巴西橡胶树HbMago3的克隆和表达分析  

Cloning and Expression Analysis of HbMago3 from Hevea brasiliensis

作　　者：杨子平[1,3,4] 郭冬[1,2,4] 李辉亮 朱家红[1,2,4] 王颖 彭世清[1,2] Yang Ziping;Guo Dong;Li Huiliang;Zhu Jiahong;Wang Ying;Peng Shiqing(Key Laboratory of Biology and Genetic Resources of Tropical Crops(Ministry of Agriculture and Rural Affairs),Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101;Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province,Hainan Institute for Tropical Agricultural Resources,Haikou,571101;Zhanjiang Key Laboratory of Tropical Crop Genetic Improvement,South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,524091;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences,Sanya,572025)

机构地区：[1]中国热带农业科学院热带生物技术研究所,农业农村部热带作物生物学与遗传资源利用重点实验室,海口571101 [2]海南热带农业资源研究院,海南省热带农业生物资源保护与利用重点实验室,海口571101 [3]中国热带农业科学院南亚热带作物研究所,湛江市热带作物遗传改良重点实验室,湛江52409 [4]中国热带农业科学院三亚研究院,三亚572025

出　　处：《分子植物育种》2023年第24期8005-8012,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31970620,32171827);中央级公益性科研院所基本科研业务费专项(1630052022-009)共同资助。

摘　　要：为了在基因组水平鉴定和分析巴西橡胶树中Mago基因家族的数量、结构和功能,本研究对巴西橡胶树Mago基因家族的数量、基因结构、染色体分布和蛋白特性进行了系统鉴定和分析;进一步克隆鉴定新的Mago基因,分析其组织表达水平和对激素(茉莉酸和乙烯)的响应。本研究从reyan7-33-97中鉴定了4个Mago基因,分别命名为HbMago1、HbMago2、HbMago3和HbMago4。HbMago2和HbMago3的可变剪切事件发生在第二个外显子,CDS存在66个碱基的差异,导致蛋白短了22个氨基酸、等电点低0.35;HbMago4可变剪切事件发生在5端的非翻译区,CDS和蛋白序列无差异。4个基因序列均包含3个外显子,HbMago1、HbMago2和HbMago4含有2个内含子,HbMago3含有3个外显子。4个基因序列分布在两条scaffold(NW-018745965.1和NW-018746420.1)上的4个位点。进化分析结果表明4个基因聚类在同一个进化枝条上,为单进化起源;橡胶树在进化过程中获得3个Mago基因而丢失1个基因。克隆了HbMago3,半定量结果表明HbMago3在根、胚、芽和花中未表达,在树皮、胶乳和叶中表达,其在胶乳中的表达量最高;实时荧光定量PCR结果表明,HbMago3在根、树皮、叶、花和胶乳中均有表达,但其在胶乳中表达水平最高,茉莉酸和乙烯可诱导HbMago3表达。结果为进一步研究HbMago3的功能提供基础。To identify and analyze the quantity,structure,and function of the Mago gene family in the Hevea brasiliensis at a genomic scale,this study systematically identified and analyzed the quantity,gene structure,chromosome distribution,and protein characteristics of the Mago gene family.Further cloned and identified the novel Mago gene,analyzed its tissue expression levels,and its expression profiles under hormone(jasmonic acid and ethylene) treatment.The results showed that we identified four Mago genes,named HbMago1,HbMago2,HbMago3,and HbMago4,from the rubber cultivar reyan7-33-97.The alternative splicing events of HbMago2 and HbMago3 occurred in the second exon,resulting in a difference of 66 nucleotides in the coding sequence(CDS),which resulted in the protein being 22 amino acids shorter and having an isoelectric point 0.35 lower.The alternative splicing event of HbMago4 occurred in the 5' untranslated region,with no difference in sequences of CDS and protein.All four gene sequences contained three exons,with HbMago1,HbMago2,and HbMago4 containing two introns,and HbMago3 containing three exons.The four genes were distributed at four locations on two scaffolds(NW-018745965.1and NW-018746420.1).Evolutionary analysis results indicated that the four genes clustered on the same evolutionary branch,indicating a single origin.During the evolution of rubber trees,three Mago genes were acquired while one gene was lost.The study cloned HbMago3,and semi-quantitative results showed that it was not expressed in roots,embryos,buds,and flowers,but expressed in bark,latex,and leaves,with the highest expression level in latex.Real-time fluorescence quantitative PCR results showed that HbMago3 was expressed in roots,bark,leaves,flowers,and latex,with the highest expression level in latex.Jasmonic acid and ethylene could induce the expression of HbMago3.These results provide a foundation for further investigating the function of HbMago3 in Hevea brasiliensis.

关 键 词：巴西橡胶树 Mago Nashi蛋白 表达分析 茉莉酸 乙烯 

分 类 号：S794.1[农业科学—林木遗传育种]