Physiol Mol Biol Plants (January–March 2016) 22(1):153–161

DOI 10.1007/s12298-016-0347-1

RESEARCH ARTICLE

Determining the most effective traits to improve

saffron (Crocus sativus L.) yield
Mahdi Bayat 1 & Mehdi Rahimi 2 & Mehdi Ramezani 3

Received: 23 November 2015 / Revised: 7 March 2016 / Accepted: 14 March 2016 / Published online: 21 March 2016
# Prof. H.S. Srivastava Foundation for Science and Society 2016

Abstract To determine the effective traits to improve saffron weight interred to the regression model and had the highest
yield, a split plot design based on RBCD was done in positive direct and indirect effects on saffron yield. Mainly, it
Mashhad region in Iran for three years (2012–2014). The re- can be derived that the implementation of correct farm man-
sults showed that all traits except number of daughter corm, agement including appropriate sowing date, saffron ecotypes,
fresh weight of daughter corm and dry leaf weight had low proper density, bigger and higher quality saffron corm can
general heritability. Results of genotypic and phenotypic co- play an important role in improving yield components and
efficients of variation and genetic advance demonstrated that subsequently increasing saffron yield.
the majority of traits had a low diversity and the selection did
not have any effect in improving the traits. As a result, the best Keywords Cluster analysis . Genetic advance . Heritability .
way to increase saffron yield is improvement of farm manage- Phenotypic and genotypic path analysis
ment. It was also found that saffron yield had the highest
phenotypic and genotypic correlations with fresh and dry
weight of daughter corm and dry and fresh flower weight. Introduction
Therefore, the efforts to improve these traits will increase saf-
fron yield. According to the present study 5-Jun to 5-Jul was Saffron (Crocus sativus L.) is the oldest plant species which is
found to be the best sowing date for planting saffron. Also, the expensive (Özdemİr et al. 2006; Gómez-Gómez et al. 2012).
Mashhad and Torbat ecotypes were the best ecotypes in this The vegetative propagation is through corms and it is ster-
study. Phenotypic and genotypic path analysis showed that in ile triploid plant (Turhan et al. 2007; Gómez-Gómez et al.
the first step three traits number of daughter corm, fresh flower 2012). Saffron is known for its aromatic properties and
weight and flower number and in the second step traits fresh flavoring in the food industry, use in confectionery
weight of daughter corm, dry flower weight and dry leaf and liquor industries (Gómez-Gómez et al. 2012). There is
increasing interest in saffron because of anticancer properties
and its application in medicine (Magesh et al. 2006;
* Mehdi Rahimi Chryssanthi et al. 2009; Dalezis et al. 2009; Siracusa et al.
[email protected] 2010). Iran is one of the most important countries in the world
where saffron is produced (Jalali-Heravi et al. 2010).
The mature and bigger corms in the previous studies have
1
Young Researchers and Elite Club, Mashhad Branch, Islamic Azad shown more flowers and daughter corms (Molina et al. 2005;
University, Mashhad, Iran
De Juan et al. 2009). Thus, one of the main goals in the
2
Department of Biotechnology, Institute of Science and High production of saffron is to achieve bigger corms (Omidbaigi
Technology and Environmental Sciences, Graduate University of
Advanced Technology, End of Haft Bagh-e-Alavi Highway
2005). In addition, Turhan et al. (2007), Renau-Morata et al.
Knowledge Paradise, 7631133131, P.O.Box: 76315-117, (2012) and Mollafilabi (2004) concluded that corm size is an
Kerman, Iran important factor to determine the presence or absence of
3
Young Researchers and Elite Club, Ahvaz Branch, Islamic Azad flowers even if the corm does not reach to its original size
University, Ahvaz, Iran and produce flowers. Amirnia et al. (2013) and Siracusa
154 Physiol Mol Biol Plants (January–March 2016) 22(1):153–161

