不同细胞质的金针菇菌丝差异蛋白分析  

Comparative Proteomic Study on Flammulina filiformis Mycelia under Different Cytoplasmic Environments

作　　者：宋淋浩 卫银 武晨剑 常明昌[1,2] 孟俊龙 刘靖宇[1,2] SONG Linhao;WEI Yin;WU Chenjian;CHANG Mingchang;MENG Junlong;LIU Jingyu(College of Food Science and Engineering,Shanxi Agricultural University,Taigu,Shanxi 030801,China;Collaborative Innovation Center of Quality and Efficiency of Loess Plateau Edible Fungi in Shanxi,Shanxi Agricultural University,Taigu,Shanxi 030801,China)

机构地区：[1]山西农业大学食品科学与工程学院,山西太谷030801 [2]黄土高原食用菌提质增效协同创新中心,山西太谷030801

出　　处：《食用菌学报》2020年第2期8-16,共9页Acta Edulis Fungi

基　　金：山西省自然科学基金面上项目(201801D121240);山西省煤基重点科技攻关项目(FT2014-03-01);山西省重点研发计划重点项目(201603D21106)。

摘　　要：以黄色金针菇(Flammulina filiformis)单核菌株Y33和白色金针菇单核菌株W8为供试菌株,采用双向核迁移技术,获得一对具有相同细胞核、不同细胞质的双核菌株Y8-33和W8-33(简称异质双核菌株)。采用菌落形态观察、显微镜检查与分子标记对这两个双核菌株进行验证分析,并以菌株W8-33菌丝样品为对照,采用双向凝胶电泳技术对金针菇异质双核菌株菌丝的蛋白质组进行分析。结果表明:异质双核菌株Y8-33和W8-33菌丝均具有锁状联合结构,在菌落形态上呈现明显差异,但ISSR分子标记的PCR扩增产物具有很高的相似性。在Y8-33菌丝样品中共鉴定出17个差异蛋白,包括8个上调蛋白、9个下调蛋白;其中,14-3-3蛋白、微管蛋白、谷胱甘肽-S-转移酶、果糖二磷酸醛缩酶、ARF/SAR蛋白家族以及热激蛋白HSP90、HSP70和HSS1差异蛋白可能在由于受体细胞不同所导致的不同细胞质环境下,对金针菇的生长生理起到重要的调控作用。Using the bi-directional nuclear migration technique, two monokaryotic strains of Flammulina filiformis, a yellow monokaryotic strain Y33 and a white monokaryotic strain W8, were used to generate a pair of dikaryotic alloplasmic strains with the same nucleus. The resultant dikaryotic alloplasmic strains, Y8-33 and W8-33, differed in cytoplasm color. Y8-33 had yellow cytoplasm, whereas W8-33 had white cytoplasm. Y8-33 and W8-33 were first verified in terms of colony morphology, microscopic structures, and ISSR molecular markers, and then compared for their mycelial proteomes by two-dimensional gel electrophoresis(2-DE). The results showed that clamp connection structures were found in hyphae of both strains. Although Y8-33 and W8-33 formed morphologically different colonies, their PCR products using ISSR(inter-simple sequence repeat) molecular markers were highly consistent. Compared with W8-33, Y8-33 contained a total of 17 deferentially expressed proteins(DEPs), of which 8 were up-regulated and 9 were down-regulated. Some of these proteins were identified as 14-3-3 protein, tubulin, glutathione-S-transferase, fructose-bisphosphate aldolase, ARF/SAR protein family, and heat shock proteins HSP90, HSP70 and HSS1. The above-mentioned DEPs may play important roles in regulating growth physiology of F. filiformis under different cytoplasmic environments.

关 键 词：金针菇 双向核迁移 双向电泳 

分 类 号：S646.15[农业科学—蔬菜学]