Studies On Identification Of Bt Transgenic-cotton Resistance To Insects And Inheritance Of Resistance

In this thesis, we identified K6 and K19 from 7030 transformed insecticidal gene Cry1Aa by methods of PCR, Bioassay and Kana. We got high-quality cotton DNA through the method of SDS. In PCR, we had concluded the best reaction system and reaction procedure: total volume 20ul,buffer 2ul, MgCl 2.0mmol/L,TaqDNA polymerase 0.2u,dNTP 0.3mmol/L,primer1 and primer2 0.25pg/ul,template DNA 3000×.We had researched the affects of MgCl to PCR: Mg2+ is very important in PCR which concentration is affected by EDTA, template DNA, primers, dNTP ect. When the concentration of Mg2+ is low, the product of PCR is small; but when it is high, other ampliation will come up. We had researched affects of TaqDNA polymerase: when it is increasing, the product of PCR is increasing; but when the Mg2+ is low, the product will reduce. When the cotton was transformed, its isozyme system and livingness will be changed. We have studied inheritance of resistance K6 and K19 .It shows that the resistance of K6 and K19 to bollworm was controlled by one pair of dominant gene. The insecticidal gene between K6 and K19 is non-allelic.We have studied the relation between K6,K19 and R19,SHANXI94-24,ZHONGXIN94, too. It shows that the insecticidal gene between K6 ,K19 and R19,SHANXI94-24,ZHONGXIN94 is non-allelic.