CAJ | 학술논문

为了验证中国林业微生物保藏管理中心库藏链格孢属(AlternariaNees)菌种资源分类地位的准确性，探索一种链格孢属真菌的快速复核鉴定方法，采用ITS序列分析、NCBI blast以及系统发育分析等方法，对中国林业微生物保藏管理中心库藏链格孢属的36个种及未定种的375个菌株进行分子复核鉴定以及系统发育关系研究。结果表明：研究的375个菌株中，有21个菌株的分类地位不准确，其中有9株菌的ITS序列与链格孢属菌种的同源性在92%~96%之间，另外12株菌的ITS序列与链格孢属菌种的同源性在60%~85%之间。基于ITS序列构建的NJ树显示，29个小孢子种的37个生物型形成了一个稳定的分支，而6个大孢子种与外群Ulocladiumsp.聚在一个分支。本研究中，ITS序列分析可以用来区分和鉴定形态差别明显的大孢子种，但无法区分亲缘关系较近的29个小孢子种。
위료험증중국임업미생물보장관리중심고장련격포속(AlternariaNees)균충자원분류지위적준학성，탐색일충련격포속진균적쾌속복핵감정방법，채용ITS서렬분석、NCBI blast이급계통발육분석등방법，대중국임업미생물보장관리중심고장련격포속적36개충급미정충적375개균주진행분자복핵감정이급계통발육관계연구。결과표명：연구적375개균주중，유21개균주적분류지위불준학，기중유9주균적ITS서렬여련격포속균충적동원성재92%~96%지간，령외12주균적ITS서렬여련격포속균충적동원성재60%~85%지간。기우ITS서렬구건적NJ수현시，29개소포자충적37개생물형형성료일개은정적분지，이6개대포자충여외군Ulocladiumsp.취재일개분지。본연구중，ITS서렬분석가이용래구분화감정형태차별명현적대포자충，단무법구분친연관계교근적29개소포자충。
In order to identify cultures of Alternaria Nees from China forestry culture collection center at molecular level by ITS sequence analysis. The internal transcribed spacer (ITS) region DNA fragment of 375 strains were amplified and sequenced by universal primers ITS1 and ITS4. And all the sequences of the 375 strains in present study were compared with the ITS sequences of related Alternaria species available in the GenBank database by BLAST search. Phylogenetic analysis was performed using the neighbor-joining method based on Paup 4.0 b10. ITS sequencing and NCBI BLAST results showed that: classification status of 21 strains from 375 trains were wrong, in which 9 strains had 92%-96% similarities with the reference strains, and the other 12 strains had 60%-85% similarities with the reference strains. The phylogenetic tree showed that 29 species were formed one cluster with 100% boot strap support, and the other seven species were divided into 6 clusters. The results indicated that:the ITS sequence could not be used to the separation of 29 species of Alternaria, but the other 6 species could be differentiated each other.