梅花PmERF4基因的克隆及其表达分析
Cloning and expression of PmERF4 gene in Prunus mume

为阐明梅花（Prunus mume）ERF基因的功能，采用RT PCR技术从梅花品种‘雪梅’中克隆获得1个PmERF4基因。生物信息学分析表明该基因cDNA全长序列为842 bp，完整开放阅读框（ORF）为690 bp，编码229个氨基酸。氨基酸序列比对发现，PmERF4蛋白包含1个AP2/ERF保守结构域。系统进化分析结果显示，梅花PmERF4蛋白与李亚科李属植物的ERF蛋白高度同源。利用荧光定量PCR（qRT PCR）检测PmERF4基因在非生物胁迫和激素处理下的表达，结果表明：PmERF4基因受低温、氧化胁迫的诱导，但响应程度不同，它能够持续且强烈地响应低温胁迫，对甘露醇、MeJA、SA处理不敏感，而高盐和ABA则抑制其表达；PmERF4基因可能在梅花低温胁迫应答中发挥重要作用。
ERF (ethylene response factor) transcription factor plays a critical role in response to environmental stress and hormonal signal transduction in plants.To elucidate the function of ERF gene in Prunus mume,the PmERF4 gene was cloned from a Prunus mume ‘Xuemei’ by RT PCR.The full length cDNA sequence of PmERF4 was 842 bp with an open reading frame (ORF) of 690 bp,encoding 229 amino acids.The deduced amino acid sequence alignment of PmERF4 contained a conserved AP2/ERF domain.The result of phylogenetic analysis showed the PmERF4 was highly homologous to other proteins in Prunus genus.qRT PCR was conducted to examine the expression patterns of PmERF4 under abiotic stress and hormone treatment.The results showed that PmERF4 was induced by cold and H2O2 stress.The induction levels of the gene were different between cold and H2O2 stress.A continuous and strong response of PmERF4 was triggered by cold.PmERF4 was insensitive to mannitol,MeJA and SA treatment and its expression was inhibited by NaCl and ABA.It is indicated that PmERF4 from Prunus mume may be involved in response to cold and H2O2 stress.