Transformation of Cotton Using a gus Gene and Study of its Inheritance and Expression in Second Generation (T1) of Transgenic Cotton



Production of transgenic cotton (Gossypium hirsutum L.) was studied utilizing Agrobacterium-mediated strain LBA4404 and pBI121 plasmid. This plasmid contains gus as a reporter gene driven by the CaMV35S promoter and nptII gene driven by NOS as selection marker. In this study, the inheritance of gus gene was evaluated using selective media, PCR and GUS staining in the second genertaion (T1) of transgenic cotton obtained from those plants in which the expression of gus gene was confirmed in different tissues (pollen and fiber). Seed samples from transgenic lines (T0) along with non transgenic control seeds were employed. The ?2 test indicated that the inheritance of gus gene in A3, A4 and A7 lines follows Mendilian ratio for single gene segregation (3:1). PCR amplification was carried out to test the presence of nptII gene in the second generation. The results revealed the presence of nptII gene in all the GUS positive transgenic plants except in one. The results confirmed the stability of gus gene in the second generation.