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August 17-18, 2017 | Toronto, Canada
ANNUAL BIOTECHNOLOGY CONGRESS
Ann Biol Sci, 2017
ISSN: 2348-1927
Genetic transformation of locally developed two cotton varieties [CRSP1 and CRSP2] with Bt and
glyphosate tolerant genes
Shafique Ahmed
University of the Punjab, Pakistan
E
xpression of the transgene with a desirable character in
crop plant is the ultimate goal of transgenic research.
Transformation of two Bt genes namely Cry1Ac and Cry2A
cloned as separate cassette under 35S promoter in pKHG4
plant expression vector was done by using shoot apex cut
method of Agrobacterium. Molecular confirmation of
putative transgenic cotton plants for Cry1Ac, Cry2A and
GT gene was done through PCR and ELISA. Transformation
efficiency of CRSP-1 and CRSP-2 was calculated to be 1.2
and 0.8% for Cry1Ac while 0.9 and 0.6% for Cry2A and 1.5
and 0.7% for GTG respectively. CRSP-1 was found to adopt
natural environment (acclimatized) earlier than CRSP-
2 when exposed to sunlight for one month. Expression
of Cry1Ac, Cry2A and GTG was found to be 1.2, 1 and 1.3
ng/μl respectively for CRSP-1 as compared to CRSP-2
where expression was recorded to be 0.9, 0.5 and 0.9 ng/
μl respectively. FISH analysis of the transgenic CRSP-1 and
CRSP-2 demonstrated the presence of one and two copy
numbers respectively. Similarly, the response of CRSP-1
against Glyphosate @1900 ml/acre was far better with
almost negligible necrotic spot and efficient growth after
spray as compared to CRSP-2 where some plants were found
to have necrosis and negative control where the complete
decay of plant was observed after seven days of spray assay.
Similarly, almost 100% mortality of 2nd instar larvae of
Heliothis armigera was recorded after three days in CRSP-1
as compared to CRSP-2 where insect mortality was found
to be less than 90%. Quantitatively speaking non transgenic
plants were found with 23–90% leaf damage by insect, while
CRSP-1 was with less than 5% and CRSP-2 with 17%. Taken
together CRSP1 was found to have better insect control and
weedicide resistance along with its natural ability of genetic
modification and can be employed by the valuable farmers
for better insect control and simultaneously for better
production.
e:
shafiq@cemb.edu.pkAnn Biol Sci, 2017, 5:3
DOI: 10.21767/2348-1927-C1-003