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Molecular Plant Breeding 2013, Vol.5, No.5, 24
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CGG GTC TGC AAG C 3' (Forward primer); 5' GTT
GGA GAG CCG AGC ACC ACT G 3' (Reverse
primer). Temperature profile used in the amplification
consisted of pre-incubation at 94°C for 5 min leading
to 35 cycles of melting at 94°C for 1 min, annealing at
65°C for 45 sec and synthesis at 72°C for 45 sec
followed by an extension at 72°C for 10 min. After
amplification, 10 µl of the product was used for
electrophoretic analysis on 1.0% agarose gels and
detected by ethidium bromide staining and
photographed under ultraviolet light.
2.6 Insect bioassay
Bioassay was carried out with the leaves of T
0
plants
and non-transformed (control) plants using the
neonate larvae of Helicoverpa armigera. Leaves
collected from the putative transgenic and
non-transformed (control) plants were placed on a wet
filter paper in sterile petridishes. Five larvae were
released into each petridish and observations were
made every 24 h on the mortality rate of the larvae.
After 3 days, insect mortality per cent was calculated
as follows
2.7 Statistical analysis
The experiment was carried out in a completely
randomized design. Data were analyzed using
standard ANOVA procedures and the difference
between the treatments means were compared using
the Fisher’s Least Significant Difference test (LSD).
Author contributions
There is a paucity of research investigations in the life of
people and plants. I have made a very small and humble
attempt to reduce a fraction of it. I wish to share my
contributions as an author of this research report. The main
goal of gene transfer in cotton is the development of bollworm
resistant varieties especially with
Bt
crystal toxin producing
(cry) genes. As only Coker varieties were found to respond
better for gene transfer, most of the desirable genes are
introduced initially into Coker and backcrossed into other
varieties later. The objective of the study was to transform the
Gossypium hirsutum
cultivar Coker 310 with cry2Ab gene
using the
Agrobacterium
system for enhancing resistance to
Lepidopteran insects. Good results were obtained which have
been interpreted in this research report. I also express the depth
of my sense of gratitude to my advisory committee members
(co-authors 2 and 3) for their indefatigable guidance, explicit
and unaccountable help rendered for this investigation.
Acknowledgement
We gratefully acknowledge ICGEB, New Delhi for providing
us the
Agrobacterium strain
LBA4404 harbouring plasmid pBI121
(containing the cry2Ab gene) for the purpose of sole scientific
research.
References
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petridish
in
placed
larvae of
number
Total
larvae
dead of
Number
(%)
Mortality