四川省雅江松茸菌的分离与系统发育

Isolation and phylogenetic study of Tricholoma spp. isolated from Yajiang, Sichuan, China

对外生菌根真菌-松茸的纯培养条件进行了探讨,并从采集自四川省雅江县的松茸子实体中获得了10株松茸菌的纯培养物;分别以NS1和NS6 ,NS1和NS8,ITS4和ITS5为引物,对分离获得的松茸菌进行了18S r DNA PCR- RFL P和ITS PCR-RFL P分析,结果显示,用Alu I,H ae III,H inf I和Msp I四种限制性内切酶,这些松茸菌株的18S r DNA、ITS片段的酶切图谱完全相同;代表菌E7的ITS序列分析结果表明,本研究分离的松茸菌与Tricholoma matsutake的菌株在系统发育上高度同源。

The culturing condition for isolating ecotomycorrhizal fungus Tricholoma spp. was performed, and 10 matsutake isolates were got from Yajiang, Sichuan Province. Using NS1 and NS6, NS1 and NS8, ITS4 and ITS5 primer pairs, the 18S rDNA and ITS fragments of these matsutake strains was amplified, and digested by AluI, HaeIII, HinfI and MspI, respectively. The results showed that these strains had the same restriction patterns for each restriction enzyme, which showed that the homology among these strains were very high. Furthermore, the ITS sequence of representative strain E7 was analyzed, and compared with the data in the Genebank, and then the phylogenic tree was constructed. The results showed that the ITS sequence of E7 was the same as that of T. matsutake KBF20T07 which was isolated from South Korea, and the ITS sequence of E7 had very high similarity to those of the most other matsutake strains in the Genebank. The phylogenetic analysis showed that E7 and the matsutake strains from Genebank were in the same phylogenic branch, which suggested that, from the taxonomic point of view, these matsutake strains isolated from Yajiang belonged to Tricholoma matsutake.