Quantitative Phosphoproteomic Analysis Reveals Potential Regulatory Mechanisms of Early Fruit Enlargement in Pecan (Carya illinoinensis).
Quantitative Phosphoproteomic Analysis Reveals Potential Regulatory Mechanisms of Early Fruit Enlargement in Pecan (Carya illinoinensis).
复制标题
DOI：
10.1021/acs.jafc.2c08876
复制
发表时间：
2023-03
影响因子：
6.1
通讯作者：
Zhengfu Yang;Tao Qin;H. Jin;Jiani Wang;Caiyun Li;Kean-Jin Lim;Zhengjia Wang
中科院分区：
文献类型：
--
作者：
Zhengfu Yang;Tao Qin;H. Jin;Jiani Wang;Caiyun Li;Kean-Jin Lim;Zhengjia Wang
山核桃(Carya Illinoinensis)是一种很受欢迎的坚果。果实发育经历缓慢生长、快速膨大、核心硬化和籽粒成熟期。然而，人们对山核桃授粉后如何启动果实发育和扩大知之甚少。在这项研究中，我们首次利用无标记磷酸蛋白质组定量技术对山核桃果实发育早期的潜在磷酸化位点和蛋白质进行了大规模鉴定。从授粉后5周和9周未授粉雌蕊和果实中的1953个含1311个磷蛋白的磷酸肽中，共鉴定出2155个亚磷酸酶。其中699个非冗余磷酸蛋白被差异磷酸化(DP)。此外，还分析了油菜素内酯(BR)中DP蛋白的磷酸化强度和生长素信号转导，并研究了CiBZR1的功能。在拟南芥中，异源表达CiBZR1导致BR反应表型叶片和果实卷曲，种子增大。亚细胞定位和转录激活活性分析表明，CiBZR1分布于胞核和胞浆中，具有转录活性。当两个亚磷酸盐突变时，CiBZR1S201P、S205G完全转移到细胞核，而转录活性保持不变。综上所述，我们的数据揭示了广泛的磷蛋白，为全面剖析山核桃果实早期发育潜在的翻译后调控机制奠定了基础。
Pecan (Carya illinoinensis) is a popular tree nut. Its fruit development undergoes slow growth, rapid expansion, core hardening, and kernel maturation stages. However, little is known about how pecan initiates fruit development and enlargement after pollination. In this study, we performed the first large-scale identification of potential phosphorylation sites and proteins at early development of pecan fruit by a label-free phosphoproteomic quantification technique. A total of 2155 phosphosites were identified from 1953 phosphopeptides covering 1311 phosphoproteins in unpollinated pistils and fruits at 5 and 9 weeks after pollination. Of these, 699 nonredundant phosphoproteins were differentially phosphorylated (DP). Furthermore, the phosphorylation intensity of DP proteins in brassinolide (BR) and auxin signaling were analyzed, and the function of CiBZR1 was investigated. Ectopic expression of CiBZR1 resulted in BR response phenotypes with curled leaves and fruit, while enlarged seed size in Arabidopsis. Subcellular localization and transcriptional activation activity assay demonstrated that CiBZR1 distributed in both the nucleus and cytoplasm with transcriptional activity. When two phosphosites mutated, CiBZR1S201P,S205G moved to the nucleus completely, while the transcriptional activity remained unchanged. Taken together, our data reveal extensive phosphoproteins and lay a foundation to comprehensively dissect the potential post-translational regulation mechanism of early development of pecan fruit.