| 乌头叶面细菌、真菌DGGE分析条件的建立与优化 |
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| 引用本文： | 李娜,胡亮,王婷,欧洪,王光志.乌头叶面细菌、真菌DGGE分析条件的建立与优化[J].中国实验方剂学杂志,2017,23(4):58-61. |
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| 作者姓名： | 李娜 胡亮 王婷 欧洪 王光志 |
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| 作者单位： | 成都中医药大学 中药材标准化教育部重点实验室, 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 成都 611137,成都中医药大学 中药材标准化教育部重点实验室, 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 成都 611137,成都中医药大学 中药材标准化教育部重点实验室, 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 成都 611137,成都中医药大学 中药材标准化教育部重点实验室, 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 成都 611137,成都中医药大学 中药材标准化教育部重点实验室, 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 成都 611137 |
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| 基金项目： | 国家自然科学基金项目（30901962）；四川省教育厅重点项目（15ZA0098） |
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| 摘 要： | 目的:筛选并优化适用于分析乌头叶面细菌、真菌多样性的变性梯度凝胶电泳(DGGE)实验条件。方法:采用Touch Down-PCR技术扩增乌头叶面细菌16 S r DNA V3区域和真菌18 S r DNA~5.8 S r DNA序列,利用该扩增产物对乌头叶面细菌、真菌DGGE电泳条件进行优化。采用DGGE垂直电泳法筛选凝胶变性梯度,时间间隔法优化电泳时间。结果:乌头叶面细菌DGGE最佳分析条件为变性剂梯度30%~65%(Gel 10%),电泳温度60℃,电压120 V,电泳时间7 h。乌头叶面真菌DGGE最佳分析条件为变性剂梯度25%~45%(Gel 8%),电泳温度60℃,电压120 V,电泳时间10 h。结论:利用以上优化条件能有效分离乌头叶面细菌16 S r DNA V3区及真菌18 S r DNA~5.8 S r DNA序列,为乌头叶面细菌,真菌的群落结构PCR-DGGE分析提供可靠的技术支撑。
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| 关 键 词： | 乌头 叶面 细菌 真菌 变性梯度凝胶电泳 |
| 收稿时间： | 2016/7/14 0:00:00 |
Establishment and Optimization of DGGE Analysis Methods for Phylloplane Bacteria and Fungi of Aconitum carmichaeli |
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| LI N,HU Liang,WANG Ting,OU Hong and WANG Guang-zhi.Establishment and Optimization of DGGE Analysis Methods for Phylloplane Bacteria and Fungi of Aconitum carmichaeli[J].China Journal of Experimental Traditional Medical Formulae,2017,23(4):58-61. |
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| Authors: | LI N HU Liang WANG Ting OU Hong and WANG Guang-zhi |
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| Institution: | Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China,Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China,Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China,Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China and Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China |
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| Abstract: | |
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| Keywords: | Aconitum carmichaeli phylloplane bacteria fungi denaturing gradient gel electrophoresis |
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