SCAR Makers and Multiplex PCR-Based Rapid Molecular Typing of Lentinula edodes Strains |
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Authors: | Xueqian Wu Haibo Li Weiwei Zhao Lizhong Fu Huazheng Peng Liang He Junwen Cheng Hailong Wei Qingqi Wu |
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Institution: | (1) Biotechnology, Zhejiang Forestry Academy, Liuhe road 399, Hangzhou, 310023, Zhejiang Province, China;(2) Zhejiang Provincial Key Laboratory of Biological and Chemical Utilization of Forest Resource, Hangzhou, 310023, Zhejiang Province, China;(3) Northeast Agricultural University, Harbin, 150030, China; |
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Abstract: | Lentinula edodes is the second most important cultivated mushroom worldwide, the most commercial strains have been identified only through
traditional phenotypic analysis. In this study, a simple rapid PCR-based molecular method was developed for distinguishing
commercial strains of L. edodes by developing specific sequence characterized amplified region (SCAR) markers and establishing multiplex PCR assays with
the SCAR primers. Derived from the randomly amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP)
techniques, 10 informative SCAR markers were generated from 10 polymorphic RAPD and SRAP bands. The differences in SCAR phenotypes
among different strains made these SCAR markers potentially useful to characterize 6 strains and identify them from other
studied strains. Moreover, different SCAR phenotypes also made the other 17 studied strains to be divided into four distinguishable
groups. The multiplex PCR assays were further established for the joint use of some SCAR markers efficiently. Compared with
some identification methods reported previously, the special feature of this new molecular method is technically rapid and
convenient in the practical use and suitable for analyzing large numbers of samples. Thus, the simple rapid PCR-based molecular
method can be used as a helpful assistant tool for the lentinula industry. To our knowledge, this study is the first to describe
a development of a new SCAR maker-based multiplex PCR assay for rapid molecular typing of edible mushroom. |
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