Chen Yun, Ji Fangfang, Xie Hong, Liang Jiansheng, Zhang Jianhua
College of Bioscience and Biotechnology, Key Laboratory of Crop Genetics and Physiology of Jiangsu Province, Yangzhou University, Yangzhou 225009, People's Republic of China.
Plant Physiol. 2006 Jan;140(1):302-10. doi: 10.1104/pp.105.069872. Epub 2005 Dec 16.
The regulator of G-protein signaling (RGS) proteins, recently identified in Arabidopsis (Arabidopsis thaliana; named as AtRGS1), has a predicted seven-transmembrane structure as well as an RGS box with GTPase-accelerating activity and thus desensitizes the G-protein-mediated signaling. The roles of AtRGS1 proteins in Arabidopsis seed germination and their possible interactions with sugars and abscisic acid (ABA) were investigated in this study. Using seeds that carry a null mutation in the genes encoding RGS protein (AtRGS1) and the alpha-subunit (AtGPA1) of the G protein in Arabidopsis (named rgs1-2 and gpa1-3, respectively), our genetic evidence proved the involvement of the AtRGS1 protein in the modulation of seed germination. In contrast to wild-type Columbia-0 and gpa1-3, stratification was found not to be required and the after-ripening process had no effect on the rgs1-2 seed germination. In addition, rgs1-2 seed germination was insensitive to glucose (Glc) and sucrose. The insensitivities of rgs1-2 to Glc and sucrose were not due to a possible osmotic stress because the germination of rgs1-2 mutant seeds showed the same response as those of gpa1-3 mutants and wild type when treated with the same concentrations of mannitol and sorbitol. The gpa1-3 seed germination was hypersensitive while rgs1-2 was less sensitive to exogenous ABA. The different responses to ABA largely diminished and the inhibitory effects on seed germination by exogenous ABA and Glc were markedly alleviated when endogenous ABA biosynthesis was inhibited. Hypersensitive responses of seed germination to both Glc and ABA were also observed in the overexpressor of AtRGS1. Analysis of the active endogenous ABA levels and the expression of NCED3 and ABA2 genes showed that Glc significantly stimulated the ABA biosynthesis and increased the expression of NCED3 and ABA2 genes in germinating Columbia seeds, but not in rgs1-2 mutant seeds. These data suggest that AtRGS1 proteins are involved in the regulation of seed germination. The hyposensitivity of rgs1-2 mutant seed germination to Glc might be the result of the impairment of ABA biosynthesis during seed germination.
最近在拟南芥(Arabidopsis thaliana;命名为AtRGS1)中发现的G蛋白信号调节(RGS)蛋白,具有预测的七跨膜结构以及具有GTP酶加速活性的RGS框,从而使G蛋白介导的信号脱敏。本研究调查了AtRGS1蛋白在拟南芥种子萌发中的作用及其与糖和脱落酸(ABA)的可能相互作用。使用在拟南芥中编码RGS蛋白(AtRGS1)和G蛋白α亚基(AtGPA1)的基因中携带无效突变的种子(分别命名为rgs1-2和gpa1-3),我们的遗传证据证明了AtRGS1蛋白参与种子萌发的调节。与野生型哥伦比亚-0和gpa1-3相比,发现不需要进行分层,后熟过程对rgs1-2种子萌发没有影响。此外,rgs1-2种子萌发对葡萄糖(Glc)和蔗糖不敏感。rgs1-2对Glc和蔗糖的不敏感性不是由于可能的渗透胁迫,因为当用相同浓度的甘露醇和山梨醇处理时,rgs1-2突变体种子的萌发表现出与gpa1-3突变体和野生型相同的反应。gpa1-3种子萌发对ABA过敏,而rgs1-2对外源ABA不太敏感。当内源性ABA生物合成受到抑制时,对ABA的不同反应在很大程度上减弱,外源ABA和Glc对种子萌发的抑制作用明显减轻。在AtRGS1过表达体中也观察到种子萌发对Glc和ABA的过敏反应。对活性内源性ABA水平以及NCED3和ABA2基因表达的分析表明,Glc显著刺激萌发的哥伦比亚种子中的ABA生物合成并增加NCED3和ABA2基因的表达,但在rgs1-2突变体种子中则不然。这些数据表明AtRGS1蛋白参与种子萌发的调节。rgs1-2突变体种子萌发对Glc的低敏感性可能是种子萌发期间ABA生物合成受损的结果。