et al. (2010) reported that the environment and corm prove- contained 8 culture lines with 3 m length and 25 cm distances
nance have a significant impact on the flower number and from another line (the plot area was 6 m2). Corm distances on
stigma yield. Also researchers such as Molina et al. (2004; lines were 8 cm and 15 cm deep (density was 50 corms m−2).
2005), Gresta et al. (2009) and Maggi et al. (2011) indicated To avoid marginal effects and minimize errors, plots were
that climatic conditions and environments (e.g., temperature, located beside each other by 50 cm distances. To enhance
soil water content) significantly changed quantitative accuracy, margins considered as two lines at the beginning
and qualitative traits of saffron. and at the end of plots as well as 50 cm at two another sides
To identify important traits affecting the yield of plants, we of plots. All agronomical operations (ecotypes and infield
used many multivariate statistical techniques such as cluster preparation, traits measuring, and experimental design) in
analysis, correlation, path analysis, factor analysis, and factor three years were similar, and at the end of each year, the infield
components (Leilah and Al-Khateeb 2005). The former stud- was plowed and corms were harvested for measuring. Rainfed
ies showed that the performance of path analysis to identify conditions met the water requirements at the start of growth
the relationships between the traits is better than correlation (October to November), and plants were drip irrigated from
coefficients (Kozak and Kang 2006; Bahraminejad et al. 2011; December to April.
Darvishzadeh et al. 2011). Correlation coefficient and path Data were collected on the following 13 characters in each
analysis have been used by several researchers (Talebi et al. pot: Flower Number (FN), Fresh Flower Weight (mg) (FFW),
2007; Behradfar et al. 2009) to determine interrelationships Dry Flower Weight (mg) (DFW), Fresh Stigma Weight (mg)
between quantitative characters. (FSW), Dry Stigma Weight (mg) (DSW), Number of daughter
Considering the above facts, this research was undertaken Corm (NDC), Fresh Weight of daughter Corm (mg) (FWDC),
to study the importance of different traits through the estima- Dry Weight of daughter Corm (mg) (DWDC), Length Leaf
tion of genetic parameters, correlation coefficients, cluster (LL), Fresh Leaf Weight (cm) (FLW), Dry Leaf Weight (mg)
analysis and path analysis. The reported information will help (DLW), Biomass (BIO), Harvest Index (HI) and Stigma Yield
to predict the effective traits to improve saffron yield. per m2 (mg m−2) (SY). Then recorded data were analyzed by
using SAS ver. 9.12 and SPSS ver. 21 programs. The coeffi-
cients of variability, heritability and genetic advance as well as
Materials and methods correlation coefficient were estimated according to Johnson
et al. (1955a; 1955b). The cluster analysis based on Ward’s
To determine the effective traits to improve saffron yield, a split method was also used to classify saffron ecotypes. Path coef-
plot experiment based on RBCD was carried out in Mashhad ficients were estimated according to Ehsanzadeh et al. (2004),
region. Sowing dates (05-May, 05-Jun, 05-Jul, 05-Aug. 05-Sep., where saffron yield was kept as the dependent variable and
05-Oct) used as main plots and different saffron ecotypes other contributing traits as independent variables.
(Mashhad, Torbat-jam, Gonabad and Birjand) as sub plot.
Mashhad region is located in 36′15″ latitude, 59′28″ longitude,
and 985 m above the sea level and the soil in the experimental
field was silt-clay with pH 7.8. In this study, different ecotypes Results and discussion
of saffron were evaluated in three distinct years (2011–012,
2012–13 and 2013–14) so that at the end of each year, we drew Analysis of variance
out pervious corms and cultivated new corms in the same farm.
Four saffron samples from different regions of Iran, namely tra- The results of the variance analysis (results are not shown)
ditional saffron production areas, were studied in this work. showed that there were significant differences between years,
Samples 1, 2, and 3 were obtained from the regions of sowing dates, saffron ecotypes, and their interactions with
Mashhad, Torbat-Jam, and Gonabad in Razavi Khorasan prov- respect to all traits. This result enforced that selection of
ince of Iran. Sample 4 was obtained from the region of Birjand in accurate sowing date and appropriate saffron ecotype are
the south Khorasan province of Iran. To prevent Fusarium and important and necessary in each region. Some of the earlier
Penicillium infestations, corms were dipped in a prochloraz so- studies on saffron ecotypes such as the finding of Anastasaki
lution (0.1 %) and dried under forced ventilation for 5–7 h to et al. (2010) on 250 saffron ecotypes and Maggi et al. (2011)
remove the surface water. on 28 authentic saffron samples reported significant differ-
The weight of saffron corms was used in this experiment ences between saffron ecotypes. According to the present
was 10 to 12 g. In each year after preparing the field in April, study 5-Jun to 5-Jul was known the best sowing date for
75 kg ha−1 pure nitrogen, 75 kg ha−1 pure phosphorous, planting saffron. Also, the Mashhad and Torbat ecotypes were
50 kg ha−1 pure potassium were used. The cultivation prac- the best ecotypes in this study. Finally, the saffron corms with
tices used were those commonly used for this crop, and an 10 g and more recommended for cultivation and the corms
organic fertilizer (mature manure) was applied. Each plot with 6 g and smaller are not recommended.
Physiol Mol Biol Plants (January–March 2016) 22(1):153–161 155

The significant differences between traits in given years con- indicated that all studied traits except NDC (CVp= 25.83, CVg=
firmed that studied traits were significantly affected by the en- 21.97) had insignificant variation. In addition, genetic advance
vironment and weather conditions. Thus, the improvement of based on trait mean (Table 1) indicated that genetic advance for
farm management can be effective in improving the traits and all traits except NDC (GAm= 32.89) had a low value. These
subsequently in increasing the saffron yield. Lage and Cantrell results, in total, showed that there is very little variation between
(2009), Molina et al. (2005) and Gresta et al. (2009) reached to saffron ecotypes with respect to all traits. Therefore, the selection
this conclusion that the temperature is one of the environmental for improving of the next saffron generation did not have great
factor to control of growth and flowering in Crocus species. progress. Therefore, the best way to increase the saffron yield is
the improvement of farm management (including proper sowing
date, selecting bigger corms, fertilization, appropriate density,
Genetic parameters
etc.). Sanghera and Kashyap (2012) reported that when genetic
variability within a plant population is low, the possibility of
Heritability is a statistic used in breeding and genetics works
further yield enhancement are scanty. Rubio-Moraga et al.
that estimates how much variation in a phenotypic trait in a
(2009) reported that all saffron accessions appear identical
population is due to genetic variation among individuals in that
clones, not only because of morphological characters but also
population. The response to selection depends on the heritability
at a molecular level. These data strongly suggested that saffron
of the trait and help the plant breeder in breeding program
is a monomorphic species (Caiola et al. 2004).
(Sanghera and Kashyap 2012). The results of the general heri-
tability for studied agronomical traits (Table 1) showed that
traits FN, DSW, FSW, DWDC, HI and SY had the lowest Phenotypic and genotypic correlation coefficients
general heritability (11–28 %) and traits DFW, FFW, LL and
BIO had moderate heritability (44–48 %). As a result, these Measuring the correlation coefficient is a good idea to plant
traits were influenced by the environmental and climatic condi- breeders for produce high yielding cultivars (Mohammadi
tions considerably. On the other hand, it was found that traits et al. 2003). In the present study, in order to study the relation-
NDC, FWDC, and DLW had a high general heritability (65– ships between saffron yield and yield components, phenotypic
72 %) and were slightly affected by the environmental and and genotypic correlation coefficients were calculated. The cor-
climatic conditions. Therefore, we can introduce these traits as relation results (Table 2) indicated that in most cases, the geno-
important criteria for selection of suitable saffron ecotypes. typic correlation is larger than the phenotypic correlation. This
Genetic variability gives a good view of the genetic diversity result was predictable and logical. Because the effects of
of different traits to breeder for use in breeding programs (Singh environmental factors were eliminated in calculation of
et al. 2003). Measuring the genetic diversity is needed to start genotypic correlation coefficient, the relationships were
plant breeding programs in different plants (Ali and Khan 2007). estimated more accurately and realistically. Singh et al. (2003)
Genotypic and phenotypic coefficients of variation (Table 1) stated that genetic correlations are more important than

