异源表达苹果属MdNAC56-L基因的克隆及功能  

Cloning and functional exploration of the heterologous expression of MdNAC56-L in Malus plants

作　　者：唐艳 杨乔木 田佶 TANG Yan;YANG Qiaomu;TIAN Ji(College of Plant Science and Technology/Beijing Key Laboratory of New Technology in Agricultural Application,Beijing 100026,China)

机构地区：[1]北京农学院植物科学技术学院/农业应用新技术北京市重点实验室,北京100026

出　　处：《北京农学院学报》2024年第3期6-10,共5页Journal of Beijing University of Agriculture

基　　金：国家林业和草原局—第三批林草科技青拔人才项目资助(202320132004)。

摘　　要：【目的】探明苹果中MdNAC56-L基因在叶绿素生物合成通路中发挥的功能。【方法】以‘王林’苹果(Malus domestica‘Orin’)为试验材料,通过RNA提取和反转录得到cDNA,克隆得到MdNAC56-L基因。构建MdNAC56-L的过表达载体及沉默载体,在野生型烟草中分别瞬时过表达和沉默MdNAC56-L基因,通过对转基因叶片进行表型分析、采用分光光度法测定叶绿素含量以及使用荧光定量PCR法鉴定叶绿素降解途径关键基因NtPAO表达情况,初步解析MdNAC56-L基因的生物学功能。【结果】与野生型烟草相比,过表达MdNAC56-L基因的烟草叶片中,叶绿素含量显著降低,叶绿素降解途径关键基因NtPAO表达量显著增高,沉默MdNAC56-L基因的烟草叶片中,叶绿素含量无显著性升高,叶绿素降解途径关键基因NtPAO表达量无显著性降低。【结论】MdNAC56-L可能参与到植物叶绿素生物合成通路中正调控叶绿素的降解。[Objective]To investigate the function of the MdNAC56-L gene in the chlorophyll biosynthesis pathway in apple.[Methods]We used‘Orin’apple(Malus domestica‘Orin’)as the experimental material.MdNAC56-L gene was cloned by RNA extraction and reverse-transcription.Constructing overexpression vectors and silencing vectors for MdNAC56-L,and instantaneously overexpressing and silencing the MdNAC56-L gene in wild-type tobacco.Phenotypic analysis was performed on transgenic leaves,chlorophyll content was measured using spectrophotometry,and the expression of the key gene NtPAO in the chlorophyll degradation pathway was identified using fluorescence quantitative PCR.The biological function of the MdNAC56-L gene was preliminarily analyzed.[Results]The results showed that compared with wild-type tobacco,tobacco leaves overexpressing the MdNAC56-L gene showed a significant decrease in chlorophyll content and a significant increase in the expression of the key gene NtPAO for chlorophyll degradation pathway.Tobacco leaves silencing the MdNAC56-L gene showed no significant increase in chlorophyll content and no significant decrease in the expression of the key gene NtPAO for chlorophyll degradation pathway.[Conclusion]MdNAC56-L may be involved in the positive regulation of chlorophyll degradation in the chlorophyll synthesis pathway.

关 键 词：苹果 叶绿素合成 荧光定量 基因克隆 

分 类 号：S661.1[农业科学—果树学]