| 橡胶树尖孢炭疽菌绿色荧光蛋白（GFP）标记转化株的获得 |
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| 引用本文： | 林春花,刘先宝,蔡吉苗,李超萍,李继锋,黄贵修.橡胶树尖孢炭疽菌绿色荧光蛋白（GFP）标记转化株的获得[J].热带作物学报,2009,30(10):1495-1500. |
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| 作者姓名： | 林春花 刘先宝 蔡吉苗 李超萍 李继锋 黄贵修 |
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| 作者单位： | 农业部热带农林有害生物入侵监测与控制重点开放买验室,中国热带业科学环境与植物保护研究所,海南儋州,571737 |
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| 基金项目： | 中国热带环植所中央级公益性科研究院所基本科研究所基本科研业务费专项(No.2009hzslJO14).中国热科院科技基金课题资助项目 |
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| 摘 要： | 建立了根癌农杆菌介导转化橡胶树尖孢炭疽菌(Collectotrichum acutatum)获得T-DNA插入突变体的体系:在尖孢炭疽菌孢子浓度10~6个/mL、农杆菌浓度OD_(600)=0.15～0.2后在200μmol/mL的乙酰丁香酮(AS)诱导6 h,共培养48 h.转化效率可达150～300个/10~6个分生孢子.获得的转化子通过PCR检测绿色荧光蛋白基因、Southern 杂交验证、分生孢子的荧光观察,结果表明:被测转化子基因组中确实整合了目的片段,成功获得了橡胶树尖孢炭疽菌菌株CHY-1绿色荧光蛋白标记转化子.
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| 关 键 词： | 根癌农杆菌 尖孢炭疽菌 遗传转化 绿色荧光蛋白 |
Collectotrichum acutatum GFP Tagged transformants Generated from Agrobacterium tumefaciens-mediated Insertional Mutagenesis |
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| Lin Chunhu,Liu Xianbao,Cai Jimiao,Li Chaoping,Li Jifeng and Huang Guixiu.Collectotrichum acutatum GFP Tagged transformants Generated from Agrobacterium tumefaciens-mediated Insertional Mutagenesis[J].Chinese Journal of Tropical Crops,2009,30(10):1495-1500. |
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| Authors: | Lin Chunhu Liu Xianbao Cai Jimiao Li Chaoping Li Jifeng and Huang Guixiu |
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| Institution: | Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests;Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests;Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests;Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests;Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests;Environment and Plant Protection Institute, CATAS/Ministry of Agriculture Key Laboratory for Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests |
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| Abstract: | Colletotrichum acutatum isolate CHY-l(the causal agent of anthracnose of rubber tree) was transformed by Agrobacterium tumefaciens-mediated transformation. The isolate CHY -1 with the concentration of 10~6 spores/mL were co-cultured with A. tumefaciens at OD600=0.15～0.2 for 48 hours in presence of acetosyringone(200 μmol/mL) , and 150～300 transformants were generated per 10~6 conidia. The transformants were identified by PCR, southern blot analysis of GFP cassette, and green fluorescence observation with CLSM. All results confirmed that the target genes were successfully integrated into the genome of C. acutatum. |
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| Keywords: | T-DNA Agrobacterium tumefaciens C acutatum Genetic transformation T-DNA green fluorescent protein |
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