Zeng Xin, Li Yong-Fang, Mahalingam Ramamurthy
Department of Biochemistry and Molecular Biology, 246 Noble Research Center, Oklahoma State University, Stillwater, OK, 74078, USA.
Planta. 2014 May;239(5):1015-25. doi: 10.1007/s00425-014-2035-0. Epub 2014 Feb 7.
Arabidopsis nudix hydrolase 7 (Atnudt7) mutants exhibit reduced seed germination phenotype following after-ripening. The role of AtNUDT7 in seeds and during early stages of imbibition was examined. Seeds of Atnudt7-1 and Col-0 following 3 days of imbibition were used to profile changes in NADH- and ADP-ribose pyrophosphohydrolase enzyme activities, expression of nudix family genes closely related to AtNudt7, and AtNUDT7 protein levels. Changes in pyridine nucleotides, phytohormones, reactive oxygen species and poly(ADP-ribose) levels in after-ripened seeds and 1 day after imbibition were also analyzed. Changes in AtNUDT7 gene expression, protein levels and enzyme activities in WT seeds and during early stages of imbibition were correlated. Atnudt7-1 seeds lacked NADH pyrophosphohydrolase activity that led to very high catabolic redox charge. Abscisic acid (ABA) levels were higher in Atnudt7-1 mutant while salicylic acid, gibberellic acid, and reactive oxygen species (ROS) levels were higher in WT seeds. In Atnudt7-1, there was excess ROS accumulation 1 day after imbibition. PAR levels were significantly higher in Atnudt7-1 mutant when compared to WT during imbibition. Based on these observations, we conclude NADH pyrophosphohydrolase activity conferred by AtNUDT7 is important for NAD:NADH homeostasis in seeds. Perturbations to this key redox couple alter ABA and ROS levels in the seeds that in turn lowers germination.
拟南芥Nudix水解酶7(Atnudt7)突变体在种子后熟后表现出种子萌发率降低的表型。研究了AtNUDT7在种子及吸水早期阶段的作用。用吸胀3天的Atnudt7 - 1和Col - 0种子分析NADH和ADP - 核糖焦磷酸水解酶活性的变化、与AtNudt7密切相关的Nudix家族基因的表达以及AtNUDT7蛋白水平。还分析了后熟种子及吸胀1天后吡啶核苷酸、植物激素、活性氧和聚(ADP - 核糖)水平的变化。野生型种子及吸水早期阶段AtNUDT7基因表达、蛋白水平和酶活性的变化具有相关性。Atnudt7 - 1种子缺乏NADH焦磷酸水解酶活性,导致分解代谢氧化还原电荷非常高。Atnudt7 - 1突变体中脱落酸(ABA)水平较高,而野生型种子中水杨酸、赤霉素和活性氧(ROS)水平较高。在Atnudt7 - 1中,吸胀1天后ROS积累过多。与野生型相比,Atnudt7 - 1突变体在吸胀过程中聚(ADP - 核糖)(PAR)水平显著更高。基于这些观察结果,我们得出结论,AtNUDT7赋予的NADH焦磷酸水解酶活性对于种子中NAD:NADH的稳态很重要。对这一关键氧化还原对的干扰会改变种子中的ABA和ROS水平,进而降低种子萌发率。