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 Beijing 100700, China Hubei University of Chinese Medicine Wuhan 430065, China.
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2021 Apr;46(8):1910-1919. doi: 10.19540/j.cnki.cjcmm.20210127.101.
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.
药用植物内生菌的研究主要依赖于传统的培养和分离方法。由于其具有促进宿主生长、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,药用植物内生菌逐渐受到广泛关注。然而,通过将传统培养分离结果与高通量测序结果进行比较发现,传统方法分离出的菌株并非药用植物真正的优势内生菌。传统方法的盲目性和随机性导致在培养基选择、培养时间以及物种间相互作用等方面缺乏标准。相反,高通量测序技术是近几十年来发展起来的一种新兴分子生物学技术。由于其高分辨率水平和无需培养的特点,它可用于深入分析环境微生物的群落结构和多样性。因此,我们提出了利用高通量测序技术指导药用植物内生菌传统培养和分离的策略。首先,通过高通量测序技术全面了解药用植物的内生菌结构和多样性,明确宿主的优势内生菌。然后根据各优势内生菌的特点设计或查询适合其生长的培养基进行培养和分离。最后,研究分离菌株的功能。该方法可防止研究人员遗漏宿主的重要功能菌株,扩大药用植物内生菌的研究范围,便于对药用植物内生菌进行深入挖掘和利用。