THIS SEMINAR HIGHLIGHTS THE ROLE OF GENETIC ENGINEERING AND BIOTECHNOLOGY FOR ACHIEVING THE SUSTAINABLE GOAL FOR 21ST CENTURY AGRICULTURE

1. TARA SINGH RAWAT
Ph.D. Scholar,
PMBB, IGKV, Raipur (C.G.)
SEMINAR
ON

2. CONTENT:
 INTRODUCTION
 SUSTAINABLE DEVELOPMENT
 SOLVING THE PROBLEM OF NITROGEN
 ENGINEERING PHOTOSYNTHESIS
 WHAT CRISPR CAN DO !
 EXPLORING PLANT MICROBIOME
POTENTIAL
 CURRENT SCENARIO OF BIOTECH CROP
AND ITS IMPACT
 CONCLUSION 3/23/2019

3. INTRODUCTION
 DETEORATING HUMAN AND ECOSYTEM HEALTH is a characteristics of an
unsustainable development.
 Populations of mammals, birds, fish, reptiles, and amphibians have, on
average, declined in size by 60 percent in just over 40 years (Living
Planet Report, 2018).
 The biggest drivers of current biodiversity loss are overexploitation
and agriculture, both linked to continually increasing human
consumption.
 The WRI report on Creating a Sustainable Food Future estimates that
we need about 70% more food in 2050 than we have today in order to
feed 9.6 billion world population.
 Given the interconnectivity between the health of nature, the well-
being of people and the future of our planet, BIOTECHNOLOGICAL
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4. THREE PILLARS OF SUSTAINABLE
DEVELOPMENT
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5. BIOTECHNOLOGICAL SOLUTION TO
NITROGEN PROBLEM
 Cereal crop productivity depends heavily on N
fertilization.
 The industrial production and use of nitrogenous
fertilizer involves significant environmental and
economic costs..
 Exploring the plant inherent capability to use N
efficiently and exploiting biological nitrogen fixation
will reduce the use of synthetic fertilizer.
 Strategies to generate N2-fixing cereals are being
pursued but barriers of nitrogenase O2 sensitivity and
biosynthetic complexity must be overcome.
Nox
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6. Global nutrients (N+P2O5+K2O) consumption
World fertilizer trend and outlook to 2018, FAO
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7. Regional and subregional share of world
increase/decrease in nitrogen
fertilizer consumption, 2014-2018
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8. N fertilizer use affects cereal crop production
and NOx gas emissions
Curatti et al., 2014,
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9. Engineering crop nutrient efficiency
 Globally 122 mMTons of nitrogenous fertilizer produced every year
costing US$41Billions.
 Improving crop nutrient efficiency can improve agricultural
sustainability by increasing yield while decreasing input costs and
harmful environmental effects.
 Application of excess fertilizer can generate and maintain high yields
but also increases input costs and contributes to environmental
damage through eutrophication, soil acidification and air pollution.
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10. Aspects of nutrient efficiency in crops
 Improving nutrient acquisition efficiency
through root modifications, such as
optimizing root architecture (i.e. topsoil
foraging vs. steep, cheap, deep
architecture), enhancing nutrient
transporters, secreting root exudates,
associating with arbuscular mycorrhizal
fungi (AMF) and/or rhizobial bacteria
(forming nodules).
 Improving nutrient utilization efficiency
via enhancing transport for soluble
nutrient translocation and remobilization,
liberating stored nutrients for
remobilization, and improving metabolic
efficiency during natural senescence
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11. Trade off between dwarf trait vs. nutrient use
efficiency
Wang et al, 2018
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13. Candidate
genes for
improving
nutrient
acquisition
efficiency in
crop plants
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14. Candidate
genes for
improving
nutrient
utilization
efficiency
in crop
plants
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15. Liu et al., 2018
Allen Good, 2018
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16. Nitrogen fixation activity in diazotrophs and
engineered strains
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17. Expression of 15 Nif proteins in the mitochondrial matrix of N. benthamiana.
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18. Achieving Sustainability in Crop
Productivity Through Engineering
Photosynthesis
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19. INGINEERING PHOTOSYNTHESIS
 Photosynthesis is a major driver of life on the Earth
 Naturally occurring photosynthesis is a highly inefficient process in terms of
conversion of light energy into biomass
 Processes limiting photosynthetic efficiency have been understood to an extent
that allows a step change in our ability to manipulate light energy assimilation
into carbon gain
 Therefore, future increases in the grain yield potential of our major crops may
depend largely on increasing the efficiency of photosynthesis
 Here is a insights into the nature of current limitations on photosynthesis and
potential targets that can be used for crop improvement
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20. Photosynthesis: An overview and major
limitations
Balint Jakli et al., 20163/23/2019

