colour development

1. WELCOME
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COLLEGE OF HORTICULTURE, BAGALKOT 587 104
Presented by:
Gurumurthy N
Dept. of Vegetable Science
UHS16PGM789
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3. Content
Introduction
Pigments in Vegetables : Types & Distribution
Genetics involved in colour development in
Vegetables
Breeding Approaches for Colour Improvement in
Vegetables
Case studies
Research Achievements
Conclusion
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4. Nutritionists of WHO-FAO suggested that vegetables are essential for
balanced diet.
VAD is recognized as a serious public health problem in India.
It is estimated that 25% of the 15 million blind people globally are
from India.
Colourful vegetables - nutritional , aesthetic and medicinal value.
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5. Photosynthesis
Encourage pollination
Higher nutritional value Higher consumer preference
What are the significance of colours in vegetables?
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6. Pigment class Subgroups Typical colors
Carotenoids Carotenes, lycopene and
xanthophylls
Orange, yellow,
red.
Flavonoids Anthocyanins;
flavonols
Purple, blue,
Red.
Betalains β-cyanins and
β-xanthins
Red, orange, pink.
Chlorophylls a and b Green
Pigments in vegetable crops: Types & Distribution
(Chen,2015)
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7. Schematic colour spectrum of Carotenoids, Anthocyanins,
Betalains and Chlorophylls
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8. Carotenoids in Human Nutrition
Significance of Provitamin A Carotenoids in Developing
Countries
Fertility and Reproductive Success
Prevention of Oxidative Stress and Inflammation
Vision and Diseases of the Eye
Cognitive Decline and Alzheimer’s Disease
Cancer Prevention
Obesity, Cardiovascular Disease, and Diabetes
CAROTENOIDS
(Chen,2015)
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10. CAROTENOIDS RICH VEGETABLES
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11. Antioxidant Properties
Cardiovascular Diseases Control
Role in Obesity
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FLAVONOIDS
(Chen, 2015)
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Water-soluble polyphenolic compounds

