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Optimization of a Sequence-Related Amplified Polymorphism (SRAP) Amplification System for Lentinula edodes

食用菌学报 2006年第4期 16-20页

作者： FU Li-zhong 1,2, WEI Hai-long 1,2, LI Hai-bo 1,2, WU Qing-qi 1,2, WU Da-feng 2, WU Xue-qian 1,2* 1 Biotechnology Research Institute, zhejiang academy of forestry science, Hangzhou 310023,China 2Lishui Edible Fungi Research and Development Center,zhejiang Essence Fungi Development Co., Ltd,Lishui,zhejiang 323000,China

A stable Sequence-Related Amplified Polymorphism （SRAP） molecular marker system for Lentinula edodes has been developed and optimized using genomic DNA extracted from fungal mycelium with sodium dodecyl sulfate-cetyltrimethylammonium bromide （SDS-CTAB）. A large number of amplification products and good reproducibility was achieved using a 20 μL reaction system consisting of 2 μL 10×Reaction Buffer（containing 1.5 mM Mg 2+）, 25 ng genomic DNA, 200 μM dNTP mixture, 1.0 mM Mg 2+, 0.4 μM forward and reverse primer （Me1 and Em16）, 1.5 U Taq polymerase, and ddH_ 2O to volume.

关键词： Lentinula edodes Molecular marker Optimization of amplification protocol SRAP

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rDNA ITS Sequence Analysis and Molecular Phylogeny of Four Fungal Strains Assigned to the Family Russulaceae

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食用菌学报 2007年第2期 29-32页

作者： FU Lizhong1, 2, LI Haibo1, 2, WEI Hailong1, 2, WU Qingqi1, 2, WU Dafeng2, WU Xueqian1, 21 Biotechnology Institute, zhejiang academy of forestry science, Hangzhou, zhejiang 310023,China 2Lishui Edible Fungi Research and Development Center,zhejiang Essence Fungi DevelopmentCompany Ltd,Lishui,zhejiang 323000,China

Internal transcribed spacer (ITS) sequences of four edible and medicinal fungi belonging to the Russulaceae, collected from the Lishui mountainous area of zhejiang Province, were cloned and sequenced. Comparisons with sequence data in GenBank were undertaken using BLAST, and the four ITS sequences, together with 15 reference sequences obtained from GenBank, were used to construct a phylogenetic tree. rDNA ITS sequences of the four strains ranged from 673 to 766 bp in length and between 47.4% and 49.48% in GC content. Phylogenetic data indicated strain R57 to be Russula cyanoxantha. Differences in the size and GC content of ITS sequences from strains R32 and R57 (R. cyanoxantha) and strain L37 (Lactarius sanguifluus) were significant, whereas the corresponding differences between strains R32 and R57 were relatively small. A close kinship, with only small detectable differences in their respective ITS sequences, existed between R. mustelina, R. virescens and R. parazurea. A similar situation applied to strains of L. sanguifluus and L. salmonicolor.

关键词： Russulaceae rDNA Internal Transcribed Spacer Sequence analysis Phylogeny Macrofungi

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