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以柑橘属寒敏感植物佛手‘青皮’品种叶片cDNA为模板,通过RT-PCR与RACE扩增,获得一个1160bp的乙烯应答因子(ethylene response factor,ERF)基因的cDNA序列,其基因编码区共999bp,含有一个保守的AP2/EREBP结构域,与拟南芥AtERF6为同源基因,命名为CmsERF6(GenBank登录号为HQ698835)。4℃低温处理佛手和枳(柑橘属近缘种耐寒植物)植株,测定其寒胁迫生理指标(电解质渗出率、丙二醛含量)。使用实时荧光定量PCR,对佛手和枳叶中的ERF6表达含量进行测定分析,结果表明:低温对佛手与枳的ERF6表达均具有诱导作用,且表达量的变化趋势存在明显的差异。尤其在枳中,ERF6的表达较对照组上升约4000倍,且电解质外渗率的变化与ERF6基因表达量变化具有一致性,说明ERF6基因参与低温胁迫应答。
The cDNA sequence of 1160bp ethylene response factor (ERF) gene was obtained by RT-PCR and RACE amplification using leaf cDNA of the genus Asparagus, , Contains a conserved AP2 / EREBP domain and is homologous to Arabidopsis AtERF6 and is named CmsERF6 (GenBank accession number is HQ698835). 4 ℃ low temperature bergamot and trifoliate orange (citrus species cold-tolerant plants) plants were measured cold stress physiological indicators (electrolyte exudation rate, malondialdehyde content). Using real-time fluorescence quantitative PCR, ERF6 expression in bergamot and citrus leaved leaves was measured and analyzed. The results showed that low temperature induced the expression of ERF6 in bergamot and trifoliate orange fruit, and there was a significant difference in the expression trend of ERF6. Especially in the trifoliate orange, the expression of ERF6 was about 4000 times higher than that of the control group, and the change of electrolyte exudation was consistent with the change of ERF6 gene expression, indicating that ERF6 gene is involved in the response to low temperature stress.