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在建立以银腺杨(84K)叶片为外植体的再生系统基础上,通过农杆菌介导法把Cry I Ac和API双价抗虫基因导入银腺杨(84K)基因组中。转化杨树叶片,在含有卡那霉素的培养基上诱导不定芽和诱导生根,获得了400株卡那霉素抗性转化再生植株。抗性植株经PCR检测,有70株呈阳性。通过Southern杂交和ELISA检测进一步证明Cry IAc和API双价抗虫基因已整合到银腺杨(84K)基因组中,拷贝数1~2个,并得到了表达。转化植株用杨扇舟蛾幼虫进行饲虫试验,结果表明昆虫幼虫的死亡率高达60.0%~80.0%。同时,存活幼虫的生长发育也受到了明显抑制。本研究结果为杨树抗虫育种提供了新的种质资源。
Based on the establishment of a regeneration system using the leaves of Ginkgo biloba (84K) as explants, the Cry I Ac and API bivalent insect-resistant genes were introduced into the genome of Agropyron giganteum (84K) via Agrobacterium-mediated transformation. Poplar leaves were transformed, adventitious buds were induced on kanamycin-containing medium and rooting was induced, and 400 kanamycin-resistant transformed and regenerated plants were obtained. The resistant plants were detected by PCR, 70 were positive. Southern hybridization and ELISA tests further proved that the Cry IAc and API bivalent insect-resistant genes have been integrated into the genome of the Ginkgo biloba (84K) with 1-2 copies and expressed. Transformed plants with Helicoverpa armigera larvae insecticide tests showed that the mortality of insect larvae as high as 60.0% to 80.0%. At the same time, the survival and development of surviving larvae have also been significantly inhibited. The results of this study provide new germplasm resources for poplar insect resistance breeding.