This document discusses various approaches to biofortification through transgenic techniques. It begins by defining a transgene and biofortification. It then discusses how biofortification can be achieved through conventional breeding or transgenic approaches when nutrients do not naturally exist in crops or cannot be bred in through conventional means. Examples of transgenic biofortification projects are provided, including cassava with increased iron and provitamin A, bananas with provitamin A and iron, vitamin E maize, and golden rice with provitamin A. The document also discusses iron deficiency globally and biofortified iron rice. It concludes by discussing a "Dream-RICE" project with the goal of developing rice fortified with multiple micronutrients through

1. An Assignment on
Transgenic for Bio fortification
Biotechnology for Crop Improvement
(GP 509)
Speaker:
Damor Kalpeshkumar M.
Reg. No : 04-AGRMA-01983-19
Dr. Kapil K. Tiwari
Assistant Research Scientist
Bioscience Research Centre
S.D. Agricultural University
Submitted to:

2. What is transgene ???
A transgene is a gene or genetic material that
has been transferred naturally, or by any of a
number of genetic engineering techniques from
one organism to another.

3. What is Biofortification
• Biofortification is the development of nutrient-dense
staple crops using conventional breeding or non
conventional practices, without sacrificing agronomic
performance and important consumer-preferred traits.
• Greek word “bios” means “life” and Latin word
“fortificare” means “make strong”.

4.  Biofortification can be achieved through conventional plant
breeding, where parent lines with high vitamin or mineral levels
are crossed over several generations to produce plants that have
the desired nutrient and agronomic traits.
 Transgenic approaches are advantageous when the nutrient does
not naturally exist in a crop (for example, provitamin A in rice)
or when sufficient amounts of bioavailable micronutrients
cannot be effectively bred into the crop.
 However, once a transgenic line is obtained, several years of
conventional breeding are needed to ensure that the transgenes
are stably inherited and to incorporate the transgenic line into
varieties that farmers prefer.
 While transgenic breeding can sometimes offer micronutrient
gains beyond those available to conventional breeders, many
countries lack legal frameworks to allow release and
commercialization of these varieties.
 To attain higher levels of provitamin A, zinc, and iron content in
crops where genetic variation for these traits has not been
identified, Harvest Plus, its partners, and other organizations
have explored transgenic approaches.

5. Global food security and
malnutrition
• Micronutrient malnutrition
because of iron and zinc
deficiencies is a serious
public health problem in
worldwide.
• 3 billion people worldwide
suffers micronutrient
deficiency.
• 840 million people are
food insecure.
• 200 million malnourished
children.
Source : WHO, 2017

6. • 1.6 billion population
suffer from Iron deficiency.
• 1 billion people reside
in iodine deficient regions.
• 400 million people have acute vitamin A
deficiency.
• Malnutrition accounts ~30 million
death/year.
• 2.5 billion world population suffer from Zinc
deficiency.
Source : WHO, 2017

7. *Micronutrient deficiencies (vitamin A, iron and zinc): WHO data for children under 5
(As of 28 Jan 2010).
Fig: 1 Global Prevalence of Micronutrient Deficiencies
Low
Moderate
High
No data
7

8. National scenario
• About 80% pregnant women, 52% of non-
pregnant women, and 74% of children are in
iron deficiency.
• About 52% of children below 5 year are zinc
deficient.
• World Bank data indicates that India has one of
the world’s highest demographics of children
suffering from malnutrition
• India’s Global Hunger Index India ranking of
67 the 80.

9. Importance of Biofortified Crops in
improving Food and Nutrition Security
Compared with conventional (non-biofortified) crops,
biofortified crops have:
Increase foods
available in homes
 Better agronomic
characteristics
• Greater: yields, resistance
to pests, tolerance to stresses
 Higher nutritional
concentration
• More: iron, zinc, beta-
carotene and/or tryptophan
and lysine
Increase the intake of
these nutrients
Improve nutrition
security
Improve food
security

10. Genetic Bio-fortified Crops
Source : Harvest Plus Programme

11. Source : ICRISAT

13. BioCassava Plus:
• The BioCassava Plus (BC+) program genetically engineers
cassava with increased levels of iron and provitamin A.
• Additional traits addressed by BC+ include increased shelf
life, reduced cyanide levels, and improved disease resistance.
• The first field trials for a provitamin A biofortified cassava
began in 2009, followed by trials for high-iron cassava .
• Delivery of the biofortified crops is expected in 2017.
• Retention and bioavailability of transgenic cassava are similar
to the findings of HarvestPlus on conventional biofortification
research.

