2025

Specialized Metabolites

Julian Verdonk
[email protected]
@julianverdonk
Horticulture and Product Physiology
 Why are F&V healthy
• “F&V are not unhealthy”
• Contain a lot of water: 85-98%
• Essential Vitamins (C, E, K, B9, provitamin A)
• (Poly)phenols (flavonoids such as anthocyanin, ...)
• Carotenoids (lycopene, carotene, lutein, zeaxanthin)
• Glucosinolates
• Other bioactive components (…)
• Many phytochemicals (including Vit C, E, pro A) are
strong antioxidants
• These compounds are often specialized metabolites

2
 Specialized Metabolites
Secondary Metabolites – Natural Products
“organic compounds that do not participate directly in growth and
development”

α-pinene

Pine (Pinus L.)

 Specialized Metabolites
Secondary Metabolites – Natural Products
“organic compounds that do not participate directly in growth and
development”

often uniquely found in specific taxonomy groups

secondary biological function – pigment / fragrance / defense
usefull applications – dyes / medicine / drugs / toxins
function of many remain unknown

α-pinene

Pine (Pinus L.)

 Specialized Metabolites
Secondary Metabolites – Natural Products
“organic compounds that do not participate directly in growth and
development”

often uniquely found in specific taxonomy groups

secondary biological function – pigment / fragrance / defense
usefull applications – dyes / medicine / drugs / toxins
function of many remain unknown

in contrast: “primary metabolites”

i.e., phytosterols / acyl lipids / nucleotides / amino acids / organic acids
found in all plants
perform metabolic roles that are essential and usually evident
 Organic Chemistry

• Plant Biologists
• initially considered these chemicals insignificant
secondary metabolites or natural products
• Organic chemists:
• stimulated development of separation techniques,
spectroscopic approaches to structure elucidation, and
synthetic methodologies since 1850s
• prompted by their great utility as dyes, polymers, fibers,
glues, oils, waxes, flavoring agents, perfumes and drugs

http://www.ivyroses.com/Chemistry/Images/336x280_ChemicalBottles.jpg http://www.victoria.ac.nz/scps/research/research-groups/natural-products/images/npc-samples.jpg
 Current focus of the field
Current focus of this field towards the search for new
drugs, antibiotics, insecticides, and herbicides

Aspirin

Salicylic Acid

Eugenol
Willow (Salix alba)

http://www.sivz.com/images/uploads/2012/03/Clove-Oil.jpg ; https://chairish-prod.freetls.fastly.net/image/product/sized/a1526f10-4646-
4533-ac63-84a38a255b7a/1950s-bayer-pill-advertising-sign-3790?aspect=fit&height=1600&width=1600 ; http://media-cache-
ec0.pinimg.com/736x/e9/dd/44/e9dd44bb235063c9bad41cd711549fe5.jpg
 Role(s) in plants

context of ecological interactions:

• protections against herbivory and microbial infection
• attractants for pollinators and seed-dispersing animals
• allopathic agents (allelochemicals that influence the competition among plant species)

Plants are excellent

biochemists!!
http://3.bp.blogspot.com/_S-nLQyZf48Y/TK6LkqE2QlI/AAAAAAAAAAU/h-
B8k5SQ_BU/s1600/abelha+cheia+de+polen+bee+full+of+food.jpg ;
https://upload.wikimedia.org/wikipedia/commons/thumb/1/15/Caterpillar_feeding_on_leaf_-
_20140906_121127_%28cropped%29.jpg/500px-Caterpillar_feeding_on_leaf_-_20140906_121127_%28cropped%29.jpg
 Trichomes
Store metabolites
• Defense
• Quality Turner et al., 2000

https://potguide.com/media/76238700/what-are-trichomes.jpg
Three Main Chemical Families
Menthol

Terpenoids / Isoprenes
• >25.000 secondary compounds Mentha arvensis

• Formed from isopentenyldiphosphate (IPP)

Three Main Chemical Families
Menthol

Terpenoids / Isoprenes
• >25.000 secondary compounds Mentha arvensis

• Formed from isopentenyldiphosphate (IPP)

Alkaloids
• ~12.000 secondary compounds
• Contain one or more nitrogen atoms
• Biosynthesized from amino acids
Three Main Chemical Families
Menthol

Terpenoids / Isoprenes
• >25.000 secondary compounds Mentha arvensis

• Formed from isopentenyldiphosphate (IPP)

Alkaloids
• ~12.000 secondary compounds
• Contain one or more nitrogen atoms
• Biosynthesized from amino acids

Phenolic compounds
• ~8000 secondary compounds
• formed from the shikimate or the malonate/acetate pathway
 Terpenoids
Tomato colour: lycopene (a carotenoid)
Menthol, Limonene, Linalool, Pinene, Eucalyptol
Camphor
Cannabinoids
limonene
(c) compound interest blog
α-pinene

Pine (Pinus L.)

