| 农杆菌介导油菜黑胫病菌遗传转化 |
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| 引用本文： | 宋培玲,皇甫海燕,史志丹,燕孟娇,郝丽芬,皇甫九茹,贾晓清,郭晨,杨永青,李子钦. 农杆菌介导油菜黑胫病菌遗传转化[J]. 中国油料作物学报, 2021, 43(2): 293-299. DOI: 10.19802/j.issn.1007-9084.2019294 |
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| 作者姓名： | 宋培玲 皇甫海燕 史志丹 燕孟娇 郝丽芬 皇甫九茹 贾晓清 郭晨 杨永青 李子钦 |
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| 作者单位： | 1. 内蒙古自治区农牧业科学院植物保护研究所，内蒙古呼和浩特，010031；2. 内蒙古大学生命科学学院，内蒙古呼和浩特，010021 |
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| 基金项目： | 内蒙古自治区农牧业创新基金项目(2017CXJJN15);国家重点研发计划(2018YFD0200907);内蒙古自治区农牧业科学院院青年创新基金(2017QNJJNO2)。 |
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| 摘 要： | 为优化建立农杆菌介导的油菜黑胫病菌遗传转化技术体系,以携带潮霉素磷酸转移酶基因的pCH-sGFP为载体,农杆菌LBA4404为介体,对油菜黑胫病菌的分生孢子进行转化.油菜黑胫病菌的最佳转化体系为:分生孢子浓度106个/mL,农杆菌OD600值0.6,乙酰丁香酮浓度200μmol/mL,pH 5.8,25℃,共培养4 d...
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| 关 键 词： | 油菜黑胫病菌 遗传转化 转化子 |
Agrobacterium tumefaciens-mediated transformation of Leptosphaeria biglobosa |
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| SONG Pei-ling,HUANGFU Hai-yan,SHI Zhi-dan,YAN Meng-jiao,HAO Li-fen,HUANGFU Jiu-ru,JIA Xiao-qing,GUO Chen,YANG Yong-qing,LI Zi-qin. Agrobacterium tumefaciens-mediated transformation of Leptosphaeria biglobosa[J]. Chinese Journal of Oil Crop Sciences, 2021, 43(2): 293-299. DOI: 10.19802/j.issn.1007-9084.2019294 |
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| Authors: | SONG Pei-ling HUANGFU Hai-yan SHI Zhi-dan YAN Meng-jiao HAO Li-fen HUANGFU Jiu-ru JIA Xiao-qing GUO Chen YANG Yong-qing LI Zi-qin |
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| Affiliation: | 1. Plant Protection Institute, Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China;2. College of Life Science, Inner Mongolia University, Hohhot 010021, China |
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| Abstract: | To optimize and establish Agrobacterium tumefaciens-mediated transformation system of Leptosphaeria biglobosa, A. tumefaciens strain LBA4404 carrying plasmid pCH-sGFP was used as transforming carrier, and conidia of L. biglobosa as transformation recipients. Hygromycin B phosphotransferase gene(hyg) and green fluorescent protain gene(gfp) were transformed into L. biglobosa. The optimal transformation system of L. biglobosa was established. Under the optimal condition of 106 spores/mL, A. tumefaciens OD600=0.6, 200 μmol/mL acetosyringone,pH 5.8,co-cultivation at 25℃ for 4 d, the transformation efficiency could get 120-161 transformants. When selecting 26 transformants randomly for successive cultures, all of them were still resisitant to hygromycin after 5 generations grown on PDA without hygromycin. Green fluorescence was visible in hypha and conidia of the transformants.PCR amplification and Southern blot showed T-DNA insertion in L. biglobosa. |
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| Keywords: | Leptosphaeria biglobosa genetictransformation transformant |
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