采用表面消毒法,从浙江天目山野生山胡椒[Lindera glauca (Sieb. et Zucc.) Blume]的茎、叶和树皮中分离出内生真菌,基于 ITS 序列分析进行分类鉴定;并以内生真菌的分离率、定殖率、分离频率、多样性指数( H’)及相似性系数为指标,分析了山胡椒内生真菌的菌群组成及多样性。 结果显示:在 26 株山胡椒样株的 728 块组织块中共分离得到 328 株内生真菌(茎、叶和树皮中分别有 161、40 和 127 株);共鉴定出 44 个分类单元(茎、叶和树皮中各有19、18 和 28 个),其中 25 个分类单元鉴定到种、17 个鉴定到属、2 个鉴定到科,ITS 序列的 GenBank 登录号从JF502420 至 JF502462。 在 44 个分类单元中,有 40 个分类单元属于子囊菌(310 株),存在于山胡椒的各个部位;仅有 4 个分类单元属于担子菌(18 株),且仅存在于茎和树皮中。山胡椒茎、树皮和叶中内生真菌的定殖率分别为65% 、60% 和 15% ,分离率分别为 0. 77、0. 61 和 0. 19;叶和树皮中内生真菌的多样性指数均为 2. 63,远大于茎(H’ =1. 83)。 山胡椒内生真菌的优势属为 Phomopsis、Paraconiothyrium、Phoma 和 Colletotrichum,大量存在于叶、茎和树皮中。 山胡椒茎与树皮、茎与叶及叶与树皮间内生真菌的相似性系数分别为 0. 27、0. 19 和 0. 18,显示树皮和叶之间以及树皮和茎之间内生真菌的组成极不相似。 研究结果表明:山胡椒体内存在大量的内生真菌,其茎、叶和树皮的内生真菌菌群组成具有一定程度的多样性和差异性,且内生真菌的分布具有组织特异性。

By the surface sterilization method, endophytic fungi were isolated from stem, leaf and bark of wild Lindera glauca ( Sieb. et Zucc.) Blume from Tiammu Mountain of Zhejiang Province, and were identified and classified according to ITS sequence analysis result. Taking isolation rate ( IR ),colonization rate(CR), relative frequency (RF), Shannon-Wiener biodiversity index (H’) and similarity coefficient ( Cs ) as indexes, the composition and diversity of endophytic fungal community were analyzed. The results show that 328 strains of endophytic fungi (161 strains in stem, 40 strains in leaf and 127 strains in bark) are obtained from 728 tissues of 26 individuals of L. glauca. 44 taxa are identified (19 taxa in stem, 18 taxa in leaf and 28 taxa in bark), in which, 25 taxa are identified to species, 17 taxa to genus and 2 taxa to family. The accession numbers of their ITS sequences in GenBank are from JF502420 to JF502462. In all of 44 taxa, 40 taxa belong to Ascomycota (310 strains) which exist in all parts of L. glauca, and 4 taxa belong to Basiodiomycota (18 strains) which only exist in stem and bark of L. glauca. Endophytic fungal CR in stem, bark and leaf of L. glauca is 65% , 60% and 15% , IR is 0. 77, 0. 61 and 0. 19, respectively. Endophytic fungal H’ in leaf and bark both is 2. 63, which is obviously higher than that in stem (H’ = 1. 83). The dominant genera of endophytic fungi in L. glauca are Phomopsis, Paraconiothyrium, Phoma and Colletotrichum, most of them exist in leaf, stem and bark. Cs value of endophytic fungi between stem and bark, stem and leaf, leaf and bark is 0. 27, 0. 19 and 0. 18, respectively, which shows the composition of endophytic fungi between bark and leaf or bark and stem is not extremely similar. It is indicated that there are a lot of endophytic fungi in L. glauca, the composition of endophytic fungal community in stem, leaf and bark has a certain diversity and difference, and the distribution of endophytic fungi has the tissue specificity.