Chen Shuangchen, Zhao Hongjiao, Wang Mengmeng, Li Jidi, Wang Zhonghong, Wang Fenghua, Liu Airong, Ahammed Golam J
College of Forestry, Henan University of Science and TechnologyLuoyang, China.
Department of Plant Science, Tibet Agriculture and Animal Husbandry CollegeLinzhi, China.
Front Plant Sci. 2017 Jun 30;8:1109. doi: 10.3389/fpls.2017.01109. eCollection 2017.
Ubiquitination is a common regulatory mechanism, playing a critical role in diverse cellular and developmental processes in eukaryotes. However, a few reports on the functional correlation between E3 ubiquitin ligases and reactive oxygen species (ROS) or reactive nitrogen species (RNS) metabolism in response to stress are currently available in plants. In the present study, the E3 ubiquitin ligase gene (Adi3 Binding E3 Ligase) was introduced into tomato line Ailsa Craig via -mediated method. Transgenic lines were confirmed for integration into the tomato genome using PCR. Transcription of in various transgenic lines was determined using real-time PCR. Evaluation of stress tolerance showed that T generation of transgenic tomato lines showed only mild symptoms of chilling injury as evident by higher biomass accumulation and chlorophyll content than those of non-transformed plants. Compared with wild-type plants, the contents of AsA, AsA/DHA, GSH and the activity of GaILDH, γ-GCS and GSNOR were increased, while HO, [Formula: see text], MDA, NO, SNOs, and GSNO accumulations were significantly decreased in overexpressing plants in response to chilling stress. Furthermore, transgenic tomato plants overexpressing showed higher activities of enzymes such as G6PDH, 6PGDH, NADP-ICDH, and NADP-ME involved in pentose phosphate pathway (PPP). The transgenic tomato plants also exhibited an enhanced tolerance against the necrotrophic fungus . Tyrosine nitration protein was activated in the plants infected with leaf mold disease, while the inhibition could be recovered in gene overexpressing lines. Taken together, our results revealed a possible physiological role of in the activation of the key enzymes of AsA-GSH cycle, PPP and down-regulation of GSNO reductase, thereby reducing oxidative and nitrosative stress in plants. This study demonstrates an optimized transgenic strategy using gene for crop improvement against biotic and abiotic stress factors.
泛素化是一种常见的调控机制,在真核生物的各种细胞和发育过程中发挥着关键作用。然而,目前关于植物中E3泛素连接酶与活性氧(ROS)或活性氮(RNS)代谢在应激反应中的功能相关性的报道较少。在本研究中,通过农杆菌介导的方法将E3泛素连接酶基因(Adi3结合E3连接酶)导入番茄品种Ailsa Craig。使用PCR确认转基因株系已整合到番茄基因组中。使用实时PCR测定各种转基因株系中该基因的转录水平。胁迫耐受性评估表明,转基因番茄株系的T代仅表现出轻微的冷害症状,其生物量积累和叶绿素含量高于未转化植株。与野生型植株相比,在低温胁迫下,过表达该基因的植株中抗坏血酸(AsA)、AsA/脱氢抗坏血酸(DHA)、谷胱甘肽(GSH)的含量以及甘油醛-3-磷酸脱氢酶(GaILDH)、γ-谷氨酰半胱氨酸合成酶(γ-GCS)和亚硝基谷胱甘肽还原酶(GSNOR)的活性增加,而过氧化氢(HO)、超氧阴离子([公式:见原文])、丙二醛(MDA)、一氧化氮(NO)、亚硝基硫醇(SNOs)和亚硝基谷胱甘肽(GSNO)的积累显著减少。此外,过表达该基因的转基因番茄植株参与戊糖磷酸途径(PPP)的酶如葡萄糖-6-磷酸脱氢酶(G6PDH)、6-磷酸葡萄糖酸脱氢酶(6PGDH)、NADP-异柠檬酸脱氢酶(NADP-ICDH)和NADP-苹果酸酶(NADP-ME)的活性更高。转基因番茄植株对坏死性真菌也表现出增强的耐受性。在感染叶霉病的植株中酪氨酸硝化蛋白被激活,而在过表达该基因的株系中这种抑制作用可以恢复。综上所述,我们的结果揭示了该基因在激活AsA-GSH循环关键酶、PPP以及下调GSNO还原酶方面可能的生理作用,从而减轻植物中的氧化和亚硝化应激。本研究展示了一种利用该基因进行作物改良以对抗生物和非生物胁迫因素优化的转基因策略。