菊花乙烯响应因子CmRAP2-12的克隆与表达特性分析

Cloning,Characterization and Expression Analysis of ERF CmRAP2-12 in Chrysanthemum morifolium

以菊花(Chrysanthemum morifolium)品种‘神马’为试材,采用RT-PCR与RACE、Real-time RT-PC等技术,克隆了菊花乙烯转录因子CmRAP2-12基因全长的cDNA序列片段,Real-time RT-PCR分析了水淹胁迫对菊花CmRAP2-12基因的表达模式的影响,以期降低水淹条件下对菊花生长的迫害性影响。结果表明:得到的CmRAP2-12的cDNA序列为1 335bp;CmRAP2-12保守氨基酸的N末端含有一个具有ⅦERFs亚家族特征的氨基酸序列,且聚类到RAP2-12亚家族一类,并预测亚细胞定位在细胞核中。Real-time RT-PCR分析显示,CmRAP2-12在菊花的根、茎、叶中均有表达,并且在经过0~72h的淹水胁迫后,CmRAP2-12基因在菊花根系中的表达量均呈现一个先升高后降低的趋势,并在处理后48h达到最高值,淹水胁迫提高了菊花根系乙烯响应转录因子RAP2-12的表达水平。该试验将为进一步研究RAP2-12对提高菊花水淹低氧耐受性的调节机理与分子响应机制提供参考依据。

In this study,a full-length cDNA named CmRAP2-12 was cloned in Chrysanthemum morifolium ‘Jinba＇roots by RT-PCR and RACE,and the real time RT-PCR was used to analyze the expression pattern of the three genes.The results showed that cDNA sequence of CmRAP2-12 obtained in this study was 1 335 bp,CmRAP2-12 conserved N-terminal amino acids contained aⅦ ERFs subfamily characteristic amino acid sequence,and clustered to RAP2-12 subfamily.CmRAP2-12 was predicted subcellular localization in the nucleus.Real-time RT-PCR analysis showed that CmRAP2-12 was all expressed in root,stem and leaf in Chrysanthemum.CmRAP2-12 expression levels presented a first increased and then decreased trend,after waterlogging stress 0-72 hours.It was treated after 48 hours reached the highest value.The expression levels of ethylene response transcription factor RAP2-12 was increased in Chrysanthemumroots and affected by hypoxic response when suffering from waterlogging stress.This study would provide the theoretical basis for further research RAP2-12 to improve chrysanthemum flooding tolerance to hypoxia and molecular mechanisms regulating response mechanism.