【作者】 李南羿； 罗志姣； 刘春滟； 蔡为明； 金群力；

【Author】 LI Nan-yi~(1,2),LUO Zhi-jiao~3,LIU Chun-yan~4,CAI Wei-ming~1,JIN Qun-li~1 (1.Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021;2.Zhejiang Forestry University, Hangzhou 311300;3.College of Life Science,Henan Agricultural University,Zhezhou 450002;4.College of Life Science, Anhui Agricultural University,Hefei 230036)

【机构】 浙江省农业科学院园艺研究所； 浙江林学院； 河南农业大学生命科学院； 安徽农业大学生命科学院；

【摘要】 设计简并引物克隆了PPO基因cDNA片段,并根据获得的序列信息用cDNA末端快速扩增(RACE)方法从双孢蘑菇中克隆到了两个PPO同源基因的cDNA全长,将其命名为AbPPO3和AbPPO4,分别编码576和611个氨基酸,具有多酚氧化酶的Cu结合保守结构域。同源性分析表明,4bPPO3编码的氨基酸序列与其他真菌PPO同源蛋白的相似性约为21～50%,AbPPO4与其他真菌PPO蛋白的相似性约为20～63%。系统进化树分析表明,AbPPO3,AbPPO4和已克隆的双孢蘑菇AbPPO2具有较近的进化关系。半定量分析表明,在双孢蘑菇子实体的不同发育时期,AbPPO3和AbPPO4在子实体刚开始形成时表达量最低,在茶杯期即商品菇开始采摘时表达量上升,至子实体完全成熟时即大开伞期表达量一直很高。双孢蘑菇4℃储藏时,AbPPO3和AbPPO4在第1天可检测到微量表达,第2～4天表达均较强,之后至第7天表达量下降。这表明,AbPPO3和AbPPO4可能与双孢蘑菇褐变相关。更多还原

【Abstract】 Using primers designed on the basis of sequence homologies in the copper-binding domains for fungal PPO,two PPO homologue gene cDNAs were cloned from Agaricus bisporus.AbPPO3 and AbPP04 encoded, resperctively,576 and 611 amino acids,containing copper-binding domains.AbPPO3 shows 21~50%identity to that of fungal origin,and AbPPO4 shows 20～63%identity to that of fungi origin.Alignment and phylogenetic analysis of PPO sequences of animal,plant,fungal and bacterial origin indicated that AbPPO3 and AbPPO4 is more closely related to AbPPO2,the homologue gene cloned from A.bisporus.During the defferent development stage of fruit body,the expression of AbPPO3 and AbPPO4 was at lowest level at"pin head"stage,then increased at"cup"stage,and at high level until the"flat"stage.During the storage at 4℃,the transcripts of AbPPO3 and AbPPO4 lowered on day 1 then increased between day 2 to day 4 and later decreased until day 7.This indicated that AbPPO3 and AbPPO4 may play a very important role in A.bisporus browning.更多还原