21. Photosynthesis: An overview and major
limitations
Balint Jakli et al., 20163/23/2019

22. Engineering photosynthesis: Different
Strategies
A number of routes are being studied to improve photosynthesis in crop plants. Douglas et al., 2017
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23. Optimizing Response To Changes In Light-use
Efficiency
 Major losses of energy conversion during plant biomass formation occur
during light absorption and the photochemical reactions.
 Maximum efficiency of energy conversion during plant biomass formation is
estimated to be 3–5% (Mark Stitt, 2013)
 NON-PHOTOCHEMICAL QUENCHING (NPQ)- NPQ is the process by which
plants dissipate excess light as heat when they receive more than they are
capable of using.
 This protective dissipation continues for many minutes and reduces
photosynthesis cost up to 20% of their potential yield.
 Dramatic increases in tobacco growth rate in the field were obtained
recently by improving the rate of relaxation of photoprotection (Johannes et
al., 2016)
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24. (Johannes et al., 2016)
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25. 3/23/2019

26. Engineering Rubisco catalysis
 Slow CO2-fixation rate and poor specificity for CO2 over O2 have made it a key engineering target to
improve photosynthesis in crop plants
 Despite having good understanding of the reaction mechanism, engineering efforts have yet to
produce the holy grail of a “super Rubisco”, as efforts to modify one aspect of its catalytic
biochemistry typically come at a cost to another.
Tansley et al., 2016
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27. Transformation of Tobacco with Rubisco from
cyanobacterium Synechococcus elongatus (Se7942)
 Knocking out the native
tobacco gene encoding
the large subunit of
rubisco by inserting the
large and small subunit
genes of the se7942
enzyme, in combination
with either the
corresponding se7942
assembly chaperone,
rbcx, or an internal
carboxysomal protein,
ccmm35
Lin et al., 2014
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30. RATE LIMITING ENZYME
Calvin-Benson cycle optimization
 Under many conditions, reductions in the activities of enzymes catalyzing highly regulated,
effectively irreversible reactions, Glyceraldehyde-3-phosphate dehydrogenase (GAPDH),
Fru bisphosphatase (FBPase), and Phosphoribulokinase (PRK), had little impact on carbon
assimilation.
 In contrast a small reductions in the enzyme Sedoheptulose-1,7-bisphosphatase (SBPase)
resulted in a decrease in CO2 fixation and growth. (Raines, 2003; Stitt et al., 2010).
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32. Introducing the C4 cycle in C3 crops
 The C4 photosynthetic pathway accounts for 25% of primary productivity on
the planet despite being used by only 3% of species.
 C4 plants are higher yielding than C3 plants, efforts are underway to introduce
the C4 pathway into the C3 crop rice.
 Induced chloroplast and mitochondrial development in rice vascular sheath
cells have been achieved through constitutive expression of maize GOLDEN2-
LIKE genes.
 This suite of traits reflects that seen in ‘‘proto-Kranz’’ species, and, as such, a
key step toward engineering C4 rice has been achieved (Peng et al., 2017)3/23/2019

33. Chloroplasts in Both Bundle Sheath and Mestome Sheath Cells of
rice Lines that Constitutively Express ZmG2 Accumulate
Photosynthetic Enzymes
Transmission electron micrographs of minor
veins in leaf 7 of wild-type (WT) (A) and
ZmUbipro:ZmG2 transgenic (B).
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34. Chloroplast Development in Vascular Sheath Cells Is Accompanied by
Increased Mitochondrial Volume and Plasmodesmatal Frequency
Arrows, mitochondria;
arrowheads, plasmodesmata; BS, bundle sheath;
M, mesophyll; MS, mestome sheath; VB, vascular
bundle; C, chloroplast. Scale bars, 500 nm.
mestome sheath bundle sheath
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35. Constitutive Expression of ZmG2 or ZmGLK1
Delays Flowering but Has No Impact on Yield
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36. Studies performed to improve the photosynthetic rate of C3 plants and know the
role of different genes in improving photosynthesis via transgenic approach
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37. Role of CRISPR Technology
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38. DEALING WITH PESTICIDE ABUSE
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39. Gene Drive: stimulating biased inheritance of a desired gene
MCR can be used to propagate a genetic modification rapidly through generations. It might
be used to eradicate a population of disease-carrying mosquitoes and insect pest.
3/23/2019
Heidi Ledford, 2015

40. 3/23/2019
Gene Drive: stimulating biased inheritance of a desired gene
MCR can be used to propagate a genetic modification rapidly through generations. It might
be used to eradicate a population of disease-carrying mosquitoes and insect pest.
3/23/2019
Heidi Ledford, 2015
CRISPR-Cas9 gene drive approach to invasive species control would be
based on a laboratory strain with a deleterious trait (e.g., distorted sex
ratio, reduced fertility, chemical sensitivity) being mass-reared and
released into the field in sufficient numbers for the engineered mutation to
spread and control the target population within a desired time frame.

