1. GENETICALLY MODIFIED CROPS
PRESENT STATUS AND FUTURE PROSPECTS
Speaker-Tabassum
I. d. no
MSc. (Genetics& Plant Breeding)

2. WHAT IS A GM CROP?
A crop whose genetic constitution has been altered by the insertion of a gene from another organism by the use of genetic engineering, is called a GM crop or transgenic crop.
The gene which is transferred is called transgene. .

3. GENETIC ENGINEERING OR RECOMBINANT DNA TECHNOLOGY
A recombinant DNA is a vector molecule into which a DNA segment of interest has been inserted.
A vector is a DNA molecule that is capable of independent replication when introduce into a suitable organism.
It is also called gene [DNA] cloning.

4. HISTORY OF GM CROPS
First GM plant was produce in 1983, an antibiotic resistant tobacco plant.
China was the first country to allow commercialization of transgenic plants, introduced a virus resistant tobacco in 1992, which was withdrawn in 1997.
In 1994, US approved flavr savr tomato for sale in US.
In 1995, Bt potato, canola, Bt cotton was approved.
In 1996, the first GM maize producing a Bt Cry protein was approved.
In 2000, the first two GM rice varieties both with herbicide-resistance, called LLRice60 and LLRice62, were approved in the United States.
In 2000, vitamin A enriched golden rice was introduced.
In India, Bt cotton was released in 2002.
In China, insect-resistant GM rice have been approved for food, feed, and cultivation in 2009.
Till 2013,a total of 36 countries have granted regulatory approvals for biotech crops, involving 27 GM crops and 336 GM events.

5. PIONEERS
One of the founders of modern plant biotechnology. in 1997, demonstrated the presence of a fragment of Agrobacterium Ti plasmid DNA in nuclear DNA of crown gall tissue. She got world food prize 2013.
MARY DELL CHILTON
Credited with discovery of Ti plasmid. First person to isolate tumor inducing principle. With Jeff Schell, he discovered gene transfer mechanism between Agrobacterium & plant. Winner of world food prize 2013.
MARC VAN MONTAGU
Worked on interaction between plants & soil bacteria. Executive vice president & chief technology officer at monsanto. In 1983, first to develop GM plant & launch first GM crop in Soybean & Cotton in US. winner of world food prize 2013.
ROB FRALEY

6. HOW TO DEVELOP A GM CROP? 6 Steps-
Production of a DNA fragment to be cloned.
Insertion of the DNA fragment into a suitable vector.[construction of recombinant DNA].
Introduction of the recombinant DNA into a suitable host.
Transformation of host cell and selection of transformed host cells.
Identification of clones having genes of interest and isolation of gene.
Introduction of foreign gene into a plant and regeneration of transformed plant having new foreign gene.

7. GENE CLONING
4 COMPONENTS-
ENZYMES
VECTOR
DNA FRAGMENT
SELECTION

8. CUTTING OF DNA RESTRICTION ENDONUCLEASES
First restriction enzyme was isolated from haemophilus influenzae in1970.
3 TYPES-
[1] Type I endonucleases
[2] Type II endonucleases
[3] Type III endonucleases
Type II is widely used in genetic engineering.

9. TYPE I
TYPE II
TYPE III
Complex nucleases with three different subunits
Simple enzyme with single polypeptide
Having two different subunits
Function simultaneously as an endonuclease & a methylase
Separate endonuclease and methylase activities
One unit for site recognition & modification, other is for nuclease action
Require ATP, Mg ion & S-adenosyl methionine as cofactor
Require Mg ion as cofactor
Require ATP, Mg ion as cofactor
Recognition site is 15 bp in length & cleavage site is 1000 bp away
Exhibit high ATPase activity
Recognize a specific nucleotide sequence (palindromic sequence) & cut DNA at this point only
Asymmetric recognition sites
Cleave DNA at specific(nonpalindromic) sequence
Lack of ATPase activity
Show specificity for the recognition site but not for the cleavage site
More specific in function
Cleave product are uniform
Not specific in function
Cleave product are not uniform

10. CONSTRUCTION OF RECOMBINANT DNA
LIGASE
VECTOR
FEATURES OF A VECTOR-
ORI
MARKER GENE
UNIQUE CLEAVAGE
SUITABLE CONTROL
ELEMENT

11. SOME COMMON VECTORS pBR322 PLASMIDS YEAST PLASMID VECTOR COSMIDS
PUC VECTOR
BACTERIOPHAGE VECTOR
PHAGEMIDS
YAC AND BAC

12. INTRODUCTION OF RECOMBINANT DNA INTO HOST CELL
Once a recombinant DNA is constructed, it is transferred into a host cell for cloning.
Generally E. coli is used as host cell.
Transformation of host cell and selection of host cells transformed by recombinant DNA.
If the plasmid has been chosen carefully it is possible to select transformed bacterial cells.

