The document summarizes a seminar on the role of genetic engineering in crop biofortification. It discusses methods of biofortification including genetic and agronomic approaches. A key example provided is the development of "Golden Rice" through genetic engineering by introducing genes that complete the biosynthesis pathway for beta-carotene, a precursor for vitamin A production. The document also discusses enhancing vitamin E in maize through overexpressing a gene involved in tocotrienol biosynthesis, resulting in large increases in vitamin E content.

1. Department of Genetics & Plant Breeding
C.S. Azad University of Agriculture & Technology, Kanpur 208002
Uttar Pradesh, INDIA

2. A Course Seminar
on
Role of genetic engineering in crop bio-fortification
Speaker :Jaydev Kumar, Id. No. :CA 8608/12
Course No.: GPB 609, Cr. :1(1+0)
Date : 05-04 -2014, Time : 03:00 P.M.
Course Instructors: Dr. D.N. Bharadwaj
Dr. R.P. Vyas
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

3. Highlights
1. What is Crop Bio fortification ?
2. Importance of crop bio-fortification
3. Methods of Biofortifocation
a. Genetic bio fortification
b. Agronomical bio fortification
4. A way use for enhance pro-vitamin A in Rice
a. Why require to enhance Pro vit. A in rice
b. Flow chart to explain a way for understanding of rice bio-fortification
5. Enhancement of Vitamin E in crop plant via crop biofortification
a. Sources of Vitamin E : Tocotrienols
b. Most suitable example: Maize
6. Advantages and disadvantages
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

4. What is bio-fortification
Bio-fortification:
Greek word “bios” means “life” and Latin word “fortificare” means
“make strong”. Food fortification or enrichment is the process of adding
micronutrients (essential trace elements and vitamins) to food.
Crop bio-fortification:
•Crop Bio-fortification is the idea of breeding crops to increase their nutritional value.
•Bio-fortification differs from ordinary fortification because it focuses on making plant
foods more nutritious as the plants are growing, rather than having nutrients added to
the foods when they are being processed.
•This is an improvement on ordinary fortification when it comes to providing nutrients
for the rural poor, who rarely have access to commercially fortified foods.
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

5. Some important view regarding bio-
fortification
• Global food system is failing to deliver adequate quantities of healthy, nutritionally balanced
food, especially to the resource-poor underprivileged people leading to micronutrient
malnutrition.
• The malnutrition of minerals (Fe, Zn) and vitamin A are major food-related primary health
problem among populations of the developing world including India where there is a heavy
dependence on cereal-based diets and limited access to meat, fruits and vegetables.
• Vitamin A deficiency (VAD) alone is significant from the public health point of view
resulting in over 330,000 child deaths every year, and about 57% of preschoolers and their
mothers having subclinical VAD leading to increased morbidity and risk of mortality. While
beneficial supplementation of vitamin A is currently being addressed through sponsored
nutrition programmes, they are not sufficient in covering the affected populations.
• Advancement in genetic engineering, results a significant increase in the total carotenoids and
β-carotene levels in the transgenic events. Provitamin A enrichment in rice crops could have a
significant impact on the nourishment and nutrient interactions by playing a major role in the
bioavailability and metabolic efficiency in the affected populations.
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

6. Importance of crop bio-fortification
Bio-fortification for important crop plants through biotechnological
applications is a cost-effective and sustainable solution for alleviating
VAD, etc.,. Some points present here to clearly identified role of crop bio
fortification …….
To overcome the mal-nutritions in human beings
To increment of nutritional quality in daily diets
To improvement of plant or crop quality, and increment of variability in
germplasm
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

7. Methods of crop bio-fortification
Genetic bio fortification
Agronomical bio fortification
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

8. Genetic bio-fortified crops
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

9. Conventional Plant Breeding to
Develop Biofortified Crops
Drought-tolerant
Bean
High-iron Bean
X
Drought-tolerant,
high-iron beans

10. Genetic engineering & bio fortification of
Rice
• Genetic engineering is the obvious alternative to
enhance the β-carotene levels in crop plants.
• The development of the ‘golden rice’ proved that, it is
possible to redirect a complete biosynthetic pathway of
carotenoids by genetic engineering of multiple genes
encoding key enzymes of the pathway.
• So, Golden Rice is such a bio-fortified crop.
• A example of Golden Rice was developed in the year
2000.
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

11. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA
• Addition of 2 genes in rice genome will complete the
biosynthetic pathway:
1. Phytoene synthase (psy): derived from daffodils
(Narcissus pseudonarcissus). Psy is a transferase enzyme
involved in the biosynthesis of carotenioids. It catalyzes
the conversion of GGPP to phytoene.
2. Lycopene cyclase (crt1)- isolated from soil bacteria
Erwina uredovora.
3. Produce enzymes and catalysts for the synthesis of
carotenoids in the endosperm of rice.
How Does It Work?

12. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

13. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA
Grain of rice represent
high Vit. A, developed
through genetic
engineering
Grain of rice lack of Vit.
A

14. The Golden Rice Solution
IPP (Isopentenyl pyrophosphate)
Geranylgeranyl diphosphate
Phytoene
Lycopene
-carotene
(vitamin A precursor)
Phytoene synthase
Phytoene desaturase
Lycopene-beta-cyclase
ξ-carotene desaturase
Daffodil gene
Single bacterial gene;
performs both functions
Daffodil gene
-Carotene Pathway Genes Added
Vitamin A
Pathway
is complete
and functional
Golden
Rice
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

15. 3.0-9.1g/g,DHhomozygouslinesdeveloped
Datta K et al PBJ, 2003/2005,2006
Parkhi et al MGG, 2005,2006
Rai et al 2003,2006
Ye et al Science, 2000
Painie et al Nature Biotech, 2005
Golden Rice (BR29) developed at IRRI is now in Bangladesh soil
Syngenta-Golden Rice (GR2) is now in field at Louisiana, USA
CommercialrightofGRremainswithSyngenta
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

16. Sources of Vitamin E : Tocotrienols
 Primary sources of vitamin E are derived from plants. Tocopherols and Tocotrienols
are plastid localised molecules.
 Oil seeds are richest source of vitamin E, having total tocol levels ranging from 330
to 2,000 µg per gram. Tocotrienols are the primary form of vitamin E in seed
endosperm of most monocots, including cereals, such as wheat, rice, and barley.
 Tocotrienols are found in the seed endosperm of a limited number of dicots, such
as tobacco and found rarely in vegetative tissues of plants.
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

17. Vitamin E- Maize
The biosynthesis of the tocotrienol and tocopherol
forms of vitamin E is initiated by prenylation of
homogentisate.
HGGT (homogentisate geranylgeranyl
transferase)catalyzes an analogous reaction to HPT
(homogentisate phytyl transferase), only it is highly
specific for GGDP whereas HPT uses Phytyl
Diphosphate as its prenyl substitute.
Results from the expression of barley HGGT in
transgenic plants suggest that this enzyme has strong
substrate specificity for geranyl geranyl
diphosphate, rather than phytyldiphosphate.
over-expressed in maize seeds, leading to a 20-fold
increase in tocotrienol level, which translated to an
eight-fold increase in total tocols (tocopherols and
tocotrienols) (Cahoon et al, 2003).
Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

18. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

19. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA

20. Department of Genetics & Plant Breeding
C.S.A.U.A.&T., Kanpur 208002 U.P., INDIA