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    基于rDNA PCR-RFLP分析的側(cè)耳屬分子系統(tǒng)學研究


    【發(fā)布日期】:2004-11-16  【來源】:
    【核心提示】:PhylogeneticRelationshipsofPleurotusspeciesBasedonrDNAPCR-RFLPAbstract:28SrDNA5'half,ITSandIGS1region

    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)學研究

    馬富英  羅晶  羅信昌 

    摘 要:對側(cè)耳屬18個種52個菌株以及雙孢蘑菇香菇和亞側(cè)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(cè)耳屬ITS擴增片段長680 bp,亞側(cè)耳700 bp,雙孢蘑菇和香菇720 bp,側(cè)耳屬IGS1擴增片段長度發(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.對擴增片段分別用7種限制性內(nèi)切酶酶切后進行聚類分析,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); 聚合酶鏈式反應(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)大學農(nóng)業(yè)微生物國家重點實驗室,華中農(nóng)業(yè)大學應(yīng)用真菌研究所,武漢,430070) 

         羅晶(華中農(nóng)業(yè)大學農(nóng)業(yè)微生物國家重點實驗室,華中農(nóng)業(yè)大學應(yīng)用真菌研究所,武漢,430070) 

         羅信昌(華中農(nóng)業(yè)大學農(nóng)業(yè)微生物國家重點實驗室,華中農(nóng)業(yè)大學應(yīng)用真菌研究所,武漢,430070) 

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    [2]Bunyard B A, Nicholson M S,Royse D J. A systematic assessment of Morchella using RFLP analysis of the 28S ribosomal RNA gene. Mycologia, 1994,86(6):762~772

    [3]Moncalvo J M, Wang H H, Hseu R S. Phylogenetic relationships in Ganoderma inferred from the internal transcribed spacers and 25S ribosomal DNA sequences. Mycologia, 1995, 87(2): 223~238

    [4]Bunyard B A, Nicholson M S, Royse D J. Phylogeny of the genus Agaricus inferred from restricition analysis of enzymarically amplified ribosomal DNA. Fungal Genetics and Biology, 1996, 20: 243~253

    [5]Ma F Y, Luo X C. PCR-based restriction analysis of internal transcribed spacers of nuclear ribosomal DNA in the genus Pleurotus. Mycosystema, 2002, 21(3): 356~362

    [6]White T J, Bruns T D, Lee S B. Amplication and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis N, Gelfand J, White T J, eds. PCR Protocols: A Guide to Methods and Applications. New York: Academic Press, 1990. 315~322

    [7]Georgiev O I, Nikolaev N, Hadjiolov A A, et al. The structure of the yeast ribosomal RNA genes. 4. Complete sequence of the 25 S rRNA gene from Saccharomyces cerevisae. Nucleic Acids Res, 1981, 9(24): 6953~6958

    [8]Walker W, Doolittle W. Redividing the basidiomycetes on the basis of 5S rRNA sequence. Nature, 1982, 299: 723~724

    [9]Ma F Y, Luo X C. Phylogeny of Pleurotus inferred from PCR-RFLP of 28S rDNA. Journal of Huazhong Agricultural University, 2002,21(3): 201~205

    華中農(nóng)業(yè)大學學報

    JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY

    2004 Vol.23 No.1

     
     
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