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| 根癌农杆菌介导柱花草炭疽菌遗传转化体系的优化 | |
| 引用本文： | 胡彩平,郑金龙,高建明,蔡志英,黄贵修,习金根,张世清,陈河龙,易克贤. 根癌农杆菌介导柱花草炭疽菌遗传转化体系的优化[J]. 热带作物学报, 2013, 34(6): 1007-1012 |
| 作者姓名： | 胡彩平 郑金龙 高建明 蔡志英 黄贵修 习金根 张世清 陈河龙 易克贤 |
| 作者单位： | 1. 海南大学农学院,海南海口 570228;中国热带农业科学院环境与植物保护研究所,海南海口 571101 2. 中国热带农业科学院环境与植物保护研究所,海南海口,571101 3. 中国热带农业科学院热带生物技术研究所,海南海口,571101 |
| 基金项目： | 国家自然科学基金项目“柱花草炭疽菌致病相关基因的克隆及功能分析”（No. 31072076）；海南大学211工程建设项目。 |
| 摘 要： | 选取含有氯嘧磺隆抗性标记的ILV1基因和报告基因GFP二元载体的农杆菌AGL-1,采用单因子和双因子方法研究根癌农杆菌AGL-1介导柱花草炭疽菌CH008转化过程中各主要因素对转化率的影响,优化构建ATMT转化柱花草胶孢炭疽菌体系条件,使转化效率达到300 ～400个转化子/106个炭疽孢子,即根癌农杆菌AGL-1浓度OD600=0.8,炭疽菌分生孢子浓度为1x106个/mL,AS浓度为100 μmol/L,诱导时间为6h,共培养温度和时间分别为25 ℃和4d.经PCR检测都有GFP片段,并能够表达绿色荧光蛋白.这为进一步开展炭疽菌对柱花草的致病机理研究提供了依据,优化转化体系有利于后续突变体库的扩大、筛选突变体和致病功能基因的研究. |
| 关 键 词： | 柱花草 炭疽菌 遗传转化 |
Optimization of Genetic Transformation System of Stylo Anthracnose Mediated by Agrobacterium tumefaciens |
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| HU Caiping,ZHENG Jinlong,GAO Jianming,CAI Zhiying,HUANG Guixiu,XI Jingen,ZHANG Shiqing,CHEN Helong and YI Kexian. Optimization of Genetic Transformation System of Stylo Anthracnose Mediated by Agrobacterium tumefaciens[J]. Chinese Journal of Tropical Crops, 2013, 34(6): 1007-1012 | |
| Authors: | HU Caiping ZHENG Jinlong GAO Jianming CAI Zhiying HUANG Guixiu XI Jingen ZHANG Shiqing CHEN Helong YI Kexian |
| Affiliation: | 1 Hainan University2 Environment and Plant Protection Institute, CATAS;Environment and Plant Protection Institute, CATAS;Institute of Tropical Bioscience and Biotechnology, CATAS;Environment and Plant Protection Institute, CATAS;Environment and Plant Protection Institute, CATAS;Environment and Plant Protection Institute, CATAS;Institute of Tropical Bioscience and Biotechnology, CATAS;Institute of Tropical Bioscience and Biotechnology, CATAS;Environment and Plant Protection Institute, CATAS |
| Abstract: | In this study, an Agrobacterium tumefaciens AGL-1 containing binary vector with ILV1 and GFP as report genes was used for transformation of Colletotrichum gloeosporioides. In order to gain higher transformation efficiency, the major facts affecting transformation efficiency was analyzed by a one-factor experiment and a two-factor experiment. The optimization of genetic transformation system of C. gloeosporioides was established, and we optimized the conditions of transformed system. The transformation efficiency was up to 300 to 400 transformants per 106 conidia. All the transformants tested had GFP gene comfirmed by PCR, and GFP signals were also detected. The study provided a basis for further study on the pathogenesis of C. gloeosporioides, optimized transformation system offered highly efficient means for generating an ATMT library of C. gloeosporioides. And it would help to analyze the function of pathogenicity gene of this pathogen. |
| Keywords: | Stylosanthes guianensias SW Colletotrichum gloeosporioides Genetic transformation |
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