Transgenics Showing Herbicide Resistance

Transgenic
showing Herbicide
Resistance
ANKIT
INTRODUCTION
• Transgenic crops are plants used in agriculture, the
DNA of which has been modified using genetic
engineering methods. In most cases, the aim is to
introduce a new trait to the plant which does not
occur naturally in the species.
• Herbicides, also commonly known as weed killers, are

chemical substances used to control unwanted
plants(Weeds).
• Weeds not only compete with crops for water,

nutrients, sunlight, and space but also harbor insects
and diseases; clog irrigation and drainage systems; Weeds controlled with herbicide
undermine crop quality; and deposit weed seeds
into crop harvests. If left uncontrolled, weeds can
reduce crop yields significantly.
Weed control practices
The tandem technique of soil-tilling and herbicide
application is an example of how farmers control
weeds in their farms.
• Unfortunately, tillage leaves valuable topsoil exposed to wind and
water erosion, a serious long-term consequence for the
environment. For this reason, more and more farmers prefer
reduced or no-till methods of farming.
• Similarly, many have argued that the heavy use of herbicides has
led to groundwater contaminations
HERBICIDES AND
TRANSGENIC SHOWING
HERBICIDE RESISTANCE
HISTORY - HERBICIDE
• When 2,4-D was commercially
released in 1946, it triggered a
worldwide revolution in agricultural
output and became the first
successful selective herbicide.
• It allowed for greatly enhanced

weed control in wheat, maize
(corn), rice, and similar cereal grass
2,4-D, the first chemical herbicide, was discovered crops, because it kills dicots
during the Second World War. (broadleaf plants), but not most
monocots (grasses). The low cost of
2,4-D has led to continued usage
today, and it remains one of the
most commonly used herbicides in
the world
The triazine family of herbicides, which includes atrazine, were
introduced in the 1950s; they have the current distinction of being the
herbicide family of greatest concern regarding groundwater
contamination.
Glyphosate (Roundup) was introduced in 1974 for nonselective

weed control. Following the development of glyphosate-
resistant crop plants, it is now used very extensively for selective
weed control in growing crops.
Paraquat , Picloram, Imazamox, Dicamba, Gluphosinate etc. are

some other herbicides used commercially.
Glyphosate
• A complementary DNA (cDNA) done encoding EPSP synthase was isolated
from a complementary DNA library of a glyphosate-tolerant Petunia hybrid cell
line (MP4-G) that overproduces the enzyme.
• This cell line was shown to overproduce EPSP synthase messenger RNA as a
result of a 20-fold amplification of the gene.
• A chimeric EPSP synthase gene was constructed With the use of the cauliflower
mosaic virus 35S promoter to attain high level expression of EPSP synthase and
introduced into petunia cells.
• Transformed petunia cells as well as regenpri4ted transgenic plants were

tolerant to glyphosate.
1986, Shah et al.

• CaMV 35S promoter
directs constitutive high
level expression of
foreign genes in plants,
they hypothesize that
glyphosate tolerance in
the transformed petunia
plants results from the
overproduction of EPSP
synthase.
• The results of subcellular

fractionation studies in
pea and petunia have
shown that most of the
EPSP synthase enzyme
activity is localized in
chloroplasts which
represent a major site of
aromatic amino acid
biosynthesis.
1986, Shah et al.

• Glyphosate resistance is also achieved by the
introduction of either Agrobacterium gene CP4 that
codes for Glyphosate-insensitive version of plant
enzyme, EPSP-synthase,
• Or, gox gene from Achromobacter, which codes for

glyphosate oxidoreductase in the breakdown of
glyphosate.
• The herbicide bromoxynil (3,5-dibromo-4-
hydroxybenzonitrile) is a photosynthetic
(photosystem II) inhibitor in plants.
• A gene, bxn, encoding a specific nitrilase

that converts bromoxynil to its primary
metabolite 3,5-dibromo-4-hydroxybenzoic
acid, was doned from the natural soil
bacterium Klebsiella ozaenae.
1988, STALKER et al.

• For expression in plants, the bxn gene was placed under control of a light-regulated
tissue-specific promoter, the ribulose bisphosphate carboxylase small subunit.
• Transfer of this chimeric gene and expression of a bromoxynil-specific nitrilase in

leaves of transgenic tobacco plants conferred resistance to high levels of a
commercial formulation of bromoxynil.
• The results presented indicate a successfull approach to obtain herbicide resistance

by introducing a novel catabolic detoxification gene in plants.

The trait is dominant and heritable, and the enzyme levels obtained in
the primary transformants relate to bromoxynil resistance levels observed
in the succeeding generation.
2,4-D, the first chemical herbicide
• Transgenic lines resistant to the herbicide 2,4- dichlorophenoxyacetic

acid (2,4-D) were developed in mustard (Brassica juncea), a major
oilseed crop grown in more than six million hectares of land in North
India.
• The developed construct contained the tfdA gene, encoding the

enzyme 2,4-D monooxygenase, cloned downstream to the 35S promoter
along with a leader sequence from RNA4 of alfalfa mosaic virus (AMV
leader sequence), for improved expression of the trans- gene in plant
cells
2002, Pental et al.

