摘要
利用RACE和电子克隆技术相结合的方法从野生红肉苹果(Malus pumila var.niedzwetzkyana Schneid)中克隆了1个MYB类转录因子的全长序列,利用生物信息学方法对其进行分析,同时进行了亚细胞定位分析以及表达特性的研究。结果表明:该基因全长为1 020 bp,编码340个氨基酸;其核苷酸序列与葡萄MYBPA1同源性最高,命名为MpMYBPA1,其N端含2个约有55个氨基酸组成的MYB特征结构域。亚细胞定位分析结果表明MpMYBPA1蛋白定位在细胞核中。荧光定量PCR结果表明:MpMYBPA1基因在根、茎、叶、果皮中均有表达,果皮中表达量最大。在20℃处理4 d后该基因的表达量得到了上调,而在30℃高温条件下,该基因的表达上调不明显,说明低温有利于该基因的表达,并且该基因受ABA调控。推测MpMYBPA1基因可能参与苹果的着色过程。
In this study, Malus pumila var. niedzwetzkyana Schneid was employed as the experimental material. The full-length sequence of one MYB transcription factor named MpMYBPA1 was cloned by RACE and silico cloning technology from Malus pumila var. niedzwetzkyana Schneid. Along with the bioinformatics analysis and subcellular localization to the MYB protein, the gene expression research on MpMYBPA1 was conducted. The result showed that the full length of this gene from apple was 1 020 bp,which was to code 340 amino acids, and with a highest homology with MYBPA1 from Vitis viniferna. This gene was named MpMYBPA1, which had two MYB HTH DNA-binding domains with 55 amino acids in parathormone-N re. It was proved from the subcellular localization that MpMYBPA1 protein was located in nucleus. Through quantitative real-time PCR,the expression of MpMYBPA1 was observed in different organs such as root, stem,leaf and peel ,with a highest level in peel, which might be influenced by temperature and ABA. After 4 days with 20 ℃ treatment, the expression level rose, however, the increase of the gene expression was not significant with 30 ℃ treatment,which might suggest that low temperature should be positive to the expression of MYBPA1. It could be concluded from the research results that MpMYBPA1 might have been involved in the process of coloration for apple.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2012年第2期32-38,共7页
Journal of Nanjing Agricultural University
基金
江苏省科技支撑计划项目(BE2008316
BE2011415)
关键词
红肉苹果
MpMYBPA1
克隆
表达分析
： Malus pumila var. niedzwetzkyana Schneid
MpMYBPA1
clone
expression analysis
