Slides from the webinar on "Plant Genome Engineering for Agriculture, Food and Nutrition" delivered by Dr. K C Bansal, as part of the Shri Vaishnav Institute of Science webinar series. Dr. K C Bansal is a former director of the Indian National Bureau of Plant Genetic Resources (ICAR) and current board member of the Global Plant Council.

1. National Bureau of Plant Genetic Resources (ICAR)
Pusa, New Delhi, India
K C Bansal
kcbansal27@gmail.com
Plant Genome Engineering for Agriculture, Food and Nutrition
Former Director

2. Human and Societal Needs
• Food availability, affordability
• Nutritionally rich diet
• Clean and green environment
• Farmers’ welfare
Challenges
o Climate change
o Shrinking per capita land and water
o Expanding biotic and abiotic stresses
o Sustainability concerns with ever-increasing
population
K C Bansal

3. 290 to 375- 400 Mt
Source: ICAR Vision 2050K C Bansal

4. Sustainable Development Goals - 2030
End Hunger, Achieve Food
Security and Improved
Nutrition and Promote
Sustainable Agriculture
K C Bansal

5. Daruma
(Japanese semi-dwarf) X
Fultz
(U.S. winter wheat, high yield)
Fultz-Daruma
(semi-dwarf, high yield)
Locals
(adapted to
U.S. Northwest)
X
X Turkey Red
(U.S. winter, high yield)
Norin 10
(semi-dwarf, winter, high yield)
(Dr Gonziro Inazuka in 1935)
Gaines
(semi-dwarf, winter,
U.S. adpted)
X Local Strains
New Wheats
(semi-dwarf, high yielding, adaptable,
rust-resistant, fast-maturing, spring)
Utilization of Plant Genetic Resources
Gave Birth to Green Revolution
WHEAT/GREEN REVOLUTION

6. Lost genes and alleles can be recovered only by going back to the
wild ancestors of our cultivated crop species
Importance of Wild Species in Food and Nutrition
Science,1997K C Bansal
Seed/gene
banks harbor
living seed
collections,
which act as
source of
genes for
improving
agricultural
crops.

7. Conventional breeding –Time consuming, genetic drag of
negative alleles from wild species, sexual incompatibility
Molecular breeding – Molecular marker-based, hastens
the process of conventional breeding
Genetic engineering – Specific genes introduced,
overcome the barrier of sexual incompatibility
Genome editing – Precise genome-wide gene alterations
or insertions possible
Approaches currently available
K C Bansal

8. Fine structure of a gene
mRNA
Protein
Metabolites
P H E N O T Y P E
K C Bansal

9. GUS
GUS
Regulation of Cell-specific Gene Expression
Upstream
regulatory
sequences
Promoter
Coding sequence
Coding sequence
GUS: β-glucuronidase Bansal et at 1992 PNAS, USA, 89: 3654
K C Bansal

10. DroughtControl Salt
Abiotic Stress-inducible Gene Expression
GENE CONSTRUCT
Promoter Coding region Ter
GUS
GUS: β-glucuronidase
K C BansalDalal et al 2009

11. Gene
Discovery
Genetic
Transformation
Transgenic
Development
Evaluate
Transgenics
Molecular
Breeding
Comme-
rcialization
Gene
Promoter
Gene
construct
Nuclear
Plastid
Crop
Elite
cultivar
Biosafety
Agronomy
Field Trials
Backcross
Breeding
Variety
Development
Farmers
Consumers
K C Bansal

12. FOREIGN GENE
LB RB
A Gene is introduced into a plant cell and
whole plant is regenerated from that cell
Transgenic Plant
K C Bansal

13. Development of Transgenic Tomato
Different steps
Transgenic Plants
K C Bansal

14. Stacking three late blight resistance genes from
wild species into African highland potato varieties
confers complete field resistance to late blight
Plant Biotechnology Journal (2019)
17, pp. 1119–1129
A Gene from a wild species
- Solanum bulbocastanum confers
broad spectrum resistance to
potato late blight
Song et al 2003, PNAS
Non-GM GM
Transgenic GM Katahdin and
control Non-GM plants
inoculated with P. infestans.
Disease symptoms were
recorded 7 days after
inoculation.
RB gene
Gene from wild potato
species to
cultivated potato
variety
K C Bansal

