Functional identification of two Glycerol-3-phosphate Acyltransferase5 homologs from Chenopodium quinoa

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science Pub Date : 2024-11-07 DOI:10.1016/j.plantsci.2024.112313

Zhen Wang , Yuxin Liu , Haodong Huang , Zhifu Zheng , Shiyou Lü , Xianpeng Yang , Changle Ma

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引用次数: 0

Abstract

Glycerol-3-phosphate acyltransferase5 (GPAT5) is the key enzyme in suberin biosynthesis in Arabidopsis, tomato and Sarracenia purpurea. However, little is known about whether GPAT5 function is conserved in halophytes. In this study, we identified two GPAT5 homologs, CqGPAT5a and CqGPAT5b, in Chenopodium quinoa, the typical halophyte. Using RT-qPCR, we found that CqGPAT5a and CqGPAT5b were highly expressed in quinoa roots and rapidly induced by high salt stress. CqGPAT5a and CqGPAT5b were localized to the endoplasmic reticulum and found to have glycerol-3-phosphate acyltransferase activity using yeast complementation assays. Compared with CqGPAT5b, CqGPAT5a showed relatively weaker function and less protein abundance when expressed in yeast, Arabidopsis or Nicotiana benthamiana. Subsequently, we identified a serine (S) to leucine (L) variation in the CqGPAT5a protein sequence (S251L) compared with CqGPAT5b, located in the connecting region between the second and third transmembrane domains. Site-directed mutagenesis together with yeast mutant complementation and transient expression in tobacco demonstrated that this variation significantly affected CqGPAT5a activity and protein abundance. These findings expand our understanding of GPAT5 and provide new evidence that GPAT5 may be functionally conserved in halophytes.

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藜麦中两种甘油-3-磷酸酰基转移酶5同源物的功能鉴定

甘油-3-磷酸酰基转移酶5（GPAT5）是拟南芥、番茄和紫云英中单宁生物合成的关键酶。然而，人们对 GPAT5 的功能在卤叶植物中是否保守知之甚少。在这项研究中，我们在典型的卤叶植物藜属植物中发现了两个 GPAT5 同源物，即 CqGPAT5a 和 CqGPAT5b。利用 RT-qPCR 技术，我们发现 CqGPAT5a 和 CqGPAT5b 在藜麦根部高表达，并在高盐胁迫下快速诱导。CqGPAT5a 和 CqGPAT5b 定位于内质网，并通过酵母互补试验发现它们具有甘油-3-磷酸酰基转移酶活性。与 CqGPAT5b 相比，CqGPAT5a 在酵母、拟南芥或烟草中表达时功能相对较弱，蛋白丰度也较低。随后，我们发现与 CqGPAT5b 相比，CqGPAT5a 蛋白序列中存在一个丝氨酸（S）到亮氨酸（L）的变异（S251L），位于第二和第三跨膜结构域之间的连接区域。定点突变以及酵母突变体互补和烟草中的瞬时表达表明，这一变异显著影响了 CqGPAT5a 的活性和蛋白质丰度。这些发现拓展了我们对 GPAT5 的了解，并提供了 GPAT5 在卤叶植物中可能具有功能保守性的新证据。

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来源期刊

Plant Science 生物-生化与分子生物学

CiteScore

9.10

自引率

1.90%

发文量

322

审稿时长

33 days

期刊介绍： Plant Science will publish in the minimum of time, research manuscripts as well as commissioned reviews and commentaries recommended by its referees in all areas of experimental plant biology with emphasis in the broad areas of genomics, proteomics, biochemistry (including enzymology), physiology, cell biology, development, genetics, functional plant breeding, systems biology and the interaction of plants with the environment. Manuscripts for full consideration should be written concisely and essentially as a final report. The main criterion for publication is that the manuscript must contain original and significant insights that lead to a better understanding of fundamental plant biology. Papers centering on plant cell culture should be of interest to a wide audience and methods employed result in a substantial improvement over existing established techniques and approaches. Methods papers are welcome only when the technique(s) described is novel or provides a major advancement of established protocols.