转Cry1A基因海岛棉的获得和鉴定

The Acquisition of Transgenic Gossypium barbadense with Cry1A Gene and Its Identification

进行农杆菌介导转抗虫基因试验,以期选育抗虫性优良的海岛棉新品系(品种)。本实验在海岛棉体细胞胚再生体系的基础上,将携带Cry1A基因的表达载体pBI121通过农杆菌介导法转化到‘新海33号’的胚性愈伤组织中。经培养基筛选、PCR检测、RT-PCR检测、抗虫性鉴定等筛选抗虫后代。共获得12棵抗性植株。PCR检测结果显示,这12棵抗性植株均能扩增出了大小相符的片段,检测结果呈阳性;RT-PCR结果显示,12棵转基因植株目的基因已经整合入基因组,且能反转录出mRNA;Bt-Cry1Ab/Ac转基因试纸条检测结果表明这12棵转基因植株含有目的蛋白;抗虫性鉴定结果证实了这些转基因植株对棉铃虫幼虫具有一定的抗性。本研究成功将抗虫基因导入至海岛棉的体内,获得了具有抗虫性的植株,其结果对转基因抗虫海岛棉新品种的培育具有一定的参考价值。

Agrobacterium mediated transgenic insect-resistant gene test is conducted in order to select new island cotton lines(varieties)with excellent insect resistance.On the basis of embryonic regeneration system of island cotton somatic cells,the expression vector pBI121 with Cry1 A gene was transferred to the embryogenic callus of‘Xinhai 33’by Agrobacterium-mediated method.The insect-resistant offspring were screened by the culture medium screening,PCR detection,RT-PCR detection and insect-resistance identification.12 resistant plants were obtained.The PCR result revealed that the 12 resistant plants could amplify the expected-size DNA fragments,and the test results were positive.RT-PCR result showed that the target gene had been integrated into 12 transgenic plant genomes,and could reverse transcription of mRNA.The test result of BtCry1 Ab/Ac transgenic paper indicated that the 12 transgenic plants contained the target protein.Transgenic plants had certain resistance to cotton bollworm larvae by insect-resistant identification.This study successfully transforms insect-resistant genes into island cotton,obtains the insect-resistant plants,and the results have some reference value for the breeding of new transgenic insect-resistant island cotton varieties.