棉花FLA基因家族的全基因组鉴定及GhFLA05在棉纤维发育中的功能分析  

Genome-Wide Identification of Cotton FLA Gene Family and Functional Analysis of GhFLA05 in Cotton Fiber Development

作　　者：唐丽媛[1] 蔡肖[1] 王海涛 李兴河 张素君[1] 刘存敬[1] 张建宏[1] TANG LiYuan;CAI Xiao;WANG HaiTao;LI XingHe;ZHANG SuJun;LIU CunJing;ZHANG JianHong(Institute of Cotton,Hebei Academy of Agriculture and Forestry Sciences/Key Laboratory of Cotton Biology and Genetic Breeding in Huanghuaihai Semiarid Area,Ministry of Agriculture and Rural Affairs/National Cotton Improvement Center Hebei Branch,Shijiazhuang 050051)

机构地区：[1]河北省农林科学院棉花研究所/农业农村部黄淮海半干旱区棉花生物学与遗传育种重点实验室/国家棉花改良中心河北分中心,石家庄050051

出　　处：《中国农业科学》2023年第23期4602-4620,I0004-I0011,共27页Scientia Agricultura Sinica

基　　金：河北省农林科学院基本科研业务费(2021070205);河北省农林科学院科技创新专项课题(2022KJCXZX-MHS-1,2023KJCXZX-MHS-11)。

摘　　要：【背景】伴随着棉纺织工艺水平的提升和人们对高品质纺织品的追求,提升棉花纤维品质日益重要。类成束阿拉伯半乳糖蛋白(fasciclin-like arabinogalactan proteins,FLAs)在棉纤维起始发育、次生壁合成等过程中可能具有重要作用。【目的】通过对棉花FLA基因家族进行全面鉴定与分析,研究该家族成员的共性特征及特异性表达模式,为FLA在棉纤维发育中的功能研究提供参考。【方法】根据棉花全基因组数据,使用HMMER 3.0对棉花FLA基因家族成员进行鉴定,并通过Pfam、Smart等软件进一步确认。使用ExPASy、TMHMM分析蛋白理化性质及跨膜结构域,应用MEGA、MCScanX、GSDS、MEME、TBtools、Jalview等工具进行进化树构建、染色体定位、共线性分析和蛋白保守结构域序列比对等。通过转录组数据分析陆地棉FLA基因在不同组织中的表达情况。利用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测GhFLAs在不同纤维品质材料的胚珠及不同发育时期纤维中的表达差异。利用病毒诱导的基因沉默(virus induced gene silencing,VIGS)技术验证GhFLA05的功能。【结果】在陆地棉、海岛棉、亚洲棉和雷蒙德氏棉全基因组中分别鉴定出41、40、20和21个FLA家族成员,系统进化树显示,棉花FLA蛋白可以分为4个群组。进一步对陆地棉FLA家族蛋白进行分析,41个成员均具有1—2个AGP-like糖基化区域和1—2个类成束蛋白结构域(fasciclin-like domain,FAS),其中,37个含有信号肽(signal peptide,SP),25个含有糖基化磷脂酰肌醇(glycosylphosphatidy linositol anchored protein,GPI)锚定信号,基因结构和基序组成在各组中相对保守。亚细胞定位显示,GhFLA05_D可能定位在细胞质的内质网,呈聚集状颗粒,GhFLA18_A和GhFLA22在细胞膜/壁、细胞质和细胞核中均有表达。转录组测序结果表明,Group A和Group B中的FLA蛋白主要在纤维中高表达,可能�【Background】It is of great importance to improve the quality of cotton fiber to meet the improvement of cotton textile production and the pursue of people for high quality cotton.Fasciclin-like arabinogalactan proteins(FLAs)play an important role in the initial development of cotton fibers and secondary wall synthesis.【Objective】Comprehensive identification and analysis of cotton FLA gene family members to reveal their common characteristics and specific expression patterns,provided a reference for the function study of FLAs in cotton fiber development.【Method】According to the whole genome data of cotton,members of FLA gene family were identified by HMMER3.0 and further verified by online softwares of Pfam and Smart.Physical and chemical properties and transmembrane domains of these proteins were analyzed by ExPASy and TMHMM.Phylogenetic tree construction,chromosome localization,collinearity analysis and protein conserved domain sequence alignment were conducted and displayed using GSDS,MCScanX,MEGA,MEME,TBtools and Jalview.Expression of FLA genes in different tissues were analyzed by cotton transcriptome data.Expression differences of GhFLAs in different developmental stages of ovules and fibers between different fiber quality materials was analyzed by quantitative real-time polymerase chain reaction(qRT-PCR).Function of GhFLA05 was verified by virus induced gene silencing(VIGS).【Result】A total of 41,40,20 and 21 FLA family members were identified in G.hirsutum,G.barbadense,G.arboreum and G.raimondii,respectively.The phylogenetic tree showed that cotton FLA proteins could be divided into four groups.Gene structure and motif composition were relatively conserved in each group.Further analysis of FLA proteins in Gossypium hirsutum showed that all 41 FLA members had 1-2 AGP-like glycosylation regions and 1-2 fasciclin-like domains(FAS),37 of which contained signal peptide(SP)and 25 contained glycosylphosphatidylinositol anchored protein(GPI)anchoring signals.Subcellular localization showed that GhFL

关 键 词：棉花 FLA 纤维发育 基因家族 表达分析 

分 类 号：S562[农业科学—作物学]