Eucalyptol
(1,3,3-Trimethyl-2-oxabicyclo[2,2,2]octane)

Camphor
(1,7,7-Trimethylbicyclo[2.2.1] heptan-2-one)
https://cdn.forestessentialsindia.com/blog/wp-content/uploads/2020/02/Camphor.png
Two main branches lead to the isoprene unit IPP
Mevalonate Acetate (MVA) pathway generates
Farnesyl diphosphate (FPP)
• Precursor for sesquiterpenes (C15), triterpenes (C30), and polyterpenes
• produced in the cytosolic and endoplasmic reticulum (ER) compartments

Methyl-erythritol-phosphate (MEP) pathway generates

Geranyl diphosphate (GPP) and geranylgeranyl diphosphate (GGPP)
• Precursors for monoterpenes (C10), diterpenes (C20), tetraterpenes
(C40), and certain prenylated quinones
• originate largely, if not exclusively, in the plastids

Van Schie et al., 2006

 Hemiterpenes (C5): photosynthetic tissues

Monoterpenes (C10): flower volatiles / essential oils from herbs

Gibberellic acid
(C20)
Abscisic acid
(C15)

Sesquiterpenes (C15): essential oils / phytoalexins / Abscisic acid

Brassinolide
(C30)
Phytol (C20)
Diterpenes (C20): phytol (Chlorophyll sidechain) / GA / resin acids of confiers & legumes

Triterpenes (C30): brassinosteroids / oleanolic acid (grapes)

Tetraterpenes (C40): caretenoid pigments Meroterpenes (Mixed): taxol

Natural rubber (Cx)

Polyterpenes (C>40): rubber / prenylated quinone electron carriers
Rubulotta, G., and Quadrelli, E.A. (2019).
Chapter 11 - Terpenes: A Valuable Family of Compounds for the Production of Fine Chemicals. In Studies in Surface Science and Catalysis, S. Albonetti, S. Perathoner, and E.A. Quadrelli, eds. (Elsevier), pp. 215–229.
Alkaloids
• Plant extracts used for potions and poisons for centuries
• Papaver somniferum (poppy) can be traced back to 1400 BC in Eastern
medeterenian
Rauwolfia serpentina
• Rauwolfia serpentina (Sarpagandha root) used in India since 1000 BC
• (used for large range of ailments)
• Use of medicinal plants spread westwards
• Socrates was executed by drinking the extract of Conium maculatum (Poison
Hemlock)
• Cleopatra used extract of Hyoscyamus to dilate her pupils (Atropine)
 Alkaloids
Opium
• Theriak: mix of mainly opium, dried snake meat and wine (~70 plants)
• Analysis of individual components of Opium led to the identification
of morphine, named for Morpheus, god of dreams in Greek mythology
• Morphine isolated 1806 by the German Pharmacist Friedrich Sertürner

Alkaloids: chemicals that influence Geopolitics

• opium wars between Britain and China
• current efforts to eradicate production of heroin (derived from
morphine) and cocaine
 Alkaloids
• 20% of flowering plants produce alkaloids
• Each in a unique, defined pattern
• Catharanthus roseus (Madagascar periwinkle) produces more than 100 monoterpenoid
indole alkaloids

• Why would a plant invest so much of its Nitrogen and energy in synthesizing
these compounds?
• Mainly defense and communication:
• Toxicity, repelling feeding (learning involved, more useful than killing)
• Attraction, pollination
• Dispersion
• Warning (i.e., attracting predators for feeding insects)
 Biosynthesis
• Precursor-feeding experiments in the 1950s established that
alkaloids are in most cases formed from amino acids (Trp, Tyr,
Phe, Lys, Arg)
• Either alone or in combination with a terpenoid-type moiety
• Of most of the 12.000 known compounds it is unknown (!!)
• But some well investigated systems serve as examples
• Often, a general precursor has a species-specific step that
produces a species-specific unique compound
i.e., S -reticuline

Kilgore, M.B., Holland, C.K., Jez, J.M., and Kutchan, T.M.

