金针菇Ⅱ类过氧化物酶基因在子实体发育与胁迫应答过程的表达特征

Characteristics of Class II Peroxidase Gene Expression During Fruiting Body Development and Stress Response in Flammulina filiformis

Ⅱ类过氧化物酶(ClassⅡperoxidase,CⅡp)是活性氧代谢的重要抗氧化物酶,参与机体氧化应答过程。基于金针菇全基因组数据鉴定到两个Ⅱ类过氧化物酶基因,并分别命名为FfCⅡp1、FfCⅡp2,采用在线网站软件对基因进行生物信息学分析,利用实时荧光定量PCR技术分析基因在金针菇不同子实体组织和胁迫应答中的表达模式。结果显示,FfCⅡp1全长1638 bp,包含一个1131 bp的完整开放阅读框,编码376个氨基酸。FfCⅡp2全长1410 bp,包含一个1032 bp的完整开放阅读框,编码343个氨基酸。保守结构域及进化树分析表明,FfCⅡp1和FfCⅡp2分别归属于锰过氧化物酶和通用过氧化物酶家族。RT-qPCR结果表明,两个FfCⅡp基因均在伸长期菌柄中高表达,且在快速伸长区段显著上调;损伤和氧化胁迫处理均能诱导FfCⅡp1和FfCⅡp2上调表达,FfCⅡp2的上调幅度明显高于FfCⅡp1。以上结果表明,金针菇两个CⅡp家族基因参与菌柄伸长与胁迫应答过程,且FfCⅡp2对损伤和氧化胁迫更为敏感。

Class II peroxidase(CIIp)is an important antioxidant enzyme in active oxygen metabolism,and participates in the oxidative response of organism.Based on the genomic data of Fammulina filiformis,two Class II peroxidase genes were identified and named as FfCIIp1 and FfCIIp2,respectively.Online software was used for bioinformatics analysis of the genes,and real-time fluorescence quantitative PCR was used to analyze the expression patterns of genes in different fruit body tissues and stress responses of F.filiformis.The results showed that the full length of FfCIIp1 was 1638 bp,including a complete open reading frame of 1131 bp,encoding 376 amino acids.FfCIIp2 was 1410 bp in length,containing a complete open reading frame of 1032 bp and encoding 343 amino acids.Conserved domain and phylogenetic analysis showed that FfCIIp1 and FfCIIp2 belonged to the manganese peroxidase and generic peroxidase families,respectively.RT-qPCR results showed that both FfCIIp genes were highly expressed in the stipe during the elongation stage,and significantly up-regulated in the stipe of fast elongation region.FfCIIp1 and FfCIIp2 were up-regulated by both injury and oxidative stress,and the up-regulated range of FfCIIp2 was significantly higher than FfCIIp1.These results indicate that two CIIp family genes are involved in stipe elongation and stress response,and FfCIIp2 is more sensitive to injury and oxidative stress of F.filiformis.