橡胶树HbWRI转录因子的克隆与表达分析

Cloning and Expression Analysis of Transcription Factor HbWRI from Hevea brasiliensis

脂质是橡胶树胶乳的主要非胶组分,与胶乳代谢和天然橡胶质量形成密切相关,但其代谢调控相关研究欠缺。WRINKLED(WRI)是植物AP2转录因子家族成员,在促进碳源从糖流向油脂合成中发挥了重要作用。本研究从橡胶树胶乳中克隆到3个HbWRI基因家族成员,分别为HbWRI1,HbWRI2和HbWRI3,并对其进行生物信息学分析、不同组织部位表达分析以及亚细胞定位分析。结果表明,HbWRI1,HbWRI2和HbWRI3的编码序列全长分别为1128、1122 bp和1050 bp,分别编码了375、373和349个氨基酸。HbWRI1-3蛋白具有2个60~70个氨基酸大小的AP2保守结构域,第1个AP2结构域中包含1个VYL基序和1个14-3-3蛋白结合基序。生信分析预测HbWRI1-3均无跨膜结构域,无信号肽,为疏水性蛋白,且含有多个磷酸化位点。亚细胞定位分析表明,HbWRI1-3蛋白定位于细胞核上。不同组织部位表达分析表明,HbWRI1、HbWRI2和HbWRI3基因均在橡胶树胶乳中表达量最高,且在胶乳中HbWRI3是家族成员中表达量最高的基因,暗示HbWRI1-3可能参与了胶乳脂质合成调控。该研究为进一步探索HbWRI1-3转录因子在橡胶树胶乳代谢及影响天然橡胶质量形成中的作用奠定基础。

Lipid is the main non-rubber component of Hevea brasiliensis,and it is closely related to latex metabolism and natural rubber quality formation.However,few reseaches were found on its metabolic regulation.WRINKLED(WRI),a member of the plant AP2 transcription factor family,plays an important role in promoting the flow of carbon sources from sugar to lipid synthesis in plants.In this study,three transcription factors HbWRI1,HbWRI2 and HbWRI3 of the HbWRI gene family were successfully cloned from the latex of H.brasiliensis,respectively.And their bioinformatics analysis,expression analysis of different tissue sites and subcellular localization analysis were performed.The results showed that the encoding sequences of HbWRI1,HbWRI2 and HbWRI3 were 1128,1122 and 1050 bp,encoding 375,373 and 349 amino acids.HbWRI1-3 protein has two AP2 conserved domains with 60-70 amino acids.The first AP2 domain contains a VYL motif and a 14-3-3 protein-binding motif.Bioinformatics analysis predicted that HbWRI1-3 protein had no transmembrane domains,no signal peptide,hydrophobic protein and multiple phosphorylation sites.Subcellular localization analysis showed that these protein were located in the nucleus.The expression level of HbWRI1,HbWRI2 and HbWRI3 were the highest in latex.The expression level of HbWRI3 gene in latex was the highest in HbWRI1-3.It is speculated that HbWRI1-3 may be involved in the regulation of latex metabolism such as lipid and rubber biosynthesis.This study lays a foundation for further exploring the effects of HbWRI1-3 on the regulation of latex metabolism and the quality of natural rubber.