转修饰豇豆胰蛋白酶抑制剂基因(sck)抗虫甜椒植株的获得

OBTAINING INSECT-RESISTANT TRANSGENIC SWEET PEPPER TRANSFORMED WITH MODIFIED CpTI GENE(sck)

以 12～ 14d苗龄的甜椒带柄子叶为外植体 ,经根癌农杆菌 (Agrobacteriumtumefaciens)介导 ,将修饰的豇豆胰蛋白酶抑制剂基因 (sck)导入甜椒杂交种“中椒 5号”和常规种“茄门”中 ,在对甜椒转化系统和芽丛诱导茎伸长的条件进行优化研究后 ,获得了卡那霉素抗性植株 ,最高转化频率为 16 .7% .卡那霉素抗性筛选、PCR检测和Southernblot杂交均证实 ,nptII基因和sck基因已整合进甜椒基因组中 .室内离体叶片饲虫和田间自然抗虫性鉴定进一步证明 ,转基因植株对棉铃虫 (HeliothisarmigeraHubner)具有一定抗性 .图 4表 2参

Modified CpTI (cowpea trypsin inhibitor) gene ( sck ) was successfully transferred into sweet pepper varieties 'Zhongjiao No. 5' and 'Qiemen' mediated by Agrobacterium tumefaciens . Kanamycin resistant plants were obtained from transformed cotyledons with petiole of 12～14 d seedling when the transformation system and the condition of inducing stem elongation were optimized. It was confirmed that sck gene was integrated into sweet pepper genome DNA by kanamycin resistant screening, PCR analysis and Southern blot assay. Insect tolerance of the transgenic plants to cotton bollworm was observed in laboratory and in field, which showed some transgenic plants had strong resistance to this pest. Fig 4, Tab 2, Ref 13