生防链霉菌Men-myco-93-63遗传转化体系的建立和优化*

摘要/Abstract

摘要： 摘要：玫瑰黄链霉菌Men-myco-93-63是分离自马铃薯疮痂病自然衰退土壤中的一株拮抗菌。该菌株及其发酵液对棉花黄萎病菌、瓜类白粉病菌等多种重要的植物病原菌具有很强的抑制作用，有良好的生防应用潜力。分别以基因整合型质粒pSET152和基因破坏型质粒pKC1139为出发质粒，ET12567(PUZ8002, pSET152/ pKC1139 ) 为供体，Men-myco-93-63孢子和菌丝体为受体，选取MS、PDA、TSB琼脂培养基、燕麦培养基为不同的培养基进行接合转移试验。结果表明MS培养基为接合转移的最适培养基，经验证，已成功地将质粒pSET152/ pKC1139转入到了Men-myco-93-63 中。以孢子为受体时，孢子预萌发条件为50℃热激10 min,.37℃温育2.5 h，转化效率是10-7~10-6。以菌丝体为受体时，转化效率是10-7~10-6，但菌丝培养过程中容易出现污染。另外，抗生素的覆盖时间对接合转移效率的影响较明显，16-18 h覆盖效果最好。

关键词: 烤烟, 烤烟, 早熟品种, 引种 PVH19

Abstract: Abstract：Streptomyces roseoflavus Men-myco-93-63 was isolated from potato scab (S. scabies) decline soil and its fermentation can inhibit many phytopathogenic fungi and control related important plant diseases such as cotton verticillium wilt (Verticillium dahliae) and cumber powdery mildew (Sphaerotheca fuliginea Poll). The genetic integrate plasmid pSET152 and genetic damage plasmid pKC1139 used as the starting plasmid, ET12567 (PUZ8002, pSET152) and ET12567 (PUZ8002, pKC1139) used as the donor, Men-myco-93-63 spores and hyphae used as the receptor, respectively, the system for the conjugal transfer of pSET152 or pKC1139 from Escherichia coli in to Men-myco-93-63 has been developed. Different medium, such as MS, PDA, TSB agar, oats medium were selected for conjugal transfer, respectively. The results showed that MS medium is fit for conjugal transfer between Men-myco-93-63 and ET12567 (PUZ8002 ), and it was confirmed that plasmid pSET152 and pKC1139 had been successfully transferred to the Men-myco-93-63, respectively. When spores were used as the receptor, the pre-conditions of the germination of spores were 50 ℃ heat shock 10 min, 37 ℃ incubated 2.5 h in MS medium, and the conversion efficiency was 10-7 to 10-6. If the mycelium were uesd as the receptor, the conversion efficiency was 10-7 to 10-6, but the mycelium pollution can not be avoided. In addition, the coverage time on antibiotics effected the joint transfer efficiency significantly, 16 to18 hours was best.

沈凤英,李亚宁,刘力强,吴伟刚,刘大群. 生防链霉菌Men-myco-93-63遗传转化体系的建立和优化*[J]. 中国农学通报, 2009, 25(13): 197-201.

Shen Fengying, Li Yaning, Liu Liqiang,Wu Weigang, Liu Daqun. Optimized of the Conjugal Transfer System of Stremptomyces roseoflavus Men-myco-93-63[J]. Chinese Agricultural Science Bulletin, 2009, 25(13): 197-201.

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