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突触结合蛋白4主要在韧皮部表达,并参与拟南芥的非生物胁迫耐受性。

SYNAPTOTAGMIN 4 is expressed mainly in the phloem and participates in abiotic stress tolerance in Arabidopsis.

作者信息

Kumar Ajay, Krausko Miroslav, Jásik Ján

机构信息

Department of Experimental Plant Biology, Institute of Botany, Plant Science and Biodiversity Centre, Slovak Academy of Sciences, Bratislava, Slovakia.

出版信息

Front Plant Sci. 2024 Jul 1;15:1363555. doi: 10.3389/fpls.2024.1363555. eCollection 2024.

Abstract

Plant synaptotagmins structurally resemble animal synaptotagmins and extended-synaptotagmins. Animal synaptotagmins are well-characterized calcium sensors in membrane trafficking, and extended-synaptotagmins mediate lipid transfer at the endoplasmic reticulum-plasma membrane contact sites. Here, we characterize , which belongs to the six-member family in Arabidopsis. Fluorometric GUS assay showed that the promoter was strongest in roots and the least active in rosettes and cauline leaves, which was confirmed by qPCR. In seedlings, promoter activity was influenced by several factors, such as plant growth regulators, mannitol, sucrose, polyethylene glycol and cold. GUS histochemistry revealed promoter activity in the phloem of all organs and even almost exclusively in sieve element precursors and differentiating sieve elements. Accordingly, the SYT-GFP fusion protein also accumulated in these cells with maximal abundance in sieve element precursors. The protein formed a network in the cytoplasm, but during sieve tube differentiation, it deposited at the cell periphery and disappeared from mature tubes. Using photoconvertible fluorescence technology, we showed that a high abundance of SYT4 protein in meristematic protophloem cells was due to its extensive synthesis. SYT4 protein synthesis was interrupted in differentiating sieve elements, but protein degradation was also reduced. In addition to phloem, the fusion protein was detected in shoot and root stem cell niche as early as the late heart stage of the embryo. We isolated and molecularly and biologically characterized five T-DNA insertion alleles and subjected them to phenotype analysis. The allele with the C2B domain interrupted by an T-DNA insertion exhibits increased sensitivity to factors such as auxins, osmotics, salicylic acid, sodium chloride, and the absence of sucrose in the root growth test.

摘要

植物突触结合蛋白在结构上类似于动物突触结合蛋白和延伸突触结合蛋白。动物突触结合蛋白是膜运输中特征明确的钙传感器,而延伸突触结合蛋白在内质网 - 质膜接触位点介导脂质转移。在这里,我们对拟南芥中属于六成员家族的[具体名称未给出]进行了表征。荧光GUS分析表明,[具体名称未给出]启动子在根中活性最强,在莲座叶和茎生叶中活性最低,qPCR证实了这一点。在幼苗中,启动子活性受多种因素影响,如植物生长调节剂、甘露醇、蔗糖、聚乙二醇和低温。GUS组织化学显示[具体名称未给出]启动子在所有器官的韧皮部都有活性,甚至几乎仅在筛管分子前体和正在分化的筛管分子中具有活性。相应地,SYT - GFP融合蛋白也在这些细胞中积累,在筛管分子前体中丰度最高。该蛋白在细胞质中形成网络,但在筛管分化过程中,它沉积在细胞周边并从成熟筛管中消失。使用光转换荧光技术,我们表明分生组织原生韧皮部细胞中高丰度的SYT4蛋白是由于其大量合成。SYT4蛋白合成在正在分化的筛管分子中被中断,但蛋白质降解也减少。除了韧皮部,早在胚胎的心形后期就可以在茎尖和根尖干细胞龛中检测到融合蛋白。我们分离并对五个[具体名称未给出]T - DNA插入等位基因进行了分子和生物学表征,并对它们进行了表型分析。在根生长试验中,C2B结构域被T - DNA插入打断的等位基因对生长素、渗透压、水杨酸、氯化钠以及蔗糖缺乏等因素表现出更高的敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/30f7/11246894/5ca6ff4b4dab/fpls-15-1363555-g001.jpg

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