• Southern analysis of T0 transgenics confirmed six out of 24 transgenics to be single copy
events, from both the flanks of T-DNA.
• Selfed progeny derived from single copy tfdA lines germinated normally and rooted in
medium containing 2,4-D at concentrations as high as 2.5 mg l–1 compared to the
wild-type seedlings which did not root even at a concentration of 0.5 mg l–1.
• The four transgenic lines tested in the study were resistant to herbicide concentration of
500 mg l–1.
• The available transgenic lines can be used for testing the potential of 2,4-D in weed
control including the control of parasitic weeds (Orobanche spp) of mustard and for
low-till cultivation of mustard.
2002, Pental et al.

a, Schematic representation of T-DNA region of the tfdA construct
showing the location of EcoRI site used for genomic DNA digestion
along with the probe fragments used for the Southern analysis
(hatched boxes).
b, Southern blot of T0 transgenics showing the T-

DNA integration pattern for left T-DNA border
when probed with nosPr–nptII–pA cassette.
• Transgenics in lanes 2, 4 and 7 represent single

copy events. Transgenics in lanes 1 and 8
represent the same transformation event.
Lane 10 is untransformed negative control
2002, Pental et al.

Glyphosate resistance crop represents
more than 80% of the 120 million ha of
Transgenic crops grown annually
worldwide. (James, 2008)
As of 2009, Five different types of GR

crop grown in united states
2015, Powles et al.
Farmers have rapidly adopted GR
crops technology because of three
main reasons: cost savings, better
weed management, and simplicity
of use.
2015, Powles et al.
Figure 3. Comparison of US tillage practices in glyphosate resistant (GR) and non
GR soybean (left) and cotton (right) from 2002 through 2006 as a percentage
of hectares planted
2015, Powles et al.

Glyphosate resistance weeds
Evolution of Glyphosate
Resistance weeds :
• Strong and Persistent Glyphosate selection pressure
• Reduction in Herbicide Diversity
• Adoption of No-Tillage(Reduced weed control

Diversity)
2015, Powles et al.

• The first evolved GR weed reported in a GR crop(2001) was Conzya Canadensis US
soyabeans fields(Van Gessel,2001). In the few Years since this first report this weed
now infest at least 2 million ha of GR crops in United states.
2015, Powles et al.

Misuse and misapplication
Agent Orange was a herbicide blend used by the British military
during the Malayan Emergency and the U.S. military during the
Vietnam War between January 1965 and April 1970 as a
defoliant.
It was a 50/50 mixture of the n-butyl esters of 2,4,5-T and 2,4-D.
Because of TCDD contamination in the 2,4,5-T component, it led

to 3 million Vietnamese people suffering health problems, one
million birth defects.
After use of Agent Orange, 24% of the area of Vietnam was

defoliated.
Conclusion:
• Glyphosate resistance crop represents more than 80% of the
120 million ha of Transgenic crops grown annually worldwide.
• Over the past three Decades, Herbicide Resistance Crops

resulted into a promising weed control management systems.
• Farmers are Definetly getting profit after adopting the

Transgenics showing Herbicide Resistance.
• New Technology are required to maintain the weed control as

it is today.
Refrences
• DILIP M. SHAH et al. (1986) Engineering Herbicide Tolerance in Transgenic Plant.
SCIENCE, VOL. 233.
• DAVID M. STALKER et al. (1988) Herbicide Resistance in Transgenic Plants Expressing a

Bacterial Detoxification Gene. REPORTS
• Deepak Pental et al. (2002) Development of 2,4-D-resistant transgenics in Indian

oilseed mustard (Brassica juncea). RESEARCH COMMUNICATIONS.
• Dhirendra Fartyal et al. (2018) Developing dual herbicide tolerant transgenic rice
plants for sustainable weed management. SCIENtIFIC RepoRtS | (2018) 8:11598 |
DOI:10.1038/s41598-018-29554-9
• www.Wikipedia.com
• Stephen O. Duke, Powles (2015) Glyphosate-Resistance Crops and Weeds: Now and
in the future. AgBioForum
Thank you

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Transgenic

• Herbicides, also commonly known as weed killers, are

• Weeds not only compete with crops for water,

• It allowed for greatly enhanced

Glyphosate (Roundup) was introduced in 1974 for nonselective

Paraquat , Picloram, Imazamox, Dicamba, Gluphosinate etc. are

• Transformed petunia cells as well as regenpri4ted transgenic plants were

1986, Shah et al.

• The results of subcellular

1986, Shah et al.

• Or, gox gene from Achromobacter, which codes for

• A gene, bxn, encoding a specific nitrilase

1988, STALKER et al.

• Transfer of this chimeric gene and expression of a bromoxynil-specific nitrilase in

• The results presented indicate a successfull approach to obtain herbicide resistance

1988, STALKER et al.

• Transgenic lines resistant to the herbicide 2,4- dichlorophenoxyacetic

• The developed construct contained the tfdA gene, encoding the

2002, Pental et al.

2002, Pental et al.

b, Southern blot of T0 transgenics showing the T-

• Transgenics in lanes 2, 4 and 7 represent single

2002, Pental et al.

As of 2009, Five different types of GR

2015, Powles et al.

• Strong and Persistent Glyphosate selection pressure

• Reduction in Herbicide Diversity

• Adoption of No-Tillage(Reduced weed control

2015, Powles et al.

2015, Powles et al.

It was a 50/50 mixture of the n-butyl esters of 2,4,5-T and 2,4-D.

Because of TCDD contamination in the 2,4,5-T component, it led

After use of Agent Orange, 24% of the area of Vietnam was

• Over the past three Decades, Herbicide Resistance Crops

• Farmers are Definetly getting profit after adopting the

• New Technology are required to maintain the weed control as

• DAVID M. STALKER et al. (1988) Herbicide Resistance in Transgenic Plants Expressing a

• Deepak Pental et al. (2002) Development of 2,4-D-resistant transgenics in Indian

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