(2016). Identification of a Noroxomaritidine Reductase
with Amaryllidaceae Alkaloid Biosynthesis Related
Activities. J Biol Chem 291, 16740–16752.
 Phenolic compounds
“acidic” hydroxyl or phenolic group Salicylic acid

Eugenol
Phenyl ring

• Phenolic compounds were essential for the adaptation of plants to land

• Cell wall structural roles: Lignin, Lignan, etc.
• Various other roles
• Defending plants
• Wood durability (bark)
• Flower color
• Flavors (taste & odor)
Flavonoids (huge group of 4500 compounds)
Subclassed into
anthocyanins (pigments),
Proanthocyanidins or tannins (feeding deterrents)
Isoflavonoids (defensive products, signaling
molecules)

Volatile Benzenoids/Phenylpropanoids (VBPs)

Pollinator attractant
Aroma/flavour
 Lignin
reinforce cell walls, huge
polymers

Lignan (dimers/oligomers)
defend against pathogens or
act as antioxidants in flowers,
fruits, seeds, nuts etc.

Suberin
is part of tissues that for
protective barriers for
desiccation in cork, bark, potato
‘Lignin’ skin

‘Suberin’
Coumarins, Furanocoumarins
and stilbenes
Appetite suppressing
Anti bacterial and fungal,
protect against herbivory

Rosmarinic acid
Biosynthesis

• Very well understood

• Derived from Phenylalanine or Tyrosine
• Phenylpropanoids Phenyl ring and Propane (C3)
• All primary derived from
• Phenylpropanoid
• Phenylpropanoid-acetate
• Shikimate Pathway and Aromatic Amino Acid Pathway
 The Shikimate Pathway
• 7 steps to Chorismate
• Only in plants and bacteria
• Occurs in the plastids
• EPSP synthase target for Glyphosate (Roundup)

Maeda, H., and Dudareva, N. (2012). The shikimate pathway and aromatic amino Acid 25
biosynthesis in plants. Annu Rev Plant Biol 63, 73–105.
 The AAA Pathway
Aromatic Amino Acids
• Tryptophan
• Tyrosine
• Phenylalanine
• Essential Amino Acids!
Animals must eat plants to get
Phe Tyr and Trp!

Maeda, H., and Dudareva, N. (2012). The shikimate pathway and aromatic amino Acid 26
biosynthesis in plants. Annu Rev Plant Biol 63, 73–105.
 Tyr

Phenylpropanoids Phe

• From Phe/Tyr network of parallel reactions

• O-methyltransferases
• CoA ligases – oxireductase – alcohol
dehydrogenase
• Matrix towards all chemical subclasses
• Lignin/lignans
• Flavonoids
• Others…
Lignin
• Located in secondary cell wall
• Lignin synthesis mostly
nonenzymatic process

28
 • 2nd part details on pigments
Flavonoids

YAHIA FIGURE 12.7

Biosynthesis pathway for flavonoid production in plants. CHS, chalcone synthase; CHI, chalcone
isomerase; F3H, flavonone 3-hydroxylase; F3'H, flavonoid 3'-hydroxylase; FNS, flavone synthase;
FLS, flavonol synthase; DFR, dihydroflavonol 4-reductase; LAR, leucoanthocyanidin reductase;
ANS, anthocyanidin synthase; UFGT, UDP-glucose: flavonoid 3-O-glucosyltransferase
Volatile Benzenoids/Phenylpropanoids (VBPs)

• Pollination syndrome
• Defense and Allopathic compounds
• Volatile Organic Compounds
• Biosynthesis from phenylpropanoids

30
• Matrix towards
• Flavonoids
• Lignin Shikimate flavonoids
• VBPs pathway
• Chlorogenic acid
• Rosmarinic acid
• Etc. Chlorogenic
Acid

AAA
Rosmarinic
Acid

VBPs lignin

Rosmarinic acid
 Combined and interlinked biosynthesis pathways
Phenylpropanoids Pathway(s) Terpenoid Pathway
• Metha spp.
• Menthol
• Rosmarinic acid
Important Antioxidant
Plant name RA conc. (mg g-1)
Mentha spp. 19.3 – 58.5
Rosmarinus officinales 7.2
Salvia spp. 4.3 – 39.9
Thymus spp. 0 – 23.5 Menthol

