Molecular Plant Breeding, 2015, Vol.6, No.23, 1
-6
4
Figure 3 Glyphosate gene’s (350 bp) amplification with gene specific primers
M: 1 kb plus DNA ladder; 1, 7: negative control; 2-6: positive GTG transgenic FH-114 plants; 8-12: CRSP -2 GTG positive plants.
Figure 4 Graphical representation of quantification of Cry1Ac,
Cry2A and GTG for both cultivars. Each line is the average of
three plants.
Figure 5 After six days, weeds died in both field of FH-114 and
CIM-598.
A (FH-114): plants were healthier no mortality was observed; B
(CIM-598): mortality of cotton plants were observed along
with weeds shown as red.
genetics of both cultivars for insect and weeds trait
and FH-114 may be descendent from germplasm
which is more dominant and have the resistance
capacity to insects and weeds.
3 Conclusions
FH-114 harboring double BT and GTG gene holds
good potential to combat with serious problems of
insects and weeds and may be good assets for local
farmers and National breeders to develop further good
varieties using this material against insects and weeds
which can pave their role in boosting up economy of
Pakistan as compared with CIM-598.
4 Materials and Methods
4.1 Plant material
Two cotton varieties FH-114 and CIM-598 were
transformed with Cry1Ac+Cry2A along with
cp4EPSPS gene. The seeds of cotton varieties were
received from Cotton Research Station Multan,
Pakistan. Concentrated H
2
SO
4
was used for delinting
while sterilization of seeds was done with 5% HgCl
2
and 10% SDS. Seeds were then allowed to germinate
at 30
o
C in incubator.
4.2 Transformation of the Bt and glyphosate gene
into cotton
Cry1Ac+Cry2A and cp4EPSPS genes were transf-
ormed in FH-114 and CIM-598 according to Rao (Rao
et al., 2011). Two constructs one with the double Bt
and other for the cp4EPSP gene were constructed for
cotton transformation under the control of the
CaMV35S promoter and NOS terminator sequence.
