园艺学报 ›› 2013, Vol. 40 ›› Issue (6): 1071-.

• 蔬菜 • 上一篇    下一篇


郭勤卫,李 季,崔 利,张停林,Kere George Mbira,陈劲枫*   

  1. (南京农业大学园艺学院,葫芦科作物育种与种质创新实验室,南京 210095)
  • 收稿日期:2013-01-18 修回日期:2013-04-12 出版日期:2013-06-25 发布日期:2013-06-25

Cloning and Expression Analysis of Cucumber CsARF10 Genes Subfamily

GUO Qin-wei,LI Ji,CUI Li,ZHANG Ting-lin,Kere George Mbira,and CHEN Jin-feng*   

  1. (College of Horticulture,Laboratory of Cucurbit Genetics and Germplasm Enhancement,Nanjing Agricultural University,Nanjing 210095,China)
  • Received:2013-01-18 Revised:2013-04-12 Online:2013-06-25 Published:2013-06-25

摘要: 利通过蛋白序列同源比对和PCR技术从黄瓜中克隆获得了3条生长素响应因子ARF10基因序列,分别命名为CsARF10a、CsARF10bCsARF10c。系统发育进化树分析发现CsARF10a与番茄SlARF10基因进化距离较近,而CsARF10bCsARF10c与拟南芥AtARF10相似性更高;启动子分析显示CsARF10家族基因具有多样化的激素应答元件,荧光定量PCR结果也证明黄瓜幼苗在不同NAA浓度处理以及其他激素的处理下,CsARF10家族基因呈现不同的表达模式;荧光定量PCR结果还显示CsARF10家族基因在黄瓜幼苗的根、茎、叶及雄花中都具有较高的转录水平;CsARF10a、b、c在黄瓜果实发育的早期具有相同的表达趋势,但CsARF10a的转录水平较高,而CsARF10bCsARF10c表达量较低。

关键词: 黄瓜, CsARF10, 基因克隆, 序列分析, 表达分析

Abstract: Three ARF10 genes(Auxin-Respond-Factor)named CsARF10aCsARF10b and CsARF10c,were screened from the cucumber genome database by protein homology BLAST and cloned from 8419s-1 by polymerase chain reaction(PCR). In present study,phylogenetic reconstruction indicated that the CsARF10a has closer relation with SlARF10,while CsARF10b and CsARF10c have higher similarity with AtARF10. Promoter prediction showed that the promoters consist of diversified multiple hormone response elements. The real-time quantitative PCR(RT-qPCR)analysis proved that the expression of cucumber ARF10 genes could be regulated by different phytohormones or concentrations of 1-naphthylacetic acid(NAA)and performed different expression patterns. It was also seen that CsARF10 gene subfamily had high transcriptional levels either in roots,stems,leaves or in male flowers;In additional,the qPCR results showed that although the expression of CsARF10 genes have similar transcriptional trend,the CsARF10a had high expression level while mRNA of CsARF10b and CsARF10c were relatively low.

Key words: cucumber, CsARF10, gene cloning, homologous analysis, expression analysis