药用真菌猪苓共生的蜜环菌种类研究

Investigation of Symbiotic Armillaria species with Chinese Traditional Medicinal Fungus Polyporus umbellatus

目的对国内猪苓产区与猪苓共生的蜜环菌种类进行探究。方法经PDA平板培养后长出健壮的蜜环菌根状菌索。运用IGS测序技术对分离的22株蜜环菌菌株的纯培养菌种进行分子鉴定,并基于蜜环菌的IGS序列进行核酸序列数据库Gen Bank同源序列比对、构建系统发育树。结果本实验获得的猪苓菌核组织分离的22株蜜环菌菌株为其纯培养菌种。基于IGS的系统发育分析表明,与猪苓菌核共生的蜜环菌分别属于Armillaria cepistipes,A.gallica,A.ostoyae以及A.mellea 4个种,其中A.cepistipes,A.gallica分别分离得到10株,为优势种。结论猪苓共生对蜜环菌物种专一性较低,与相关书籍及文献报道的与猪苓共生蜜环菌为单一A.mellea种的结论不符。与猪苓共生的蜜环菌多样性及不同种类蜜环菌对猪苓菌核生长发育的影响需进一步研究。

OBJECTIVE To identify symbiotic Armillaria species with Polyporus umbellatus. METHODS Armillaria was cul- tured on PDA pahes. To determine whether the strains were pure culture, the internal intergenic spacer （IGS） region were sequenced. Furthermore, the NCBI BLAST program was used to search similar sequences in the GenBanksequence database for the IGS sequence of the species on homology, and a phylogenetic tree was constructed based on neighbor-joining method. RESULTS All isolates were similar in colony morphology and grew well in PDA medium with well-developed rhizomorphs. The isolated 22 strains were pure culture of the fungus. Phylogenetic analysis based on IGS sequence indicated that the symbiotic Armillaria species with Polyporus umbellatus be- longed to A. cepistipes, A. gallica, A. ostoyae, and A. mellea. CONCLUSION In the present study, phylogenetic analysis based on molecular method was carried out for the symbiotic Armillaria species with P. umbellatus for the first time. The results suggest that Ar- millaria species have no specificity with P. umbellatus, which is different from the previous reports. Further research will be done on the growth of P. umbellatus affected by different kinds of Armillaria species.