State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai'an, 271018, China.
State Key Laboratory of Crop Biology, College of Life Science, Shandong Agricultural University, Tai'an, 271018, China.
Plant Cell Rep. 2017 Dec;36(12):1985-1994. doi: 10.1007/s00299-017-2221-1. Epub 2017 Oct 14.
Expression of TaRUB1 gene in Arabidopsis thaliana elevates the level of disease-related genes in response to pathogen invasion through the accumulation of callose, necrotic cells, and the outbreak of ROS. Ubiquitin (Ub) and ubiquitin-like proteins are highly conserved in sequence and can covalently bind and modify many intracellular proteins which can be recognized and degraded by 26S proteasome. Post-translational modification of proteins has become a hot research spot today. In the previous study, a cDNA of related-to-ubiquitin protein belonged to ubiquitin-like proteins, whose spatial structure comprised Ub and NEDD8, was obtained from wheat SN6306 by suppression-subtractive hybridization and was named TaRUB1. TaRUB1 is induced by wheat powdery mildew and significantly upregulated in resistant wheat SN6306. In this study, heterologous expression of TaRUB1 in A. thaliana was used to study the function of this gene in response to pathogen Pseudomonas syringae pv. Tomato DC3000 (Pst DC3000). Transgenic A. thaliana showed relatively fewer disease symptoms, accompanied by common inhibition of living body parasitic defense responses, accumulation of more callose and reactive oxygen species (ROS), and concentrated cell death, simultaneously antioxidant enzyme activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase were higher than those in wild-type (WT) plant after infection with Pst DC3000. Meanwhile, hypersensitive cell death, which was possibly ROS burst, was also observed in transgenic A. thaliana. By quantitative reverse transcription-polymerase chain reaction analysis, some marker genes for hypersensitive response showed significantly higher transcriptional expression level in transgenic A. thaliana, which activates system-acquired resistance, than that of WT plants. Heterologous expression of TaRUB1 can significantly enhance resistance to Pst DC3000 in A. thaliana, suggesting that TaRUB1 is related to plant disease resistance.
TaRUB1 基因在拟南芥中的表达通过积累胼胝质、坏死细胞和活性氧(ROS)的爆发,提高了对病原体入侵的疾病相关基因的水平。泛素(Ub)和类泛素蛋白在序列上高度保守,可以共价结合并修饰许多细胞内蛋白质,这些蛋白质可以被 26S 蛋白酶体识别和降解。蛋白质的翻译后修饰已成为当今研究的热点。在之前的研究中,通过抑制差减杂交从小麦 SN6306 中获得了一种与泛素蛋白相关的 cDNA,属于类泛素蛋白,其空间结构由 Ub 和 NEDD8 组成,被命名为 TaRUB1。TaRUB1 被小麦白粉菌诱导,在抗性小麦 SN6306 中显著上调。在这项研究中,通过在拟南芥中异源表达 TaRUB1 来研究该基因对病原体丁香假单胞菌 pv.番茄 DC3000(Pst DC3000)的反应功能。转基因拟南芥表现出相对较少的疾病症状,同时伴随着生物体寄生防御反应的常见抑制、更多胼胝质和活性氧(ROS)的积累以及集中的细胞死亡,同时超氧化物歧化酶、过氧化物酶、过氧化氢酶和抗坏血酸过氧化物酶的抗氧化酶活性在感染 Pst DC3000 后高于野生型(WT)植物。同时,在转基因拟南芥中也观察到可能由 ROS 爆发引起的超敏性细胞死亡。通过定量逆转录聚合酶链反应分析,一些与超敏反应相关的标记基因在转基因拟南芥中的转录表达水平明显高于 WT 植物,从而激活了系统获得性抗性。TaRUB1 的异源表达可以显著提高拟南芥对 Pst DC3000 的抗性,表明 TaRUB1 与植物抗病性有关。