12. Anthocyanins
Flavones
Flavonols
Isoflavones
Distribution of flavonoids in vegetables
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13. Betalains
Betalains
β-cyanins
β-xanthin
Betalains are a class of water-soluble pigments that are found only in the
order Caryophyllales.
Betalains differ from anthocyanins in the chemical structures but share
similarities to anthocyanins in the colour spectra, biological functions.
(Chen, 2015)
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14. Betalains
Swiss Chard
M.
Spinach
Amaranthus
Beet
Distribution of Betalain in vegetables
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15. Breeding Approaches For Colour Improvement In
Vegetable Crops
Selection
Mutagenesis
Hybridization
Interspecific hybridization
Somaclonal Variation
Genetic engineering
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16. Most of the colour varieties are developed by this method
Selected on the basis of phenotype
Carrot cream colour variety Pusa Kulfi content high lutein and Pusa
Ashita have high anthocyanin are the two Classical example
Pusa Kulfi Pusa Asita
Selection
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17. Mutagenesis
Carotenoid mutants in Pepper & Tomato
Species Mutant gene Phenotype References
Tomato r (yellow flesh) Yellow fruit colour Fray and
Grierson,1993
delta Orange fruit colour Ronen et al.,1999
tangerine Orange fruit colour Isaacson et al.,2002
Beta Orange fruit colour Ronen et al.,2000
Pepper y (yellow) Yellow fruit colour Lefebvre et al., 1998
c2 Yellow fruit colour Thorup et al., 2000
(Farre et al.,2010)
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18. Hybridization
High-Carotene Cucumber Germplasm
Early Orange Mass 400, Early Orange Mass 402, Late Orange Mass 404
Developed by crosses between U.S. pickling cucumber lines
(Cucumis sativus L. var. sativus) and the orange-fruited Xishuangbannan
cucumber lines (C. sativus L. var. xishuangbannanesis Qi et Yuan)
Fig. Cross sections of four mature fruits of
LOM 404 high carotene cucumber (15-25
mg.kg-1).
(Simon and Navazio, 1997)
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19. Vertical Section of Early Orange Mass 402 Of well matured single fruit
(Simon and Navazio, 1997)
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20. Case Study-1
Objective
 Selection of high temperature tolerant inbreds along with enhanced
nutritional value and yield
(Saha et al., 2016)
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21. Materials and Methods
 38 Inbreds of tropical carrot were evaluated for different Nutritional traits
for four consecutive years (2009-2012).
 Well developed and matured carrot roots of all inbreds were harvested on
same day. The roots were properly cleaned with tap water to remove the
dust particles.
 Estimation of Anthocyanins
 Total Carotenoids and Lycopene were extracted in acetone
 TSS is measured by Refractometer.
 UPGMA (Unweighted pair-group method with arithmetic averaging
algorithm) was used for establishing cluster to search natural groupings
among the genotypes for morphological and biochemical parameters.
(Saha et al., 2016)
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23. (Saha et al., 2016)
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24. INFERENCE
 Maximum anthocyanin was noted in IPC-126 which is dark purple carrot
inbred followed by IPC-020
 Maximum total carotenoid in IPC-124 followed by IPC-11.
 Optimum temperature required for proper growth and good colour
development varies from 15.5-21.1˚C.
(Saha et al., 2016)
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25. Case Study -2
To develop Carotenoids and Anthocyanin rich transgenic sweet potato with
high antioxidant capacity.
To characterize carotenoid contents and antioxidant activity in transgenic
sweet potato
Objective
(Park et al., 2015)
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26. Materials and Methods
Plant materials and expression vectors
Take Anthocyanin rich sweet potato cultivar ‘Sinzami’(SZM) as wild type
(WT).
Under the control of the cauliflower mosaic virus (CaMV) 35S promoter
was constructed.
Agrobacterium -mediated transformation
Fig. A -Schematic diagram of the T-DNA region of the IbOr constructs used for
plant transformation. LB and RB, left and right T-DNA borders
respectively; 35S Pro- 35S promoter; gray boxes-cloning site of
gateway vector systems; FLAG, FLAG octapeptide.
(Park et al., 2015)
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27. Fig.B - Phenotypes of storage roots and
leaves of three IbOr transgenic and
wild-type (WT) lines.
Fig.C -Expression of IbOr in leaves
and storage roots
Results and Discussion
B
C
(Park et al., 2015)
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28. Table-1: Carotenoids contents in the storage roots of transgenic
and wild-type sweet potato plants (μg /g DW).
Fig.D - Analysis of total Anthocyanin contents.
Continued….
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29. Fig.- F Photograph of aerial plant
parts and storage roots
Fig.- E. Average yields of storage roots
E F
(Park et al., 2015)
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30. INFERENCE
In conclusion, Anthocyanin-rich transgenic sweet potato plants with
high Carotenoids were successfully generated
Carotenoids contents and antioxidant activity in storage roots were
characterized
Transgenic plants is expected to contribute to human health by
increasing the contents of Carotenoids
Transgenic sweet potato show tolerance to environmental stresses .This
would be beneficial for sustainable agriculture on marginal lands
worldwide.
(Park et al., 2015)
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31. Case Study -3
To enrich the Anthocyanin content of the fruits of a cultivated tomato cultivar
To enhance the antioxidant value of tomato fruits
objectives
(Maligeppagol et al., 2013)
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32. Materials and Methods
Two gene - delila and rosea from Antirrhinum majus
Transformation of tomato with pGAntho construct was carried out
by Agrobacterium-mediated gene transformation
Vectors : pTZ57R/T and pBI121
Figure1 . Schematic diagram of the pGAntho gene construct in pGreen
II.
Gene cloning and vector construction
(Maligeppagol et al., 2013)
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33. Results and Discussion
Transgenic tomato plants accumulating high amounts (70–100
fold) of Anthocyanin in the fruit were developed
Figure 2. Biochemical analysis of tomato fruit.
Anthocyanin content; Antioxidant capacity.
WT- Wild type; C-Commercial; V-Vector and pGAntho, Transgenic
tomato. (Maligeppagol et al., 2013)
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34. Figure 2. Biochemical analysis of tomato fruit.
Total carotenoid content; Lycopene content.
WT, Wild type; C, Commercial; V, vector and pGAntho, Transgenic tomato.
Continued….
(Maligeppagol et al., 2013)
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35. Gene Expression Analysis
Figure 3. Expression analysis of Del and Ros1 and their targets:
Relative mRNA levels of Del and Ros1 at three developmental stages
of fruit.
Relative mRNA levels of F3H and CHI induced by Del and Ros1 at
three developmental stages of fruit.
(Maligeppagol et al., 2013)
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36. Figure . Tomato fruit colour in pGAntho
(E8:Ros/Del) transgenic, WT-control and
vector control .
Comparison of ripened fruits of pGAntho,
WT-control and Vector control.
Transverse section of the ripened fruits of
pGAntho, WT-control and Vector control.
Fig.4 Fruit colour during developmental stages of the
fruit, from left green, breaker and red-ripe of WT-
control (top) and pGAntho (bottom).
(Maligeppagol et al., 2013)
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37. In conclusion, they have developed transgenic tomato producing fruits
rich in Anthocyanins
This approach has achieved high levels of Anthocyanin content
through-out the fruit
The fruit is of value for its health benefits as it is rich in Anthocyanins
along with naturally occurring Carotenoids.
In addition to the direct consumption, these fruits can also be utilized
to extract biological food colourants and Anthocyanin compounds used
as a nutraceuticals.
INFERENCE
(Maligeppagol et al., 2013)
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38. Achievements
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39. Semi-dominant, single-locus mutation .
Beta carotene range from 3 to 320 mg/ 100 gram fresh tissue
(Li et al.,2012)
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40. Cauliflower Pr gene
Anthocyanins
Purple (Pr) gene mutation is a spontaneous mutation.
single, semi dominant gene
‘Graffiti’ (Harris Seeds) cultivated in USA.
accumulated 3.75 mg - cyanidin diglucoside
(Chiu et al., 2010)
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41. CROP GENE PIGMENTS
TOMATO B β- CAROTENE
TOMATO Aft ANTHOCYANIN
TOMATO (Psy-1) CAROTENOIDS
CARROT A α-CAROTENE SYNTHESIS
CARROT K LYCOPENE SYNTHESIS
CARROT O ORANGE XYLEM
CARROT Y YELLOW XYLEM
CARROT P-1, P-2 PURPLE ROOT
CHILLI A ANTHOCYANIN
CHILLI B BETA CAROTENE
CHILLI t HIGH BETA CAROTENE
Genetic Inheritance of Vegetable crops
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*In Black Carrot, colour is governed by Digenic trait
(Hari hara ram, 2006)

