Abstract
Herbicides are the chemicals used to kill unwanted plants or weeds which can compete with main crop for light, water and
other useful nutrients. Two types of herbicides i.e. selective and non-selective are being used but the later one is more efficient
and it cannot differentiate between the crops and weeds. Hence, development of herbicide tolerant (ht) plant is an ultimate
solution to cope with the challenge of non-selective ones. Nucleotide sequence of ht bacterial gene encoding 5-
enolpyruvylshikimate-3-phosphate (EPSP) synthase was modified according to crop codon usage. The synthetic herbicide
tolerant gene was cloned in a gateway compatible plant expression vector and introduced into an Agrobacterium strain
LBA4404. Tissue culture independent in-planta transformation system was used to integrate the transgene into cotton plants
at three different stages of flower after pollination. Flower shedding was the main problem observed during inoculation.
Maximum inoculated bolls were developed at 16 hours after pollination and plantlets were screened using different doses of
roundup spray having glyphosate as an active ingredient. Successful transgene integration was verified using PCR and Southern
blot analysis in plants surviving at @ 600 mL/acre roundup herbicide. Whereas on higher dose of herbicide transgenic plant
did not manage to survive. These results showed that the cotton is successfully transformed with ht gene but the expression of
transgene is low or it may have been gone in silent mode. Gene silencing is a natural phenomenon which could be avoided by
using more efficient regulatory elements or by introducing the transgene into other genetic compartments of plant. In addition,
development of more transformation events could result in transgenic cotton plants with higher transgene expression
Shahid Nazir, Muhammad Zaffar Iqbal, Ihsan Ullah, Sajid-ur-Rahman. (2020) IN-PLANTA GENETIC TRANSFORMATION OF SYNTHETIC HERBICIDE TOLERANT GENE IN UPLAND COTTON, Pakistan Journal of Agricultural Sciences, Volume 57, Issue 6.
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