露蕊乌头内生真菌的分离与鉴定

Isolation and Identification of Fungal Endophyte in Aconitum gymnandrum

为探明甘肃露蕊乌头内生真菌的种类与种群分布情况,对甘肃天祝的露蕊乌头样品采用表面消毒法进行内生真菌分离,用形态学和ITS序列分析技术对分离得到的菌株进行种属鉴定,并通过MEGA5.05软件进行系统进化分析。结果表明,露蕊乌头内存在丰富的内生真菌,从其茎、叶、根、花和种子中分离得到的内生真菌共33株,经过鉴定,分别属于2纲、3目、3科、4属;总相对分离频率为93.94%;内生真菌对叶的侵染频率最高,茎次之,种子最少;青霉属(Penicillium sp.)在露蕊乌头植株内相对分离频率最高,是露蕊乌头内生真菌的优势菌种,且在上述组织内均存在,分布最广泛;将所得到的菌株使用MEGA 5.05软件作系统进化分析,可根据亲缘关系远近分为种群Ⅰ、种群Ⅱ和种群Ⅲ。试验结果可丰富乌头属植物内生真菌的种群多样性,也为后续露蕊乌头内生真菌的次生代谢产物研究奠定基础。

In order to investigate the species and population distribution of fungal endophyte in Aconitum gymnandrum collected from Tianzhu of Gansu province,endophytic fungal isolation was conducted by surface disinfection,species and ITS sequence analysis techniques for species identification of isolated strains,and systematic evolution analysis used by MEGA5.05 software.The results showed that there were abundant fungal endophytes in Aconitum gymnandrum,33 fungal endophytes were isolated from roots,stems,leaves,flowers and seeds,after identification,they were belonged to 2 classes,3 orders,3 families and 4 genera;the total relative isolatio frequency was 93.94%.Endophytic fungi have the highest frequency of leaf infection,the second is the stem,least in the seed;their species Penicillium sp.had the highest relative isolation frequency and was the dominant species of fungal endophytes in Aconitum gymnandrum,their species were found in all the above tissues and their distribution was the most extensive.According to phylogenetic analysis,these isolated strains were divided into populationⅠ,populationⅡand populationⅢ.These results can further enrich the diversity of fungal endophytes of Aconitum and lay a foundation for the research of secondary metabolites of endophytic fungi in Aconitum gymnandrum.