摘要
为了解HD-ZipⅠ转录因子调控梨花青苷合成的机制,以西洋梨品种‘红茄梨’为试材,克隆了1个HD-ZipⅠ家族基因PcHB12(GenBank序列号XM009363028)。通过系统进化树分析、qRT-PCR、酵母单杂交、电泳迁移率变动分析(Electrophoretic mobility shift assay,EMSA)和荧光素酶报告试验,探究PcHB12调控梨花青苷合成的作用。结果表明,PcHB12的开放阅读框为696 bp,编码231个氨基酸,预测的蛋白质分子量是26.77 kD。系统进化树分析表明,PcHB12与拟南芥AtHB12蛋白序列相似度最高。‘茄梨’中PcHB12的表达量明显高于其红色芽变品种‘红茄梨’,而花青苷含量和花青苷相关基因PcMYB10.1、PcUFGT的表达量明显低于‘红茄梨’。酵母单杂交和EMSA试验发现,PcHB12结合在PcMYB10.1启动子的MBS序列。荧光素酶试验表明PcHB12负调控PcMYB10.1启动子的转录活性。因此,PcHB12可能通过负调控PcMYB10.1的表达从而抑制梨花青苷的合成。
In order to understand the mechanism that the HD-ZipⅠtranscription factor regulates anthocyanin synthesis in pears,the European pear cultivar’Red Clapp Favorite’was used as study material to clone PcHB12(GenBank accession number:XM009363028)from the HD-ZipⅠfamily. Phylogenetic tree analysis,qRT-PCR,yeast one-hybrid assay,EMSA,and luciferase reporter assay were employed to study the regulatory function of PcHB12 on anthocyanin synthesis. The results showed that the open reading frame of PcHB12 is 696 bp,encoding 231 amino acids,and the predicted molecular weight of the protein is 26.77 kD. Phylogenetic tree analysis showed that PcHB12 has the highest similarity to the AtHB12 protein sequence of Arabidopsis thaliana. The expression level of PcHB12 in ’Clapp Favorite’ is significantly higher than that in its red bud mutation cultivar’Red Clapp Favorite’. In contrast,anthocyanin content and the expression levels of anthocyanin-related genes PcMYB10.1 and PcUFGT in ’Clapp Favorite’were significantly lower than that in’Red Clapp Favorite’. Yeast one-hybrid and electrophoretic mobility shift assay(EMSA)experiments found that PcHB12 binds to the MBS sequence in the PcMYB10.1 promoter. Luciferase experiments showed that PcHB12 negatively regulates transcription activity on the PcMYB10.1 promoter. Therefore,PcHB12 may negatively regulate PcMYB10.1 expression to inhibit anthocyanin synthesis in pears.
作者
张震
李晨智宇
张雅
李玺
王烁
王鑫玉
陈学森
冯守千
ZHANG Zhen;LI Chenzhiyu;ZHANG Ya;LI Xi;WANG Shuo;WANG Xinyu;CHEN Xuesen;FENG Shouqian(College of Horticulture Science and Engineering,Shandong Agricultural University,State Key Laboratory of Crop Biology,Tai’an,Shandong 271018,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2021年第3期456-464,共9页
Acta Horticulturae Sinica
基金
国家重点研发计划项目(2018YFD1000105)
山东省良种工程项目(2019LZGC008)
国家自然科学基金项目(31201593,31872940)。