13. IDENTIFICATION OF CLONE CARRYING DESIRED GENE
Probes.
Colony and plaque hybridization.
Immunological detection.
Southern blot analysis of cloned gene.

15. DESIGNING OF GENE FOR INSERTION
Once a gene has been isolated and cloned it must undergo several modifications before it can be effectively inserted into a plant.

16. GENE TRANSFER IN PLANTS
Movement of specific piece of DNA into the cell.
It is done by transformation process.
TRANSFORMATION- The directed desirable gene transfer from one organism to other & the subsequent stable integration & expression of a foreign gene into the genome of cell, is referred as genetic transformation.
EVENT- Desirable events are those that have transgene inserted in a part of chromosome that allow for the expression of gene without having a destructive influence on other gene of plant.

17. GENE TRANSFER METHODS VECTORLESS OR DIRECT
VECTOR MEDIATED
VECTORLESS OR DIRECT
i. Agrobacterium mediated transformation. ii. Virus mediated gene transfer.
(a) PHYSICAL METHODS. i. Particle bombardment/ microprojectile / biolistics ii. Microinjection and macroinjection. iii. Liposome mediated transformation. iv. Electroporation. v. Ultrasound mediated vi. DNA transfer via pollen (b) CHEMICAL METHODS. i.PEG mediated gene transfer. ii. Calcium phosphate co precipitation. iii. Polycation DMSO technique

19. SOME GENE TRANSFER TOOLS
GENE GUN
MACROINJECTION
MICROINJECTION
LIPOSOME MEDIATED
ULTRASOUND MEDIATED

20. SELECTION OF SUCCESSFULLY TRANSFORMED TISSUES
After the gene insertion process, plant tissues are transferred to a selective medium containing an antibiotic or herbicide, depending upon which selectable marker was used. Only plants expressing the selectable marker gene will survive. These plants will also possess the transgene of interest.

21. REGENERATION OF WHOLE PLANTS
To obtain whole plants from transgenic tissues, they are grown under controlled environmental conditions in a series of media containing nutrients and hormones, a process known as tissue culture. Once whole plants are generated and produce seed, evaluation of the progeny begins.

23. TRANSGENIC PLANT-LAB TO FIELD
Once a gene is successfully enter into a plant genome then it can be transferred into any variety by the back crossing method.
First the breeder obtain an inbred line by self pollinating the transgenic line.
At the same time, transgenic seed and seed from an elite inbred is planted nearby.
When the plants reach the proper stage they are cross pollinated.
F1 seed & an elite inbred line seed planted at the same time.F1 plant is back cross to the elite inbred. The seed is harvested, called BC1 generation.
BC1 seed & elite inbred seed is planted at the same time.[process continues 5-6 generation]
All plants screened. Final product will be transgenic line with 98% elite gene.

24. Self pollination Cross pollination Inbred line Back crossing
All plants screened. Final product will be transgenic line with 98% elite gene.

25. REGULATORY SYSTEM OF TRANSGENICS
IN INDIA
IN USA
FDA EPA APHIS
IBSC(DBT) RCGM(DBT) GEAC(MoEF) MEC(DBT & ICAR) SBCC DLC

26. ROLE OF COMPETENT AUTHORITIES
IBSC- A nodal point for interaction within the institution for biosafety regulation.
RCGM- Bring out guidelines & protocols, issue import permit, review projects.
GEAC- Responsible for approval of R & D, safety assessment, field trials, environmental release and import/ export of GM crops.
MEC- Monitoring of field trials, storage facility & documentation of records.
SBCC- Inspect, investigate, assess the damage if any
DLC-Monitor safety regulation, assess damage & report to SBCC in case of non-compliance at district level.

