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

側耳屬核rDNA轉錄間區(qū)擴增產(chǎn)物的限制性酶切分析

馬富英　 羅信昌　

摘　要：對側耳屬18個形態(tài)種52個菌株和蘑菇屬、香菇屬、亞側耳屬各1個菌株的核糖體DNA轉錄間區(qū)進行PCR特異性擴增后,用7種限制性內切酶進行酶切分析.結果表明,BamHⅠ對所有的菌株均無酶切位點;AluⅠ, Hae Ⅲ,HinfⅠ, TaqⅠ, HhaⅠ, MspⅠ可分別將52個菌株分為6, 5, 5, 4, 2, 2組,且可將金頂側耳與其它側耳區(qū)分開.基于限制性酶切多態(tài)性構建的聚類圖表明,在93%相似水平,可將所有的側耳分為七組,第一組:糙皮側耳、佛羅里達側耳、美味側耳、裂皮側耳、黃白側耳、哥倫比亞側耳、灰白側耳、白阿魏蘑、阿魏蘑及一些未定名的側耳;第二組:刺芹側耳;第三組:肺形側耳和鳳尾菇;第四組:具核側耳;第五組:鮑魚菇和囊蓋側耳;第六組:紅平菇和桃紅側耳;第七組:金頂側耳.聚類圖結果進一步表明,金頂側耳與其它側耳的關系較雙孢蘑菇和香菇與其它側耳的關系要遠.

關鍵詞：側耳,系統(tǒng)發(fā)育,聚合酶鏈式反應, 限制性片段長度多態(tài)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 (國家自然科學基金資助項目)

Author Unit：馬富英（農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室,華中農(nóng)業(yè)大學植物保護系,武漢,430070）　

　　　　　羅信昌（農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室,華中農(nóng)業(yè)大學植物保護系,武漢,430070）　

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