【摘要】 开展丛枝菌根(Arbuscular mycorrhizal,AM)真菌的群落结构与多样性研究对促进菌物多样性的保护与发掘及生态系统的可持续发展均具有重要意义,而研究方法是实现和推进相关研究的重要基础.本文首先简要回顾AM真菌的研究方法发展历程,介绍基于孢子形态鉴定的形态学方法和以聚合酶链式反应(Polymerase chain reaction,PCR)、变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)、454高通量测序为代表的分子生物学方法的检测原理和分析步骤.然后,结合本实验室利用孢子计数与鉴定、定量PCR与PCR-DGGE以及454高通量基因测序等3套检测手段解析长期不同施肥影响土壤AM真菌群落结构和多样性的研究案例,比较分析不同方法所得AM真菌群落结构与多样性的研究结果.综合来看,不同方法既有相互印证和互为补充的效果,又有不尽一致甚至互相矛盾的地方.最后,着重就AM真菌高通量测序技术的升级及与生物标记物方法、同位素标记技术等的联用提出了研究展望.更多还原

【Abstract】 Investigating the community structure and diversity of arbuscular mycorrhizal(AM) fungi is important for accelerating the protection and application of mycodiversity, and for the sustainable development of ecosystems. Nevertheless, establishment of related research methods is the key to realizing and promoting such investigations. In this paper, we first provide a very brief historical survey on research methods of AM fungi, and introduce the principles and steps of both morphological and molecular biological methods, including spore morphological identification, polymerase chain reaction(PCR), denaturing gradient gel electrophoresis(DGGE), and 454 pyrosequencing. Then, the results related to the long-term fertilization effects on soil AM fungal community structure and diversity revealed by spore calculation and identification, real-time PCR and PCR-DGGE, and 454 pyrosequencing were integrated for method comparison. Altogether, different methods could not only complement each other or have effects of mutual corroboration, but also have in consistencies and contradictions. Finally, the future development of research methods for AM fungi are proposed, including an upgrade of the pyrosequencing technique, and its combined application with biomarker methods, isotope labeling techniques, and others.更多还原