Genetic Transformation Studies in Cotton (Gossypium spp.)
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Date
2015-12
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University of Agricultural Science, Dharwad
Abstract
In the present genetic transformation study, both in vitro and in planta approaches were tried to transfer cry1Ac and cry1Acm genes. Coker-312 (Gossypium hirsutum L.) having in vitro regeneration ability, was used in in vitro transformation study. In planta transformation studies were carried out in Sahana (Gossypium hirsutum L.) and BCS 23-18-7 (Gossypium barbadanse L.) cotton varieties. In in vitro transformation studies, 230 embryogenic calli and 135 hypocotyl, (20mm) were used as explants source for each gene. In in planta transformation, eight to ten days old seedlings were used as explants. Each 90 of seedling of Sahana and 70 of BCS 23-18-7 were used in transfer of cry1Acm and cry1Ac respectively. The variable numbers of plants (26-37) of each combination were established. In these established plants, few plants from each combination (3-6 plants) were recorded as positive for PCR with gene of interest (cry1Ac and cry1Acm) and selectable marker gene (npt-II) through PCR. PCR positive plants from in vitro studies were also subjected for expression studies through RT-PCR and detection of Cry protein through Bt express strips (Amar immunodiagnostics). However absence of inheritance of these genes to next generation (T1) in both cases indicates transient expression. This may be due to non integration of foreign gene into host genome and its elimination during cell division.
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