利用mRNA差异显示技术研究香菇发育相关基因.doc

利用mRNA差异显示技术研究香菇发育相关基因 蔡为明1李南羿1*谢鸣1高中山2金群力1 1浙江省农业科学院园艺研究所 杭州 310021 2浙江大学农业与生物技术学院园艺系 杭州310029 摘 要：以香菇子实体不分化突变株和菌株L98411为材料，利用mRNA差异显示技术研究二者的基因表达差异，共分离到21条差异片段，其中个分别与泛素、ATP合成酶、锌指蛋白、GTP结合蛋白、延伸因子线粒体前体、过氧化物酶、硫酯酶、类TrpB酶、2，4-二烯酰辅酶A还原酶、糖苷水解酶、热激蛋白、疏水蛋白等已知基因有较高的同源性，这表明香菇子实体分育是一个复杂的过程，涉及到胞内转运转录调控细胞分化蛋白合成各种代谢途径中的酶抗逆反应等诸多途径的协同调控。 关键词：差异显示突变体发育基因 Identification of developmentally related genes in Lentinula edodes by mRNA differential expression CAI Wei-Ming1 LI Nan-Yi1* XIE Ming1 GAO Zhong-Shan2 JIN Qun-Li 1 1 Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China 2College of Agriculture Biotechnology, Zhejiang University, Hangzhou 310029, China Abstract：Differential-isplay reverse transcription-PCR(DDRT-PCR) was used to investigate the difference of gene expression between unfruitificative mutant and wild type strain of Lentinula edodes, and 21 cDNA fragments were obtained. Twelve fragments showed high sequence similarity to known gene products, including ubiquitin, ATP synthase, zinc finger family protein, GTP binding protein, elongation factor g1 mitochondrial precursor, peroxiredoxin, thioesterase family protein, 2,4-dienoyl-CoA reductase, pyridoxal-phosphate dependent TrpB-like enzyme, glycoside hydrolase family 65, heat shock protein and hydrophobin. The genes indicate that the fruiting body development in L.edodes is a complex process involving several pathways and signal transduction such as intracellular trafficking, transcriptional regulation, cell division, protein synthesis, metabolism, and stress response. Keywords: differential display reverse transcription-PCR, mutant, fruiting body development, gene expression 基金项目：浙江省自然科学基金（No.305631）. *通讯作者Corresponding author. Tel: 057 E-mail: linanyi2000@ 香菇是世界上最重要的栽培食用菌之一。香菇子实体原基分化形成的菌盖、菌柄、菌褶，直接决定香菇的品质和产量。香菇子实体发育相关基因的分离及其在子实体分化发育中的表达研究，是获得控制子实体发育功能基因的重要途径，对阐明香菇子实体分化发育的分子机理具有重要意义菇子实体发育是一个复杂的细胞分化过程，除交配型基因外，还需一系列基因参与结实的遗传调控(张树庭和林方灿1997)。近年来，在香菇中相继克隆到了一些与香菇发育相关的基因，其中包括在担子菌