巴西橡胶树雄性不育相关基因HbACO1的表达与生物信息学分析  

Expression and Bioinformatics Analysis of Male Sterility-Related Gene HbACO1 in Hevea brasiliensis

作　　者：党一敏 刘红东 王英[1] 高和琼[1] 庄南生[1] Dang Yimin;Liu Hongdong;Wang Ying;Gao Heqiong;Zhuang Nansheng(College of Tropical Crops,Hainan University,Haikou,570228;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]海南大学热带作物学院,海口570228 [2]中国热带农业科学院橡胶研究所,海口571101

出　　处：《分子植物育种》2023年第1期94-101,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31760216);国家天然橡胶产业技术体系项目(CARS-33-YZ2)共同资助

摘　　要：ACO1(1-aminocyclopropane-1-carboxylateoxidase1)是植物内源乙烯生物合成途径的限速酶,与花芽分化和器官衰老密切相关。本研究通过实时荧光定量技术对基因HbACO1在雌雄花、叶、胶乳和嫩皮组织的表达模式进行分析,结果显示基因HbACO1在雄花表达量最高,在雌花、叶和嫩皮中表达量低,在胶乳中几乎不表达。对雄花发育不同时期(小孢子母细胞时期,四分体期和单核小孢子时期)的HbACO1基因表达量分析表明:在不育品种(‘热研7-33-97',‘PR107’)的四分体和小孢子时期表达量明显高于在可育品种(热研‘93-114',‘天任31-45')相应时期的表达量,推测基因HbACO1可能在雄花的四分体及单核孢子期间异常高表达导致了雄花败育现象。利用mRNA原位杂交技术对HbACO1基因进行了原位表达分析,结果显示在不育品种中绒毡层细胞、花药外壁细胞和小孢子母细胞能检测出基因HbACO1的荧光信号,在可育品种中在花柱和花药外壁有荧光信号,推测HbACO1基因可能主要是在雄花的绒毡层、花药外壁和小孢子母细胞特定细胞类型中表达来调控雄花的发育,而在这些细胞类型中的异常表达可能导致了橡胶树的雄性不育。HbACO1基因的ORF长度为921bp,编码306个氨基酸,HbACO1基因编码的蛋白理论等电点为5.69,相对分子质量为34.80kD,总平均亲水性GRAVY为负值,亚细胞定位预测结果表明定位于细胞质中,且不具有信号肽及跨膜螺旋结构的亲水性蛋白,进化树显示HbACO1蛋白与木薯(Manihotesculenta)的MeACO1蛋白聚为一类,亲缘关系较近。本研究结果可为橡胶树雄性不育分子机制的深入研究以及橡胶树的育种工作提供基础数据和科学依据。ACO1(1-aminocyclopropan e-1-carboxylate oxidase 1) is a rate-limiting enzyme in plant ethylene biosynthesis pathway, which is closely related to flower bud differentiation and organ senescence. In this study, the expression pattern of HbACO1 gene in flowers, leaves, latex and young skin tissues was analyzed by real-time fluorescence quantitative technique. The results showed that HbACO1 gene was highly expressed in male flowers, low in female flowers, leaves and skin tissues, and almost not expressed in latex. Analysis of HbACO1 gene expression at different stages of male flower development(microspore mother cell stage, tetrad stage and mononuclear mi crospore stage) showed that: the expression level of HbACO1 gene in the tetrad and microspore stage of male sterile varieties(MS) was significantly higher than that in the corresponding stage of male fertile varieties(MF). It was speculated that the abnormal high expression of HbACO1 gene in the tetrad and mononuclear spore stage of male flowers might lead to male flower abortion. The in situ expression of HbACO1 gene was analyzed by mRNA-FISH technique. The results showed that HbACO1 had fluorescence signal in tapetum cells, anther outer wall cells and microspore mother cells in MS, and had fluorescence signal in style and anther outer wall of MF. It is speculated that HbACO1 gene is mainly expressed in tapetum, outer wall of anther and specific cell types of microspore mother cells to regulate the development of male flowers, and abnormal expression in these cell types may lead to male sterility in rubber trees. The ORF length of HbACO1 gene is 921 bp, encoding 306 amino acids. The theoretical isoelectric point of HbACO1 protein is 5.69, and the relative molecular weight is 34.80 kD. The total average hydrophilic GRAVY is negative. The prediction results of subcellular localization showed that the hydrophilic protein was located in the cytoplasm without signal peptide and transmembrane helix structure. The phylogenetic tree showed that HbACO1 protein and Me ACO1 p

关 键 词：巴西橡胶树 ACO1基因 荧光定量PCR mRNA-FISH技术 生物信息学分析 

分 类 号：S794.1[农业科学—林木遗传育种]