农杆菌介导的针刺法将双价抗虫基因导入水稻  

Transforming Two Insect-resistant Genes into Rice through Agrobacterium Mediation by Inoculating Needle

作　　者：张启军[1] 颜文飞[2] 夏士健[1] 宗寿余[1] 吕川根[1] 

机构地区：[1]江苏省农业科学院粮食作物研究所,江苏南京210014 [2]南京农业大学农学院,江苏南京210095

出　　处：《江西农业学报》2015年第8期6-9,15,共5页Acta Agriculturae Jiangxi

基　　金：江苏省自主创新产业技术体系(CX(12)1003);国家转基因生物新品种培育重大专项(2014ZX08001-004)

摘　　要：用含2个抗虫基因的转化质粒p CDMARUBA-Hyg的农杆菌直接针刺侵染籼稻9311和粳稻南粳45、110183的种子胚芽生长点,对存活成苗的10株9311、5株南粳45和1株110183进行常规PCR检测,阳性株分别为9株、3株和1株,混合计算,阳性率为81%。去除南粳45发芽种子的种根和幼芽后,对裸露的胚芽生长点进行针刺接种,9株存活。经对成苗的PCR检测,4株含有2个外源基因,各有2株只含有1个基因(sbk或sck)。与传统的农杆菌介导法相比,本研究的方法不需要诱导愈伤组织以及之后的一系列组织培养,基因转化时间缩短,节省了大量的人力和财力,具有简单、快速、高效之优点,是改良的水稻转基因新技术。The plasmid of pCDMARUBA-Hyg containing two exogenous insect-resistant genes （sbk and sck） was transformed into Agrobacterium EHA105, and the seed apical meristems of three rice varieties （indica rice 9311, japonica rice Nanjing 45 and 110183） were directly infected by the above strain EHA105 through needling inoculation. It was found that 10 plants of 9311, 5 plants of Nanjing 45, and 1 plant of 110183 seedlings survived. The results of conventional PCR detection showed that 9 plants of 9311, 3 plants of Nanjing 45, and 1 plant of 110183 seedlings were positive, and the total positive plants accounted for 81% of the total survived plants. After seminal root and bud were removed from the germinative seeds of Nanjing 45, its naked apical meristems were infected through needling inoculation, and 9 plants survived. PCR detection results indicated that： among the survived 9 plants, 4 plants contained divalent insect-resistant genes, 2 plants only contained gene sbk, and another 2 plants only contained gene sck. In comparison with the traditional method of Agrobacterium-mediated transformation, the present method does not need callus inducement and tissue cuhure, so it can save a large number of labors and costs. It is a simple, quick, efficient method for the transformation of rice genes.

关 键 词：水稻 胚芽生长点 转基因 抗虫 

分 类 号：S511[农业科学—作物学]