欧洲葡萄体胚诱导及农杆菌介导的遗传转化研究  

Study on Agrobacterium-mediated Transformation of Vitis vinifera ( L. ) and Plant Regeneration

作　　者：谭超[1] 张剑侠[1] 王跃进[1] 

机构地区：[1]西北农林科技大学园艺学院,农业部西北园艺植物种质资源利用重点开放实验室,陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《北方园艺》2012年第19期113-117,共5页Northern Horticulture

基　　金：国家自然科学基金资助项目(30971972);现代农业产业技术体系建设专项资金资助项目(CARS-30-yz-7);公益性行业(农业)科研专项经费资助项目(200903044)

摘　　要：以欧洲葡萄(Vitis vinifera L.)品种"无核白"(‘Thompson Seedless’)、"佳利酿"(‘Carignane’)、"红地球"(‘Red Globe’)的雄蕊、雌蕊和花蕾为外植体,对花器官体细胞胚诱导及农杆菌介导的转芪合成酶基因进行研究。结果表明:所采用的5种胚性愈伤组织诱导培养基中,MC培养基较适于雄蕊胚性愈伤组织的诱导,"无核白"、"佳利酿"和"红地球"雄蕊的胚性愈伤组织诱导率分别为1.7%、0.3%和11.8%;PIV培养基可诱导"无核白"和"佳利酿"雌蕊胚性愈伤组织形成,诱导率分别为4.0%和3.2%,而"红地球"雌蕊在MC培养基上诱导率最高,达5.6%;仅"红地球"的花蕾在MS1培养基上可诱导产生胚性愈伤组织,诱导率为0.5%。改良X6液体培养基较有利于次生胚的诱导;B培养基有利于体细胞胚萌发成苗。转基因材料经抗性筛选和植株再生,获得了2株"无核白"抗性苗,经PCR和PCR-Southern检测表明,目的基因被整合到转基因植株中。Stamen,pistils and flower buds fromVitis vinifera cvs.‘Thompson Seedless’,‘Carignane’and ‘Red Globe’were used as explants to investigate their efficience of embryonic callus induction and Agrobacterium-mediated stilbene synthase gene transformation.The results indicated that MC medium was optimum for embryonic callus induction from stamens among 5medium for embryonic callus induction.The induction rate in ‘Thompson Seedless’,‘Carignane’and ‘Red Globe’were 1.7%0.3%and 11.8%,respectively.Embryonic callus from ‘Thompson Seedless’and ‘Carignane’could be induced in PIV medium with 4.0%and 3.2%induction rates.However,the highest induction rate occurred in embryonic callus from flower buds of ‘Red Globe’in MC medium with 5.6%induction rate.Only flower buds from Redglobe could be induced in MS1medium with 0.5%induction rate.A modified X6liquid medium was suitable for secondary embryogenesis,and medium B favored germination and seedling of somatic embryos.Transformed plants were selected through resistance screening and 2resistent plants of ‘Thompson Seedless’were obtained.Results from PCR and PCR-Southern indicated that stilbene synthase gene were integrated into the genome of ‘Thompson Seedless’.

关 键 词：葡萄 花器官 体细胞胚 植株再生 转基因 

分 类 号：S663.103.6[农业科学—果树学]