胡桃楸成花相关JmCO基因的克隆和表达分析  

Cloning and Expression Analysis of JmCO Gene Related to Flowering of Juglans mandshurica Maxim

作　　者：蔡佳友 董天一 傅靖棋 范新蕊 张丽杰[1] CAI Jia-you;DONG Tian-yi;FU Jing-qi;FAN Xin-rui;ZHANG Li-jie(College of Forestry/Key Laboratory of Forest Tree Genetics,Breeding and Cultivation of Liaoning Province,Shenyang Agricultural University Shenyang,110161,China)

机构地区：[1]沈阳农业大学林学院/辽宁省林木遗传育种重点实验室,沈阳110161

出　　处：《沈阳农业大学学报》2021年第5期618-624,共7页Journal of Shenyang Agricultural University

基　　金：国家“十三五”重点研发计划项目(2017YFD0600600);辽宁省教育厅重点攻关项目(LSNZD201905);辽宁省自然科学基金重点项目(20170540801)。

摘　　要：为探讨JmCO基因在胡桃楸花发育过程中的调控作用及促进提早开花的分子机理,从胡桃楸转录组数据库中筛选到JmCO基因片段,利用JmCO基因差异片段设计上下游引物,通过PCR扩增技术克隆得到该基因的CDS序列全长,将该基因命名为JmCO。利用生物信息学软件对JmCO基因所编码蛋白质的进行聚类及序列特征、蛋白结构、系统进化分析及亚细胞定位预测等。应用RT-PCR技术分析JmCO基因在胡桃楸不同器官、组织中的表达量。结果表明:克隆得到胡桃楸成花相关JmCO基因序列全长为582bp,其编码194个氨基酸。蛋白质分子量为21569.04Da;理论等电点(pI)值为8.3;NCBI数据库Blast比对表明,胡桃楸成花相关JmCO基因与其他物种的CO基因同源性均在70%以上。qRT-PCR荧光定量表达分析得出胡桃楸JmCO基因在雌雄花芽、果实、叶片、茎中均有表达,但在雌雄花芽中表达量最高。推测该基因参与了胡桃楸花芽发育的整个过程,在成花过程中发挥了重要的调控作用。该研究结果为进一步探索胡桃楸成花的分子机理奠定了研究基础。To explore the regulatory role of JmCO gene in the flower development of Juglans mandshurica and the molecular mechanism of promoting early flowering.The JmCO gene fragment was screened from the transcriptome database of Juglans mandshurica.The upstream and downstream primers were designed by using the JmCO gene differential fragment.The full length of the CDS sequence of the gene was cloned by PCR amplification technology,and the gene was named JmCO.Using bioinformatics software,JmCO gene was analyzed for protein clustering,sequence characteristics,protein structure,phylogenetic analysis and subcellular localization prediction.RT-PCR was used to analyze the expression of jmco gene in different organs and tissues of Juglans mandshurica.The results showed that the total length of JmCO gene related to flower formation of Juglans mandshurica was 582bp,encoding 194 amino acids.The protein molecular weight is 21569.04Da.The theoretical isoelectric point pI value is 8.3.Blast comparison of NCBI database showed that JmCO gene related to flowering of Juglans mandshurica had more than 70%homology with CO gene of other species.qRT-PCR fluorescence quantitative expression analysis showed that JmCO gene was expressed in male and female flower buds,fruits,leaves and stems,but the expression was the highest in male and female flower buds.The JmCO gene related to flowering was successfully cloned from Juglans mandshurica.It is speculated that this gene is involved in the whole process of flower bud development and plays an important regulatory role in the process of flowering.The results laid a solid foundation for further exploring the molecular mechanism of flowering of Juglans mandshurica.

关 键 词：胡桃楸 JmCO基因克隆 表达分析 

分 类 号：S792.132[农业科学—林木遗传育种]