Phylogenetic Relationships of Pleurotus species Based on rDNA PCR-RFLP

Abstract：28S rDNA 5' half, ITS and IGS1 region of eighteen Pleurotus taxa, together with Agaricus bisporus, Lentinula edodes and Hohenbuehelia serotina were amplified using PCR and then digested with seven restriction endonucleases. A single uniform 1.46 kb product resulted from PCR amplification of 28S rDNA 5' half of four genera. Amplification of ITS resulted in a single 680 bp product for Pleurotus, 700 bp for H. serotina, 720 bp for L. edodes and A. bisporus. The length of IGS1 was varied: a 1.1 kb product for P. djamor and P. salmoneostramineus, a common product 1.0 kb for the other Pleurotus isolates, 700 bp for H. serotina, 1.3 kb for L. edodes and A. bisporus. Dendrogram based on PCR-RFLP of 28S rDNA 5'half, ITS and IGS1 suggested that H. serotina, L. edodes and A. bisporus could distinguish from Pleurotus, the isolates of Pleurotus were divided into five groups: P. tuber-regium, P. djamor and P. salmoneostramineus, P. abalonus and P. cystiodisus, P. citrinopileatus, the other isolates clustered together. P. abalonus and P. cystidiosus were the same species, as well as P. djamor and P. salmoneostramineus, P. pulmonarius and P. sajor-caju. rDNA PCR-RFLP was more suitable for taxonomy and phylogenetic relationships of Pleurotus.

Keywords：Pleurotus; 28S rDNA; ITS; IGS; PCR; RFLP

基于rDNA PCR-RFLP分析的側耳屬分子系統學研究

馬富英　 羅晶　 羅信昌　

摘　要：對側耳屬18個種52個菌株以及雙孢蘑菇、香菇和亞側耳各1個菌株的28S rDNA 5'端、ITS和IGS1進行PCR擴增,擴增4個屬的28S rDNA 5'端均得到一長度為1.46 kb的片段,側耳屬ITS擴增片段長680 bp,亞側耳700 bp,雙孢蘑菇和香菇720 bp,側耳屬IGS1擴增片段長度發生變異,紅平菇和桃紅側耳為1.1 kb,其他側耳為1.0 kb,亞側耳700 bp,雙孢蘑菇和香菇1.3 kb.對擴增片段分別用7種限制性內切酶酶切后進行聚類分析,結果表明雙孢蘑菇、香菇和亞側耳可與側耳分開,18種側耳分為五大類:具核側耳,紅平菇和桃紅側耳,鮑魚菇和囊蓋側耳,金頂側耳,其他側耳聚為一類,鮑魚菇和囊蓋側耳,紅平菇和桃紅側耳都只代表1個種.

關鍵詞：側耳屬; 大亞基核糖體DNA; 轉錄間區; 基因間區; 聚合酶鏈式反應; 限制性酶切分析

CLC Number：S646.1;Q78

Foundation Item：Supported by National Natural Science Foundation of China

Author Resume：Ma Fuying, female, born in 1971,Ph.D. Address:College of Life Science and Technology, Huazhong University of Science and Technology,Wuhan 430074.羅信昌,Corresponding author. E-mail: xinchangluo@mail.hzau.edu.cn

Author Unit：馬富英（華中農業大學農業微生物國家重點實驗室,華中農業大學應用真菌研究所,武漢,430070）　

　　　　　羅晶（華中農業大學農業微生物國家重點實驗室,華中農業大學應用真菌研究所,武漢,430070）　

　　　　　羅信昌（華中農業大學農業微生物國家重點實驗室,華中農業大學應用真菌研究所,武漢,430070）　

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華中農業大學學報

JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY

2004 Vol.23 No.1