Rosmarinic acid
Mentha spp

Shekarchi, M., Hajimehdipoor, H., Saeidnia, S., Gohari, A.R., and Hamedani, M.P. (2012). Comparative 32
study of rosmarinic acid content in some plants of Labiatae family. Pharmacogn Mag 8, 37–41.
 Cannabis: many interesting compounds besides CBP and THC

33
https://www.nature.com/articles/s41598-020-60172-6
 Quality and Postharvest
• Specialized metabolites have diverse protective
roles, protects against oxidative stress
• Synthesis is tightly regulated by (a)biotic factors
• ROS induction by these factors triggers defense
system
• biosynthesis of antioxidant secondary metabolites
and activation of antioxidant enzymes
• Induced production of specialized metabolites
can be desirable for health-, and quality related
benefits
• Could also reduce quality, e.g. bitterness, colour
loss, etc. YAHIA FIGURE 12.8
Increase in PCs by stress-inducing postharvest
treatments in fruits and vegetables. 34
Summary:
Specialized Metabolites
• Chemicals that protect plant against stress – antioxidant, defense and
other ecological functions
• Quality benefits (desirability and health)
• Flavour, aroma, colour, texture, antioxidant, vitamins, etc.
• Terpenoids, Alkaloids, Phenolic compounds
• Biosynthesis pathways as illustration
• Compartmentalization
• Pre-Postharvest influence biosynthesis and catabolism
• Growing Quality…

35
Specialized Metabolites
Part II- Pigments
 Plant Pigments
• Specialized metabolites!
• Natural compounds that provide colour to plants
• Function for plants:
• Communicate: defence, attraction, warning
• Pollination, seed dispersal, repulsion
• Photosynthesis and associated processes
• Protect against damage caused by UV and visible light

38
 Plant Pigments
• Specialized metabolites!
• Natural compounds that provide colour to plants
• Function for us:
• Important quality features… for commodities
• Harvest index
• Connection of visible pigments to health-related features
• Natural food colouring
• Health benefits due to antioxidant and anti-inflammatory
properties
• Cosmetic products, skin care, anti aging and UV protection

Yahia FIGURE 11.2

Absorption spectra (in acetone) of Chl a (1), Chl b (2), lutein (3), and
cyanidin-3-galactoside (4). Lutein is a carotenoid in the xanthophyll
subgroup, and cyanidin-3-galactoside is a commonly seen
anthocyanin. In the anthocyanin absorption spectrum, a longwave
maximum in the green region (λmax5535 nm) is shown, the short-
wave part below 400 nm is not presented. 39
Chlorophyll
Carotenoids
Flavonoids
Betalains

https://www.georgeperry.co.uk/images/P/orangepepper.jpg; https://cdn.shopify.com/s/files/1/1380/2059/products/Beet-Root-Ruby-
Red_large.jpg?v=1598078262 ; https://cdn.shopify.com/s/files/1/0102/8043/5812/products/watermelon_300x300.png?v=1563408141 ;
https://www.lekkervanbijons.be/sites/default/files/styles/image_on_detailpage/public/ps_image/istock_aubergine.jpg?itok=bSr_QuF- 40
Chlorophylls

41
Chlorophylls

• Well known green pigments

• Primary photosynthetic pigments in plants
• Fresh fruits and green vegetables are rich in Chl
• Food additive (green colorant) used in a variety of foods and
beverages
• Generally, Chl comprises 0.6% - 1.6% on a dry weight basis
• large variations exist in plants
• Capture light to drive photosynthesis

This Photo by Unknown Author is licensed under CC BY-SA

42
Chlorophylls

• Capture light to drive photosynthesis

• Light energy -> Chemical energy
• Creates carbohydrates out of thin air (and light)
• Absorb red and blue light
• In higher plants two forms: Chlorophyll a and b
• Lower plants, algae and cyanobacteria have
other types as well

© Wikimedia Commons / Daniele Pugliesi, M0tty

43
 Chlorophylls
• Cyclic tetrapyrrole ring
• Chl a blue-green Chl b yellow-green
• Long hydrophobic phytol (C20 Diterpene) that anchors the protein to the
pigment binding proteins in thylakoid membrane
• Both highly susceptible to degradation during processing: food colour
changes

44
Chlorophylls

• Synthesized in chloroplasts
• Breakdown is enzymatic process, 2 main
pathways
• Removal of phytate, Mg, and oxidative
cleavage
• Oxidative degradation of Chl through
hydroperoxide and radical formation by
oxidation of lipids

