杉木OFP基因家族的表达模式及亚细胞定位  

Expression Pattern and Subcellular Localization of the OFP Gene Family in Chinese Fir(Cunninghamia lanceolata)

作　　者：应威洋 胡现铬 黄华宏[1] 林二培[1] 童再康[1] YING Wei-Yang;HU Xian-Ge;HUANG Hua-Hong;LIN Er-Pei;TONG Zai-Kang(The State Key Laboratory of Subtropical Silviculture,Zhejiang Agricultural and Forestry University,Lin'an 311300,China)

机构地区：[1]浙江农林大学亚热带森林培育国家重点实验室,临安311300

出　　处：《农业生物技术学报》2023年第8期1610-1622,共13页Journal of Agricultural Biotechnology

基　　金：浙江省农业新品种选育项目(2021C02070-8);“十三五”国家重点研究计划林业资源培育及高效利用技术创新重点专项(2017YFD0600201)。

摘　　要：卵形家族蛋白(OVATE family protein,OFP)是植物特有的一类调控植物形态建成和次生壁发育的转录因子,目前在杉木(Cunninghamia lanceolata)中尚未见该基因家族的相关报道。本研究基于全长转录组数据,鉴定杉木OFP基因家族成员,进行序列和进化树构建分析,并检测其在不同组织和器官中的表达模式及编码蛋白的亚细胞定位情况。结果显示,鉴定到6个杉木OFP基因,均包含典型的OVATE结构域且不含内含子,分散于5个亚组中。实时荧光定量PCR显示,6个ClOFPs的表达模式不尽相同,其中ClOFP1在杉木幼叶中的表达量最高,在茎中的表达量呈现随木质化程度提高而下降的趋势;ClOFP2在雌球花中的表达量最高,ClOFP3在根中的表达最高且2个基因在低木质化茎(S1)中的表达量较在其他茎中高;ClOFP4在雌、雄球花和叶片中的表达量普遍较高;ClOFP5在雄球花中的表达量最高,且在茎中的表达量随木质化程度提高而上升;ClOFP6在茎段、雌球花中的表达量相对较高。此外,所有ClOFPs基因在叶片中的表达呈现随叶片成熟程度增加而减弱的趋势。亚细胞定位结果显示,ClOFP1蛋白定位在细胞壁上,ClOFP2/3蛋白定位在细胞膜上,ClOFP4/5/6蛋白定位在细胞核中,表明杉木ClOFPs基因很可能具有不同的调控途径。因此,ClOFPs基因在杉木的球花、叶和茎等器官的发育以及木质素合成方面具不同的调控作用。研究结果为深入研究杉木器官形态建成和木材形成提供理论依据,同时也为杉木分子育种提供潜在的基因资源。The OVATE family protein(OFP)is a plant-specific regulatory protein that plays a significant role in regulating plant morphogenesis and secondary cell wall formation.However,little information about the OFP gene family in Chinese fir(Cunninghamia lanceolata)is available.In this study,ClOFP genes were identified based on the full-length transcriptome.Bioinformatics characteristics and evolutionary relationships were analyzed using online service.Besides,relative expression patterns of all members in different tissues and organs were determined by real time fluorescence quantitative PCR(qRT-PCR).Furthermore,the subcellular localizations of all proteins were determined by transient transfection assays.The results showed that 6 ClOFP genes were identified from RNA-seq data,with a typical OVATE domain and contained no introns.The phylogenetic analysis revealed that ClOFP were distributed in 5 subgroups.In addition,relative expression patterns of different tissues and organs varied differently.ClOFP1 was strongly expressed in young leaves and the relative expressions in stems were approximately decreased with an increase of lignification.ClOFP2 dominantly expressed in female cone.ClOFP3 strongly expressed in root.The relative expression of ClOFP2/3 in low lignified stem(S1)was higher than that in others.ClOFP4 dominantly expressed in female cone,male cone and leaves.ClOFP5 was strongly expressed in male cone and the relative expressions of ClOFP5 in stems were approximately increased with an increase of lignification.ClOFP6 was strongly expressed in stems and female cone.Moreover,the relative expressions of ClOFPs in leaves were decreased with an increase in maturity.The results of subcellular localization showed the localization of ClOFP1 in the cytoplasm,ClOFP2/3 in the plasma membrane and nuclear localization of ClOFP4/5/6.This suggested that ClOFPs might involve in different regulatory pathways in Chinese fir.Hence,ClOFPs might involve in regulating the development of cone,leaf,stem and other organs,as well as lign

关 键 词：杉木 卵形家族蛋白(OFP)基因家族 基因克隆 表达模式 亚细胞定位 

分 类 号：S791.27[农业科学—林木遗传育种]