Jiao Yuntong, Xu Weirong, Duan Dong, Wang Yuejin, Nick Peter
College of Horticulture, Northwest A & F University, Yangling 712100, Shaanxi, People's Republic of China Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Yangling 712100, Shaanxi, People's Republic of China State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A & F University, Yangling, Shaanxi 712100, People's Republic of China.
Molecular Cell Biology, Botanical Institute 1, Karlsruhe Institute of Technology, Kaiserstr. 2, D-78133 Karlsruhe, Germany.
J Exp Bot. 2016 Oct;67(19):5841-5856. doi: 10.1093/jxb/erw351. Epub 2016 Oct 4.
Stilbenes are central phytoalexins in Vitis, and induction of the key enzyme stilbene synthase (STS) is pivotal for disease resistance. Here, we address the potential for breeding resistance using an STS allele isolated from Chinese wild grapevine Vitis pseudoreticulata (VpSTS) by comparison with its homologue from Vitis vinifera cv. 'Carigane' (VvSTS). Although the coding regions of both alleles are very similar (>99% identity on the amino acid level), the promoter regions are significantly different. By expression in Arabidopsis as a heterologous system, we show that the allele from the wild Chinese grapevine can confer accumulation of stilbenes and resistance against the powdery mildew Golovinomyces cichoracearum, whereas the allele from the vinifera cultivar cannot. To dissect the upstream signalling driving the activation of this promoter, we used a dual-luciferase reporter system in a grapevine cell culture. We show elevated responsiveness of the promoter from the wild grape to salicylic acid (SA) and to the pathogen-associated molecular pattern (PAMP) flg22, equal induction of both alleles by jasmonic acid (JA), and a lack of response to the cell death-inducing elicitor Harpin. This elevated SA response of the VpSTS promoter depends on calcium influx, oxidative burst by RboH, mitogen-activated protein kinase (MAPK) signalling, and JA synthesis. We integrate the data in the context of a model where the resistance of V. pseudoreticulata is linked to a more efficient recruitment of SA signalling for phytoalexin synthesis.
芪类化合物是葡萄中的主要植保素,关键酶芪合酶(STS)的诱导对于抗病性至关重要。在这里,我们通过将从中国野生葡萄刺葡萄(VpSTS)中分离的一个STS等位基因与其来自酿酒葡萄品种‘佳丽酿’(VvSTS)的同源物进行比较,探讨了利用该等位基因进行抗性育种的潜力。尽管两个等位基因的编码区非常相似(氨基酸水平上的同一性>99%),但启动子区却有显著差异。通过在拟南芥中作为异源系统进行表达,我们发现来自中国野生葡萄的等位基因能够赋予芪类化合物的积累以及对白粉病菌黄瓜白粉菌的抗性,而来自酿酒葡萄品种的等位基因则不能。为了剖析驱动该启动子激活的上游信号传导,我们在葡萄细胞培养中使用了双荧光素酶报告系统。我们发现野生葡萄启动子对水杨酸(SA)和病原体相关分子模式(PAMP)flg22的反应性增强,茉莉酸(JA)对两个等位基因的诱导作用相同,并且对诱导细胞死亡的激发子Harpin无反应。VpSTS启动子对SA的这种增强反应依赖于钙内流、RboH介导的氧化爆发、丝裂原活化蛋白激酶(MAPK)信号传导以及JA合成。我们将这些数据整合到一个模型中,该模型认为刺葡萄的抗性与更有效地募集SA信号用于植保素合成有关。