Table 1 Phenotypic and genotypic parameters of studied traits in saffron ecotypes

Traits General CV% Genetic advance

X
Heritability CVp CVg GA GAm

FN 16.84 0.15 15.78 4.12 3.15 5.14

FFW 352.11 0.48 2.52 1.75 7.55 2.14
DFW 43.14 0.48 2.80 1.94 1.02 2.36
FSW 28.09 0.28 6.03 3.22 0.85 3.02
DSW 5.11 0.15 8.39 3.23 0.11 2.18
NDC 2.30 0.72 25.83 21.97 0.76 32.89
FWDC 5.98 0.65 7.54 6.09 0.52 8.66
DWDC 3.45 0.21 15.79 7.16 0.20 5.71
LL 18.19 0.45 14.72 9.84 2.10 11.57
DLW 3.00 0.69 7.11 5.89 0.26 8.57
BIO 6498.14 0.44 10.36 6.84 516.29 7.95
HI 0.08 0.11 11.34 3.74 0.0001 2.17
SY 89.07 0.13 20.41 7.30 4.09 4.59

Abbreviations are described in Materials and Methods section

156 Physiol Mol Biol Plants (January–March 2016) 22(1):153–161

Table 2 Phenotypic (above diagonal) and genotypic (below diagonal) correlation coefficients between traits of saffron ecotypes
Traits FN FFW DFW FSW DSW NDC FWDC DWDC LL DLW BIO HI SY
FN FN 0.75 ** 0.82 ** 0.93 ** 1.08 ** 0.96 ** 0.97 ** 0.89 ** 0.78 ** 0.90 ** 1.03 ** 0.65 ** 0.76 **
FFW 0.45 ** FFW 0.71 ** 0.75 ** 0.75 ** 0.78 ** 0.83 ** 0.77 ** 0.75 ** 0.77 ** 0.72 ** 0.74 ** 0.98 **
DFW 0.70 ** 0.81 ** DFW 0.75 ** 0.75 ** 0.74 ** 0.78 ** 0.74 ** 0.70 ** 0.77 ** 0.71 ** 0.71 ** 0.98 **
FSW 0.54 ** 0.60 ** 0.63 ** FSW 0.71 ** 0.88 ** 0.76 ** 0.70 ** 0.78 ** 0.87 ** 0.72 ** 0.80 ** 0.79 **
DSW 0.71 ** 0.59 ** 0.75 ** 0.91 ** DSW 0.88 ** 0.76 ** 0.69 ** 0.77 ** 0.86 ** 0.71 ** 0.74 ** 0.73 **
NDC 0.53 ** 0.66 ** 0.67 ** 0.67 ** 0.69 ** NDC 0.84 ** 0.73 ** 0.67 ** 0.74 ** 0.69 ** 0.65 ** 0.82 **
FWDC 0.53 ** 0.66 ** 0.67 ** 0.67 ** 0.69 ** 0.99 ** FWDC 0.60 ** 0.75 ** 0.96 ** 0.70 ** 0.73 ** 1.01 **
DWDC 0.71 ** 0.60 ** 0.70 ** 0.66 ** 0.73 ** 0.54 ** 0.55 ** DWDC 0.66 ** 0.88 ** 0.66 ** 0.75 ** 0.96 **
LL 0.70 ** 0.61 ** 0.71 ** 0.66 ** 0.72 ** 0.55 ** 0.54 ** 0.98 ** LL 0.76 ** 0.66 ** 0.70 ** 0.88 **
DLW 0.72 ** 0.58 ** 0.69 ** 0.70 ** 0.79 ** 0.59 ** 0.58 ** 0.96 ** 0.96 ** DLW 0.75 ** 0.66 ** 0.87 **
BIO 0.71 ** 0.59 ** 0.70 ** 0.71 ** 0.78 ** 0.58 ** 0.59 ** 0.95 ** 0.95 ** 0.98 ** BIO 1.06 ** 0.87 **
HI 0.62 ** 0.56 ** 0.62 ** 0.62 ** 0.67 ** 0.74 ** 0.74 ** 0.71 ** 0.71 ** 0.70 ** 0.70 ** HI 0.83 **
SY 0.64 ** 0.75 ** 0.77 ** 0.65 ** 0.68 ** 0.68 ** 0.68 ** 0.66 ** 0.66 ** 0.65 ** 0.65 ** 0.64 ** SY
Abbreviations are described in Materials and Methods section
** Significant at 1% probability levels

