Yu Dongdong, Zhang Lihua, Zhao Kai, Niu Ruxuan, Zhai Huan, Zhang Jianxia
College of Horticulture, Northwest A&F UniversityYangling, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of AgricultureYangling, China; State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F UniversityYangling, China.
Front Plant Sci. 2017 Mar 7;8:297. doi: 10.3389/fpls.2017.00297. eCollection 2017.
Early responsive to dehydration () genes can be rapidly induced to counteract abiotic stresses, such as drought, low temperatures or high salinities. Here, we report on an gene () related to cold tolerance from Chinese wild accession 'Heilongjiang seedling'. The full-length cDNA is 685 bp, including a 66 bp 5'-untranslated region (UTR), a 196 bp 3'-UTR region and a 423 bp open reading frame encoding 140 amino acids. The VaERD15 protein shares a high amino acid sequence similarity with ERD15 of . In our study, was shown to have a nucleic localization function and a transcriptional activation function. Semi-quantitative PCR and Western blot analyses showed that was constitutively expressed in young leaves, stems and roots of accession 'Heilongjiang seedling' plants, and expression levels increased after low-temperature treatment. We also generated a transgenic Col-0 line that over-expressed and carried out a cold-treatment assay. Real-time quantitative PCR (qRT-PCR) and Western blot analyses showed that as the duration of cold treatment increased, the expression of both gene and protein levels increased continuously in the transgenic plants, while almost no expression was detected in the wild type . Moreover, the plants that over-expressed showed higher cold tolerance and accumulation of proline, soluble sugars, proteins, malondialdehyde and three antioxidases (superoxide dismutase, peroxidase, and catalase). Lower levels of relative ion leakage also occurred under cold stress. Taken together, our results indicate that the transcription factor was induced by cold stress and was able to enhance cold tolerance.
早期脱水响应()基因可被快速诱导以对抗非生物胁迫,如干旱、低温或高盐度。在此,我们报道了一个来自中国野生种‘黑龙江幼苗’的与耐寒性相关的基因()。该基因全长cDNA为685 bp,包括一个66 bp的5'非翻译区(UTR)、一个196 bp的3'UTR区和一个423 bp的开放阅读框,编码140个氨基酸。VaERD15蛋白与的ERD15具有高度的氨基酸序列相似性。在我们的研究中,显示具有核定位功能和转录激活功能。半定量PCR和蛋白质印迹分析表明,在‘黑龙江幼苗’植株的幼叶、茎和根中组成型表达,低温处理后表达水平升高。我们还构建了一个过表达的转基因Col-0株系并进行了冷处理试验。实时定量PCR(qRT-PCR)和蛋白质印迹分析表明,随着冷处理时间的延长,转基因植株中基因和蛋白水平的表达均持续增加,而野生型中几乎未检测到表达。此外,过表达的植株表现出更高的耐寒性以及脯氨酸、可溶性糖、蛋白质、丙二醛和三种抗氧化酶(超氧化物歧化酶、过氧化物酶和过氧化氢酶)的积累。在冷胁迫下相对离子渗漏水平也较低。综上所述,我们的结果表明转录因子受冷胁迫诱导并能够增强耐寒性。