45
 Chlorophylls
• Biosynthesis and breakdown regulated by
developmental processes and environmental
cues (e.g., light)
• During ripening of tomato, change in colour
through degradation of chlorophyll and
accumulation of carotenoids
• Mutants in this breakdown: green flesh (gf)
• Chlorophyl and carotenoids (lycopene): brownish

Lycopene
Schouten, R.E., Farneti, B., Tijskens, L.M.M., Alarcón, A.A., and Woltering, E.J. (2014). Quantifying lycopene synthesis and chlorophyll breakdown in tomato fruit using remittance VIS spectroscopy. Postharvest Biology
and Technology 96, 53–63.
46
Barry, C.S., McQuinn, R.P., Chung, M.-Y., Besuden, A., and Giovannoni, J.J. (2008). Amino Acid Substitutions in Homologs of the STAY-GREEN Protein Are Responsible for the green-flesh and chlorophyll retainer
Mutations of Tomato and Pepper. Plant Physiology 147, 179–187. https://pbgworks.org/sites/pbgworks.org/files/user/334/cherokeepurple1_0.jpg
Chlorophylls

Black Velvet Petunia

Petal Chlorophyll breakdown mutant
Green and Dark Purple
Almost Black!

47
https://www.gardencrossings.com/wp-content/uploads/2022/01/petunia_starlet_velvet_petunia_bh_petve_01.jpg
Carotenoids
Carotenoids
• Yellow, orange and red
pigments – Lipid soluble
• Synthesized in plants, algae and
some photosynthetic bacteria
• 1000 - 750 found in nature

49
http://herbal-essentials.com/2016/03/11/your-food-guide-to-healthy-skin/
 Carotenoids
• Health, nutritional quality, plant dev
• Plants:
• Photosynthetic system (light harvesting)
• Prevent photo oxidative damage by dissipating
excess energy as heat
• Plant grow and development (ABA &
strigolactones)
• Role in defence mechanisms
• Humans/Animals:
• Quality: Colour/Flavour
• Dietary source of pigmentation of many fish,
insects, crustaceans and birds
• Health, eyes, brain, Vitamin A, Vitamin E

50
https://bio1151.nicerweb.com/Locked/media/ch10/10_carotenoids-shrimp.jpg ; https://scx2.b-cdn.net/gfx/news/2016/1-howbirdsturn.jpg http://www.apsu.edu/hiattl/research
 • Branches from the plastidial
Biosynthesis terpenoid pathway
• Biosynthesis in the plastid,
essential to the integrity of
photosynthetic apparatus
• All biosynthetic steps very
well known
• Biosynthesis divided in 2
parts:
1. early lycopene yielding
Carotenes 2. α, and β-carotene and
xanthophylls

Xanthophylls

van Schie, C.C., Haring, M.A., and Schuurink, R.C. (2006). Regulation of terpenoid and benzenoid production in flowers. Curr. Opin. Plant Biol 9, 203–208.
Giuliano, G., Tavazza, R., Diretto, G., Beyer, P., and Taylor, M.A. (2008). Metabolic engineering of carotenoid biosynthesis in plants. Trends in Biotechnology 26, 139–145.
 Regulation RIN • Regulation less understood
NAC1 • Seems to be connected
more to development than
to specific regulators for
carotenoid biosynthesis
• RIN, NAC1
• Recent work has identified
new potential regulators
• MYB68
• RCP1 (MYB)
• CAR in Petunia (not
published yet)

MYB68

Sagawa, J.M., Stanley, L.E., LaFountain, A.M., Frank, H.A., Liu, C., and Yuan, Y.-W. (2016). An R2R3-MYB transcription factor regulates carotenoid pigmentation in Mimulus lewisii flowers. New Phytol 209, 1049–1057.
Zhu, F., Luo, T., Liu, C., Wang, Y., Yang, H., Yang, W., Zheng, L., Xiao, X., Zhang, M., Xu, R., et al. (2017). An R2R3-MYB transcription factor represses the transformation of α- and β-branch carotenoids by negatively
regulating expression of CrBCH2 and CrNCED5 in flavedo of Citrus reticulate. New Phytologist 216, 178–192.
 Valuable molecules
• Carotenes: Provitamin A function (α- and β-carotene)
• Eye and Brain development
• Lycopene: Antioxidant activity
• Lutein, Zeaxanthin: Blue light filtering
• Astaxanthin: Antioxidant, food colouring
• Tocopherol: Vitamin E

https://images.theconversation.com/files/296496/original/file-20191010-188807-
u4v7v7.jpg?ixlib=rb-1.1.0&q=45&auto=format&w=1200&h=1200.0&fit=crop 53
 Golden Rice psy
• transformed rice with β-carotene
biosynthesis genes: crtI
• psy (phytoene synthase)
• from daffodil (Narcissus pseudonarcissus) crtI

