辣椒细胞质雄性不育系和保持系蔗糖转化酶活性与相关基因表达分析  

Invertase Activity and Related Genes Expression Analysis in Cytoplasmic Male Sterile and Fertile Pepper(Capsicum annuum) Lines

作　　者：阮美颖[1] 万红建[1] 杨有新[2] 周国治[1] 王荣青[1] 叶青静[1] 姚祝平[1] 杨悦俭[1] 程远[1] 李志邈[1] Ruan Mei-Ying;WAN Hong-Jian;YANG You-Xin;ZHOU Guo-Zhi;WANG Rong-Qing;YE Qing-Jing;YAO Zhu-Ping;YANG Yue-Jian;CHENG Yuan;LI Zhi-Miao(Institute of Vegetables,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;College of Agriculture,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]浙江省农业科学院蔬菜研究所,杭州310021 [2]江西农业大学农学院,南昌330045

出　　处：《农业生物技术学报》2018年第12期2036-2046,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31501749和No.31772294);浙江省农业新品种选育重大科技专项(No.2016C02051);浙江省育才工程项目(No.2018YCGC005);2016年浙江省蔬菜种质资源收集;繁殖与鉴定评价(No.2016R23A60D08);物种品种资源保护费(No.111821301354052030);浙江省自然科学基金(No.LY18C150008);国家重点实验室自主设计创新课题(No.2017YFD0101902)

摘　　要：蔗糖是植物光合作用的主要同化产物。植物生殖器官对蔗糖的有效利用,在植物的受精和坐果过程中发挥重要作用。蔗糖转化酶(invertase, INV)将蔗糖水解为葡萄糖和果糖,是蔗糖代谢的关键酶,根据细胞内位置不同,INV被分成细胞质蔗糖转化酶(cytoplasmic invertase, CIN)、液泡蔗糖转化酶(vacuole invertase, VIN)和细胞壁蔗糖转化酶(cell wall invertase, CWIN)。为探究蔗糖代谢在花粉败育导致辣椒(Capsicum annuum)雄性不育过程中的相关生理分子机理,本研究分析了INV的酶活性、基因表达模式与辣椒雄性不育性状之间的关系。本研究以1对辣椒细胞质雄性不育系131A和保持系131B为实验材料,利用INV酶活性测定、辣椒基因组INV鉴定与结构分析等,以及辣椒CWIN合成相关基因qRT-PCR等方法,对不同组织以及不同发育时期花中蔗糖转化酶的活性、转化酶合成基因表达模式等指标进行了比较分析。结果表明,在大花蕾和成花时期保持系131B的CWIN酶活性显著高于不育系131A,且差异显著性分别达到极其显著(P<0.001)和极显著(P<0.01),而这一现象与辣椒CWIN合成基因CaCWIN3和CaCWIN4在保持系131B雄蕊中的高表达水平紧密相关。据此推测,辣椒CaCWIN3和CaCWIN4可能是蔗糖转化酶参与调控辣椒雄性不育的重要功能基因。本研究结果为深入研究辣椒雄性不育分子调控机理提供了方向。Sucrose is one of the main assimilates in plant photosynthesis process. The effective use of sucrose in plant reproduction plays important roles in the process of fertilization and fruit-setting. Invertase(INV), as a key enzyme of sucrose metabolism, regulate the hydrolysis of sucrose into glucose and fructose. According tothe cellular localization, INVs are further divided as cytoplasmic invertase(CIN), vacuolar invertase(VIN),and cell wall invertase(CWIN). In order to understand the physiological and molecular mechanism of sucrose metabolism involved in the process of male sterility of pepper(Capsicum annuum) induced by pollen abortion,the INV enzyme activities, INV genes expression pattern and their correlation with pepper male sterility were analyzed. A pair of pepper male sterile line 131 A and maintainer line 131 B(fertile) were used as investigating materials in this study. The parameters including INV enzyme activities, VIN and CWIN gene identification in pepper genome, and CWIN encoding gene expression pattern by qRT-PCR, were comparatively analyzed in different tissues and developmental stages of pepper flower. The results showed that the CWIN activities in the big bud and flower of the maintainer line 131 B were significantly higher than those in the male sterile line131 A(P<0.001 and P<0.01, respectively), which were closely correlated with the high expression level of pepper CaCWIN encoding genes CaCWIN3 and CaCWIN4. According to the results, CaCWIN3 and CaCWIN4 were likely the key functional genes involved in the process of male sterility of pepper. These results shed light on the further investigation of molecular regulation mechanism of capsicum male sterility.

关 键 词：蔗糖转化酶 辣椒 雄性不育 辣椒细胞壁蔗糖转化酶合成基因3(CaCWIN3) 辣椒细胞壁蔗糖转化酶合成基因4(CaCWIN4) 

分 类 号：S641.2[农业科学—蔬菜学]