phenotypic correlations because of removing environment ef- due to low estimates of variances and high covariance. Such
fects in calculating genetic correlation coefficients. In this study, results may also have arisen due to sampling errors.
it was found that the saffron yield had the positive and signif- Overall, based on the results of the correlation coefficients, we
icant correlations (phenotypic and genotypic) with all studied can conclude that traits NDC and FWDC are the most important
traits. Therefore, any improvement in yield components can traits to improve yield components and subsequently increase
have a positive effect in increasing the saffron yield. In this saffron yield. Because saffron is a perennial plant, its
study, it was also found that SY had the highest correlation with agronomic traits have high positive phenotypic and genotypic
FWDC (rg= 1.01**; rp= 0.68**), DFW (rg = 0.98**; rp = correlation with each other. Therefore, choosing an appropriate
0.77**), FFW (rg= 0.98**; rp= 0.75**) and DWDC (rg= density and bigger and higher quality saffron corms for
0.96**; rp= 0.66**), respectively. The results show that the planting will improve FN, FFW, LN and DLW and
selection of these traits is useful to improve yield. These results thus subsequently increase saffron yield in the first year and
are supported by previous findings of Gresta et al. (2009). It coming years. Turhan et al. (2007) suggested that the quality of
was also found that traits FWDC with SY, BIO with HI, FN corms such as size and emergence capacity as the number of
with BIO, FN with DSW (Table 2) had a genotypic correlation corms is important in saffron cultivation. On the other hand,
more than unity. In this way, Khanna and Singh (1975) and Molina et al. (2004) showed that the limiting factor for the
Singh and Singh (1979) stated that such estimate was probably flowering is reduced corm size.

Table 3 Phenotypic path analysis (predictors grouped according to first- and second- order variables) in studied saffron ecotypes

Traits SY NDC FFW FN TIE rp

NDC FFW FN FWDC DFW FWDC DLW DFW

NDC 0.21 0.31 0.17 0.48 0.68 **

FFW 0.14 0.47 0.14 0.28 0.75 **
FN 0.11 0.21 0.32 0.32 0.64 **
FWDC 1.0 — 0.99 **
DFW 0.66 0.14 0.14 0.81**
FWDC 0.44 0.21 0.44 0.66 **
DLW 0.46 0.26 0.26 0.72 **
DFW 0.32 0.38 0.32 0.70 **
R2 0.68 0.99 0.67 0.60

Italic numbers indicate the direct effects

Abbreviations are described in materials and methods
TIE Total of Indirect Effect, rp Phenotypic correlation coefficient
** Significant at 1% probability levels
Physiol Mol Biol Plants (January–March 2016) 22(1):153–161 157

introduce these traits as the most effective traits to improve

SY; therefore any improvement in these traits can have posi-
tive direct effects on increasing SY. Similarly, Yasin and Singh
(2010) reported that plant breeders commonly prefer yield
components that indirectly increase yield. In addition,
Bhagowati and Saikia (2003); Tuncturk and Çiftçi (2005) stat-
ed that the variables have the greatest effect on the dependent
variable considered as an early predictor variables and used
these variables to increase the dependent variable.
Saffron is a sterile plant and it is propagated through corm
Fig. 1 Phenotypic path model illustrating interrelationships among only. Thus, one of the most important ways to increase saffron
various traits contributing to saffron yield yield is the farm management improvement, especially by
applying appropriate density and choosing high quality big
Path analysis corms of saffron to increase traits FN, FFW, NDC and FWDC
and subsequently increase SY in the first year and coming years.
Path coefficient analysis is widely used in breeding programs De Juan et al. (2003) reported that the mother corm size has a
to identify direct and indirect effects of traits that affect yield significant effect on vegetative development and the production
and these results are used to increase yield of crops of daughter corms. Molina et al. (2004) also suggested that the
(Mohammadi et al. 2003; Ali et al. 2009). In the present study, corm size reduction looks to limit flowering in saffron.
path analysis was performed based on phenotypic and geno- By continuing path analysis in the second step, traits NDC,
typic correlation coefficients. In this analysis, the saffron yield FFW and FN were considered as the dependent variable and
was considered as the dependent variable and all traits were the other traits were considered as independent variables
considered as independent variables. (Table 3, Fig. 1). The results showed that FWDC was the most
effective trait on NDC and there was a high positive correla-
Phenotypic path analysis tion between them. These results indicated that corms with
more FWDC not only have higher quality but also can pro-
Phenotypic path analysis showed that traits such as number of duce more daughter corms which subsequently increase FN
daughter corm (NDC), fresh flower weight (FFW) and flower followed by SY in coming years.
number (FN) in the first step were entered into the regression It was also found that two traits DFW and FWDC with 0.66
model (Table 3, Fig. 1) and could justify more than 0.68 % of and 0.21 direct effect, respectively, had the most positive direct
saffron yield variation. These traits not only had positive direct effects on FFW (Table 3; Fig. 1). Traits DFW and FWDC not
effects on saffron yield (SY) but also had high positive indirect only had a high positive phenotypic correlation with each other
effects from each other on SY. Based on these results, we can (rp = 0.67**), but also had a high positive phenotypic correlation

