板栗花发育B类基因CmPI的克隆及功能分析  

Cloning and functional analysis of CmPI,a class B flower development gene in Chinese chestnut

作　　者：郭晓萌 杨前宇 程丽莉[1,2] 胡广隆 刘钊[4] 周广柱 兰彦平 程运河[1,2] GUO Xiaomeng;YANG Qianyu;CHENG Lili;HU Guanglong;LIU Zhao;ZHOU Guangzhu;LAN Yanping;CHENG Yunhe(Institute of Forestry and Pomology,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100093,China;Chestnut Engineering Technology Research Center,National Forestry and Grassland Administration,Beijing 100093,China;College of Forestry,Shenyang Agricultural University,Shenyang 110086,Liaoning,China;Beijing University of Agriculture,Beijing 102200,China)

机构地区：[1]北京市农林科学院林业果树研究所,北京100093 [2]国家林业和草原局板栗工程技术研究中心,北京100093 [3]沈阳农业大学林学院,沈阳110086 [4]北京农学院,北京102200

出　　处：《果树学报》2024年第7期1322-1331,共10页Journal of Fruit Science

基　　金：北京市农林科学院青年科研基金(QNJJ202315);北京市农林科学院林业果树研究所青年科研基金(LGSJJ202402);国家自然科学基金(32301624)。

摘　　要：【目的】PISTILLATA(PI)基因作为控制花器官发育的B类功能基因中的一员,在花器官发育中起到重要的作用。为探究PI基因在板栗花发育中的功能,对板栗花器官B类基因PISTILLATA(PI)进行鉴定和功能分析。【方法】在板栗基因组数据中检索PI同源基因CmPI,并克隆编码区序列。利用在线工具对CmPI进行生物信息学分析。通过荧光定量PCR分析CmPI在板栗不同组织及不同花发育时期的时空表达模式。构建35S::CmPI-GFP融合载体,瞬时转化烟草叶片,进行基因表达的亚细胞定位分析。将过表达载体35S::CmPI转入野生型拟南芥获得过表达植株,验证CmPI在花发育中的功能。【结果】克隆到的板栗CmPI基因开放阅读框长度630 bp,编码209个氨基酸,有高度保守的MADS域(MADS-box)和K域(K-box),属MADS的Ⅱ型亚家族转录因子,定位于细胞核。CmPI主要在板栗雄花中表达。在拟南芥中过表达CmPI导致萼片转变为花瓣样组织。荧光定量PCR结果表明,CmPI过表达拟南芥中除C类基因AtAG表达量显著高于野生型外,其他A类、B类、E类基因的表达量均低于野生型拟南芥。【结论】鉴定到的MADSbox基因CmPI为板栗花发育B类基因,可导致萼片花瓣化,可能是抑制雌花苞片进一步发育的关键基因。【Objective】A complete flower consists of four floral organs,including sepals,petals,stamens,and carpels from the outside to the inside.In the ABC(D)E flower development regulation model,each round of flower organ development is specifically regulated by corresponding genes.The variation of plant flowers in the process of evolution has produced a different four ring flower organ structure from the classic one.Such as chestnut,poplar,walnut,jatropha,birch are catkins,their flowers are unisexual,and do not have petals and sepals.In these plants,the petals and sepals are replaced by the bracts.The development of bracts may be co-regulated by A,B and E genes.As a typical representative of Fagaceae plants,the female and male flowers of chestnut differ greatly in morphology.The male bracts of chestnut are small and thin,and the stamens are easy to extend out during flowering.The female flowers of chestnut are usually 3 to 5 grouped together in clusters at the base of the mixed inflorescences.The female flower cluster is surrounded by numerous bracts.After the female flower is fertilized,the bracts develop into epicarp.The sexual dimorphism of chestnut bracts may also be regulated by MADS-box gene.PI gene is one of class B genes,which is involved in the development regulation of petals and stamens,and is one of the ideal genes for studying the development regulation of chestnut bracts.In this study,the Castanea mollissima variety‘Yanshan Hongli’was used as experimental material.Homology cloning,evolutionary analysis,spatio-temporal expression analysis and functional verification of the Class B flower organ development gene CmPI of C.mollissima were used to lay a foundation for the molecular regulation of bracteal dimorphism of C.mollissima.【Methods】The PI homologous gene CmPI was retrieved from the genome data of Chinese chestnut.The coding region sequence was cloned by RT-PCR.Bioinformatics analysis of the CmPI was performed using online tools.The temporal and spatial expression patterns of the CmPI in diffe

关 键 词：板栗 花器官 B类基因 基因功能验证 PISTILLATA 

分 类 号：S664.2[农业科学—果树学]