馥郁滇丁香LgFKF1基因的克隆及节律表达分析  

Cloning and Rhythmic Expression Analysis oi LgFKF1 Gene in Luculia gratissima

作　　者：万友名[1] 马宏[1] 刘雄芳[1] 张序 安静 刘秀贤[1] 李正红[1] WAN You-ming;MA Hong;LIU Xiong-fang;ZHANG Xu;AN Jing;LIUXiu-xiang;LI Zheng-hong(Research Institute of Resource Insects,Chinese Academy of Forestry,Kunming 650224,Yunnan China)

机构地区：[1]中国林业科学研究院资源昆虫研究所,云南昆明650224

出　　处：《林业科学研究》2020年第1期99-106,共8页Forest Research

基　　金：云南省科技厅重点研发项目(2018BB012);云南省技术创新人才培养对象(2016HB007);国家林业局林业公益性行业科研专项经费面上项目(201404705);中国林科院基本科研业务费专项项目(CAFYBB2017MB014)。

摘　　要：[目的]研究馥郁滇丁香LgFKF1基因的结构特点及其在特异组织中的节律表达特征,探索其在成花调控过程中的作用。[方法]采用RACE技术克隆获得馥郁滇丁香LgFKF1基因的cDNA全长序列,利用生物信息学方法对获得的基因核昔酸序列及编码蛋白质序列进行分析,利用qRT-PCR技术对特异组织的节律表达模式进行分析。[结果]序列分析表明:LgFKF1基因cDNA全长为2 271 bp,开放阅读框为1 917 bp,编码一个638个氨基酸的蛋白质;推导的氨基酸序列与扁果树、软枝黄蝉、纳塔尔倒吊笔、马利筋的FKF1蛋白具有较高同源性,达92.59%;预测LgFKF1蛋白属于不稳定的亲水性蛋白,定位在细胞核,无信号肽和跨膜区;结构主要由无规则卷曲结构、α螺旋结构、扩展长链和β-转角构成;遗传进化关系与扁果树最亲近。qRT-PCR分析表明:Zg如7在诱导光周期下处理7d时较非诱导光周期下的表达量高,而诱导光周期下处理10d后其表达量则低于非诱导光周期的表达。一天中,在各组织中均有表达,且以叶片中的相对表达量较高。LgFKF1因组织不同在不同的时间呈现出单峰和双峰表达,其中,根、芽及花蕾中均在夜间23:00出现单峰表达;茎、叶及开放的花朵中在不同时间出现双峰表达,茎在晚上20:00和凌晨5:00,叶在凌晨2:00和早上8:00,开放花朵在夜间23:00和凌晨5:00. [结论]从馥郁滇丁香中克隆获得了LgFKF1基因,其编码的蛋白序列与其它植物的FKF1具有较高的同源性。LgFKF1基因的表达受光周期影响,在诱导光周期下,不同的组织呈现出不同的表达模式,其中,仅叶片中的1个表达峰值出现在白昼,其余组织中的表达峰值均出现在夜间。在各组织中,叶片中的表达量较高。LgFKF1基因的特异组织节律表达有助于为其生物学功能的进一步研究提供参考依据。[Objective]To study the regulatory role of LgFKF1 gene during flowering process of L.gratissima,based on the analysis of its gene structural characteristics and rhythmic expression in specific tissues.[Method]The RACE technology was used to clone the full length cDNA of LgFKF1 gene.Sequence of nucleotides and code of protein were analyzed by using the software of bioinformatics.qRT-PCR was applied in the rhythmic expression analysis of the gene in specific tissues.[Result]The result of sequence analysis showed that the LgFKF1 cDNA,2271 bp in length,has a 1917 bp open reading frame encoding a putative protein with 638 amino acids.The results of sequence alignment showed that the amino acids sequence was highly homologous with the FKF1 of Kaliphora madagascariensis 9 Allamanda cathartic^Wrightia natalensis and Asclepias curassavica,with the homology of 92.59%.LgFKF1 protein that had the closest genetic relationship with K.madagascariensis was unstable hydrophilic proteins without signal peptide and transmembrane regions,and located in the nucleus,mainly consisted of alpha helix,irregular curling structure and extended long chain.The result of qRT-PCR showed that the expression of LgFKF1 under inductive photoperiod for 7 days was higher than that under non-inductive photoperiod treatment,but the expression was lower than that under non-inductive photoperiod treatment if inductive photoperiod was more than 10 days.In a day,LgFKF1 expressed in various tissues,and showed a high transcription level in leaf.For the rhythmic expression of LgFKF1 in different tissues,one peak value was appeared at 23:00 in root,leaf bud and flower bud,and two peak values were appeared at 20:00 and 5:00 in stem,2:00 and 8:00 in leaf 23:00 and 5:00 in mature flower.[Conclusion]LgFKF1 was cloned from L.gratissima,and its putative protein sequence was highly homologous with the FKF1 of other plants.The expression of LgFKF1 gene was affected by photoperiod.Under the inductive photoperiod,various tissues showed itself expression pattern.A peak v

关 键 词：馥郁滇丁香 LgFKF1基因 克隆 节律表达 

分 类 号：S718.46[农业科学—林学]