Table 4 Genotypic path analysis (predictors grouped according to first- and second- order variables) in studied saffron ecotypes

Traits SY NDC FFW FN TIE rg

NDC FFW FN FWDC DFW FWDC DLW DFW

NDC 0.49 0.68 −0.35 0.33 0.82 **

FFW 0.38 0.87 −0.27 0.11 0.98 **
FN 0.47 0.65 −0.36 1.12 0.76 **
FWDC 0.73 — 0.73 **
DFW 0.49 0.49 0.49 0.98 **
FWDC 0.38 0.63 0.38 1.01 **
DLW 0.66 0.24 0.24 0.90 **
DFW 0.51 0.31 0.51 0.82 **
R2 0.76 0.98 0.87 0.75

Italic numbers indicate the direct effects

Abbreviations are described in materials and methods
TIE Total of Indirect Effect, rg Genotypic correlation coefficient
** Significant at 1% probability levels
158 Physiol Mol Biol Plants (January–March 2016) 22(1):153–161

saffron should be considered crop management techniques

such as the planting, fertilization, irrigation, growing media,
etc. On the other hand, some of the past studies on saffron
reasoned that application of fertilization have a positive effect
on saffron yield (Behzad et al. 1990a; Behzad et al. 1990b;
McGimpsey et al. 1997).

Genotypic path analysis

In order to eliminate the environmental effects and to estimate

the relationships between saffron yield and other traits more
Fig. 2 Genotypic path model illustrating interrelationships among accurately, path analysis was conducted based on the genotypic
various traits contributing to saffron yield correlations (Table 4, Fig. 2). Genetic path analysis showed that
fresh flower weight (FFW), number of daughter corm (NDC)
with FFW (rp = 0.81** and rp = 0.66** respectively). Therefore, and flower number (FN) in the first step entered the regression
these traits were identified as the most important traits on FFW. model and justified more than 76 % of the saffron yield varia-
These results highlighted that the bigger corm of saffron which tion. Among these traits, FFW and NDC with the highest pos-
has a higher quality, affects a direct and positive effect on DFW itive direct effect (0.87 and 0.49, respectively) were identified
and FFW improvement and followed by increasing SY. as the most important traits which affect the saffron yield. On
Therefore, it can be concluded that the selection of bigger corms the other hand, trait FN had a negative direct effect on the SY
for establishing a new saffron farm is an important factor. (−0.36), but due to its very high positive indirect effects via
It also became clear that dry leaf weight (DLW) and dry traits FFW and NDC (1.12 in total) can have a positive signif-
flower weight (DFW) had the greatest positive direct effects icant effect on SY. It is notable that in this study, only consid-
on improving the flower number (FN) (Table 3). These traits ering the correlation coefficients (phenotypic and genotypic) is
also had a positive indirect effect by each other on FN (0.26 not accurate and may lead to incorrect conclusions (Dewey and
and 0.32 respectively), which managed its positive effects on Lu, 1959). As it was found previously that FN had a positive
FN, rp = 0.72** and rp = 0.70**, respectively, in total. There is and significant correlation with SY (rp = 0.64**, rg = 0.76**),
no relation between flowers and leaves in each year because the genetic path analysis showed that this trait had a negative
the flowers of saffron appearance in the beginning of growing direct effect on SY. On the other hand, the results highlighted
season when saffron leaves are very small (Schmidt et al. that path analysis based on the genetic correlation coefficients is
2007). However, if the farm management especially a proper and accurate way to identify the true relationship be-
irrigating and fertilizing is done correctly and accurately, it tween the independent and dependent variable. So we can use
will improve the growth of leaves and subsequently will this analysis as an efficient method to identify effective traits on
lead to bigger and more daughter corms followed by a the saffron yield improvement.
significant increase in saffron yield in the coming years. By continuing genotypic path analysis, it was found that
Behnia et al. (1999) and Ünal and Çavuşoğlu (2005) stated fresh weight of daughter corm (FWDC), dry flower weight
that the sterility of saffron is a limiting factor through conven- (DFW), and dry leaf weight (DLW) in the second step entered
tional plant breeding. So to increase the yield and quality of the regression model and had positive direct effects on the first

Fig. 3 Dendrogram of cluster

analysis to classify saffron
ecotypes based on Ward’s method
 Physiol Mol Biol Plants (January–March 2016) 22(1):153–161
Table 5 Variance analysis based on cluster number and mean and standard deviation of studied traits in each cluster

