CAJ | 학술논문

用含2个抗虫基因的转化质粒pCDMARUBA－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。
용함2개항충기인적전화질립pCDMARUBA－Hyg적농간균직접침자침염선도9311화갱도남갱45、110183적충자배아생장점，대존활성묘적10주9311、5주남갱45화1주110183진행상규PCR검측，양성주분별위9주、3주화1주，혼합계산，양성솔위81％。거제남갱45발아충자적충근화유아후，대라로적배아생장점진행침자접충，9주존활。경대성묘적PCR검측，4주함유2개외원기인，각유2주지함유1개기인（ sbk혹sck）。여전통적농간균개도법상비，본연구적방법불수요유도유상조직이급지후적일계렬조직배양，기인전화시간축단，절성료대량적인력화재력，구유간단、쾌속、고효지우점，시개량적수도전기인신기술。
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 EHA 105 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 culture , so it can save a large number of labors and costs .It is a simple, quick, efficient method for the transformation of rice genes .