西葫芦种皮纤维素合成酶基因CpCesA的克隆及表达分析  

Cloning and Expression Analysis of Cellulose Synthase Gene CpCesA in Testa of Cucurbita pepo L.

作　　者：赵夏云 瞿飞 杨静[1] 马关鹏 田颖 文林宏[1] Zhao Xiayun;Qu Fei;Yiang Jing;Ma Guanpeng;Tian Ying;Wen Linhong(Institute of Horticulture,Guizhou Academy of Agricultural Sciences,Guiyang,550025)

机构地区：[1]贵州省农业科学院园艺研究所,贵阳550025

出　　处：《分子植物育种》2024年第10期3148-3155,共8页Molecular Plant Breeding

基　　金：贵州省科技计划项目(黔科合支撑[2021]一般212);贵州省农业科学院2018年度学术新苗培养及创新探索专项(黔科合平台人才[2018]5768-04);贵州省高层次创新型人才培养计划“百”层次人才(黔科合平台人才[2016]5671号);贵州省蔬菜现代农业产业技术体系项目(GZCYTX2021);贵州省农业科学院项目(黔农科院种质资源[2020]10号)共同资助。

摘　　要：裸仁西葫芦因其种皮退化无需脱壳而备受关注,纤维素是植物细胞壁的主要组成成分,是影响西葫芦种皮形成的重要因素,而纤维素合成酶(CES)在植物纤维素合成过程中起关键作用。为深入探究西葫芦CpCesA基因对种皮形成的调控机制,本研究通过克隆获得CpCesA的基因序列,利用生物信息学方法分析其结构和编码蛋白的理化性质。利用实时荧光定量PCR检测种皮不同发育时期和西葫芦不同组织部位CpCesA的表达模式。通过克隆获得CpCesA开放阅读框序列全长2 952 bp,生物信息学分析表明,其编码的蛋白质在葫芦科作物中有很强的保守性,且具有纤维素合成酶家族蛋白的保守结构域,属于质膜蛋白。实时荧光定量PCR结果显示:在种子发育过程中,Cp Ces A在裸仁西葫芦种皮中的表达量比有壳西葫芦普遍显著偏低,有壳西葫芦在授粉后10 d CpCesA表达量达到最高,而裸仁西葫芦则在授粉后20 d达到最高后开始下降。不同品种种皮发育相同时期、同一品种不同发育时期Cp CesA表达量表现均不同,这表明CpCesA与西葫芦种皮的发育和形成有着密切关系。组织表达分析表明,Cp CesA在两种西葫芦叶片、茎、根、花瓣等不同组织中均有表达,茎和根中表达量高且差异极显著,具有组织表达特异性。本研究为进一步研究纤维素影响西葫芦种皮发育的机理提供参考依据。Hull-less Cucurbita pepo L.has attracted much attention because of distinctive seedcoat features.Cellulose is the main component of plant cell wall and an important factor affecting the testa formation of C.pepo.Cellulose synthase(CES)plays a key role in the synthesis of plant cellulose.In order to further explore the regulation mechanism of CES gene on testa formation of C.pepo,the gene sequence of CpCesA was obtained by cloning,and its structure and physicochemical properties of the encoded protein was analyzed by bioinformatics methods.Quantitative real-time PCR was used to detect the expression patterns of CpCesA in different tissue and different development stages of testa.The full-length CpCesA ORF sequence is 2952 bp.Bioinformatics analysis shows that CesA proteins are highly conserved in Cucurbitaceae,and CpCesA belongs to plasma membrane protein with the conserved domain of cellulose synthase family proteins.Quantitative real-time PCR results show that,during seed development,there is generally a lower expression of CpCesA in the testa of hull-less strain as compared to the hull strain.The maximum expression of CpCesA in hull-less strain is found at 10 days after pollination in contrast to 20 days after pollination for the hull strain.In the same period of testa development of different varieties,the expression levels of CpCesA are different.The relative transcript levels of CpCesA,meanwhile,are also different in different development periods of the same variety.The results indicate that CpCesA is closely related to the development and formation of testa.Tissue expression analysis shows the CpCesA transcript is present in leaves,stems,roots,petals and other different tissues of the two strains.The expression levels in stems and roots are higher and the differences are extremely significant,showing tissue specificity of expression.This study provides a reference for further study on the mechanism of cellulose affecting the testa development of C.pepo.

关 键 词：西葫芦 纤维素合成酶 基因克隆 表达分析 

分 类 号：S642.6[农业科学—蔬菜学]