Molecular Plant Breeding, 2015, Vol.6, No.23, 1
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6
1
Research Report Open Access
Analysis of Genetically modified BT and cp4EPSPS Cotton cultivars for
transformation efficiency, acclimatization, expression and toxic levels to
insects
Tahir Rehman Samiullah, Arfan Ali. , Saira Azam, Ayesha Latif, Fatima Batool, Idrees Ahmad Nasir, Tayyab Husnain
Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
Corresponding author Email
International Journal of Aquaculture, 2015, Vol.6, No.23 doi
Received: 17 Aug., 2015
Accepted: 10 Oct., 2015
Published: 29 Nov., 2015
Copyright © 2015
Shah Kevalkumar et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article
:
Samiullah T.R., Ali A., Azam S., Latif A., Batool F., Nasir I.A., and Husnain T., 2015, Analysis of Genetically modified BT and cp4EPSPS Cotton cultivars for
transformation efficiency, acclimatization, expression and toxic levels to insects, Molecular Plant Breeding, 6(23): 1-6 (doi
Abstract
The major application of Biotechnology is the transfer of desirable characteristic in the host. To attain resistance against
insects and weeds this application was employed to transfer a double Bt and a glyphsate gene into two cotton varieties, FH-114 and
CIM-598. Three genes Cry1Ac, Cry2A and Glyphosate gene were transferred through the Agrobacterium method using a plant
expression vector with genes under the control of the CaMV35S promoter and NOS terminator sequence. Confirmation of insertion
and expression of these genes in cotton plants was done through PCR and ELISA. Transformation efficiency for FH-114 and
CIM-598 was 1.2% and 0.8% for Cry1Ac, 0.9% and 0.6% for Cry2A and 1.5 and 0.7% for GTG respectively. FH-114 plants
acclimatized better than CIM-598 plants when exposed to sunlight. Cry1Ac, Cry2A and GTG proteins were 1.2,1 and 1.3 ng/µl for
FH-114 which was more than CIM-598 for all three genes. FH-114 plants were able to control better insects and weed damage when
subjected to a cotton leaf bioassay. Taken together FH-114 genetic profile was more suitable for genetic modification to control
insects and weed when compared to CIM-598.
Keywords
BT cotton; Glyphosate; Transformation; Acclimatization; Weeds; Insects
Introduction
Cotton is the most important and in-dispensable part of
human life and is the backbone of the economy and
employment in the world including Pakistan (Muzaffar
et al., 2015). The insect problem is a serious threat for
cotton causing an estimated $ 645 million a year and
yield losses (Awan et al., 2015).
Lepidopteron
insects
are the major problem causing heavy losses with
Heliothus species responsible for an estimated $216
million loss (Gutierrez et al., 2013).
Pectinophora
gossypiella,
which is the pink bollworm, is also a
serious problem on a smaller scale of the cotton
acreage planted (USA) and accounts for an estimated
$71 million in direct damage (Kumar and Kiran, 2011).
Control of these insect pests is a major expense and
therefore a problem for today’s farmers (Pimentel et al.
,
2014). On an average 6-7 insect treatments per season
are applied against these insects (Pedigo et al., 2014).
Chemical insecticide use is however limited due to
their expense, their persistence in the environment and
their escalating rate of application with decreasing
effectiveness (Popp et al., 2013).
Manual hoeing is a reliable method for removing
weeds but this technique is slow, costly and a
time-taking process (Pérez-Ruíz et al., 2014). Total
weed seed numbers in the soil was also found to rise
significantly after shifting from conventional chemical
weed control to non-chemical control (Bond and
Grundy, 2001). Hence, complete non-chemical
methods are not viable or economically sustainable
further, glyphosate (N-phosphonomethyl glycine), a
herbicide widely used as a control for weeds belongs
to herbicide group ‘glycines’. Glyphosate is a
broad-spectrum universal
used to kill annual
broadleaf weeds and grasses. Glyphosate affects the
shikimate metabolic pathway by preventing the
synthesis of 5-enolpyrovyle 3-phosphoshikimate
(EPSPS). It inhibits the synthesis of three aromatic
amino acids including tryptophan, phenylalanine and
trypsin (Yamada et al., 2009).
However biotechnology provides a tool to control
insects and weeds. In particular genetic engineering of
plants utilizing plant genes conferring disease
and weed resistance offers an alternative to conventional