15. Pepper
Gene from Pepper to Bananahypersensitivity response‐assisting protein (Hrap)
K C Bansal

16. Traditional plant breeding
Desired
Gene
X
Many genes are
transferred
Donor
Plant
Commercial
Plant Variety
New Plant
Variety
+
A single gene
is transferred
Desired Gene
Commercial
Plant Variety
Improved
Commercial
Plant Variety
Transgenic route
Desired
Gene
Donor
Species
Transgenics vs Traditional Breeding
Needs to be discovered
K C Bansal

18. K C Bansal

19. ISAAA Brief 54
GM crop plantings have increased ~113-fold since 1996, with an
accumulated area of 2.5 billion hectares, showing that biotechnology is
the fastest adopted crop technology in the world.
Global Status of GM Crops - 2018
K C Bansal

20. Transgenic Papaya Resistant to Papaya Ring Spot
Virus (PRSV): A GMO Success Story
1986: Efforts initiated develop virus resistant
transgenic papaya by transforming Hawaiian
papaya with CP gene of PRSV by Gonsalves &
coworkers.
1991: Transgenic line (55-1) resistant to PRSV
identified.
1992: Two PRSV resistant transgenic cultivars
‘SunUp’ and ‘Rainbow’ developed.
1998: License to commercialize papaya in Hawaii.
Approval in other countries: USA, Canada &
Japan
Severely PRSV affected fields in 1994 Green healthy transgenic Rainbow papaya in 1999
Source: Gonsalves et al. (2004) APSNet (Feature Story)
Gene from a virus
K C Bansal

21. GM Apple Approved with Non-Browning Phenotype
 Contains PGAS PPO suppression gene
Approved in 2015: USA & Canada (As Food for direct use or in processed
form)
Shutting down apple’s own Genes
K C Bansal
The PPO suppression transgene (PGAS) consists of 394, 457, 457 and
453 bp regions of apple PPO genes (PPO2, GPO3, APO5, pSR7,
respectively). Suppression is achieved through the formation of dsRNA
between the suppression transcript and native PPO mRNA.

22. GOLDEN RICE
Genes from daffodils, maize and bacteria
8 ug/g
25 ug/g
K C Bansal

23. • Vitamin A Deficiency is the
leading cause of child
blindness
• A source of Vitamin A reduces
child mortality by 23-34%
• The Beta-carotene in Golden Rice
is the immediate source of
Vitamin A
Potential Impact of Golden Rice
K C Bansal

24. Genes from maize and bacteria to
Banana
50% of the EAR of
Vitamin A with
consumption of
only 300 g per
person per day.
Will be
released by
2021 to have
a significant
impact in
alleviating
VAD in a
sustainable
way,
especially in
rural Uganda
K C Bansal

25. Next Generation of GM Crops for Human Health Benefits
Current Opinion in Plant Biology 2013, 16:255–260
K C Bansal

26. Global cost benefit from the production of genetically modified
(GM) crops in the year 2013. The total cost benefit comprised
of less input cost and high yield with adoption of GM crop
in the respective countries.
Global cost benefit from the production of GM
crops
K C Bansal

27. Bt CottonNon Bt Cotton
Bt Cotton Commercialized
in India - 2002
Gene from Bacteria
K C Bansal

28. Adoption of Bt Cotton in India
(2002 - 2017)
K C Bansal

29. Impact of Bt Cotton
Share of India in World Cotton Market Increased, 2002 to 2016
Source: ICAC, 2016; USDA, 2016
Non-Bt Cotton
Bt Cotton
Ginners and Textile Mills Prefer Bt Cotton
K C Bansal

30. Fruit & Shoot Borer Resistant Transgenic (Bt) Brinjal
*Approval of Bt Brinjal Event EE1 in Bangladesh (2013)
Source: ISAAA, 2014
In India, Bt Brinjal under moratorium since 2010
K C Bansal

31. Traits necessary to be introduced through GM route
• Insect resistance – cotton, brinjal, chickpea,
pigeon pea and other crops
• Herbicide tolerance – cotton, maize, rice, wheat
and other crops
• Disease resistance (virus, bacterial, fungal) –
potato, tomato, banana, rice, chickpea, cotton
• Nutritional quality –rice, wheat and other crops
• Abiotic stress tolerance – Drought, salinity, cold
• Increased productivity – Yield components, early
flowering, hybrid production
8/6/2020
K C Bansal

32. BRL-I Trial, Kumher
BRL-I Trial, Navgaon
Crossability Study at Bawana
BRL-I Trial, Sri Ganganagar
GM Mustard Hybrid in India
Pending approval since 2017
Seed Yield of GM Mustard Hybrid DMH-11= 27- 37% more

