| 三七根腐病原菌毁坏柱孢霉分子定量检测方法及其应用 |
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| 引用本文： | 吴照祥,郝志鹏,曾燕,郭兰萍,黄璐琦,王勇,陈保冬.三七根腐病原菌毁坏柱孢霉分子定量检测方法及其应用[J].微生物学通报,2015,42(3):598-607. |
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| 作者姓名： | 吴照祥 郝志鹏 曾燕 郭兰萍 黄璐琦 王勇 陈保冬 |
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| 作者单位： | 1. 中国科学院生态环境研究中心 城市与区域生态国家重点实验室 北京 100085,1. 中国科学院生态环境研究中心 城市与区域生态国家重点实验室 北京 100085,2. 中国药材公司 北京 100097,3. 道地药材国家重点实验室 中国中医科学院中药资源中心 北京 100700,3. 道地药材国家重点实验室 中国中医科学院中药资源中心 北京 100700,4. 文山学院 文山三七研究院 云南 文山 663000,1. 中国科学院生态环境研究中心 城市与区域生态国家重点实验室 北京 100085 |
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| 基金项目： | 国家自然科学基金项目(No. 41101245)；国家科技支撑计划项目(No. 2012BAI29B02) |
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| 摘 要： | 【目的】建立一种快速准确的三七根腐病病原真菌毁坏柱孢霉(Cylindrocarpon destructans)的分子定量检测方法，探讨毁坏柱孢霉与植株生长和AM真菌侵染之间的数量效应关系。【方法】根据GenBank登录的毁坏柱孢霉rDNA基因IGS序列片段，设计特异性引物对CDU2和CDL2b，利用含有SYBR Green I的实时荧光定量PCR建立毁坏柱孢霉定量检测方法，检测三七根际土壤毁坏柱孢霉rDNA基因IGS片段拷贝数，并分析其与植株生物量和AM真菌侵染之间的关系。【结果】发病植株根际土壤毁坏柱孢霉rDNA基因IGS片段拷贝数显著高于健康植株。三七根际土壤中毁坏柱孢霉数量与植株地上部生物量以及菌根侵染强度呈显著负相关(P＜0.05)，与根系生物量以及根内丛枝丰度相关性不显著。【结论】基于实时定量PCR技术建立的毁坏柱孢霉的分子定量方法能够有效反映三七根际土壤中毁坏柱孢霉的数量及其与植株生长和AM真菌侵染的关系。
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| 关 键 词： | 三七，根腐病，毁坏柱孢霉，定量PCR，AM真菌 |
Molecular quantification of Cylindrocarpon destructans in the rhizosphere of Panax notoginseng for predicting plant growth response |
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| WU Zhao-Xiang,HAO Zhi-Peng,ZENG Yan,GUO Lan-Ping,HUANG Lu-Qi,WANG Yong and CHEN Bao-Dong.Molecular quantification of Cylindrocarpon destructans in the rhizosphere of Panax notoginseng for predicting plant growth response[J].Microbiology,2015,42(3):598-607. |
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| Authors: | WU Zhao-Xiang HAO Zhi-Peng ZENG Yan GUO Lan-Ping HUANG Lu-Qi WANG Yong and CHEN Bao-Dong |
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| Institution: | 1. State Key Laboratory of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China,1. State Key Laboratory of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China,2. China National Group Corporation of Traditional and Herbal Medicine, Beijing 100097, China,3. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,3. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,4. Wenshan Institute of Sanchi Ginseng, Wenshan, Yunnan 663000, China and 1. State Key Laboratory of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China |
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| Abstract: | Objective] To establish a fast and accurate quantification method for Cylindrocarpon destructans causing root rot disease in the rhizosphere of Panax notoginseng, and to reveal the correlation of C. destructans abundance with plant growth and AM fungal colonization. Methods] Based on the rDNA IGS sequence of C. destructans obtained from GenBank, specific primer pair CDU2 and CDL2b was designed. Molecular quantification of C. destructans by real-time PCR with SYBR Green I was established. IGS fragment copies of C. destructans in the rhizosphere of Panax notoginseng in field was detected with the newly established method and its relationships with plant biomass and AM fungal colonization were statistically analyzed. Results] C. destructans was more abundant in rhizosphere of infected P. notoginseng than that of healthy plants (P<0.05). IGS fragment copies of C. destructans was negatively correlated with shoot biomass and mycorrhizal colonization rate, but the correlation between IGS fragment copies of C. destructans and root biomass and arbuscule abundance were insignificant. Conclusion] The quantification method for C. destructans based on real-time PCR could well reflect the population dynamics of C. destructans in the rhizosphere of P. notoginseng in relation to plant growth and AM fungal colonization. |
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| Keywords: | Panax notoginseng Root rot disease Cylindrocarpon destructans Real-time PCR AM fungi |
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