摘要
为高效地分离闽楠内生真菌,以福建成熟闽楠林下自生闽楠苗木为研究对象,利用高通量培养法获得闽楠内生真菌并研究其生物多样性,研究不同稀释比例对内生真菌分离的影响并筛选最佳分离浓度。结果共获得656株内生真菌,经内转录间隔区(ITS)序列分析,鉴定为35种和12个未知种,未知种比例达25.53%。其中,黑腐皮壳科未知种(Valsaceae sp.,代表菌株P28)为分离频率最高的内生真菌,分离频率为12.04%。闽楠根、茎和叶片内生真菌辛普森指数均超过0.80,说明闽楠内生真菌多样性高;根部内生真菌多样性最高(香农指数2.304),其次为茎(香农指数2.234),最低的为叶(香农指数2.128)。有9种内生真菌的ITS序列相似度低于97%,共147株,占全部内生真菌株数的22.41%。从不同稀释比例的组织悬浮液中获得的内生真菌的数量和种类均存在差异,未经稀释的原液中分离出的内生真菌数量和种类均是最多的,共获得265株36种,分离种类占分离数量的13.58%;稀释16倍的组织悬浮液中分离出的内生真菌总数最少,共获得64株15种,但是分离种类占分离数量的23.44%。在一定范围内,随着稀释比例的增大,高通量培养法单位分离数量获得的种类增加。
Plant endophytic fungi demonstrate species and biological function diversity.The endophytic fungi of Phoebe bournei were isolated to enrich microbial resources,screen for growth-and resilience-promoting stains,and estimate the level of diversity of endophytic fungi.Samples from the roots,stems,and leaves of healthy P.bournei trees were collected and used to isolate the endophytic fungi using a high-throughput culturing method.The biodiversity of endophytic fungi was then calculated.The effects of different dilution ratios on the separation of endophytic fungi were assessed to determine the optimal dilution ratio.Results:a total of 656 endophytic fungi were obtained,and 35 known species and 12 unknown species were identified by internal transcribed spacer(ITS)sequence analysis.The proportion of unknown species was 25.53%.An unknown species of the family Valsaceae,represented by strain P28,was the most frequently isolated endophytic fungi,with an isolation frequency of 12.04%.All Simpson index values of endophytic fungi from P.bournei roots,stems,and leaves exceeded 0.8,indicating a high level of diversity.The highest level of diversity of endophytic fungi was found in the roots,with a Shannon index value of 2.304,followed by values of 2.234 in the stems and 2.128 in the leaves.The ITS sequence similarity values of nine species of endophytes were lower than 97%,including 147 strains,which accounted for 22.41%of all endophytes.The number and species of endophytic fungi isolated at different dilution ratios varied.The largest number and species of endophytic fungi were isolated from an undiluted stock solution.This included 265 strains and 36 species,which accounted for 13.58%of the isolated species.Only 64 strains,identified as 15 species,were isolated from the suspension and diluted 16 times.This was the lowest number among the treatments but still accounted for 23.44%of the isolated strains.Within a certain range,as the dilution ratio increased,the number of species identified from certain isolated strains increas
作者
陈少如
许晓晨
赵文宝
王倩倩
龙宇
张清华
CHEN Shaoru;XU Xiaochen;ZHAO Wenbao;WANG Qianqian;LONG Yu;ZHANG Qinghua(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)
出处
《森林与环境学报》
CSCD
北大核心
2022年第5期544-551,共8页
Journal of Forest and Environment
基金
国家自然科学基金项目“假单胞菌新种NP-1抗真菌物质鉴定及生物合成分子机制研究”(31901312)。
关键词
闽楠
内生真菌
高通量培养法
多样性
Phoebe bournei
endophytic fungi
high-throughput culturing
diversit
