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小麦鸟氨酸氨基转移酶(TaOAT)编码基因的克隆与分子特性分析。

Cloning and molecular characterization of Triticum aestivum ornithine amino transferase (TaOAT) encoding genes.

机构信息

Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, People's Republic of China.

School of Veterinary and Life Sciences, Murdoch University, Perth, WA, 6150, Australia.

出版信息

BMC Plant Biol. 2020 Apr 29;20(1):187. doi: 10.1186/s12870-020-02396-2.

Abstract

BACKGROUND

Ornithine aminotransferase (OAT, EC:2.6.1.13), alternatively known as ornithine delta aminotransferase (δOAT), is a pyridoxal phosphate (PLP)-dependent enzyme involved in the conversion of ornithine into glutamyl-5-semi-aldehyde (GSA) and vice versa. Up till now, there has been no study on OAT in wheat despite the success of its isolation from rice, maize, and sorghum. This study focuses on identification and molecular characterization of OAT in wheat.

RESULTS

In total, three homeologous OAT genes in wheat genome were found on chromosome group 5, named as TaOAT-5AL, TaOAT-5BL, and TaOAT-5DL. Sequence alignment between gDNA and its corresponding cDNA obtained a total of ten exons and nine introns. A phylogenetic tree was constructed and results indicated that OATs shared highly conserved domains between monocots and eudicots, which was further illustrated by using WebLogo to generate a sequence logo. Further subcellular localization analysis indicated that they functioned in mitochondria. Protein-protein interactions supported their role in proline biosynthesis through interactions with genes, such as delta 1-pyrroline-5-carboxylate synthetase (P5CS) and pyrroline-5-carboxylate reductase (P5CR), involved in the proline metabolic pathway. Promoter analysis exposed the presence of several stress responsive elements, implying their involvement in stress regulation. Expression profiling illustrated that TaOAT was highly induced in the wheat plants exposed to drought or salt stress condition. Upregulated expression of TaOATs was observed in stamens and at the heading stage. A potential role of TaOAT genes during floret development was also revealed. Furthermore, the transgenic plants overexpressing TaOAT showed enhanced tolerance to drought stress by increasing proline accumulation. In addition, salt tolerance of the transgenic plants was also enhanced.

CONCLUSION

TaOATs genes were involved in proline synthesis and nitrogen remobilization because they interacted with genes related to proline biosynthesis enzymes and arginine catabolism. In addition, TaOAT genes had a role in abiotic stress tolerance and a potential role in floret development. The results of this study may propose future research in the improvement of wheat resistance to abiotic stresses.

摘要

背景

鸟氨酸氨基转移酶(OAT,EC:2.6.1.13),也称为鸟氨酸 δ 氨基转移酶(δOAT),是一种依赖吡哆醛磷酸(PLP)的酶,参与将鸟氨酸转化为谷氨酰-5-半醛(GSA)和反之亦然。尽管从水稻、玉米和高粱中成功分离出 OAT,但迄今为止,小麦中尚未对 OAT 进行研究。本研究专注于鉴定和分子表征小麦中的 OAT。

结果

在小麦基因组的 5 号染色体组上共发现了三个同源 OAT 基因,分别命名为 TaOAT-5AL、TaOAT-5BL 和 TaOAT-5DL。对 gDNA 与其相应 cDNA 进行序列比对,共获得了 10 个外显子和 9 个内含子。构建了系统发育树,结果表明 OAT 在单子叶植物和双子叶植物之间具有高度保守的结构域,这进一步通过使用 WebLogo 生成序列标志来阐明。进一步的亚细胞定位分析表明,它们在线粒体中发挥作用。蛋白质-蛋白质相互作用支持它们通过与脯氨酸代谢途径中涉及的 delta 1-吡咯啉-5-羧酸合成酶(P5CS)和吡咯啉-5-羧酸还原酶(P5CR)等基因的相互作用,在脯氨酸生物合成中的作用。启动子分析揭示了存在几种应激响应元件,这意味着它们参与了应激调节。表达谱分析表明,在暴露于干旱或盐胁迫条件下的小麦植株中 TaOAT 高度诱导。在雄蕊和抽穗期观察到 TaOATs 的上调表达。还揭示了 TaOAT 基因在小花发育过程中的潜在作用。此外,过表达 TaOAT 的转基因植物通过增加脯氨酸积累来提高对干旱胁迫的耐受性。此外,转基因植物的耐盐性也得到增强。

结论

TaOAT 基因参与脯氨酸合成和氮再利用,因为它们与脯氨酸生物合成酶和精氨酸分解代谢相关基因相互作用。此外,TaOAT 基因在非生物胁迫耐受性和小花发育方面具有作用。本研究的结果可能为提高小麦对非生物胁迫的抗性的未来研究提供建议。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8a6a/7189522/d2e3a47d4734/12870_2020_2396_Fig1_HTML.jpg

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