27. TRANSGENICS V/S CONVENTIONAL BREEDING
Gene exchange limitation.
Gene of interest with flanking sequence transferred.
Technology is quite simple.
History of safety.
More time & labour consuming.
No limitation or barrier.
Only gene of interest transferred.
Intensive technology.
New technique, it will take time to make safe history.
Less time & labour consuming.

29. OBJECTIVES TO DEVELOP A GM CROP
1. BIOTIC STRESS RESISTANCE
Insect & pest resistance
Disease resistance
Fungal, bacterial & viral resistance
2. ABIOTIC STRESS RESISTANCE
Heat, cold resistance or tolerance
Drought resistance or tolerance
Mineral stress
3.QUALITY IMPROVEMENT (Protein, oil, slow-ripening)
4.VALUE ADDITION (Vitamins, micro elements and macro elements)
5.CHEMICAL RESISTANCE Herbicide resistance

30. PRESENT STATUS

31. PRESENT STATUS OF GM CROPS

36. Country
Biotech crops
USA
Maize, Soybean, Cotton, Canola, Sugarbeet, Alfalfa,
Papaya, Squash
Brazil
Soybean, Maize, Cotton
Argentina
India
Cotton
Canada
Canola, Maize, Soybean, Sugarbeet
china
cotton, Papaya, Popular, Tomato, Sweet Peeper

38. Crop
Traits
Modification
% modified in US
% modified in world
Alfalfa
Tolerance of glyphosate or glufosinate
Genes added
In approved for sale
Apples
Delayed browning
Genes for reduced polyphenol oxidase (PPO) production
2015 approved for sale
Canola/ Rapeseed
Tolerance of glyphosate or glufosinate High laurate ,Oleic acid
87% (2005)
21%
Corn
Tolerance of herbicides glyphosate glufosinate, and 2,4-D. Insect resistance. Added enzyme, alpha amylase
Genes , some from Bt
Herbicide-resistant: 85% Bt: 76% (2013)
26%
Cotton (seed oil)
Insect resistance
Gene, some from Bt, added
Herbicide-resistant:82% Bt:75% Stacked: 71%(2013)
49%
Papaya
Resistance to the papaya ringspot virus.
Gene added
80%
Potato (food)
Resistance to Colorado beetle Resistance to potato leaf roll virus and Potato virus Y Reduced acrylamide when fried and reduced bruising
Bt cry3Ac, coat protein 0%

39. Potato (starch)
Antibiotic resistance gene,
Gene from bacteria 0%
Rice
Enriched with beta-carotene (vitamin A)
Genes from maize & a soil microorganism.
Forecast on market in 2015 or 2016
In china
Soybeans
Tolerance of glyphosate or glufosinate ,Reduced saturated fatty acids
Herbicide resistant gene , Bt crystal proteins
2013: 93%
77%
Squash (Zucchini/Courgette)
Resistance to watermelon, cucumber & zucchini/courgette yellow mosaic viruses
Viral coat protein genes
13% (2005)
Sugar beet
Tolerance of glyphosate,
Genes added
95% 9%
Sugarcane
Pesticide tolerance High sucrose
Sweet peppers
Resistance to cucumber mosaic virus
grown in China
Tomatoes
Suppression of the enzyme polygalacturonase (PG)
Antisense gene of the gene responsible for PG enzyme production
Taken off the market .
Small quantities grown in China
Wheat
Tolerance of glyphosate
unknown

41. GLOBAL IMPACT OF GM CROPS
FARM INCOME- GM crops have a positive impact on farm income due to enhance productivity & efficiency gains.
In 2012, direct global farm income benefit was $ 18.8 billion.
PESTICIDE USE- ( ) Pesticide input in field reduced by 8.8% or over 503 million kg.
On average, GM technology adoption has reduced chemical pesticide use by 37%, increased crop yields by 22%, and increased farmer profits by 68%. Yield gains and pesticide reductions are larger for insect-resistant crops. Yield and profit gains are higher in developing countries than in developed countries. (2014)

