PCR-BASED RESTRICTION ANALYSIS OF INTERNAL TRANSCRIBED SPACERS OF NUCLEAR RIBOSOMAL DNA IN THE GENUS PLEUROTUS
Abstract：The internal transcribed spacers of the nuclear ribosomal DNA of eighteen Pleurotus taxa (52 isolates), together with outgroups A. bisporus, L. edodes, H. serotina, were amplified using PCR and then digested with seven restriction endonucleases. PCR-RFLP results showed that AluI, Hae III, HinfI, TaqI, HhaI, MspI could divide 52 isolates into 6, 5, 5, 4, 2, 2 groups respectively and distinguish P. citrinopileatus from the other Pleurotus. No restriction site was observed for BamHI in all isolates. Dendrogram based on PCR-RFLP suggested that seven groups were distinguishable for Pleurotus on 93% similarity coefficient, i.e. P. ostreatus complex (including P. ostreatus, P. florida, P. sapidus, P. corticatus, P. cornucopiae, P. columbinus, P. spodoleucus, P. ferulae, P. nebrodensis and P. sp.), P. eryngii, P. pulmonarius-P. sajor-caju, P. tuber-regium, P. abalonus-P. cystiodisus, P. djamor- P. salmoneostramineus and P. citrinopileatus. The phylogenetic relationships between P.citrinopileatus and the other Pleurotus were more distant than L. edodes, A. bisporus and the other Pleurotus.
Keywords：Oyster mushroom, phylogeny, polymerase chain reaction, RFLP
侧耳属核rDNA转录间区扩增产物的限制性酶切分析
马富英 罗信昌
摘 要：对侧耳属18个形态种52个菌株和蘑菇属、香菇属、亚侧耳属各1个菌株的核糖体DNA转录间区进行PCR特异性扩增后,用7种限制性内切酶进行酶切分析.结果表明,BamHⅠ对所有的菌株均无酶切位点;AluⅠ, Hae Ⅲ,HinfⅠ, TaqⅠ, HhaⅠ, MspⅠ可分别将52个菌株分为6, 5, 5, 4, 2, 2组,且可将金顶侧耳与其它侧耳区分开.基于限制性酶切多态性构建的聚类图表明,在93%相似水平,可将所有的侧耳分为七组,第一组:糙皮侧耳、佛罗里达侧耳、美味侧耳、裂皮侧耳、黄白侧耳、哥伦比亚侧耳、灰白侧耳、白阿魏蘑、阿魏蘑及一些未定名的侧耳;第二组:刺芹侧耳;第三组:肺形侧耳和凤尾菇;第四组:具核侧耳;第五组:鲍鱼菇和囊盖侧耳;第六组:红平菇和桃红侧耳;第七组:金顶侧耳.聚类图结果进一步表明,金顶侧耳与其它侧耳的关系较双孢蘑菇和香菇与其它侧耳的关系要远.
关键词：侧耳,系统发育,聚合酶链式反应, 限制性片段长度多态性
CLC Number：Q939.96 Document ID：A
Article ID：1007-3515(2002)03-0356-0362
Foundation Item：Supported by the National Natural Science Foundation of China (国家自然科学基金资助项目)
Author Unit：马富英（农业部农业微生物重点实验室,华中农业大学植物保护系,武汉,430070）
罗信昌（农业部农业微生物重点实验室,华中农业大学植物保护系,武汉,430070）
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