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拟南芥酰基辅酶 A 结合蛋白 ACBP6 定位于韧皮部并影响茉莉酸组成。

Arabidopsis acyl-CoA-binding protein ACBP6 localizes in the phloem and affects jasmonate composition.

机构信息

School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China.

Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA.

出版信息

Plant Mol Biol. 2016 Dec;92(6):717-730. doi: 10.1007/s11103-016-0541-0. Epub 2016 Sep 19.

Abstract

Arabidopsis thaliana ACYL-COA-BINDING PROTEIN6 (AtACBP6) encodes a cytosolic 10-kDa AtACBP. It confers freezing tolerance in transgenic Arabidopsis, possibly by its interaction with lipids as indicated by the binding of acyl-CoA esters and phosphatidylcholine to recombinant AtACBP6. Herein, transgenic Arabidopsis transformed with an AtACBP6 promoter-driven β-glucuronidase (GUS) construct exhibited strong GUS activity in the vascular tissues. Immunoelectron microscopy using anti-AtACBP6 antibodies showed AtACBP6 localization in the phloem especially in the companion cells and sieve elements. Also, the presence of gold grains in the plasmodesmata indicated its potential role in systemic trafficking. The AtACBP6 protein, but not its mRNA, was found in phloem exudate of wild-type Arabidopsis. Fatty acid profiling using gas chromatography-mass spectrometry revealed an increase in the jasmonic acid (JA) precursor, 12-oxo-cis,cis-10,15-phytodienoic acid (cis-OPDA), and a reduction in JA and/or its derivatives in acbp6 phloem exudates in comparison to the wild type. Quantitative real-time PCR showed down-regulation of COMATOSE (CTS) in acbp6 rosettes suggesting that AtACBP6 affects CTS function. AtACBP6 appeared to affect the content of JA and/or its derivatives in the sieve tubes, which is consistent with its role in pathogen-defense and in its wound-inducibility of AtACBP6pro::GUS. Taken together, our results suggest the involvement of AtACBP6 in JA-biosynthesis in Arabidopsis phloem tissues.

摘要

拟南芥酰基辅酶 A 结合蛋白 6(AtACBP6)编码一种细胞溶质 10kDa AtACBP。它通过与酰基辅酶 A 酯和磷脂酰胆碱的结合赋予转基因拟南芥抗冻性,这表明其与脂质相互作用,重组 AtACBP6 具有结合活性。在此,用 AtACBP6 启动子驱动的β-葡萄糖醛酸酶(GUS)构建体转化的转基因拟南芥在维管束组织中表现出强烈的 GUS 活性。使用抗 AtACBP6 抗体的免疫电子显微镜显示 AtACBP6 定位于韧皮部,特别是在伴胞和筛分子中。此外,在胞间连丝中存在金颗粒表明其在系统运输中的潜在作用。在野生型拟南芥的韧皮部渗出物中发现了 AtACBP6 蛋白,但未发现其 mRNA。使用气相色谱-质谱法进行脂肪酸分析显示,12-氧代顺式,顺式-10,15-植物二烯酸(cis-OPDA)作为茉莉酸(JA)前体增加,而 acbp6 韧皮部渗出物中的 JA 和/或其衍生物减少与野生型相比。定量实时 PCR 显示 acbp6 莲座丛中的 COMATOSE(CTS)下调表明 AtACBP6 影响 CTS 功能。AtACBP6 似乎影响筛管中 JA 和/或其衍生物的含量,这与其在病原体防御和 AtACBP6pro::GUS 的伤口诱导中的作用一致。总之,我们的结果表明 AtACBP6 参与了拟南芥韧皮部组织中 JA 的生物合成。

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