Zhao Ying, Cui Yifan, Huang Shiyu, Yu Jingyao, Wang Xinyu, Xin Dawei, Li Xin, Liu Yonghui, Dai Yuxin, Qi Zhaoming, Chen Qingshan
College of Agriculture, Northeast Agricultural University, Harbin, China.
Key Lab of Maize Genetics and Breeding, Heilongjiang Academy of Agricultural Sciences, Harbin, China.
Front Plant Sci. 2020 Feb 26;11:214. doi: 10.3389/fpls.2020.00214. eCollection 2020.
Glucose-6-phosphate dehydrogenase (G6PDH) is known as a critical enzyme responsible for nicotinamide adenine dinucleotide phosphate (NADPH) generation in the pentose phosphate pathway (PPP), and has an essential function in modulating redox homeostasis and stress responsiveness. In the present work, we characterized the nine members of the G6PDH gene family in soybean. Phylogenic analysis and transit peptide prediction showed that these soybean G6PDHs are divided into plastidic (P) and cytosolic (Cy) isoforms. The subcellular locations of five GmG6PDHs were further verified by confocal microscopy in mesophyll protoplasts. The respective genes had distinct expression patterns in various soybean tissues and at different times during seed development. Among them, the - were strongly expressed in roots, developing seeds and nodules, while the transcripts of were mainly detected in green tissues. In addition, the activities and transcripts of were dramatically stimulated by different stress treatments, including salt, osmotic and alkali. Notably, the expression levels of a cytosolic isoform (GmG6PDH2) were extraordinarily high under salt stress and correlated well with the G6PDH enzyme activities, possibly implying a crucial factor for soybean responses to salinity. Enzymatic assay of recombinant proteins expressed in showed that the enzyme encoded by had functional NADP-dependent G6PDH activity. Further analysis indicated overexpression of gene could significantly enhance the resistance of transgenic soybean to salt stress by coordinating with the redox states of ascorbic acid and glutathione pool to suppress reactive oxygen species generation. Together, these results indicate that might be the major isoform for NADPH production in PPP, which is involved in the modulation of cellular AsA-GSH cycle to prevent the oxidative damage induced by high salinity.
葡萄糖-6-磷酸脱氢酶(G6PDH)是戊糖磷酸途径(PPP)中负责生成烟酰胺腺嘌呤二核苷酸磷酸(NADPH)的关键酶,在调节氧化还原稳态和应激反应中具有重要作用。在本研究中,我们对大豆中G6PDH基因家族的九个成员进行了表征。系统发育分析和转运肽预测表明,这些大豆G6PDHs分为质体(P)和胞质(Cy)亚型。通过共聚焦显微镜在叶肉原生质体中进一步验证了五个GmG6PDHs的亚细胞定位。各个基因在大豆不同组织和种子发育的不同时期具有不同的表达模式。其中,[具体基因]在根、发育中的种子和根瘤中强烈表达,而[具体基因]的转录本主要在绿色组织中检测到。此外,[具体基因]的活性和转录本受到盐、渗透和碱等不同胁迫处理的显著刺激。值得注意的是,一种胞质亚型(GmG6PDH2)在盐胁迫下的表达水平异常高,并且与G6PDH酶活性密切相关,这可能意味着它是大豆对盐度响应的关键因素。对在[具体表达系统]中表达的重组蛋白进行酶活性测定表明,[具体基因]编码的酶具有依赖NADP的G6PDH活性。进一步分析表明,[具体基因]的过表达可以通过与抗坏血酸和谷胱甘肽池的氧化还原状态协同作用来抑制活性氧的产生,从而显著增强转基因大豆对盐胁迫的抗性。总之,这些结果表明,[具体基因]可能是PPP中NADPH产生的主要亚型,参与调节细胞的AsA-GSH循环以防止高盐度诱导的氧化损伤。