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应激调节的拟南芥 GAT2 是一种低亲和力 γ-氨基丁酸转运体。

Stress-regulated Arabidopsis GAT2 is a low affinity γ-aminobutyric acid transporter.

机构信息

Institute of Plant Sciences, Plant Physiology, University of Bern, Altenbergrain 21, 3013 Bern, Switzerland.

Institute of Plant Sciences, Chemical Ecology, University of Bern, Altenbergrain 21, 3013 Bern, Switzerland.

出版信息

J Exp Bot. 2024 Oct 30;75(20):6295-6311. doi: 10.1093/jxb/erae321.

Abstract

The four-carbon non-proteinogenic amino acid γ-aminobutyric acid (GABA) accumulates to high levels in plants in response to various abiotic and biotic stress stimuli, and plays a role in C:N balance, signaling, and as a transport regulator. Expression in Xenopus oocytes and voltage-clamping allowed the characterization of Arabidopsis GAT2 (At5g41800) as a low affinity GABA transporter with a K0.5GABA ~8 mM. l-Alanine and butylamine represented additional substrates. GABA-induced currents were strongly dependent on the membrane potential, reaching the highest affinity and highest transport rates at strongly negative membrane potentials. Mutation of Ser17, previously reported to be phosphorylated in planta, did not result in altered affinity. In a short-term stress experiment, AtGAT2 mRNA levels were up-regulated at low water potential and under osmotic stress (polyethylene glycol and mannitol). Furthermore, AtGAT2 promoter activity was detected in vascular tissues, maturating pollen, and the phloem unloading region of young seeds. Even though this suggested a role for AtGAT2 in long-distance transport and loading of sink organs, under the conditions tested neither AtGAT2-overexpressing plants, atgat2 or atgat1 T-DNA insertion lines, nor atgat1 atgat2 doubleknockout mutants differed from wild-type plants in growth on GABA, amino acid levels, or resistance to salt and osmotic stress.

摘要

四碳非蛋白氨基酸γ-氨基丁酸(GABA)在植物中响应各种非生物和生物胁迫刺激而积累到高水平,并在 C:N 平衡、信号转导以及作为运输调节剂方面发挥作用。在非洲爪蟾卵母细胞中的表达和电压钳位允许对拟南芥 GAT2(At5g41800)进行表征,其为低亲和力 GABA 转运体,K0.5GABA~8 mM。l-丙氨酸和丁胺是其他底物。GABA 诱导的电流强烈依赖于膜电位,在强负膜电位下达到最高亲和力和最高转运速率。先前报道在植物中 Ser17 被磷酸化,其突变并未导致亲和力改变。在短期胁迫实验中,AtGAT2 mRNA 水平在低水势和渗透胁迫(聚乙二醇和甘露醇)下上调。此外,在血管组织、成熟花粉和幼种子的韧皮部卸载区检测到 AtGAT2 启动子活性。尽管这表明 AtGAT2 在长距离运输和汇器官装载中的作用,但在测试的条件下,AtGAT2 过表达植物、atgat2 或 atgat1 T-DNA 插入系以及 atgat1 atgat2 双敲除突变体在 GABA、氨基酸水平或对盐和渗透胁迫的抗性方面与野生型植物没有差异。

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