HE Xi-qiang,XU Heng-jian.Study on Regeneration System and Transgenic Impact Factors of Chinese Cabbage[J].HEILONGJIANG AGRICULTURAL SCIENCES,2013,(09):9-13.
大白菜再生体系和转基因影响因素的研究
- Title:
- Study on Regeneration System and Transgenic Impact Factors of Chinese Cabbage
- 文章编号:
- 1002-2767（2013）09-0009-05
- 分类号:
- S634.303
- 文献标志码:
- A
- 摘要:
- 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-1Cb+200mg.L-1 Cef。
- Abstract:
- 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 establish genetic transformation of Chinese cabbage by Agrobacterium-mediated,which transfered T-DNA containing 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 Agrobacterium dilution was 8 and 16 times,the best common cultivation time was 2 d,the optimal antibiotic combination was 400 mg.L-1Cb+200 mg.L-1Cef.
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备注/Memo
收稿日期：2013-04-17
第一作者简介：贺玺强（1988-），男，山东省日照市人，硕士，从事植物分子生物学研究。E-mail：813088248@qq.com。
通讯作者：徐恒戬(1966-),男，山东省莒南县人，博士，副教授，从事植物生物学及农业生物技术教学与研究。E-mail:xhj310@163.com。