42. (2012) increase in farm income (1996-2012 )increase in farm income
Global farm income benefits from growing GM crops, (US$ million)
Changes in the use of herbicides and insecticides in GM crops globally,
GM Trait
(2012) increase in farm income
( )increase in farm income
HT Soybean
4,797.9
37,008.6
HT maize
1,197.9
5,414.7
HT cotton
147.2
1,371.6
HT canola
481.0
3,664.4
IR maize
6,727.8
32,317.2
IR cotton
5,331.3
36,317.2
Others
86.3
496.7
Totals
18,769.4
116,590.4
GM Trait
Change in volume of AI used (million kg)
Change in field EIQ impact (million field EIQ/ha units)
% change in AI use on GM crops
HT Soybean
-4.7
-6,654
-0.2
HT maize
-203.2
-6,025
-9.8
HT canola
-15.0
-509
-16.7
HT cotton
-18.3
-460
-6.6
IR maize
-57.6
-2,215
-47.9
IR cotton
-205.4
-9,256
-25.6
HT sugar beet
+1.3 -1
+29.3
Totals
-503.1
-25,121
-8.8

43. EXPERIENCES OF DEVELOPED COUNTRIES
US- Enhanced farm income from biotech crops by 53.1 billion $ from
An estimated cost savings by farmers planting HT soybean was 71.3 $/ha in 2012.
Annual total farm income benefit from HT soybean increase from 5 million $ (1996) to 6.07 billion $ in 2012.
In US, by 2014, 94% of the planted area of soybeans, 96% of cotton and 93% of corn were genetically modified varieties.
CANADA- Net increase in farm income from GM crops by 4.9 billion $ from , out of which 12.2 million $ by HT maize farmers in 2012.
HT canola boosted total canola production by 11% in Adopters of biotech canola earned 446 million $ in 2012.

44. EXPERIENCES OF DEVELOPING COUNTRIES
INDIA- From , 1097 Bt cotton hybrid approved for planting.
96% of the countries cotton area is now covered by Bt hybrid (11.57 mha)
Cotton output now from 13 million bales (2002) to 40 million bales in 2014.
In 2013, India rank first in Bt cotton production, yield increase by 31%, reduced insecticide spraying by 39%, increase in profit by 88%.(US $ 250/ha)
Cotton yield increase from 300 kg/ha to 500 kg/ha.
In , India’s largest cotton producing state was Gujarat, key supplier of cotton to export market.
CHINA- In China, insect-resistant GM rice have been approved for food, feed, and cultivation in 2009.
Rice is most important crop in china, accounting 28% of the world total production.
GM rice yield is 6 to 9% higher as compared to conventional varieties, required 80% less pesticide input.
54 % OF TOTAL GLOBAL GM CROP AREA IS NOW BEING GROWN IN DEVELOPING COUNTRIES.

45. NO. OF EVENT OF SOME GM CROPS
EVENTS
COTTON 54
APPLE 2
MAIZE
138
CANOLA (B. napus) 32
SOYBEAN 30
FLAX 1
RICE 7
POLISH CANOLA (B. rapa) 4
SUGARBEET 3
ROSE
POTATO 42
SQUASH
PAPAYA
SWEET PEPPER
MELON
POPLAR
EGGPLANT
PLUM
BEAN
PETUNIA
TOMATO 11
ALFALFA
WHEAT

46. STEARIDONIC ACID (SDA) OMEGA-3 SOYBEANS
Currently in Phase IV of Monsanto’s R&D Pipeline
Science has shown consuming more long-chain fatty acids, such as omega-3s, can help support a healthy heart
These soybeans have been enriched with steridonic acid (SDA), which in the body converts to heart-healthy1 eicosapentaenoic acid (EPA), one of three omega-3 fatty acids used by the body
A genetically modified cassava under development offers lower Cyanogen glucosides and enhanced protein and other nutrients (called BioCassava)

47. PRESENT STATUS IN INDIA

48. GM CROP EVENTS APPROVED IN INDIA
Event Name and Code
Cotton : 6 Events
Trade Name
Name: BNLA Code: not available
not available
Name: Event Code: not available
JK 1
Name: GFM Cry1A Code: GTL-GFM311-7
Name: MLS Code: not available
Name: MON Code: MON
Bollgard II™ Cotton
Name: MON Code: MON-ØØ531-6
Bollgard™ Cotton, Ingard™

