杉木ClCRY2基因克隆及表达特性分析  

Cloning and Expression Analysis of ClCRY2 Gene in Cunninghamia lanceolata

作　　者：郭胜周 廖文海 李婧姝 邓煜钊 曹光球[1,2,3] 曹世江 GUO Shengzhou;LIAO Wenhai;LI Jingshu;DENG Yuzhao;CAO Guangqiu;CAO Shijiang(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Chinese Fir Engineering Technology Research Center of National Forestry and Grassland Administration,Fujian Agriculture and Forestry University,Fuzhou 350002,China;University Key Laboratory of Forest Stress Physiology,Ecology and Molecular Biology of Fujian Province,Fuzhou 350002,China)

机构地区：[1]福建农林大学林学院,福州350002 [2]国家林业草原杉木工程技术研究中心,福州350002 [3]林木逆境生理生态及分子生物学福建省高校重点实验室,福州350002

出　　处：《四川农业大学学报》2024年第3期620-627,共8页Journal of Sichuan Agricultural University

基　　金：“十四五”国家重点研发计划:杉木大径级无节良材培育技术(2021YFD2201302)。

摘　　要：【目的】通过克隆杉木隐花色素CRY2基因,研究其在不同组织和不同光质处理下的表达特性,为探索CRY2基因在杉木生长发育过程中的调控作用提供参考。【方法】以1 a生杉木优良无性系061扦插苗为试验材料,基于课题组杉木全长转录组测序的结果,利用逆转录聚合酶链式反应(RT-PCR)技术,设计基因特异性引物,从无性系杉木苗嫩叶中克隆CRY2基因,命名为ClCRY2(登录号为PP400318)。对其进行生物信息学分析,并利用实时荧光定量PCR技术分析其表达情况。【结果】生物信息学分析结果显示,杉木的ClCRY2基因CDs区全长2 151 bp,共编码716个氨基酸,与拟南芥中AtCRY2蛋白序列的同源一致性为68.93%。进化树分析发现ClCRY2蛋白与日本柳杉(Cryptomeria japonica)CjCRY2的亲缘性最近。ClCRY2蛋白二级结构主要以无规则卷曲(random curls)和α-螺旋(alpha helix)为主,分别为51.26%和34.78%。ClCRY2蛋白分子式为C3 620H5 573N1 033O1 063S27,理论等电点为6.46,占比最大的氨基酸为亮氨酸(leucine)占9.8%,其次是丝氨酸(serine),占8.0%。ClCRY2蛋白为亲水性蛋白,不具有跨膜区,无信号肽。实时荧光定量PCR分析结果:ClCRY2基因在杉木苗木根茎叶中均有表达,其中茎中的表达量最高,叶中次之,根中表达量最低;蓝光能显著增加ClCRY2基因的表达。【结论】通过对ClCRY2基因所编码的蛋白的理化性质和结构的研究,推测其在杉木茎和叶片发育进程中起重要的调控作用,参与蓝光信号的传导过程。【Objective】By cloning the CRY2 gene of Cunninghamia lanceolata,the expression character⁃istics of CRY2 gene in different tissues and under different light quality treatments were studied,which provided a reference for exploring the regulatory role of CRY2 gene in the growth and development of C.lanceolata.【Method】Based on the results of our research group′s full-length transcriptome sequencing of C.lanceolata,one-year old cuttings from the superior clone 061 were employed as experimental mate⁃rials.Using reverse transcription polymerase chain reaction(RT-PCR)technology,gene-specific prim⁃ers were designed and the CRY2 gene were cloned from the tender leaves of C.lanceolata seedlings,named ClCRY2(Accession number PP400318).Furthermore,bioinformatics analysis were conducted on this gene and its expression pattern were analyzed using real-time quantitative PCR technology.【Result】 The bioinformatics analysis results showed that the full length of the CDs region of the ClCRY2 gene in C. lanceolata was 2 151 bp, encoding a total of 716 amino acids, with a homologous consistency of 68.93% with the AtCRY2 protein sequence in Arabidopsis. Phylogenetic tree analysis revealed that the ClCRY2 protein was most closely related to CjCRY2 in Japanese cedar (Cryptomeria japonica). The sec⁃ ondary structure of ClCRY2 protein was mainly dominated by random coils and alpha helix, accounting for 51.26% and 34.78%, respectively. The molecular formula of ClCRY2 protein is C3 620H5 573N1 033O1 063S 27, with a theoretical isoelectric point of 6.46. The largest proportion of amino acids was leucine, accounting for 9.8%, followed by serine, accounting for 8.0%. ClCRY2 protein was a hydrophilic protein that did not have a transmembrane region and had no signal peptide. Real time quantitative PCR analysis results showed: ClCRY2 gene was observed to express in the roots, stems, and leaves of C. lanceolata seedlings, with the highest expression level in the stem, followed by the leaves, and the lowest expression level in

关 键 词：杉木 隐花色素 ClCRY2基因 生物信息学 表达分析 

分 类 号：S722.8[农业科学—林木遗传育种]