中国中药杂志

2021, v.46(08) 1910-1919

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基于高通量测序和传统培养分离方法的药用植物内生菌资源研究策略
Research strategies for endophytes in medicinal plants based on high-throughput sequencing and traditional culture and isolation methods

王红阳;康传志;王升;蒋待泉;彭政;徐扬;杜用玺;张燕;刘大会;郭兰萍;
WANG Hong-yang;KANG Chuan-zhi;WANG Sheng;JIANG Dai-quan;PENG Zheng;XU Yang;DU Yong-xi;ZHANG Yan;LIU Da-hui;GUO Lan-ping;State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences;Hubei University of Chinese Medicine;Guangdong Pharmaceutical University;

摘要(Abstract):

药用植物内生菌的研究主要依赖传统培养分离的方法,因其具有促生长、提高抗逆性、促进药用活性成分积累或直接产生药用活性成分等功能,逐渐受到广泛关注。然而,通过比较传统培养分离和高通量测序的研究结果发现,用传统方法分离的菌株并不是药用植物真实的优势内生菌。传统培养分离方法具有一定的盲目性和随机性导致在培养基的选择、培养时间、物种之间相互作用等方面缺乏标准。相反,高通量测序技术是近十几年来发展起来的新兴分子生物学技术,因其分辨水平高、不需要培养等特点,可以对环境微生物的群落结构和多样性进行深入分析。因此,该文提出用高通量测序技术指导传统培养分离药用植物内生菌的策略。首先,借助高通量测序技术对药用植物内生菌结构和多样性进行整体了解,确定宿主的优势内生菌;然后根据各优势内生菌的特性设计或查询适合其生长的培养基进行培养分离;最后对分离菌株进行功能研究。高通量测序指导传统培养分离药用植物内生菌可以避免研究者漏筛对宿主重要的功能菌株,扩大药用植物内生菌的研究范围,有利于药用植物内生菌的深入挖掘和利用。
The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.

关键词(KeyWords): 高通量测序技术;传统培养分离方法;药用植物;内生菌
high-throughput sequencing technology;traditional culture and isolation methods;medicinal plants;endophytes

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划项目(2017YFC1700701,2017YFC1700704,2018YFD0201100);; 国家自然科学基金项目(81891014,81874337);; 中国中医科学院重点领域研究项目(ZZ10-027);; 中央本级重大增减支项目(2060302);; 国家发改委标准化项目(ZYBZH-C-HLJ-17,ZYBZH-C-GD-07);; 现代农业产业技术体系建设专项(CARS-21);; 中央级公益性科研院所基本科研业务费专项(ZZ13-036-2)

作者(Author): 王红阳;康传志;王升;蒋待泉;彭政;徐扬;杜用玺;张燕;刘大会;郭兰萍;
WANG Hong-yang;KANG Chuan-zhi;WANG Sheng;JIANG Dai-quan;PENG Zheng;XU Yang;DU Yong-xi;ZHANG Yan;LIU Da-hui;GUO Lan-ping;State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences;Hubei University of Chinese Medicine;Guangdong Pharmaceutical University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20210127.101

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