33. Coordinators: NK Singh and KC Bansal

34. Sundaresha et al (2019), Functional Plant Biology https://doi.org/10.1071/FP17299
RBKJ lines were developed using Kufri Jyoti as
female parent and the RB-transgenic Katahdin
event SP951 as the male parent
Non-GM GM
GM
Non-GM
Transfer of the RB gene by crossing a
specific RB-transgenic event with well
adapted Indian potato cultivar Kufri
Jyoti
RB gene
K C Bansal
Late-blight Resistant Potato

35. K C Bansal
Drought and Salinity Tolerant Tomato

36. RCGM Contained Open Field Trial of Transgenic Mustard for Event Selection
Wild Type Transgenic
6.4 Permission to conduct Biosafety Research Level-1 (BRL-1) trials on
transgenic mustard (Brassica juncea L) varieties namely Pusa Jaikisan and
Varuna containing osmotin gene (event Omb5-B) confers tolerance to drought
stresses in Brassica juncea during first week of November, 2010 by
National Research Centre on Plant Biotechnology, IARI Campus, New Delhi
Decision taken in the 103rd meeting of the
Genetic Engineering Appraisal
Committee (GEAC) held on 29.9.2010.
Wild Type Transgenic
Drought Tolerant Mustard

37. Drought Tolerant Rice
TRANSGENIC
WILD TYPE
T WT
Indica rice cv. Pusa Sugandh 2
Indica rice cv. Samba Mahsuri

38. Under cold stress, significantly longer root length, higher biomass
Cold
Control
OsPYL10 is the first
functionally characterized
ABA receptor from a
monocot, Rice cv N22)
(FN556370) 2009
Lenka et al 2018K C Bansal

40. Kaybonnet
Homozygous lines
Swarna
Swarna
MTU1010
BPT 5204
ASD 16
ADT43
IR64
IR36
GoldenSwarna Introgression Breeding for Development of Golden Rice in India
K C Bansal
Golden Indian Rice
Source: Dr AK Singh, IARI

41. GM Crops under Regulatory Evaluation in India
Crops Trait
Brinjal, cabbage, cauliflower, maize, cotton,
okra, rice, sorghum, sugarcane, tomato
Insect Resistance
Cotton, groundnut, papaya, potato, tomato,
watermelon
Virus resistance
Maize, cotton Herbicide tolerance
Maize, cotton and rice Herbicide tolerance +
Insect resistance
Chickpea, groundnut, mustard, sorghum Drought tolerance
Rice Yield enhancement
Tomato Delayed ripening
Mustard, rice Male sterility, female
inbred lines

42. Health Safety Assessment Environmental Safety Assessment
 Compositional Analysis
 Alteration in Nutrient Composition of Leaf
and Seed
 Alteration in Toxin/ Anti-Nutrient
Composition in Leaf and Seed
 Toxicity Potential
 Expression Levels of Introduced Protein
 Acute Oral Toxicity of Purified Protein
 Sub-Chronic Toxicity with edible parts
 Allergenicity Potential
 Bioinformatics Analysis of Proteins
 Pepsin Digestibility
 Thermal Stability
 Weediness Potential
 Seed Germination & Speed of Seed Germination
 Seedling Vigour & Seed Size
 Long Continuous Seed Production
 Pod Shattering
 Crossability and Gene Flow
 Extent of Cross Pollination with related species
 Extent of Cross Pollination with other plant species
 Alteration in Pollen Viability
 Alteration in Pollen Production
 Effect on Soil Microflora
 Alteration in abundance (CFU/gm)
 Alteration in predominant species in the region
 Effect on Pests, Diseases and Beneficial
Insects
 Change in the susceptibility for Insets and diseases
 Change in predator abundance
 Change in receptibility towards honeybees & any
toxicity to honey bees
All these tests are in conformity
with Biosafety Regulations and
Guidelines
Bio-safety Assessment of GM Crops
TESTS PERFORMED TO ADDRESS ENVIRONMENTAL AND HEALTH CONCERNS

43. o 123 Nobel Laureates supported biotechnology and condemned critics
o International bodies: UN FAO, IFPRI, G20 to eradicate hunger and
malnutrition in 16 years or less through modern tools of plant breeding
o US National Academies of Sciences, Engineering and Medicine,
reported that GM crops are as safe or safer than conventional crops
o National Academy of Agricultural Sciences, India supports GM crops
GM crops are as safe or safer than
conventionally-bred crops
K C Bansal

44. 8/6/2020
Chloroplast genome
engineering
– Inserting a foreign
gene at a targeted
location in the
chloroplast genome
Foreign gene
K C Bansal
Chloroplast Genome Engineering

