Chen Chao, Sun Xiaoli, Duanmu Huizi, Zhu Dan, Yu Yang, Cao Lei, Liu Ailin, Jia Bowei, Xiao Jialei, Zhu Yanming
Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, P.R. China.
Agronomy College, Heilongjiang Bayi Agricultural University, Daqing, P.R. China.
PLoS One. 2015 Nov 9;10(11):e0141888. doi: 10.1371/journal.pone.0141888. eCollection 2015.
Calcium, as the most widely accepted messenger, plays an important role in plant stress responses through calcium-dependent signaling pathways. The calmodulin-like family genes (CMLs) encode Ca2+ sensors and function in signaling transduction in response to environmental stimuli. However, until now, the function of plant CML proteins, especially soybean CMLs, is largely unknown. Here, we isolated a Glycine soja CML protein GsCML27, with four conserved EF-hands domains, and identified it as a calcium-binding protein through far-UV CD spectroscopy. We further found that expression of GsCML27 was induced by bicarbonate, salt and osmotic stresses. Interestingly, ectopic expression of GsCML27 in Arabidopsis enhanced plant tolerance to bicarbonate stress, but decreased the salt and osmotic tolerance during the seed germination and early growth stages. Furthermore, we found that ectopic expression of GsCML27 decreases salt tolerance through modifying both the cellular ionic (Na+, K+) content and the osmotic stress regulation. GsCML27 ectopic expression also decreased the expression levels of osmotic stress-responsive genes. Moreover, we also showed that GsCML27 localized in the whole cell, including cytoplasm, plasma membrane and nucleus in Arabidopsis protoplasts and onion epidermal cells, and displayed high expression in roots and embryos. Together, these data present evidence that GsCML27 as a Ca2+-binding EF-hand protein plays a role in plant responses to bicarbonate, salt and osmotic stresses.
钙作为最广泛被接受的信使,通过钙依赖信号通路在植物应激反应中发挥重要作用。类钙调蛋白家族基因(CMLs)编码Ca2+传感器,并在响应环境刺激的信号转导中发挥作用。然而,直到现在,植物CML蛋白的功能,尤其是大豆CMLs的功能,在很大程度上仍不清楚。在这里,我们分离出一种野生大豆CML蛋白GsCML27,它具有四个保守的EF-手型结构域,并通过远紫外圆二色光谱将其鉴定为一种钙结合蛋白。我们进一步发现,GsCML27的表达受碳酸氢盐、盐和渗透胁迫诱导。有趣的是,GsCML27在拟南芥中的异位表达增强了植物对碳酸氢盐胁迫的耐受性,但在种子萌发和早期生长阶段降低了对盐和渗透胁迫的耐受性。此外,我们发现GsCML27的异位表达通过改变细胞离子(Na+、K+)含量和渗透胁迫调节来降低耐盐性。GsCML27的异位表达还降低了渗透胁迫响应基因的表达水平。此外,我们还表明,GsCML27定位于整个细胞,包括拟南芥原生质体和洋葱表皮细胞的细胞质、质膜和细胞核,并在根和胚中高表达。总之,这些数据表明,GsCML27作为一种Ca2+结合的EF-手型蛋白在植物对碳酸氢盐、盐和渗透胁迫的反应中发挥作用。