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基于rDNA PCR-RFLP分析的侧耳属分子系统学研究

2004-11-16 00:000

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分析的侧耳属分子系统学研究

马富英  罗晶  罗信昌 

摘 要:对侧耳属18个种52个菌株以及双孢蘑菇香菇和亚侧耳各1个菌株的28S rDNA 5'端、ITS和IGS1进行PCR扩增,扩增4个属的28S rDNA 5'端均得到一长度为1.46 kb的片段,侧耳属ITS扩增片段长680 bp,亚侧耳700 bp,双孢蘑菇和香菇720 bp,侧耳属IGS1扩增片段长度发生变异,红平菇和桃红侧耳为1.1 kb,其他侧耳为1.0 kb,亚侧耳700 bp,双孢蘑菇和香菇1.3 kb.对扩增片段分别用7种限制性内切酶酶切后进行聚类分析,结果表明双孢蘑菇、香菇和亚侧耳可与侧耳分开,18种侧耳分为五大类:具核侧耳,红平菇和桃红侧耳,鲍鱼菇和囊盖侧耳,金顶侧耳,其他侧耳聚为一类,鲍鱼菇和囊盖侧耳,红平菇和桃红侧耳都只代表1个种.

关键词:侧耳属; 大亚基核糖体DNA; 转录间区; 基因间区; 聚合酶链式反应; 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.罗信昌,Corresponding author. E-mail: xinchangluo@mail.hzau.edu.cn

Author Unit:马富英(华中农业大学农业微生物国家重点实验室,华中农业大学应用真菌研究所,武汉,430070) 

     罗晶(华中农业大学农业微生物国家重点实验室,华中农业大学应用真菌研究所,武汉,430070) 

     罗信昌(华中农业大学农业微生物国家重点实验室,华中农业大学应用真菌研究所,武汉,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

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[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

华中农业大学学报

JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY

2004 Vol.23 No.1

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