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Volume 15, Issue 5 (9-2013)
Abstract
Transgene integration and expression in host plant is quite unpredictable and is considered as the major problem in plant transformation. The variation in transgene copy number in transgenic plants influences the expression level and is one of such complication. In many plant species, the analysis of transgenic plants has shown that independent transgenic plants have one to many copies of transgenes. This study focused on molecular characterization of difference in copy number of transgenes and its impact on expression level on mRNA basis. Four advanced transgenic lines of phytochrome B were analyzed for the integration of the gene. These transgenic lines were taken out on the basis of difference in copy number as determined by Southern blot analysis and Fluorescence in situ hybridization (FISH) for transgene expression. Results taken by both real time PCR and Northern blot analysis determined high expression in Line QCC11 having two copies of transgenes in homozygous condition while the least expression was seen in lines QCC10 showing three copy number in heterozygous condition as multiple copies can be incorporated from one to few insertion sites.
Volume 22, Issue 6 (11-2020)
Abstract
Genetic manipulation to get desirable characters in cash crops like cotton remains the prime objective of crop biotechnology. To produce transgenic plants resistant against bollworms, newly emerged embryos of MNH-786 cotton variety were transformed with Agrobacterium tumefaciens strain LBA4404 harboring the plasmid pKHG4 vector containing Cry1Ac+Cry2A genes under the control of CaMV35S promoter. The integration and expression of these genes were evaluated by PCR, Florescent In-Situ Hybridization (FISH), and ELISA, respectively. Out of 700 putative transgenic cotton plants, 10 plants (1.03% transformation efficiency) showed the presence of genes Cry1Ac+Cry2A through PCR analysis. In vitro, insect feeding bioassay was done for estimation of mortality percentage of Helicoverpa armigera. Insect mortality rate and morphological characteristics of Bt cotton were analyzed by phenotypic correlation, path coefficient regression and covariance to evaluate the advantage of transgenic technology in numerical terms. Statistical analysis indicated significant positive correlation between insect mortality and cotton seed yield. Helicoverpa armigera mortality data produced a directly proportional relation with cotton seed yield. The results of this study support the improvement of cotton defense mechanism against insects and natural competitors through genetic modification.
