秀珍菇染料脱色过氧化物酶基因的克隆及表达分析  

Cloning and Expression Analysis of PpDypA Gene of Pleurotus pulmonarius

作　　者：王伟科[1] 陆娜[1] 林佳瑶 陈观平[2] WANG Wei-ke;LU Na;LIN Jia-yao;CHEN Guan-ping(Hangzhou Academy of Agricultural Sciences,Hangzhou 310000,China;Zhejiang Academy of Traditional Chinese Medicine,Hangzhou 310012,China)

机构地区：[1]杭州市农业科学研究院,浙江杭州310000 [2]浙江省中医药研究院,浙江杭州310012

出　　处：《中国食用菌》2023年第6期56-61,68,共7页Edible Fungi of China

基　　金：浙江省食用菌新品种选育重大科技专项(2021C02073);杭州市农科院科技创新项目(2022HNCT-03)

摘　　要：通过对经低温刺激(10℃)和恒温(26℃)培养的秀珍菇菌丝体进行高通量转录组测序,利用生物信息学分析筛选获得秀珍菇染料脱色过氧化物酶全长基因(命名为PpDypA)。为了探究PpDypA在低温刺激后菌丝体中的功能与表达情况,克隆基因后进行生物信息学分析,结果表明该基因全长为1515 bp,编码504个氨基酸,其蛋白相对分子质量约54.8 kDa。氨基酸序列系统进化分析结果显示,染料脱色过氧化物酶家族具有很高的同源性,PpDypA编码蛋白与紫孢侧耳亲缘关系最近。经实时荧光定量PCR检测,该基因在秀珍菇经过低温处理后表达量显著上升,推测其可能经低温诱导表达,促使菌丝体启动抗氧化防御系统缓解低温逆境胁迫,且促进木质素降解从而为原基形成提供营养和能量。To obtain the full-length of dye-decolorizing peroxidase gene of Pleurotus pulmonarius(named PpDypA)and to analyze the function and expression of PpDypA in the process of primordial formation of P.pulmonarius after low(10℃)and room(26℃)temperature stimulation.The full-length of PpDypA was isolated through RNA-seq,in order to investigate the function and expression of PpDypA in mycelium after low-temperature stimulation.The results showed that the full-length of PpDypA is 1515 bp in length.The deduced amino acid sequence of PpDypA has 504 amino acid residues which form a 54.8 kDa polypeptide.Phylogenetic analysis of the amino acid sequences showed that the DypA family has a high degree of homology,and the protein encoded by the PpDypA was more related to the Pleurotus sapidus.After real-time qPCR,the expression of the gene increased significantly after low-temperature treatment,and it was hypothesized that its expression may be induced by low temperature,which prompted the mycelium will activate the antioxidant defense system to alleviate the low-temperature adversity stress,and promote the degradation of lignin so as to provide nutrients and energy for the formation of the primordium.

关 键 词：秀珍菇 染料脱色过氧化物酶基因 克隆 序列分析 表达分析 

分 类 号：S646.1[农业科学—蔬菜学]