14. Four strategies for the enhancement of protein
levels in cassava storage roots.
(a) Expression of a vacuolar linamarase (VL lines). The expression of a
vacuolar-targeted linamarase increased the deglycosylation of linamarin
leading to an increased assimilation of cyanide into amino acids.
(b) Coexpression of a vacuolar linamarase and the novel storage protein
sporazein (SVL lines). The SVL lines, similar to the VL lines showed
increased free amino acid levels, however, in difference to the VL lines
the SVL lines showed a two fold increase in protein levels in the roots.
(c) Expression of zeolin. The chimeric storage protein zeolin expressed
under the control of the root specific patatin promoter resulted in a
fourfold increase in root storage proteins in older cassava roots.
(d) Over expression of hydroxynitrile lyase (HNL lines). The
overexpression of HNL in storage roots resulted in a twofold to
threefold increase in protein content in younger cassava roots.
• The HNL enzyme is targeted to the apoplast.

15. Vitamin A and Iron Bananas:
• Queensland University of Technology and the National
Agricultural Research Organization of Uganda are
developing transgenic provitamin A and iron bananas
for Uganda.
• Bananas with up to 20ppm provitamin A have been
developed and trials have commenced in Uganda .
Provitamin A bananas released in 2019.
• A human bioavailability study using transgenic
provitamin A banana began in late 2013.
• High-iron bananas are not yet ready for use in human
trials.

16. Iron Wheat:
• Efforts to increase iron concentrations in wheat by
conventional breeding have not been successful, and there are
currently no iron-biofortified wheat varieties available for
farmers.
• Whole wheat grain contains approximately 30 ppm iron, of
which only 5% is estimated to be bioavailable.
• It is estimated that wheat requires an additional 22 ppm iron in
the whole wheat grain, for a total concentration of 52 ppm
iron, to adequately biofortify a wheat-based diet with iron.

17. Vitamin E- Maize
 HGGT catalyzes an analogous reaction to
HPT, only it is highly specific for GGDP
whereas HPT uses PDP as its prenyl
substitute.
 Results from the expression of barley HGGT
in transgenic plants suggest that this enzyme
has strong substrate specificity for
geranylgeranyl diphosphate, rather than
phytyl diphosphate.
 Expression of HGGT enzyme in tobacco calli
and Arabidopsis leaves resulted in
accumulation of Vitamin E antioxidants in the
form of tocotrienols ,principally as γ-
Tocotrienols, and generated little or no
change in the content of Tocopherols (Cahoon
et al, 2003)
 Barley HGGT gene was 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).

18. Golden rice
• Golden Rice was first developed at the Swiss Federal Institute of
Technology and the University of Freiburg, Germany.
• The inventors donated the technology for public sector research and
development and farmers’ use, free of charge, in developing countries.
• This effort was assisted by Syngenta who arranged, for humanitarian
purposes, royalty-free access to intellectual property for a number of key
technologies used in Golden Rice held by several biotechnology
companies.
• These arrangements allow the International Rice Research Institute
(IRRI) and others to develop Golden Rice on a non-for-profit basis.
• In parallel, Golden Rice product development was furthered by Syngenta
as part of their then-commercial pipeline.
• Transgenic events with higher levels of provitamin A, up to 37 ppm in a
U.S. variety (GR2 events), were produced and were then donated for use
by the Golden Rice Network when Syngenta decided not to pursue the
trait as a commercial product.

19. BR29
• In 2005, a team of researchers
at Syngenta produced Golden
Rice 2.
• They combined the phytoene
synthase gene from maize with
crt1 from the original golde n
rice.
• Golden rice 2 produces 23
times more carotenoids than
golden rice (up to 37 µg/g),
and preferentially accumulates
beta-carotene (up to 31 µg/g of
the 37 µg/g of carotenoids).
Golden Rice 2

21. Essential Minerals: Iron
Iron deficiency is the most widespread micronutrient
deficiency worldwide.
Approx. 30% of world population suffers from
serious nutritional problems caused by insufficient
intake of iron (WHO 1992).
It is the important constituent of hemoglobin, the
oxygen carrying component of blood, and also a part
of myoglobin that helps muscle cells to store oxygen.
It is present in food in both inorganic (ferric and
ferrous) and organic (heme and nonheme) forms.
Highly bioavailable heme iron is derived primarily from
animal source.

22. Biofortified iron rice
1. High iron and enhanced carotenoids/beta-carotene rice
2. Reduced content of phytate in rice grains
Mutational
breeding
Transgenic plant
strategy
Screening for iron-
rich rice varieties
Increased bioavailabillity of Fe and Zn

23. Value added Dream-RICE
• High iron rice (after polishing)
Provitamin A rice
Other micronutrient-rich rice
Development of Value added rice for both
favorable and unfavorable ecosystems.
combination of high yield with value-added rice

24. THANKS