S.O.V Df FN FFW DFW FSW DSW NDC FWDC DWDC LL DLW BIO HI SY

Year 2 432.9** 129.4 2.6 49.1 1.6 4.3** 17.2** 5.4** 115.4* 0.04 6,017,265.3** 0.00233** 5973.2*
Erorr1 6 8.5 51.8 1.2 13.0 0.4 0.1 0.1 0.1 12.7 0.02 98,980.9 0.00013 784.5
Month 5 1440.5** 1057.8** 122.5** 121.6** 12.8** 9.3** 37.1** 24.8** 401.2** 1.62** 38,381,266.5** 0.00077** 67,660.8**
Year × Month 10 46.5** 112.8 1.7 5.2 0.2 0.5** 1.3** 0.7** 11.7* 0.04* 670,373.2** 0.0001 1089.8
Erorr2 30 7.7 64.8 0.8 2.7 0.5 0.1 0.1 0.2 5.0 0.02 208,471.5 0.0002 533.8
Ecotype 1 130.7** 7315.0** 124.9** 128.2** 4.3** 32.9** 23.9** 9.8** 432.1** 5.87** 31,106,643.5** 0.00158** 9002.4**
Year × Ecotype 5 8.3 60.1 1.9 4.9 0.2 0.1 0.7* 0.4 17.3* 0.14** 300,020.3 0.00016 569.2
Ecotype × Month 2 9.7 528.8** 3.1 18.9** 0.6* 0.1 0.4 0.1 1.2 0.15* 246,244.9 0.00003 730.8
Year × Ecotype × Month 10 11.5 47.9 1.2 2.6 0.1 0.4* 1.1** 0.8** 18.1** 0.01 800,243.8** 0.00006 523.4
Erorr3 144 15.3 46.4 1.1 2.1 0.1 0.2 0.3 0.3 7.1 0.03 351,981.6 0.00006 454.6
Cluster
1 17.62 ± 6.9 357.93 ± 5.9 43.90 ± 1.7 28.86 ± 2.5 5.25 ± 0.7 2.69 ± 32.1 6.31 ± 0.6 3.67 ± 58.5 19.61 ± 1.2 3.16 ± 0.9 6877.63 ± 954.3 0.08 ± 3.8 95.53 ± 9.2
2 16.06 ± 6.7 346.29 ± 5.5 42.38 ± 2.0 27.31 ± 1.5 4.97 ± 0.5 1.91 ± 29.1 5.65 ± 0.6 3.24 ± 56.1 16.78 ± 1.1 2.83 ± 0.8 6118.65 ± 802.9 0.08 ± 3.9 82.62 ± 14.9
Total 16.84 ± 6.9 352.11 ± 9.3 43.14 ± 2.0 28.09 ± 2.2 5.11 ± 0.6 2.30 ± 36.3 5.98 ± 0.7 3.45 ± 59.7 18.19 ± 1.2 3.00 ± 0.9 6498.14 ± 907.4 0.08 ± 4.1 89.07 ± 14.9

Abbreviations are described in Materials and Methods section

* and ** Significant at 5% and 1% probability levels, respectively

159
160 Physiol Mol Biol Plants (January–March 2016) 22(1):153–161

step traits (Table 4 and Fig. 2). Among these traits, FWDC confirmed these traits were influenced by the environment
with a positive direct effect on NDC (0.73) and FFW (0.63) significantly. Results of genotypic and phenotypic coefficients
was found to be the most influential and important trait. of variation and genetic advance also showed that most of the
Therefore, any increase in this trait such as the selection of traits had a low diversity so the selection for improving saffron
bigger and more quality corms can have a high positive effect yield in the next generation did not have a notable progress.
on yield components and consequently will increase the saf- Therefore, the improvement of farm management (such as
fron yield (SY). proper sowing date, selecting bigger and higher quality corm,
It also became evident that DFW with the direct (0.49) and fertilization, appropriate density, etc.) is the best way to increase
indirect (0.49) effects on FFW and the direct (0.31) and indi- the saffron yield. In this study, it was found that saffron yield
rect (0.51) effects on FN had the positive effects on these had the most correlation (genotypic and phenotypic) with fresh
traits. Flower weight is a trait that is directly related to the weight of daughter, dry flower weight, fresh flower weight and
quality and weight of saffron corm. Therefore, corms with a dry weight of daughter corm, respectively. The results of phe-
higher quality and more weight can produce flowers with notypic and genotypic path analysis showed that traits number
more weight and this subsequently increases the saffron yield. of daughter corm, fresh flower weight and flower number in the
first step and trait fresh weight of daughter corm, dry flower
Cluster analysis weight and dry leaf weight in the second step had the most
positive direct and indirect effects on saffron yield. As a result,
Grouping and classifying different saffron ecotypes was per- any improvement in these traits, especially in number of daugh-
