为明确药食两用植物朝天委陵菜(Potentilla supina)与深色有隔内生真菌(dark septate endophytes,DSE)之间的关系,以酸性品红染色和直接镜检观察朝天委陵菜根内DSE的定殖情况,采用常规组织分离方法分离DSE菌株并纯化培养,利用形态学和分子生物学相结合的方法鉴定DSE菌株,通过平板和液体培养了解DSE菌株的动态生长特性.结果显示:朝天委陵菜根内有DSE菌株定殖,菌丝和微菌核定殖率分别为59.2%和10.4%;从根部分离得到19株内生真菌,其中包含9株DSE,是朝天委陵菜内生真菌优势种.经形态学观察、比较和rDNA-ITS分析,将9株DSE中的CTW-9菌株鉴定为链格孢(Alternaria alternata),登录号为MT451845,该菌株在PDA平板和PDB培养液中均可正常生长并完成生活史,菌丝能形成哈氏网和类泡囊等结构.综上所述,DSE菌株在朝天委陵菜根部有较高的定殖率,CTW-9菌株的菌丝常形成哈氏网和类泡囊等特殊结构,可能有助于植物从环境中吸收和积累水分及营养物质.

In order to clarify the relationship between the medicinal and edible plant Potentilla supina with dark septate endophytes(DSE), acid fuchsin staining and direct microscopy were used to observe the colonization of DSE in the roots of P. supina. By DSE isolation and purification by conventional tissue isolation methods, DSE identification by morphology and molecular biology technique, DSE dynamic growth characteristics by plate and liquid culture was elaborated. The results showed that DSE strains colonized the roots of P. supina, the colonization rates of hyphae and microsclerotia were 59.2% and 10.4%, respectively. Nineteen endophytic fungus were isolated from the roots, including nine strains of DSE, which was the dominant population of endophytic fungi in P. supina. Among them, CTW-9 strain was identified as Alternaria alternata Keissler by morphological observation and rDNA-ITS analysis, its GeneBank accession number is MT451845. CTW-9 strain could grow and complete its life cycle normally in both PDA plate and PDB medium, in which, hyphae often could form Hartig net and vesicle-like structures. In conclusion, there were a high colonization frequency of DSE in P. supina plant roots, and Hartig net and vesicle-like structures formed by CTW-9 strain hyphae might be helpful to absorb water and- DOI:10.3969/j.issn.1000-1565.2023.06.011朝天委陵菜DSE的分离鉴定及动态生长特性吴艳清,白菁华,刘沺沺(保定学院 生物化工与环境工程学院,河北 保定 071002)摘 要:为明确药食两用植物朝天委陵菜(Potentilla supina)与深色有隔内生真菌(dark septate endophytes,DSE)之间的关系,以酸性品红染色和直接镜检观察朝天委陵菜根内DSE的定殖情况,采用常规组织分离方法分离DSE菌株并纯化培养,利用形态学和分子生物学相结合的方法鉴定DSE菌株,通过平板和液体培养了解DSE菌株的动态生长特性.结果显示:朝天委陵菜根内有DSE菌株定殖,菌丝和微菌核定殖率分别为59.2%和10.4%;从根部分离得到19株内生真菌,其中包含9株DSE,是朝天委陵菜内生真菌优势种.经形态学观察、比较和rDNA-ITS分析,将9株DSE中的CTW-9菌株鉴定为链格孢(Alternaria alternata),登录号为MT451845,该菌株在PDA平板和PDB培养液中均可正常生长并完成生活史,菌丝能形成哈氏网和类泡囊等结构.综上所述,DSE菌株在朝天委陵菜根部有较高的定殖率,CTW-9菌株的菌丝常形成哈氏网和类泡囊等特殊结构,可能有助于植物从环境中吸收和积累水分及营养物质. 关键词:深色有隔内生真菌(DSE);朝天委陵菜;链格孢;分离纯化;动态生长特性中图分类号:Q954.6 文献标志码:A 文章编号:1000-1565(2023)06-0646-07Isolation, identification and dynamic growth characteristics of DSE in Potentilla supinaWU Yanqing, BAI Jinghua, LIU Tiantian(College of Biochemical and Environmental Engineering, Baoding University, Baoding 071002, China)Abstract: In order to clarify the relationship between the medicinal and edible plant Potentilla supina with dark septate endophytes(DSE), acid fuchsin staining and direct microscopy were used to observe the colonization of DSE in the roots of P. supina. By DSE isolation and purification by conventional tissue isolation methods, DSE identification by morphology and molecular biology technique, DSE dynamic growth characteristics by plate and liquid culture was elaborated. The results showed that DSE strains colonized the roots of P. supina, the colonization rates of hyphae and microsclerotia were 59.2% and 10.4%, respectively. Nineteen endophytic fungus were isolated from the roots, including nine strains of DSE, which was the dominant population of endophytic fungi in P. supina. Among them, CTW-9 strain was identified as Alternaria alternata Keissler by morphological observation and rDNA-ITS analysis, its GeneBank accession number is MT451845. CTW-9 strain could grow and complete its life cycle normally in both PDA plate and PDB medium, in which, hyphae often could form Hartig net and vesicle-like structures. In conclusion, there were a high colonization frequency of DSE in P. supina plant roots, and Hartig net and vesicle-like structures formed by CTW-9 strain hyphae might be helpful to absorb water and- 收稿日期:2022-10-05 基金项目:河北省高等学校科学研究计划项目(QN2021401);河北省应用技术研究会项目(JY2021099) 第一作者:吴艳清(1982—),男,河北河间人,博士,保定学院高级实验师,主要从事微生物资源开发及应用研究.E-mail:wuyanqing@bdu.edu.cn第6期吴艳清等:朝天委陵菜DSE的分离鉴定及动态生长特性nutrition from environment for plants.