龙眼DlAGO7基因克隆与亚细胞定位及表达分析  

Cloning,Subcellular Localization and Expression Analysis of DlAGO7 Gene in Dimocarpus longan

作　　者：陈荣珠[1] 王小平[1] 林美珍[1] 赖钟雄[2] Chen Rongzhu;Wang Xiaoping;Lin Meizhen;Lai Zhongxiong(College of Pharmacy,Zhangzhou Health Vocational College,Zhangzhou,363000;Institute of Horticultural Biotechnology,Fujian Agriculture and ForestryUniversity,Fuzhou,350002)

机构地区：[1]漳州卫生职业学院药学院,漳州363000 [2]福建农林大学,园艺植物生物工程研究所,福州350002

出　　处：《分子植物育种》2025年第2期380-386,共7页Molecular Plant Breeding

基　　金：漳州市自然科学基金项目(ZZ2021J16);漳州卫生职业学院高学历人才科研支持专项(ZWYGXL202101);漳州卫生职业学院科技创新团队培育项目(Kjcx-3;Kjcx-5);福建农林大学科技创新专项(CXZX2017189)共同资助。

摘　　要：Argonaute7(AGO7)在植物叶和花发育过程中发挥着重要的调控作用,而AGO7蛋白如何参与调控龙眼体胚的发育却未见报道。为了深入研究龙眼AGO7基因的功能与调控作用,本研究基于龙眼基因组数据库,采用RT-PCR法克隆龙眼DlAGO7基因,对其进行生物信息学分析,构建瞬时融合表达载体,明确其亚细胞定位,最后,利用实时荧光定量仪检测其在龙眼体胚不同阶段的相对表达情况。克隆获得DlAGO7的CDS序列为3060 bp,编码1019个氨基酸;所编码氨基酸序列含有PAZ和Piwi高度保守结构域;blastp序列比对分析表明,DlAGO7蛋白与其他植物的AGO7蛋白具有高度同源性;系统进化发育树分析显示,DlAGO7蛋白与甜橙AGO7蛋白亲缘关系最近,与单子叶植物玉米和水稻的亲缘关系最远;由亚细胞定位研究可知,龙眼DlAGO7基因为核定位基因;RT-qPCR分析表明,DlAGO7在龙眼球形胚阶段显著表达;克隆获得DlAGO7基因的5′端上游启动子序列长度为1746 bp,启动子分析共预测出9类顺式作用元件。由此推测,DlAGO7基因可能在龙眼球形胚阶段发挥着重要的调控作用,该基因主要在细胞核中发挥作用。以上研究结果可为龙眼的分子育种提供理论基础。Argonaute7(AGO7)played a conserved role in the regulation of plant leaf and flower development.However,little is known about how AGO7 is involved in the regulation of longan somatic embryos.To deeply investigate the function and regulatory role of the AGO7 gene in longan,in this study,based on the longan genome database,the DlAGO7 gene of longan was cloned by RT-PCR method.Bioinformatics analysis was conducted,a transient fusion expression vector was constructed and determine its subcellular localization.Finally,the relative expression of it at different stages of longan somatic embryos was detected using real-time fluorescence quantitative PCR.The full length of CDS sequence of DlAGO7 was 3060 bp,encoding 1019 amino acids,containing PAZ and Piwi conserved domains.blastp analysis showed that DlAGO7 protein had high homology with other plant,and phylogenetic tree analysis showed that it was closely related to AGO7 protein of Citrus sinensis and the farthest related to rice and maize.According to the subcellular localization,DlAGO7 gene was localized in the nucleus.RT-qPCR analysis showed that DlAGO7 was significantly expressed in the globular embryo stage of longan.The sequence length of the 5'upstream promoter of DlAGO7 gene was 1746 bp,and promoter analysis predicted 9cis-acting elements.It is speculated that DlAGO7 may be mainly involved in the globular embryo stage of longan,which functions mainly in the nucleus.These results can provide a theoretical basis for molecular breeding of Dimocarpus longan.

关 键 词：龙眼体胚发生 AGO7基因 启动子序列 亚细胞定位 

分 类 号：S66[农业科学—果树学]