formed using all traits based on UPGMA method by cluster ter corm and fresh weight of daughter corm can have a positive
analysis. The results of analysis (Fig. 3) showed that four effect on increasing the saffron yield. Cluster analysis showed
saffron ecotypes were grouped in two clusters. To determine that saffron ecotypes grouped in two clusters. Ecotypes
the validity of clustering, the variance analysis based on the Mashhad and Torbat-Jam (cluster I), compared with ecotypes
cluster numbers was used. Variance analysis results (Table 5) Birjand and Gonabad (cluster II) was more compatible with the
showed significant differences (p < 0.01) between clusters. climatic conditions of Mashhad region.
These results confirmed the validity of cluster analysis and
the dendrogram was also cut in the appropriate position. Compliance with ethical standards
To evaluate and compare the differences between the clus-
ters, the mean and standard deviation of traits in each of the Conflict of interest The authors declare that they have no conflict of
clusters were calculated separately (Table 5). The results interest.
showed that cluster I and II were ranked as the first and second
cluster, respectively. The results showed that the trait means of
cluster I (Mashhad and Torbat-Jam ecotypes) were much
higher than cluster II (Birjand and Gonabad ecotypes).
Consequently, ecotypes Mashhad and Torbatjam, as the best References
ecotypes, had the maximum compatibility with the weather
conditions of Mashhad. Ehsanzadeh et al. (2004) in their study Ali MA, Khan IA (2007) Assessment of genetic variation and inheritance
mode of some metric traits in cotton (Gossypium hirsutum L.). J
on ten saffron ecotypes in Shahrcord region reported that there Agric Social Sci 3:112–116
were high significant differences between ecotypes. These Ali MA, Nawab NN, Abbas A, Zulkiffal M, Sajjad M (2009) Evaluation
researches, in total, stated that ecotypes Shahrcord, Birjand of selection criteria in Cicer arietinum L. using correlation coeffi-
and Gaean had the highest yield in Shahrcord region and cients and path analysis. Aust J Crop Sci 3:65–70
Amirnia R, Bayat M, Gholamian A (2013) Influence of corm provenance
weather conditions had significant effects on saffron yield
and sowing dates on stigma yield and yield components in saffron
and yield components. (Crocus sativus L.). Turkish Journal Of Field Crops 18:198–204
Anastasaki E, Kanakis C, Pappas C, Maggi L, Del Campo C, Carmona
M, Alonso G, Polissiou M (2010) Differentiation of saffron from
Conclusions four countries by mid-infrared spectroscopy and multivariate analy-
sis. Eur Food Res Technol 230:571–577
Bahraminejad A, Mohammadi-Nejad G, Abdul Khadir M (2011) Genetic
The results of this study showed that there were significant Diversity Evaluation of Cumin (‘Cumin cyminum’L.) Based on
differences between sowing dates and saffron ecotypes with Phenotypic Characteristics. Aust J Crop Sci 5(3):304–310
respect to all studied traits. Thus, the accurate selection of the Behnia M, Estilai A, Ehdaie B (1999) Application of fertilizers for in-
appropriate sowing date and saffron ecotype in each region are creased saffron yield. J Agron Crop Sci 182:9–15
Behradfar A, Gorttapeh AH, Zardashty MR, Talat F (2009) Evaluation
considerably important and essential factors. The results of the correlated traits for seed and oil yield in sunflower (Helianthus
general heritability showed that the majority of studied traits annuus L.) through path analysis in under condition relay cropping.
had a low and or moderate general heritability, which res. J Biol Sci 4:82–85
Physiol Mol Biol Plants (January–March 2016) 22(1):153–161 161

