马尾松PmSWEET基因的克隆、亚细胞定位及表达分析  

Cloning,subcellular localization and expression analysis of PmSWEET gene in Pinus massoniana

作　　者：梁大曲 石长双 涂晶晶 徐刚 吴峰 LIANG Daqu;SHI Changshuang;TU Jingjing;XU Gang;WU Feng(Institute for Forest Resources and Environment of Guizhou,Key Laboratory of Forest Cultivation in Plateau Mountain of Guizhou Province,Guizhou University,Guiyang 550025,China;College of Forest,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学贵州省森林资源与环境研究中心,贵州省高原山地林木培育重点实验室,贵阳550025 [2]贵州大学林学院,贵阳550025

出　　处：《植物生理学报》2022年第2期447-457,共11页Plant Physiology Journal

基　　金：贵州省科技厅计划项目(黔科合平台人才[2018]5261);国家自然科学基金(31660201);贵州省一流学科建设项目(GNYL-[2017]007)。

摘　　要：SWEETs蛋白是一类糖转运蛋白,通过调控糖类物质的跨膜运输影响植物生长发育。为深入研究PmSWEET在马尾松(Pinus massoniana)短枝腋芽休眠解除过程中的作用,通过克隆得到PmSWEET基因的CDS序列并进行生物信息学分析,同时分析其亚细胞定位情况及在马尾松短枝腋芽休眠解除过程中的表达规律。结果表明:PmSWEET完整CDS序列共687 bp;编码228个氨基酸,含有2个保守结构域(跨膜螺旋域,MtN3)和7个跨膜区域。PmSWEET与杨树和拟南芥的SWEET16、SWEET17基因进化关系最近。原生质体瞬时表达分析显示,PmSWEET蛋白主要定位于液泡膜。截顶处理后,PmSWEET基因在短枝上的针叶及腋芽中的表达模式均发生了变化,其中截顶处理1 d内,PmSWEET的表达量在短枝腋芽内显著提高;处理2 d后,PmSWEET在针叶中的表达量开始大幅升高,第4天达到最大值。上述结果表明PmSWEET基因参与了马尾松短枝腋芽休眠解除的调控。SWEETs protein are a kind of sugar transporter that affect plant growth and development by regulating the transmembrane transport of sugars.To reveal the function of PmSWEET in the dormancy release of axillary buds of Pinus massoniana,the CDS sequence of PmSWEET was cloned,as well as the bioinformatic analysis and subcellular localization of the gene.At the same time,the expression in needles and buds under decapitation treatments was analyzed.The results showed that the complete CDS of PmSWEET was 687 bp,encoded 228 amino acids,contained two conserved domains(transmembrane helical domain,MtN3),and seven transmembrane domains.Moreover,PmSWEET was most closely related to the SWEET16 and SWEET17 genes in Poplar and Arabidopsis.Transient expression assay in protoplast showed that PmSWEET was localized on vacuolar membrane.Furthermore,the expression of PmSWEET in axillary buds was significantly increased on the first day of treatments;after two days,the expression level in needles began to increase sharply and reached the maximum on the 4 th day.These results indicated that the Pm SWEET was involved in the regulation of axillary bud release of the dwarf shoot in P.massoniana.

关 键 词：马尾松 PmSWEET基因 腋芽 休眠解除 亚细胞定位 

分 类 号：S791.248[农业科学—林木遗传育种]