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(cè)耳屬分子系統(tǒng)學(xué)研究

馬富英　 羅晶　 羅信昌　

摘　要：對(duì)側(cè)耳屬18個(gè)種52個(gè)菌株以及雙孢蘑菇、香菇和亞側(cè)耳各1個(gè)菌株的28S rDNA 5'端、ITS和IGS1進(jìn)行PCR擴(kuò)增,擴(kuò)增4個(gè)屬的28S rDNA 5'端均得到一長(zhǎng)度為1.46 kb的片段,側(cè)耳屬ITS擴(kuò)增片段長(zhǎng)680 bp,亞側(cè)耳700 bp,雙孢蘑菇和香菇720 bp,側(cè)耳屬IGS1擴(kuò)增片段長(zhǎng)度發(fā)生變異,紅平菇和桃紅側(cè)耳為1.1 kb,其他側(cè)耳為1.0 kb,亞側(cè)耳700 bp,雙孢蘑菇和香菇1.3 kb.對(duì)擴(kuò)增片段分別用7種限制性內(nèi)切酶酶切后進(jìn)行聚類分析,結(jié)果表明雙孢蘑菇、香菇和亞側(cè)耳可與側(cè)耳分開,18種側(cè)耳分為五大類:具核側(cè)耳,紅平菇和桃紅側(cè)耳,鮑魚菇和囊蓋側(cè)耳,金頂側(cè)耳,其他側(cè)耳聚為一類,鮑魚菇和囊蓋側(cè)耳,紅平菇和桃紅側(cè)耳都只代表1個(gè)種.

關(guān)鍵詞：側(cè)耳屬; 大亞基核糖體DNA; 轉(zhuǎn)錄間區(qū); 基因間區(qū); 聚合酶鏈?zhǔn)椒磻?yīng); 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(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物國(guó)家重點(diǎn)實(shí)驗(yàn)室,華中農(nóng)業(yè)大學(xué)應(yīng)用真菌研究所,武漢,430070）　

　　　　　羅晶（華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物國(guó)家重點(diǎn)實(shí)驗(yàn)室,華中農(nóng)業(yè)大學(xué)應(yīng)用真菌研究所,武漢,430070）　

　　　　　羅信昌（華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物國(guó)家重點(diǎn)實(shí)驗(yàn)室,華中農(nóng)業(yè)大學(xué)應(yīng)用真菌研究所,武漢,430070）　

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華中農(nóng)業(yè)大學(xué)學(xué)報(bào)

JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY

2004 Vol.23 No.1