CAJ | 학술논문

为培育抗旱白菜品种，以大白菜品种二包头为试验材料，以大白菜的带柄子叶为转化受体，研究农杆菌介导的大白菜抗旱遗传转化体系的建立。结果表明：种子消毒的方法为先用75％的酒精浸泡30 s ，再用0．1％氯化汞浸泡10 min ，出芽率较高且无菌；最佳分化培养基为MS＋6-BA 2．5 mg·L-1＋NAA 0．6 mg·L-1，不定芽分化率为30％；最佳生根培养基为MS＋NAA 0．4 mg·L-1，根多且粗壮。大白菜的遗传转化使用农杆菌介导的叶盘转化法，将含有抗旱基因 T59的 T-DNA转入到外植体中去，通过 Hyg筛选，获得了抗性苗。转基因试验研究了多种因素对农杆菌转化的影响，其中最佳的农杆菌稀释倍数为8倍和16倍，最佳的共培养时间为2 d ，最佳的脱菌抗生素组合为400 mg·L-1 Cb＋200 mg·L-1 Cef。
위배육항한백채품충，이대백채품충이포두위시험재료，이대백채적대병자협위전화수체，연구농간균개도적대백채항한유전전화체계적건립。결과표명：충자소독적방법위선용75％적주정침포30 s ，재용0．1％록화홍침포10 min ，출아솔교고차무균；최가분화배양기위MS＋6-BA 2．5 mg·L-1＋NAA 0．6 mg·L-1，불정아분화솔위30％；최가생근배양기위MS＋NAA 0．4 mg·L-1，근다차조장。대백채적유전전화사용농간균개도적협반전화법，장함유항한기인 T59적 T-DNA전입도외식체중거，통과 Hyg사선，획득료항성묘。전기인시험연구료다충인소대농간균전화적영향，기중최가적농간균희석배수위8배화16배，최가적공배양시간위2 d ，최가적탈균항생소조합위400 mg·L-1 Cb＋200 mg·L-1 Cef。
In order to develop drought-resistant varieties of cabbage ,taking Chinese cabbage Erbaotou as test material and cotyledon petioles as transformation receptor ,the establishment of genetic transformation system of Chinese cabbage drought-resistant were studied by Agrobacterium-mediated .The results showed that seed disinfection method was soaking with 75% ethanol for 30 s first ;and then soaking with 0 .1% mercuric chloride for 10 minutes ,the budding rate could be higher and sterile .The best differential medium was MS + 6-BA 2 .5 mg·L-1 +NAA 0 .6 mg·L-1 ,differentiation rate of adventitious bud was 30% .The best rooting medium was MS+NAA 0 .4 mg·L-1 ,the roots were thick and strong .Leaf disc transformation method was used to es-tablish genetic transformation of Chinese cabbage by Agrobacterium-mediated ,which transfered T-DNA contai-ning drought-resistant gene T59 into explants ,resistance seedlings were acquired by Hyg screening .Effect of several factors on Agrobacterium-mediated transformation was studied through transgenic test ,the following conclusions could be reached :the best A grobacterium dilution was 8 and 16 times ,the best common cultivation time was 2 d ,the optimal antibiotic combination was 400 mg·L-1 Cb+200 mg·L-1 Cef .