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PpybZIP43 通过激活 PpySPS3 表达促进梨果实中蔗糖的合成,并与 PpySTOP1 相互作用。

PpybZIP43 contributes to sucrose synthesis in pear fruits by activating PpySPS3 expression and interacts with PpySTOP1.

机构信息

College of Horticulture, Nanjing Agricultural University, Nanjing, China.

College of Life Science and Technology, Guangxi University, Nanning, China.

出版信息

Physiol Plant. 2022 May;174(3):e13732. doi: 10.1111/ppl.13732.

Abstract

Sucrose is an important factor affecting sweetness and flavor in pear fruits, but the molecular mechanism of sucrose synthesis regulation is relatively unknown. Here, we characterized a transcription factor gene from pear (Pyrus pyrifolia Nakai cv. "Hosui") fruits, PpybZIP43, and found that the transient overexpression of PpybZIP43 in pear fruits significantly increased the sucrose content and the relative expression level of sucrose phosphate synthase genes (PpySPS3 and PpySPS8). Subcellular localization analysis in tobacco leaves showed that PpybZIP43 was localized in the nucleus. Yeast one-hybrid, electrophoretic mobility shift assay (EMSA), and dual-luciferase reporter assays indicated that PpybZIP43 was able to activate the expression of PpySPS3 by binding specifically to the G-box (CACGTG) element in the promoter. The protein-protein interaction assays using yeast two-hybrid, bimolecular fluorescence complementation (BiFC), firefly luciferase complementation imaging (LCI), and glutathione S-transferase (GST) pull-down demonstrated that PpybZIP43 could directly interact with PpySTOP1 to form a transcription complex. This study is helpful for understanding the molecular basis of sucrose synthesis and accumulation in pear fruits and provides candidate genes for breeding.

摘要

蔗糖是影响梨果实甜度和风味的重要因素,但蔗糖合成调控的分子机制尚不清楚。本研究从梨(Pyrus pyrifolia Nakai cv. "Hosui")果实中鉴定了一个转录因子基因 PpybZIP43,发现瞬时过表达 PpybZIP43 可显著提高梨果实中的蔗糖含量和蔗糖磷酸合成酶基因(PpySPS3 和 PpySPS8)的相对表达水平。烟草叶片的亚细胞定位分析表明,PpybZIP43 定位于细胞核。酵母单杂交、电泳迁移率变动分析(EMSA)和双荧光素酶报告基因分析表明,PpybZIP43 能够通过特异性结合启动子中的 G-Box(CACGTG)元件来激活 PpySPS3 的表达。酵母双杂交、双分子荧光互补(BiFC)、萤火虫荧光素酶互补成像(LCI)和谷胱甘肽 S-转移酶(GST)下拉实验证实,PpybZIP43 可以与 PpySTOP1 直接相互作用形成转录复合物。本研究有助于理解梨果实中蔗糖合成和积累的分子基础,并为品种选育提供了候选基因。

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