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Expression of TaWRKY44 a wheat WRKY gene in transgenic tobacco confers multiple abiotic stress tolerances

机译:TaWRKY44(小麦WRKY基因)在转基因烟草中的表达赋予多种非生物胁迫耐受性

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摘要

The WRKY transcription factors have been reported to be involved in various plant physiological and biochemical processes. In this study, we successfully assembled 10 unigenes from expressed sequence tags (ESTs) of wheat and designated them as TaWRKY44–TaWRKY53, respectively. Among these genes, a subgroup I gene, TaWRKY44, was found to be upregulated by treatments with PEG6000, NaCl, 4°C, abscisic acid (ABA), H2O2 and gibberellin (GA). The TaWRKY44-GFP fusion protein was localized to the nucleus of onion epidermal cells, and TaWRKY44 was able to bind to the core DNA sequences of TTGACC and TTAACC in yeast. The N-terminal of TaWRKY44 showed transcriptional activation activity. Expression of TaWRKY44 in tobacco plants conferred drought and salt tolerance and transgenic tobacco exhibited a higher survival rate, relative water content (RWC), soluble sugar, proline and superoxide dismutase (SOD) content, as well as higher activities of catalase (CAT) and peroxidase (POD), but less ion leakage (IL), lower contents of malondialdehyde (MDA), and H2O2. In addition, expression of TaWRKY44 also increased the seed germination rate in the transgenic lines under osmotic stress conditions while exhibiting a lower H2O2 content and higher SOD, CAT, and POD activities. Expression of TaWRKY44 upregulated the expression of some reactive oxygen species (ROS)-related genes and stress-responsive genes in tobacco under osmotic stresses. These data demonstrate that TaWRKY44 may act as a positive regulator in drought/salt/osmotic stress responses by either efficient ROS elimination through direct or indirect activation of the cellular antioxidant systems or activation of stress-associated gene expression.
机译:据报道,WRKY转录因子参与了多种植物的生理和生化过程。在这项研究中,我们成功地从小麦的表达序列标签(EST)组装了10个单基因,并将其分别命名为TaWRKY44-TaWRKY53。在这些基因中,发现亚类I基因TaWRKY44通过PEG6000,NaCl,4°C,脱落酸(ABA),H2O2和赤霉素(GA)处理而被上调。 TaWRKY44-GFP融合蛋白位于洋葱表皮细胞的核中,并且TaWRKY44能够与酵母中TTGACC和TTAACC的核心DNA序列结合。 TaWRKY44的N端显示转录激活活性。 TaWRKY44在烟草植物中的表达赋予了干旱和耐盐性,转基因烟草表现出更高的存活率,相对含水量(RWC),可溶性糖,脯氨酸和超氧化物歧化酶(SOD)含量以及过氧化氢酶(CAT)和过氧化物酶(POD),但离子泄漏(IL)较少,丙二醛(MDA)和H2O2含量较低。此外,在渗透胁迫条件下,TaWRKY44的表达还提高了转基因品系的种子发芽率,同时具有较低的H2O2含量和较高的SOD,CAT和POD活性。 TaWRKY44的表达上调了渗透胁迫下烟草中一些活性氧(ROS)相关基因和胁迫响应基因的表达。这些数据表明,TaWRKY44可以通过直接或间接激活细胞抗氧化剂系统或激活与压力相关的基因表达来有效清除ROS,从而在干旱/盐/渗透胁迫响应中充当正调控因子。

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