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、CsARF10b和CsARF10c。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 CsARF10b 和 CsARF10c 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 CsARF10b 和 CsARF10c 表达量较低。
Three ARF10 genes (Auxin-Respond-Factor) named CsARF10a, CsARF10b 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 CsARFIOa has closer relation with SIARFIO, while CsARFIOb and CsARFIOc have higher similarity with AtARFIO. 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 ARFIO genes could be regulated by different phytohormones or concentrations of 1-naphthylacetic acid (NAA) and performed different expression patterns. It was also seen that CsARFIO 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 CsARFIO genes have similar transcriptional trend, the CsARFIOa had high expression level while mRNA of CsARFIOb and CsARFIOc were relatively low.
出处
《园艺学报》
CAS
CSCD
北大核心
2013年第6期1071-1080,共10页
Acta Horticulturae Sinica
基金
国家重点基础研究发展计划‘973’项目(2012CB113904)
国家‘863’项目(2012AA100202)
教育部博士点基金项目(20120097122237)
南京农业大学青年科技创新基金项目(KJ2012013)