Behzad S, Razavi M, Mahajeri M (1990a) The effect of mineral nutrients Leilah A, Al-Khateeb S (2005) Yield analysis of canola (Brassica napus
(NPK) on saffron production. International Symposium on L.) using some statistical procedures. Saudi J Bio Sci 12:103–113
Medicinal and Aromatic Plants, XXIII IHC 306:426–430 Magesh V, Singh JPV, Selvendiran K, Ekambaram G, Sakthisekaran D
Behzad S, Razavi M, Mahajeri M (1990b) The effect of various amount (2006) Antitumour activity of crocetin in accordance to tumor inci-
of ammonium phosphate and urea on saffron production. dence, antioxidant status, drug metabolizing enzymes and histopath-
International Symposium on Medicinal and Aromatic Plants, ological studies. Mol Cell Biochem 287:127–135
XXIII IHC 306:337–339 Maggi L, Carmona M, Kelly SD, Marigheto N, Alonso GL (2011)
Bhagowati R, Saikia M (2003) Character association and path coefficient Geographical origin differentiation of saffron spice (Crocus sativus
analysis for yield attributes in open pollinated and hybrid true potato L. stigmas)–preliminary investigation using chemical and multi-
seed populations. Crop Research-HISAR- 26:286–290 element (H, C, N) stable isotope analysis. Food Chem 128:543–548
Caiola MG, Caputo P, Zanier R (2004) RAPD analysis in Crocus sativus McGimpsey J, Douglas M, Wallace A (1997) Evaluation of saffron
L. accessions and related crocus species. Biol Plant 48:375–380 (Crocus sativus L.) production in New Zealand. N Z J Crop Hortic
Chryssanthi D, Dedes P, Lamari F (2009) Crocetin, the active metabolite Sci 25:159–168
of crocins, inhibits growth of breast cancer cells and alters the gene Mohammadi S, Prasanna B, Singh N (2003) Sequential path model for
expression pattern of metalloproteinases and their inhibitors in the determining interrelationships among grain yield and related char-
cell line MDA-MB-231. 3rd international symposium on saffron acters in maize. Crop Sci 43:1690–1697
forthcoming challenges in cultivation research and economics, Molina R, Valero M, Navarro Y, Garcıa-Luis A, Guardiola J (2004) The
Krokos, Kozani, Greece, p. 58 effect of time of corm lifting and duration of incubation at inductive
Dalezis P, Papageorgiou E, Geromichalou E, Geromichalus G (2009) temperature on flowering in the saffron plant (Crocus sativus L.). Sci
Antitumor activity of crocin, crocetin and safranal on murine P388 Hortic 103:79–91
leukemia bearing mice. 3rd International symposium on saffron Molina R, Valero M, Navarro Y, Guardiola J, Garcia-Luis A (2005)
Forthcoming challenges in cultivation research and economics. Temperature effects on flower formation in saffron (Crocus sativus
Krokos, Kozani, Greece L.). Sci Hortic 103:361–379
Darvishzadeh R, Maleki HH, Sarrafi A (2011). Path Analysis of the Mollafilabi A (2004). Experimental findings of production and echo
Relationships between Yield and Some Related Traits in Diallel physiological aspects of saffron (Crocus sativus L.). Acta Hortic
Population of Sunflower (‘Helianthus annuus’ L.) under Well- 650:195–200
Watered and Water-Stressed Conditions. Omidbaigi R (2005) Effect of corms weight on quality of saffron (Crocus
De Juan J, Moya A, López S, Botella O, López H, Munoz R (2003) sativus Linn.). Natural Product Radiance 4:193–194
Influence of the corm size and the density of plantation in the yield Özdemİr C, Baran P, Akyol Y (2006) The morphology and anatomy of
and the quality of the production of corms of Crocus sativus L. In: Crocus flavus Weston subsp. flavus (iridaceae). Turk J Bot 30:175–
ITEA (Información Técnica Económica agraria). Vegetal (España), 180
Producción Renau-Morata B, Nebauer S, Sánchez M, Molina R (2012) Effect of corm
size, water stress and cultivation conditions on photosynthesis and
De Juan JA, Córcoles HL, Muñoz RM, Picornell MR (2009) Yield and
biomass partitioning during the vegetative growth of saffron
yield components of saffron under different cropping systems. Ind
(Crocus sativus L.). Ind Crop Prod 39:40–46
Crop Prod 30:212–219
Rubio-Moraga A, Castillo-López R, Gómez-Gómez L, Ahrazem O
Ehsanzadeh P, Yadollahi AA, Maibodi AM (2004) Productivity, growth
(2009) Saffron is a monomorphic species as revealed by RAPD,
and quality attributes of 10 Iranian saffron accessions under climatic
ISSR and microsatellite analyses. BMC Research Notes 2:189
conditions of Chahar-Mahal Bakhtiari. Central Iran Acta Hortic:
Sanghera GS, Kashyap SC (2012) Genetic parameters and selection in-
183–188
dices in F3 progenies of hill rice genotypes. Notulae Scientia
Gómez-Gómez L, Trapero-Mozos A, Gómez MD, Rubio-Moraga A, Biologicae 4:110–114
Ahrazem O (2012) Identification and possible role of a MYB tran- Schmidt M, Betti G, Hensel A (2007) Saffron in phytotherapy: pharma-
scription factor from saffron (Crocus sativus). J Plant Physiol 169: cology and clinical uses. Wien Med Wochenschr 157:315–319
509–515 Singh B, Goswami A (2014) Correlation and path coefficient analysis in
Gresta F, Avola G, Lombardo G, Siracusa L, Ruberto G (2009) Analysis okra (Abelmoschus esculentus). Indian J Agr Sci 84(10):1262–1266
of flowering, stigmas yield and qualitative traits of saffron (Crocus Singh S, Yadav H, Shukla S, Chatterjee A (2003) Studies on different
sativus L.) as affected by environmental conditions. Sci Hortic 119: selection parameters in opium poppy (P. somniferum L.). Journal of
320–324 Medicinal and Aromatic Plant Sciences 25:380–384
Jalali-Heravi M, Parastar H, Ebrahimi-Najafabadi H (2010) Self- Siracusa L, Gresta F, Avola G, Lombardo GM, Ruberto G (2010)
modeling curve resolution techniques applied to comparative anal- Influence of corm provenance and environmental condition on yield
ysis of volatile components of Iranian saffron from different regions. and apocarotenoid profiles in saffron (Crocus sativus L.). J Food
Anal Chim Acta 662:143–154 Compos Anal 23:394–400
Johnson HW, Robinson H, Comstock R (1955a) Estimates of genetic and Talebi R, Fayaz F, Jelodar N-AB (2007) Correlation and path coefficient
environmental variability in soybeans. Agron J 47:314–318 analysis of yield and yield components of chickpea (Cicer arietinum
Johnson HW, Robinson H, Comstock R (1955b) Genotypic and pheno- L.) under dry land condition in the west of Iran. Asian J Plant Sci 6:
typic correlations in soybeans and their implications in selection. 1151–1154
Agron J 47:473–483 Tuncturk M, Çiftçi V (2005) Selection criteria for potato (Solanum
Khanna K, Singh U (1975) Correlation studies in papavier somniferum tuberosum L.) breeding. Asian J Plant Sci 4:27–30
and their bearing on yield improvement. Planta Med 28:92 Turhan H, Kahriman F, Egesel CO, Gul MK (2007) The effects of differ-
Kozak M, Kang MS (2006) Note on modern path analysis in application ent growing media on flowering and corm formation of saffron
to crop science. Communications In Biometry And Crop Science 1: (Crocus sativus L.). Afr J Biotechnol 6:2328–2332
32–34 Ünal M, Çavuşoğlu A (2005) The effect of various nitrogen fertilizers on
Lage M, Cantrell CL (2009) Quantification of saffron (Crocus sativus L.) saffron (Crocus sativus L.) yield. Akdeniz Üniversitesi Ziraat
metabolites crocins, picrocrocin and safranal for quality determina- Fakültesi Dergisi 18:257–260
tion of the spice grown under different environmental Moroccan Yasin AB, Singh S (2010) Correlation and path coefficient analyses in
conditions. Sci Hortic 121:366–373 sunflower. Journal of Plant Breeding and Crop Science 2:129–133