45. Metabolic engineering of the carotenoid pathway
in transplastomic tomato
Pal Maliga, and Ralph Bock Plant Physiol. 2011;155:1501-1510
©2011 by American Society of Plant Biologists
Wild type
Daffodil
(200 ng provitamin A g−1)
(1 mg g−1 dry weight)
K C Bansal

46. • Ability to edit native crop genes coding for important traits
1. Precision –precise, similar to natural mutations
2. Regulation – science-based – several countries have
classified genome-edited as non-GM
3. Speed – substantially faster
4. Cost – faster-speed and less onerous regulation
translates to significant cost savings
• Genome-edited crops being improved include rice, wheat,
soybean, maize, potato, tomato and other crops
New Breeding Technologies – Genome editing
K C Bansal

47. K C Bansal
Emmanuelle Marie
Charpentier
Jennifer Anne Doudna
Gene editing allows introduction of double-stranded breaks
at a specific sequence in the genome
Targeted Genome Editing
DSBs repaired by
the cell’s
nonhomologous end
joining (NHEJ)
pathway can result
in point mutations
at the target locus.

48. Non-browning Mushroom
PRODUCTS OF GENOME EDITING
K C Bansal
Crispr-Cas9 technology used to
create small deletions in a specific
gene encoding a polyphenol
oxidase (PPO)
PPO catalyses the oxidation of
polyphenols when mushroom are
exposed to air, the first step in the
production of dark melanin
pigments
The final product does not
contain any DNA from a donor
or vector organism, so it is not
covered by current regulations
in USA

49. Leaves of edited tomato
lines infected with
Oidium neolycopersici
showing full resistance
toward this pathogen
when compared to wild-
type leaves.
Langner, T., Kamoun, S., & Belhaj, K. (2018). CRISPR Crops: Plant Genome Editing Toward
Disease Resistance. Annual Review of Phytopathology, 56(1). doi:10.1146/annurev-phyto-
Disease Resistant Tomato Plants
K C Bansal
MLO = Mildew resistance locus
Genome-editing in tomato with sgRNA targeting SlMLO1
Loss-of-function mutations
of the Mildew resistance
locus o (Mlo) genes are
can protect plants from
infection by
powdery mildew fungi.

50. Increased Rice Growth and Yield by Genome Editing
the ABA Receptors
WT Genome-edited Plants
Miao et al 2018, PNASK C Bansal
WT pyl1/2/3/4/5/6 pyl 1/4/6
balance between growth and stress adaptation

51. Santosh Kumar et al. 2020K C Bansal
आई.सी.ए.आर.-भारतीयकृषिअनुसंधानसंस्थान,नईदिल्ली

52. • GM crops with improved traits needed sooner than
later in the national interest
• Potential of New Breeding Techniques such as
genome editing - CRISPR technology is enormous
• The vast potential of chloroplast genome
engineering needs to be realized
• What is needed ? A science-based and efficient
regulation system to ensure benefits of these new
technologies to transform agriculture in the interest
of farmers, consumers and the nation
K C Bansal

53. Developmental paths for the production of plant-made antibodies or vaccines against
SARS-CoV-2 virus. Transient transformation approaches allow high protein yields in the
transformed plants, which are processed to purify the target biopharmaceutical and obtain
injectable vaccines or monoclonal antibodies. Stable genetic transformation technologies
applied in edible plant species can render oral vaccine formulations (e.g. capsules or tables
containing freeze-dried leaves), which can be applied as boosting agents following a
parenteral priming.
Will plant-made
biopharmaceuticals play a role
in the fight against COVID-19?
Sergio Rosales-Mendoza
April, 2020
K C Bansal

54. V. Babu
Anjanasree
Rooz
Abhijit Das
Lakshmi
Sangram
Abhay
Monika
Dipti
Deepa
Preeti
Parul
Divya
Shiv Shanker
Geeta
Amit
Shuchi
SenthilNational Young Scientist Award to Sangram Lenka
Acknowledgements
ICAR - NRCPB,
ICAR - IARI
DST, DBT
DRDO
NPTC-ICAR

55. Thanks to Hon’ble Governor and Chief Minister, Haryana
Haryana Vigyan Ratna awarded to
Prof. K C Bansal, May 10 2017

56. 1920-1958
Franklin’s work played a catalytic role in
unravelling the double-helical structure of DNA
This webinar is dedicated to

57. THANKS
Genes from Petunia to Carnation