Zhou Shu-Mei, Kong Xiang-Zhu, Kang Han-Han, Sun Xiu-Dong, Wang Wei
State Key Laboratory of Crop Biology, College of Life Science, Shandong Agricultural University, Tai'an, Shandong, People's Republic of China.
College of Horticulture Science and Engineering, Shandong Agricultural University, Tai'an, Shandong, People's Republic of China.
PLoS One. 2015 Apr 23;10(4):e0122117. doi: 10.1371/journal.pone.0122117. eCollection 2015.
As one of the largest gene families, F-box domain proteins have been found to play important roles in abiotic stress responses via the ubiquitin pathway. TaFBA1 encodes a homologous F-box protein contained in E3 ubiquitin ligases. In our previous study, we found that the overexpression of TaFBA1 enhanced drought tolerance in transgenic plants. To investigate the mechanisms involved, in this study, we investigated the tolerance of the transgenic plants to oxidative stress. Methyl viologen was used to induce oxidative stress conditions. Real-time PCR and western blot analysis revealed that TaFBA1 expression was up-regulated by oxidative stress treatments. Under oxidative stress conditions, the transgenic tobacco plants showed a higher germination rate, higher root length and less growth inhibition than wild type (WT). The enhanced oxidative stress tolerance of the transgenic plants was also indicated by lower reactive oxygen species (ROS) accumulation, malondialdehyde (MDA) content and cell membrane damage under oxidative stress compared with WT. Higher activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD), were observed in the transgenic plants than those in WT, which may be related to the upregulated expression of some antioxidant genes via the overexpression of TaFBA1. In others, some stress responsive elements were found in the promoter region of TaFBA1, and TaFBA1 was located in the nucleus, cytoplasm and plasma membrane. These results suggest that TaFBA1 plays an important role in the oxidative stress tolerance of plants. This is important for understanding the functions of F-box proteins in plants' tolerance to multiple stress conditions.
作为最大的基因家族之一,F-box结构域蛋白已被发现通过泛素途径在非生物胁迫响应中发挥重要作用。TaFBA1编码一种包含在E3泛素连接酶中的同源F-box蛋白。在我们之前的研究中,我们发现TaFBA1的过表达增强了转基因植物的耐旱性。为了研究其中涉及的机制,在本研究中,我们研究了转基因植物对氧化胁迫的耐受性。使用甲基紫精诱导氧化胁迫条件。实时PCR和蛋白质免疫印迹分析表明,氧化胁迫处理上调了TaFBA1的表达。在氧化胁迫条件下,转基因烟草植株比野生型(WT)表现出更高的发芽率、更长的根长和更小的生长抑制。与WT相比,转基因植物在氧化胁迫下较低的活性氧(ROS)积累、丙二醛(MDA)含量和细胞膜损伤也表明其氧化胁迫耐受性增强。在转基因植物中观察到超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和过氧化物酶(POD)等抗氧化酶的活性高于WT,这可能与TaFBA1的过表达导致一些抗氧化基因的表达上调有关。此外,在TaFBA1的启动子区域发现了一些胁迫响应元件,并且TaFBA1定位于细胞核、细胞质和质膜。这些结果表明TaFBA1在植物的氧化胁迫耐受性中起重要作用。这对于理解F-box蛋白在植物对多种胁迫条件的耐受性中的功能很重要。