• crtI (carotene desaturase)

• from the soil bacterium Erwinia uredovora lcy

• lcy (lycopene cyclase)

• gene not needed, it is already being produced in
wild-type rice endosperm
This Photo by Unknown Author is licensed under CC BY
Astaxanthin
• Ketocarotenoid, made by red algae
• One plant species (Adonis)
• Valuable pigment
• Metabolic engineering Giuliano, G., Tavazza, R., Diretto, G., Beyer, P., and Taylor, M.A. (2008). Metabolic
engineering of carotenoid biosynthesis in plants. Trends in Biotechnology 26, 139–145.

• Three genes needed, not just CrtW/CrtO

astaxanthin
3,3′-dihydroxy-β,β-carotene-4,4′-dione Mann V, Harker M, Pecker I, and Hirschberg J. Metabolic engineering of astaxanthin production
in tobacco flowers. Nat Biotech. 2000:18(8):888–892. https://doi.org/10.1038/78515
Quality – Shelf life

Fruit-specific overexpression
of LYCOPENE β-CYCLASE (LCYb)
Tocopherols
(Vitamin E)
• Combined pathways
• Shikimate / Terpenoids

DellaPenna, D., and Pogson, B.J. (2006). Vitamin Synthesis in Plants: 57

Tocopherols and Carotenoids. Annual Review of Plant Biology 57, 711–738.
Flavonoids
Flavonoids
• Widest colour range; pale yellow to blue
• Red, pink, mauve, purple, blue, and violet compounds
• Water soluble pigments, stored in vacuole
• Widely distributed in seed plants

59
 Flavonoids
• Among the best characterized plant secondary metabolites
• Phenylpropanoids – Shikimate / Aromatic amino acid
• Anthocyanins, Proanthocyanins, Flavonols, Flavones,
Chalcones, Aurones
• Over 6000 different carotenoid compounds identified

60
 Flavonoids
• Plants:
• Attraction of pollinators and seeds dispersers, signalling between
plants and soil borne symbiont bacteria, defence against
microorganisms
• Human:
• Antioxidant, quality and appreciation
• pH indicator: Litmus test
• Widely used as natural food colorants
• i.e. Cyanidin acylglucosides from red cabbage and Perilla frutescence

https://sils.uva.nl/binaries/_ht_1539094845503/cardm
edium/content/gallery/onderzoek/sils/research/plant-
development-and-epi-genetics/new-phytopathologist-
blue-and-red-petunia.png
 Chemistry
• Anthocyanins, a class of flavonoids derived
ultimately from phenylalanine, are water-
soluble, synthesized in the cytosol, and
localized in vacuoles
• Basic structure C6-C3-C6
• Dozen different groups depending on
decoration
• Chalcones
• Flavones
• Flavonols
• Anthocyanins

Tanaka, Y., Sasaki, N., and Ohmiya, A. (2008). Biosynthesis of plant pigments:
anthocyanins, betalains and carotenoids. The Plant Journal 54, 733–749.
 Chemistry
• Seven subclasses
• Classification based on the oxidation degree of the central
heterocycle.
• The sites of methyl and hydroxyl groups on the other two
rings lead to various flavonoid glycoside modifications
such as glycosylation and acylation.

Santos EL, Maia BHLNS, Teixeira APF and SD, Santos EL, Maia BHLNS, and Teixeira APF and SD. Flavonoids: Classification, Biosynthesis and Chemical Ecology. . In. Flavonoids - From Biosynthesis to Human
Health. (IntechOpen). https://doi.org/10.5772/67861
Biosynthesis
very well
understood

Winkel-Shirley, B. (2001). Flavonoid Biosynthesis. A Colorful Model for Genetics, Biochemistry, Cell Biology, and Biotechnology. Plant Physiology 126, 485–493.
Regulation very well
understood

