Cytological and proteomic analysis of Ginkgo biloba L. pollen intine
Cytological and proteomic analysis of Ginkgo biloba L. pollen intine
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银杏花粉内壁的细胞学和蛋白质组学分析
DOI：
10.1016/j.hpj.2020.04.001
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发表时间：
2020
影响因子：
5.7
通讯作者：
Li Wang
中科院分区：
文献类型：
--
作者：
Weixing Li;Yunling Ye;Fangmei Cheng;Yan Lu;Biao Jin;Li Wang
花粉内壁在花粉萌发和花粉管生长过程中发挥着重要作用，但银杏的相关信息尚不清楚。我们分离并获得了G.的去壳花粉。银杏叶。使用荧光探针，我们观察到花粉内壁的纤维素荧光最强。用 JIM5 免疫标记的去酯化果胶存在于整个细胞壁中，而单克隆抗体 JIM7 识别的酯化果胶集中在一些区域。在花粉内壁的整个表面观察到用苯胺蓝进行胼胝质染色。这些结果通过傅里叶变换红外（FTIR）分析得到证实。我们还使用蛋白质组学方法在体外鉴定成熟花粉和脱核花粉（水合后 48 小时）之间的不同蛋白质。质谱分析表明，去核花粉比成熟花粉含有更多的蛋白质，包括钙调蛋白、丝氨酸羟甲基转移酶、β-半乳糖苷酶6和IV类几丁质酶。根据基因本体论（GO）分析，差异表达蛋白主要与运输、防御反应、糖代谢、能量代谢、信号转导和细胞壁形成相关。这些发现表明，大多数参与花粉萌发和花粉管生长的蛋白质是在花粉水合过程中合成的，表明花粉水合在G.花粉繁殖过程中的重要作用。银杏叶。
The pollen intine plays important roles in pollen germination and tube growth, but related information inGinkgo bilobaremains unclear. We isolated and obtained de-exined pollen fromG. biloba. Using fluorescent probes, we observed the strongest cellulose fluorescence in the pollen intine. De-esterified pectin immunolabeled with JIM5 was present throughout the entire cell wall, whereas esterified pectin recognized by the monoclonal antibody JIM7 was concentrated in some regions. Callose staining with aniline blue was observed across the entire surface of the pollen intine. These results were confirmed by Fourier Transform InfraRed (FTIR) analysis. We also used proteomic approaches to identify different proteins between mature and de-exined pollen (48 h after hydration)in vitro. Based on mass spectrometry, de-exined pollen had more proteins than mature pollen, including calmodulin, serine hydroxymethyltransferase,β-galactosidase 6, and class IV chitinase. According to Gene Ontology (GO) analysis, the differentially expressed proteins were mainly associated with transportation, defense reaction, sugar metabolism, energy metabolism, signal transduction, and cell wall formation. These findings suggest that most proteins involved in pollen germination and pollen tube growth are synthesized during pollen hydration, indicating the important role of pollen hydration in the reproductive process ofG. Biloba.