album. Front Plant Sci 6:1–17. doi:10.3389/fpls.2015.
A DC: Santalaceae) and hosts. J R Soc West Aust 85:37–42 00661

123