摘要
SOC1(SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1)是植物中的一个开花时间调节因子,属于MADS-box转录因子家族,在植物从营养生长向生殖生长的转变中发挥关键作用。SOC1参与多种生物过程,包括花器官发育、开花时间控制以及对环境刺激和激素信号的响应。本研究通过遮荫和喷施乐斯本的方式对莲雾进行催花处理,统计处理后的叶芽和花芽数量以及成花枝率,同时,利用课题组现有的莲雾转录组进行分析,筛选出调控莲雾开花的候选基因SsSOC1,并通过实时荧光定量PCR技术研究遮荫处理和喷施乐斯本药剂处理下莲雾不同时期和组织中SsSOC1表达量的差异。结果显示:经过遮荫处理的莲雾树体芽点、叶片、叶尖以及叶柄中的SsSOC1基因表达量显著增加,其中叶尖的相对表达量最高。本研究成功克隆了莲雾黑糖芭比品种的SsSOC1基因,并利用生物信息学方法对其进行启动子分析、蛋白理化性质分析、保守功能域的预测和序列比对。结果表明:SsSOC1基因的开放阅读框长度为654 bp,共编码217个氨基酸,其编码蛋白的理论等电点为9.37,分子式为C_(1076)H_(1794)N_(322)O_(334)S_(10),分子量为24.91kDa,带正电荷氨基酸残基总数(Lys+Arg)为40,带负电荷氨基酸残基总数(Asp+Glu)为32,脂肪系数为80.64,总平均亲水性系数(GRAVY)为–0.729,不稳定系数为66.17;莲雾SsSOC1蛋白结构不含β-折叠,不属于分泌蛋白与膜结构蛋白,亚细胞定位显示其主要存在于细胞核中,属于不稳定的亲水性碱性蛋白,该蛋白具有MADS_MEF2_like和K-Box保守结构域,同时隶属于MADS超级家族和K-Box超级家族,该结构域的起止氨基酸位点为第3~75位和第88~171位,符合MADS蛋白家族与K-Box家族的典型结构特征;系统进化树分析显示,莲雾SsSOC1序列与油叶蒲桃的同源性最高;通过花序侵染法将SsSOC1基因转化拟南芥,发现35S::SsSOC1表现出早花表型,莲座叶数量比野生型要少。本研究初步确定了SsSOC1基因在成花方面的重要作用,并揭示莲雾遮荫与成花相关的内在调控机理,为莲雾生产提供指导意义,以及为莲雾反季节栽培技术改良提供理论依据。
SOC1(SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1)is a flowering time regulator in plants.It belongs to the MADS-box transcription factor family and plays a crucial role in the transition from vegetative to repro-ductive growth in plants.SOC1 is involved in various biological processes,including flower organ development,flow-ering time control,and response to environmental stimuli and hormone signals.In this study,shading and application of Lorsben were used for flower induction in wax apple,and the number of leaf buds,flowers,and the rate of flowering shoots were recorded after treatment.Additionally,the existing transcriptome data of wax apple in our research group were analyzed to identify the candidate gene SsSOC1 involved in regulating flowering in wax apple.The differential expression of SsSOC1 in different stages and tissues of wax apple under shading and Lorsben treatment was studied using qPCR.The results showed that the expression of the SsSOC1 gene significantly increased in the buds,leaves,leaf tips,and petioles of wax apple treated with shading,with the highest relative expression in the leaf tips.In this study,the SsSOC1 gene from Hei Tang Ba Bi wax apple was successfully cloned,and its promoter analysis,physicochemical properties of the protein,prediction of conserved functional domains,and sequence alignment were performed using bioinformatics methods.The open reading frame of the SsSOC1 gene in wax apple was found to be 654 bp,encoding a protein of 217 amino acids.The theoretical isoelectric point of the protein was 9.37,with a molecular formula of C_(1076)H_(1794)N_(322)O_(334)S_(10) and a molecular weight of 24.91 kDa.The protein contained 40 positively charged amino acid residues(Lys+Arg)and 32 negatively charged amino acid residues(Asp+Glu).The protein had a fat coefficient of 80.64,a grand average of hydropathicity(GRAVY)of–0.729,and an instability coefficient of 66.17.The protein structure lackedβ-folds and did not belong to secretory or membrane proteins.Subcellular localization analysis indicated that the protein was mainly present in the nucleus and belonged to an unstable hydrophilic basic protein.The SsSOC1 protein in wax apple possessed conserved domains including MADS_MEF2_like and K-Box and belonged to the MADS super-family and K-Box superfamily.The analysis identified the amino acid positions 3 to 75 and 88 to 171 as the start and end sites of the conserved domains,respectively,conforming to the typical structural features of the MADS protein family and the K-Box family.The evolutionary tree showed the highest homology between the SsSOC1 sequence in wax apple and Syzygium oleosum.By floral dip transformation,the SsSOC1 gene was introduced into Arabidopsis,and it was found that the 35S::SsSOC1 line exhibited an early flowering phenotype and a reduced number of rosette leaves com-pared to the wild type.This study preliminarily elucidated the important role of the SsSOC1 gene in flowering in wax apple and attempted to reveal the intrinsic regulatory mechanism of shading-induced flowering,providing guidance for wax apple production and theoretical basis for improving out-of-season cultivation techniques.
作者
杨胤延
孙东宇
蔡昕成
胡志群
吴国麟
周碧燕
YANG Yinyan;SUN Dongyu;CAI Xincheng;HU Zhiqun;WU Guolin;ZHOU Biyan(Key Laboratory of Horticultural Crop Biology and Germplasm Creation in South China,Ministry of Agriculture and Rural Affairs/College of Horticulture,South China Agricultural University,Guangzhou,Guangdong 510642,China;Center for Modern Agricultural Development,Zhuhai,Guangdong 519000,China)
出处
《热带作物学报》
北大核心
2025年第3期553-562,共10页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目(No.32072515)
广州市重点研发计划项目(No.202206010023)
关键词
莲雾
成花
SsSOC1
基因克隆
转录活性分析
表达模式分析
wax apple
flowering
SsSOC1
gene cloning
transcriptional activity analysis
expression pattern analysis
