Hairpin-mediated down-regulation of the urea cycle enzyme argininosuccinate lyase in Agaricus bisporus
Hairpin-mediated down-regulation of the urea cycle enzyme argininosuccinate lyase in Agaricus bisporus
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DOI：
10.1016/j.mycres.2008.01.009
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发表时间：
2008-06-01
影响因子：
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通讯作者：
Burton, Kerry S.
中科院分区：
文献类型：
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作者：
Eastwood, Daniel C.;Challen, Michael P.;Burton, Kerry S.
针对栽培蘑菇双孢蘑菇，开发了双链（ds）RNA发夹介导的下调系统，并研究了尿素循环酶精氨琥珀酸裂合酶（asl）在蘑菇采后发育中的作用。构建发夹表达载体以启动as]的下调，并通过根癌农杆菌介导的转化引入双孢蘑菇中。在两个转化体中，asl 的转录物显着减少（93.1% 和 99.9%），并且发夹载体转基因序列在整个子实体发育过程中得以维持。 Southern 分析中观察到单个和多个发夹整合事件。 asl下调的转化体在收获后子实体发育过程中表现出产量和菌盖扩张降低。发夹转化菌株和对照菌株之间的尿素水平没有可检测到的差异。这是首次报道双孢蘑菇中引入 dsRNA 发夹导致基因表达减少，该技术的应用将促进蘑菇的功能研究。 (C) 2008 英国真菌学会。由爱思唯尔有限公司出版。保留所有权利。
A double-stranded (ds) RNA hairpin-mediated down-regulation system was developed for the cultivated mushroom Agaricus bisporus, and the role of the urea cycle enzyme argininosuccinate lyase (asl) in mushroom post-harvest development was investigated. Hairpin expression vectors were constructed to initiate down-regulation of as] and introduced into A. bisporus by Agrobacterium tumefaciens-mediated transformation. Transcripts of asl were significantly reduced (93.1 and 99.9%) in two transformants and hairpin vector transgene sequences were maintained throughout sporophore development. Single and multiple hairpin integration events were observed in Southern analysis. Transformants with down-regulated asl exhibited reduced yield and cap expansion during post-harvest sporophore development. There were no detectable differences in urea levels between the hairpin-transformed and control strains. This is the first report of reduced gene expression resulting from the introduction of dsRNA hairpins in A. bisporus and the applications of this technology will facilitate functional studies in the mushroom. (C) 2008 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.