49. SOYBEAN- 5 EVENT EVENT NAME AND CODE TRADE NAME
Name: A Code: ACS-GMØØ5-3
Liberty Link™ soybean
Name: A Code: ACS-GMØØ6-4
Name: CV127 Code: BPS-CV127-9
Cultivance
Name: MON87701 x MON89788 Code: MON-877Ø1-2 x MON
Intacta™ Roundup Ready™ 2 Pro
Name: MON89788 Code: MON
Genuity® Roundup Ready 2 Yield™

50. WORK & PROJECTS ON GM IN INDIA
Project Rise: Helping Farming Communities in India Produce More through Knowledge Sharing.
Bollgard Cotton: Improving Cotton and Lives Around the World.
Bt chickpea developed by Assam Agriculture University, Bt brinjal by IARI.
Some GM crops are currently in the pipeline for approval, such as Bt brinjal, Bt rice, Bt maize, GM tomato, etc.
Maharashtra govt. has given NOC for field trials to 5 GM crops. Brinjal, maize, rice, chickpea, cotton.
Punjab, Haryana, Delhi, AP also given NOC for field trials.
Since june 2011, it has been made mandatory for companies to obtain NOC from state govt. where they want to conduct field trials.

51. 2015:NOC for Bt brinjal and 4 other crops trials.

52. ADVANTAGES
Required lesser pesticide, insecticide, so environment friendly.
Disease resistant & insect resistant
Decrease cost of production & enhance farming.
Better biotic and abiotic stress tolerence.
Better shelf life.
Nitrogen Use Efficiency
Less labor requirement.
Environment (bioremediation)
Industry (biomass/fuel, lignin/paper, plastics)
Medical & veterinary application (vaccines/antigens/antibiotics, enzymes)
Have a desirable better taste, quality, yield, more nutritious.
Potentially non allergenic & conserve less natural reserve( water & energy)

54. PROBLEMS Cross pollination contaminates regular crops.
Harm to other organisms(Kill Bees and Butterflies)
Harmful for biodiversity.
Development of ‘super weeds’ & ‘super pests’.
Safety & ethical issues.
Health problems.
Political, social & consumer issues.
Less awareness.
Farmers can not harvest seeds.
It is illegal to accidentally grow a GM plant.

55. FUTURE PROSPECTS

56. [1]MAXIMUM FOOD PRODUCTION
FUTURE NEED(MILLION TONNES)
COMMODITY
2000
2010
2020
FOOD GRAINS
208.0
266.0
343.0
EDIBLE OIL
6.3
9.4
13.0
VEGETABLES
80.0
117.2
168.0
FRUITS
22.2
42.9
81.0
MILK
84.0
153.1
271.0
MEAT, FISH & EGGS
6.2
12.7
27.0
SUGAR
12.8
17.3
22.0

57. [2] INCREASE TASTE, QUALITY, NUTRITION
Rice is the main source of food for Half of the world population. Golden rice is enriched in Vitamin A so reduce the level of vitamin A deficiencies.
The World health organization has stated that iron deficiency affects 30% of the world's population, so IRRI is tried to develop iron rich & zinc rich rice varieties.
Quality increased in canola, increased flavor & life in tomato.

58. [3] RESOLVE THE ISSUES Redarding- Ethical issues
Safety & health problems
Political, social & consumers issues (Labeling issue)
Increase public awareness regarding GMOs
Research regarding to the development of super weeds & super insects & gene flow(cross pollination)
Farmer’s concern

59. ENVIRONMENTAL CONCERN
LOSS OF BIODIVERSITY AND THE CHANGE IN SOIL MICROBIAL CONDITIONS.
Biotech crops can contribute to a “sustainable intensification” strategy favoured by many science academies worldwide, which allows productivity to be increased only on the current million hectares of global crop land, thereby saving forests and biodiversity.
FARMER CONCERN. The GM crop seeds are expensive as compare to traditional seeds. We need to extend GM technology to more crops & also encourage PPP so that our farmers can get benefit.

60. CONCLUSION
As such, we should not promote widespread use of GMOs until more research has been done on long term health effects, GMO seeds are available outside of corporate control, the biological effects of gene insertion are better understood, and research confirms that the presence of GMOs will not harm the native species in an ecosystem.

61. THANK YOU