42. Open pollinated variety
Deep golden orange colour
Beta carotene : 0.1-1.20 g/100 g FW
Lycopene : 7.8-18.1 mg/100 g FW
Caro rich - Tomato
WVC -Tomato Breeding
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43. Nutritious tomato line
WVC -Tomato Breeding
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44. Interspecific Hybridization
M. L. Tomes(1958) , bred Caro-Red, a Provitamin -A rich tomato variety
 Developed by cross between common tomatoes, Lycopersicon esculentum
Mill., and the wild species, L. hirsutum Humb
Due to its orange colour it is got commercial acceptance and consumers
preference in Abroad
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45. Tissue culture derived plants show variation termed somaclonal variation
Chromosomal rearrangements are an important source of this variation
A cultivar of sweet potato ‘Scarlet’ having higher yield and disease
resistance characteristics similar to their parent but also have darker and
more stable skin colour
Somaclonal Variation
(Moyer and Collins, 1983)
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46. Pusa Rudhira
(High lycopene)
Pusa Meghali
(High β-carotene)
Pusa Asita
(High anthocyanin)
Selection
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Carrot
Pusa Vristi
(Lycopene: 405µg /100g)
Pusa Payasa
(High β-Carotene)

47. F1-Hybrids
Pusa Nayanajyoti
(High β-Carotene)
Pusa Vasuda
(Lycopene)
Kuroda
( β-Carotene)
Oxheart
(Lycopene+Anthocyanin)
Private sectors
Public Sector
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48. Arka Shresta
(β- Carotene)
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Pusa Rohini (Lycopene) Pusa Uphar (Lycopene)
F1-hybrid
Selection

49. Sree Vijaya
Beta carotene
Sree Vardhini
Carotene : 1200 IU/100g
Sree Bhadra
carotene : 972 IU/100g
Varieties - Clonal selection
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Sree Ratna
(Carotene :3500 IU/100g)
Hybrid

50. Black beauty
(Anthocyanin)
Arun
(Anthocyanin)
Salkeerti
(Lycopene)
Hybrids
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51. Punjab lal
(Capsanthin)
Arka Abhir
(Capsanthin)
Arka Neelakanta
(Anthocyanin)
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Onion(Pusa Ridhi)
(Quercetin 107.42 mg/100g)
selection

52. Pusa Sagarika
(Anthocyanin)
Chinese pink
(Anthocyanin)
Chinese rose
(Lycopene)
Chinese Red
(Lycopene)
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Radish

53. Pusa Mridula
(Lycopene)
Pusa Jamuni
(Anthocyanin )
Pusa Gulabi
(Anthocyanin)
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Radish
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Selection

54. Pumpkin
Pusa Vikas (β-carotene)
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Amaranthus
Pusa Lal Chaul (Anthocyanin)
Red Lettuce
Red oka leaf
(Lutein + zeaxanthin)
Arka Chandan
β-carotene( 3331 IU/100 g)
Mass selection
Selection

55. Now days coloured pigments play major role in human nutrition as well as
attraction of consumer.
There is lot of scope for natural colourants by pigment extraction.
Colours are major visual component in export oriented value.
Now a days Molecular markers used in easy identification of colours at early
stage and effective utilisation of that particular quality aspects.
Still the work is going on to bred the varieties with resistant
against particular biotic and abiotic factors along with colour improvement
Conclusion
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