41. Dynamics of the spread of the dsxFCRISPRh allele and effect
on population reproductive capacity.
Targeting the female-specific isoform of doublesex. Schematic
representation of the male- and female-specific dsx transcripts and the
gRNA sequence used to target the gene
Kyrou et al., 2018;
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Disease symptoms of wild-type (WT) and tamlo-aabbdd mutant plants 7 d after inoculation in planta
Micrographs of microcolony formation of Bgt on the surfaces of leaves

43. 3/23/2019
Transgenic N. benthamiana and
Arabidopsis plants develop resistance to
beet severe curly top virus (BSCTV)
Cas9 gene
gRNA gene targeting viral DNA

44. Mutations In A Subfamily Of Abscisic Acid Receptor
Genes Promote Rice Growth And Productivity
Miao et al., 2018
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45. 3/23/2019
Mutations In A Subfamily Of Abscisic Acid Receptor
Genes Promote Rice Growth And Productivity
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46. Blocking OsAAP3 improves grain yield by increasing tiller number in rice
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47. Crop improvement by inbreeding often results in fitness penalties and loss of genetic diversity.
Desirable traits can be introduced into wild species by using multiplex CRISPR–Cas9 editing
of coding sequences, cisregulatory regions or upstream open reading frames of genes
associated with morphology, flower and fruit production, and ascorbic acid synthesis
WHAT ABOUT IMPROVING WILD SPECIES….
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RESISTANT TO BROWNING- The effect is
achieved by targeting the family of genes that
encodes polyphenol oxidase (PPO), an enzyme
that causes browning. By deleting just a handful
of base pairs in the mushroom’s genome,
knocking-out one of six PPO genes — reducing
the enzyme’s activity by 30%.
EMILY WALTZ, 2016

54. EXPLORING PLANT MICROBIOME
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55. Tiny Microbes, Big Yields: enhancing food
crop production with biological solutions
Sustainable increase in farm productivity by harnessing microbial technologies is critical for delivery of
multiple Sustainable Development Goals (SDGs)
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56. Biotechnological Advances for Restoring
Degraded Land for Sustainable Development
 Rapid industrialization, urbanization, and agricultural activities during the
last few decades have resulted in the wide-scale degradation of global land
resources.
 12.2 billion hectares of global land have been severely degraded due to
pollution and other reasons.
 Biotechnological advancements (e.g., genomics, metabolomics, and proteo-
mics) can be exploited for restoring degraded lands for multipurpose envir-
onmental benefits.
 Customized (site-specific, pollutant-specific, and cost-effective) packages are
essential for successful restoration programs.
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57. Schematic Representation of Various Biotechnological
Advancements for Biomonitoring and Bioremediation of
Polluted Soils
Identifying Potential Strains for Effective Microbial Formulations
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58. Microbes based Remediation and Restoration of
Contaminated and Degraded Lands
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59. 3/23/2019
MICROBIOME BASED FORMULATION BY INDIGO
FARMA TECH GIVES PROMISSING RESULTS ON
COTTON, WHEAT CROP PRODUCTIVITY
 10-15% INCREASE IN
CROP PRODUCTIVITY
OVER UNTREATED CROP
under water stress3/23/2019

60. MICROBIOME BASED FORMULATION BY INDIGO
FARMA TECH GIVES PROMISSING RESULTS ON
COTTON, WHEAT CROP PRODUCTIVITY
 10-15% INCREASE IN
CROP PRODUCTIVITY
OVER UNTREATED CROP
under water stress
Leveraging the plant
microbiome to improve
crop yields is more and
more promising, but any
upstarts will have a hard
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61. GLOBAL SCENARIO OF BIOTECH CROP
AND ITS IMPACT
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66. Monsanto’s Transgenic Drought Tolerant Maize
Agricultural biotechnology giant Monsanto has received the green light from the US Department
of Agriculture to sell its transgenic drought-tolerant maize (corn)
MON 87460.
3/23/2019
Hybrid seed sold under this
trademark combine a novel
transgenic trait (based on the
bacterial cspB gene) with the best
of Monsanto's conventional
breeding programme
Drought Gard™ maize
was the first commercially available
transgenic (GM) drought tolerant crop
released in 2013

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69. CONCLUSION
Creating sustainable bio-
economies for the 21st century
relies on optimizing the use of
biological resources to improve
agricultural productivity and
create new bio-based products
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70. This is not Life…
It is LIFE…
National capital choked in thick Smock
Morning scene of a remote village in Uttarakhand
3/23/2019