花生油酸脱氢酶基因遗传转化体系的建立与表达分析.pdfVIP

中国农业科学 2009,42(5):1827-1832 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.05.041 花生油酸脱氢酶基因遗传转化体系的建立与表达分析 1 2 1 1 1 殷冬梅 ，杨海棠 ，台国琴 ，杨秋云 ，崔党群 1 2 （河南农业大学，郑州 450002 ； 郑州农林科学研究所，郑州 450007 ） 摘要： 【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。 关键词：花生；油酸脱氢酶基因；遗传转化；实时荧光定量PCR Expression Analysis and Establishment of Regeneration System of Oleate Desaturase Gene in Peanut 1 2 1 1 1 YIN Dong-mei , YANG Hai-tang , TAI Guo-qin , YANG Qiu-yun , CUI Dang-qun 1 2 ( Henan Agricultural University, Zhengzhou 450002; Zhengzhou Research Institute of Agriculture and Forestry, Zhengzhou 450007) Abstract: 【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 whi