枣树novel-miR16靶基因ZjTCP4鉴定及生物信息学分析

Identification and bioinformatics analysis of novel-miR16 target gene ZjTCP4 in Chinese jujube

TCPs转录因子主要通过调节细胞增殖和植物激素途径来调控植物器官发生和形态构型,探究枣树ZjTCP4转录因子的序列特征及其表达特性,以期为TCP转录因子响应干旱胁迫的机理研究提供参考,也可以为后续对枣树分子育种奠定基础。novel-miR16的二级结构具有miRNA典型的发夹结构,为枣树miR319家族成员;靶基因ZjTCP4的降解位点多数都在novel-miR16结合位点的第10和11个核苷酸的位置;生物信息学分析表明,ZjTCP4基因的全长cDNA序列为1386 bp,预测编码452个氨基酸的多肽,预测编码蛋白的等电点(pI)为6.51,分子量为49091.55 u;应该是一个膜内或膜外蛋白,为非分泌性蛋白,为亲水性蛋白质,定位于细胞核;枣树ZjTCP4基因编码的氨基酸序列与拟南芥AtTCP4最为相似,与拟南芥AtTCP17,AtTCP 18和AtTCP 12的相似性也较高;枣树novel-miR16在响应干旱胁迫的过程中上调表达,而其靶基因ZjTCP4在干旱胁迫下下调表达。枣树novel-miR16通过靶向调控ZjTCP4转录因子响应干旱胁迫,干旱胁迫下枣树novel-miR16的上调表达导致靶基因ZjTCP4(CCG006181)下调表达来提高枣树的耐旱性。

TCPs transcription factor regulates plant organogenesis and morphological configuration mainly through regulating cell proliferation and plant hormone pathways,so as to explore the sequence characteristics and expression characteristics of ZjTCP4 transcription factor of Chinese jujube,in order to provide reference for the mechanism study of TCP transcription factor response to drought stress,and lay a foundation for the subsequent molecular breeding of jujube tree.The novel-miR16 secondary structure has the typical hairpin structure of miRNA and is a member of the jujube miR319 family.Most of the degradation sites of target gene ZjTCP4 were located at the 10th and 11th nucleotides of the novel-miR16 binding sites.Bioinformatics analysis showed that the full-length cDNA sequence of ZjTCP4 gene was 1386 bp,predicted to encode 452 amino acid polypeptide,isoelectric point(pI)of 6.51,molecular weight of 49091.55 u.It should be an intramembranous or extracellular protein,a non-secretory protein,a hydrophilic protein,located in the nucleus.The amino acid sequence encoded by ZjTCP4 gene of Chinese jujube is most similar to AtTCP4 of Arabidopsis thaliana,and has high similarity to AtTCP17,AtTCP 18 and AtTCP 12 of Arabidopsis thaliana.Novel-miR16 was up-regulated in response to drought stress,while its target gene ZjTCP4 was down-regulated under drought stress.Novel-miR16 can regulate ZjTCP4 transcription factor in response to drought stress.Under drought stress,the up-regulated expression of novel-miR16 leads to down-regulated expression of target gene ZjTCP4(CCG006181)to improve drought tolerance of Chinese jujube.