MYB transcription factors

bHLH TFs
WD40

Verdonk and Quattrocchio, unpublished

Anthocyanin biosynthesis:
Specificity of the DFR and other enzymes

66
Anthocyanin biosynthesis:
Gerbera: Pink and Orange

Regina

President
Teemu Teeri

67
Anthocyanin biosynthesis:
Petunia: Red, Blue and White

68
Anthocyanin biosynthesis:
Carnation x Petunia Gene: Blue

Petunia
F3'5'H

69
Anthocyanin biosynthesis:
Petunia F3’H F3’5’H mutants are white

70
Anthocyanin biosynthesis:
Meyer 1987: Maize DFR

Meyer et al. 1987

Maize DFR

71
 Anthocyanin biosynthesis:
Elomaa 1995: Gerbera DFR
• DFR from Gerbera does an even better job
• less prone to methylation

Gerbera DFR Elomaa et al. 1995

72
Orange Petunia?

Railway station, Helsinki 2015

73
 Return of the orange petunia

Cyanidin Pelargonidin

Bonnie Orange

Teemu Teeri
Railway station, Helsinki 2016
74
Return of the orange petunia

Indian Summer Bonnie Orange African Sunset African Sunset

African Sunset African Sunset Aladdin Orange Orange color petunia

Railway station, Helsinki 2016
75
Return of the orange petunia
Woops!

77
Woops! – No more Orange Petunia’s…. (??)

78
Betalains

79
 Betalains
• Yellow-to-red colours
• Nitrogen-containing water-soluble
• Derived from Tyr (Tyrosine)
• Plants:
• Attraction of animal pollinators and dispersers
• Photoprotection, tolerance to droughts and salinity stress
• High antioxidant and free radical scavenging activities
• Human:
• Commercial food colorants and additives
• Health connection: preventative to several forms of cancer and intermediates are
important pharmaceuticals for treatment of Parkinson’s disease (L-DOPA)
• Substate substrates for other pharmaceutical agents (e.g. dopamine and
isoquinoline alkaloids such as morphine)
80
 Biosynthesis
• Two classes: betaxanthins and betacyanins
• Betaxanthins derived from betalamic acid
conjugation with amines/amino acids

• Betacyanins from condensation of

Betalamic acid with cyclo-DOPA
(cyclo-dihydroxyphenylalanine)

Timoneda, A., Feng, T., Sheehan, H., Walker‐Hale, N., Pucker, B., Lopez‐Nieves, S., Guo, R., and Brockington, S. (2019). The evolution of
81
betalain biosynthesis in Caryophyllales. New Phytologist 224, 71–85.
 Caryophyllales
• Only found in limited number of
plant lineages
• Does not coincide with
anthocyanins in nature
• Substitutes ubiquitous anthocyanin
pigmentation in those species
• Pattern of mutual exclusion
between the two pigments types
• GMOs can have both
Timoneda, A., Feng, T., Sheehan, H., Walker‐Hale, N., Pucker, B., Lopez‐Nieves, S., Guo, R., and
Brockington, S. (2019). The evolution of betalain biosynthesis in Caryophyllales. New
Phytologist 224, 71–85.
Polturak, G., Grossman, N., Vela-Corcia, D., Dong, Y., Nudel, A., Pliner, M., Levy, M., Rogachev,
I., and Aharoni, A. (2017). Engineered gray mold resistance, antioxidant capacity, and
pigmentation in betalain-producing crops and ornamentals. PNAS. 82
Lopez‐Nieves, S., Yang, Y., Timoneda, A., Wang, M., Feng, T.,
Smith, S.A., Brockington, S.F., and Maeda, H.A. (2018).
Relaxation of tyrosine pathway regulation underlies the
evolution of betalain pigmentation in Caryophyllales. New 83
Phytologist 217, 896–908.
84
85
 Summary
Chlorophyll
Carotenoids
Flavonoids
Betalains

https://www.georgeperry.co.uk/images/P/orangepepper.jpg; https://cdn.shopify.com/s/files/1/1380/2059/products/Beet-Root-Ruby-
Red_large.jpg?v=1598078262 ; https://cdn.shopify.com/s/files/1/0102/8043/5812/products/watermelon_300x300.png?v=1563408141 ;
https://www.lekkervanbijons.be/sites/default/files/styles/image_on_detailpage/public/ps_image/istock_aubergine.jpg?itok=bSr_QuF- 86
Postharvest Central
Postharvest Central

Ernst Woltering Rob Schouten

@julianverdonk
Horticulture and Product Physiology 87