花生油酸脱氢酶基因遗传转化体系的建立与表达分析

 【目的】培育高油酸的花生新种质。【方法】以根癌农杆菌为介导将含有油酸脱氢酶基因的植物双元表达载体pFGC/2nd转入花生外植体丛生芽，通过卡那霉素筛选，器官发生再生转化植株;荧光定量PCR检测转基因植株。【结果】丛生芽外植体于OD600为0.6的农杆菌菌液中浸泡8～10 min后，在MS培养基上暗培养3 d效果较好。诱导筛选培养3个月左右，共得到16株转化植株，其中11个转基因植株经PCR检测呈阳性，说明外源基因已整合进入受体植物。进一步的荧光定量PCR检测显示，有4个植株基本上不表达或表达量很低。【结论】建立花生丛生芽遗传转化体系;RNA干扰对内源的油酸脱氢酶基因产生了抑制作用。

Expression Analysis and Establishment of Regeneration System of Oleate Desaturase Gene in Peanut

【Objective】 This article aims at breeding of new materials with high oleate in peanut. (Method) Plant dual expression vector pFGC/2nd, containing oleic desaturase gene, was transferred into full bud explants of peanut via Agrobacterium tumefaciens, and trsnsgenic plants were got by screening with kanamycin. 【Result】 It was better-performing that the bud explants were dipped into Agrobacterium suspension (OD600=0.6) for 8 - 10 min and then cultured on MS medium in the dark for three days. After induction screening for three months or so, a total of 16 conversion plants were produced, of which 11 transgenic plants were positive by PCR analysis. It stated that the foreign gene had been integrated into the receptor plant. The further Real Time PCR indicated that the expression level of four transgenic plants was significantly reduced. 【Conclusion】 It is speculated that target genes maybe suppressed by dsRNA interference, and the high oleate material could be selected.