花生FUSCA3基因的克隆与表达分析  

Cloning and Expression Analysis of FUSCA3 Gene in Peanut( Arachis hypogaea L. )

作　　者：潘丽娟[1,2] 刘风珍[2] 万勇善[2] 迟晓元[1,3] 陈娜[1] 陈明娜[1] 王通[1] 王冕[1] 杨珍[1] 禹山林[1] 

机构地区：[1]山东省花生研究所,山东青岛266100 [2]山东农业大学农学院/作物生物学国家重点实验室,山东泰安271018 [3]中国农业科学院油料作物研究所/农业部油料作物生物学与遗传育种重点实验室,湖北武汉430062

出　　处：《核农学报》2015年第6期1044-1051,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家花生产业技术体系项目(CARS-14);山东省自然基金项目(ZR2011CQ036;ZR2012CQ031);国家自然科学基金项目(31000728;31100205;31200211);国家国际科技合作专项(2011DFA30930);青岛市科技计划应用基础研究项目[11-2-4-9-(3)-jch;11-2-3-26-nsh;12-1-4-11-(2)-jch];农业部油料作物生物学与遗传育种重点实验室开放课题基金(2014010)

摘　　要：为了挖掘花生品质性状相关功能基因,本研究以花生品种花育17号为试材,根据c DNA文库中已知的FUSCA3基因全长序列设计引物,通过RT-PCR克隆到该基因,命名为Ah FUSCA3。Ah FUSCA3全长为1 532 bp,开放阅读框ORF为1 134 bp,对应的基因组序列3 892 bp,由6个外显子和5个内含子组成,内含子剪接方式符合GT-AG剪接规则。根据编码区预测Ah FUSCA3编码一条378个氨基酸组成的多肽,预测分子量为42.3 k D,等电点为5.58。预测该基因编码的蛋白含有B3保守结构域,可能定位于细胞核中。蛋白序列比对和进化树分析表明,花生与大豆(Glycine max)、菜豆(Phaseolus vulgaris)、鹰嘴豆(Cicer arietinum)和苜蓿(Medicago truncatula)等豆科植物中的FUSCA3序列相似性最高,亲缘关系最近。荧光定量PCR结果显示,Ah FUSCA3基因表达量在花生荚果、种子发育的过程中呈现先增后降的趋势,推测Ah FUSCA3基因在花生胚胎形成和发育过程中发挥重要作用。本研究为阐明花生胚胎发育过程和油脂代谢机制提供了理论基础,为花生品质改良育种提供了新的基因资源。To screen functional genes related to peanut quality traits,a FUSCA3 gene was cloned from the seed of peanut（ Arachis hypogaea L. cultivar Huayu17） using RT-PCR,and was designated as Ah FUSCA3. Ah FUSCA3 c DNA was 1532 bp in length,and had an open reading frame of 1 134 bp. With 6 exons,5 introns and typical GT-AG splice sites,its genomic sequence was 3 892 bp long,encoding a protein of 378 deduced amino acid residues,with a theoretical molecular weight of 42. 3 k D and an isoelectric point（ p I） of 5. 58. The deduced protein was predicted to be located in nucleus,containing the conserved B3 domain. Multiple sequence alignments and phylogenetic analysis indicated that Ah FUSCA3 showed the highest sequence similarity and closest relatedness with FUSCA3 from leguminous species including Glycine max,Phaseolus vulgaris,Cicer arietinum,and Medicago truncatula. Expression of Ah FUSCA3 during peanut pod and seed development process,as indicated by the real time RT-PCR assay,exhibited a first increase and then decrease tendency,suggesting that Ah FUSCA3 may play an important role in peanut embryogenesis. This study provides a theoretical basis for elucidating the peanut embryogenesis and lipid biosynthesis,and prrides new gene resources for molelular breeding of peunut.

关 键 词：花生 FUSCA3基因 克隆 系统发育分析 荧光定量PCR 

分 类 号：S565.2[农业科学—作物学]