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小麦籽粒中谷氨酰胺合成酶同工酶的定位、基因表达及功能(.)

Localization, Gene Expression, and Functions of Glutamine Synthetase Isozymes in Wheat Grain (.).

作者信息

Wei Yihao, Xiong Shuping, Zhang Zhiyong, Meng Xiaodan, Wang Lulu, Zhang Xiaojiao, Yu Meiqin, Yu Haidong, Wang Xiaochun, Ma Xinming

机构信息

Collaborative Innovation Center of Henan Grain Crops, College of Agronomy, Henan Agricultural University, Zhengzhou, China.

Department of Biochemistry and Molecular Biology, College of Life Science, Henan Agricultural University, Zhengzhou, China.

出版信息

Front Plant Sci. 2021 Feb 9;12:580405. doi: 10.3389/fpls.2021.580405. eCollection 2021.

Abstract

Glutamine synthetase (GS) plays a major role in plant nitrogen metabolism, but the roles of individual GS isoforms in grains are unknown. Here, the localization and expression of individual TaGS isozymes in wheat grain were probed with TaGS isoenzyme-specific antibodies, and the nitrogen metabolism of grain during the grain filling stage were investigated. Immunofluorescence revealed that TaGS1;1, TaGS1;3, and TaGS2 were expressed in different regions of the embryo. In grain transporting tissues, TaGS1;2 was localized in vascular bundle; TaGS1;2 and TaGS1;1 were in chalaza and placentochalaza; TaGS1;1 and TaGS1;3 were in endosperm transfer cells; and TaGS1;3 and TaGS2 were in aleurone layer. GS exhibited maximum activity and expression at 8 days after flowering (DAF) with peak glutamine content in grains; from then, increased largely from reduction, glutamate dehydrogenase (GDH) aminating activity increased continuously, and the activities of GS and glutamate synthase (GOGAT) decreased, while only TaGS1;3 kept a stable expression in different TaGS isozymes. Hence, GS-GOGAT cycle and GDH play different roles in assimilation of grain in different stages of grain development; TaGS1;3, located in aleurone layer and endosperm transfer cells, plays a key role in Gln into endosperm for gluten synthesis. At 30 DAF, grain amino acids are mainly transported from maternal phloem.

摘要

谷氨酰胺合成酶(GS)在植物氮代谢中起主要作用,但单个GS同工型在籽粒中的作用尚不清楚。本文利用TaGS同工酶特异性抗体探究了小麦籽粒中单个TaGS同工酶的定位和表达,并研究了灌浆期籽粒的氮代谢。免疫荧光显示,TaGS1;1、TaGS1;3和TaGS2在胚的不同区域表达。在籽粒运输组织中,TaGS1;2定位于维管束;TaGS1;2和TaGS1;1位于合点和胎座合点;TaGS1;1和TaGS1;3位于胚乳转移细胞;TaGS1;3和TaGS2位于糊粉层。GS在开花后8天(DAF)表现出最大活性和表达,籽粒中谷氨酰胺含量达到峰值;此后,随着还原作用的大幅增加,谷氨酸脱氢酶(GDH)的胺化活性持续增加,GS和谷氨酸合酶(GOGAT)的活性下降,而在不同的TaGS同工型中只有TaGS1;3保持稳定表达。因此,GS-GOGAT循环和GDH在籽粒发育的不同阶段对籽粒同化作用发挥不同作用;位于糊粉层和胚乳转移细胞中的TaGS1;3在谷氨酰胺进入胚乳用于面筋合成过程中起关键作用。在开花后30天,籽粒氨基酸主要从母本韧皮部运输而来。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4daf/7901976/ef7e402eee95/fpls-12-580405-g0001.jpg

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