Construction Of Plant Expression Vector Containing Insect Resistant Bt Gene And Herbicide Resistant Bar Genes And Research On Transformation Of Cotton

Cotton is an important cash crop, but which was gotten the damage of pests and weeds ingrowth by the long-term. Directly. It caused the economic loss by the10%~20%of cropoutput. So the development of cotton genetic engineering will greatly promote the cultivationof cotton in pests resistance, herbicide resistance and resistance.The research firstly amplificated the bar gene of herbicide resistance and the Bt gene ofpests resistance by PCR, and then the bivalent plant expression vector pBI121-bar/Bt wasconstructed successfully. The Plant expression vector pBI121-bar/Bt and the pBI121-Bt weretransformed into the LBA4404. Then the bacteria liquid infected the embryogenic callus inxh14、xh16、xh24and xh30. Screening the PPT concentration though the resistanceembryogenic callus, it suggested that the growth of embryogenic callus was good and theeffection was obvious when the PPT concentration was in3.0mg/l. The growth of xh24wasthe worst in four kinds of different genotypes of embryogenic callus, it showed that differentgene type of PPT have different sensitivity. Screening the Cef concentration aftertransformating the pBI121-bar/Bt, it showed which can effectively prevent the pollutioncaused by agricultural bacillus and not prevent the growth of embryogenic callus when theCef concentration was800mg/l. The effective Cef concentration is in600mg/l when thepBI121-Bt gene was transformated the embryogenic callus. It indicated that the sensitivity ofthe embryogenic callus to antibiotic was different.When the concentration of PPT was in3mg/L and Cef was in800mg/L aftertransformating the pBI121-bar/Bt gene in different varieties of cotton embryogenic callus, thecallus rate was the highest than other varieties and a lesser degree of the black, but xh24,respectively.When the concentration of Kna was in50mg/L was and Cef was in600mg/Lafter transformating the Bt-4AB gene in different varieties of cotton embryogenic callus, thecallus rate of xh16was the highest than other varieties and a lesser degree of the black afterinfected by Agrobacterium, but the callus rate of xh30is the lowest.