National Key Laboratory of Cotton Bio-Breeding and Integrated Utilization/Institute of Cotton Research, Chinese Academy of Agricultural Sciences (ICR, CAAS), Anyang, Henan, 455000, China.
College of Agronomy, Hebei Agricultural University/North China Key Laboratory for Crop Germplasm Resources of Ministry of Education, Baoding, 071001, Hebei, China.
BMC Plant Biol. 2023 Oct 18;23(1):501. doi: 10.1186/s12870-023-04455-w.
The cotton industry suffers significant yield losses annually due to Verticillium wilt, which is considered the most destructive disease affecting the crop. However, the precise mechanisms behind this disease in cotton remain largely unexplored.
Our approach involved utilizing transcriptome data from G. australe which was exposed to Verticillium dahliae infection. From this data, we identified ethylene-responsive factors and further investigated their potential role in resistance through functional validations via Virus-induced gene silencing (VIGS) in cotton and overexpression in Arabidopsis.
A total of 23 ethylene response factors (ERFs) were identified and their expression was analyzed at different time intervals (24 h, 48 h, and 72 h post-inoculation). Among them, GauERF105 was selected based on qRT-PCR expression analysis for further investigation. To demonstrate the significance of GauERF105, VIGS was utilized, revealing that suppressing GauERF105 leads to more severe infections in cotton plants compared to the wild-type. Additionally, the silenced plants exhibited reduced lignin deposition in the stems compared to the WT plants, indicating that the silencing of GauERF105 also impacts lignin content. The overexpression of GauERF105 in Arabidopsis confirmed its pivotal role in conferring resistance against Verticillium dahliae infection. Our results suggest that WT possesses higher levels of the oxidative stress markers MDA and HO as compared to the overexpressed lines. In contrast, the activities of the antioxidant enzymes SOD and POD were higher in the overexpressed lines compared to the WT. Furthermore, DAB and trypan staining of the overexpressed lines suggested a greater impact of the disease in the wild-type compared to the transgenic lines.
Our findings provide confirmation that GauERF105 is a crucial candidate in the defense mechanism of cotton against Verticillium dahliae invasion, and plays a pivotal role in this process. These results have the potential to facilitate the development of germplasm resistance in cotton.
由于黄萎病,棉花产业每年都会遭受重大的产量损失,这种疾病被认为是对作物危害最大的疾病。然而,棉花黄萎病的确切发病机制在很大程度上仍未得到探索。
我们的方法涉及利用 G. australe 的转录组数据,该数据暴露于 Verticillium dahliae 感染下。从这些数据中,我们鉴定了乙烯响应因子,并通过在棉花中进行病毒诱导的基因沉默(VIGS)和在拟南芥中过表达来进一步研究它们在抗性中的潜在作用。
共鉴定出 23 个乙烯响应因子(ERFs),并在不同时间间隔(接种后 24、48 和 72 小时)分析它们的表达。其中,根据 qRT-PCR 表达分析,选择 GauERF105 进行进一步研究。为了证明 GauERF105 的重要性,我们使用了 VIGS,结果表明与野生型相比,抑制 GauERF105 会导致棉花植株受到更严重的感染。此外,与 WT 植株相比,沉默植株的茎部木质素沉积减少,表明 GauERF105 的沉默也会影响木质素含量。在拟南芥中过表达 GauERF105 证实了它在赋予对 Verticillium dahliae 感染的抗性方面的关键作用。我们的结果表明,与过表达系相比,WT 具有更高水平的氧化应激标志物 MDA 和 HO。相比之下,在过表达系中 SOD 和 POD 等抗氧化酶的活性更高。此外,过表达系的 DAB 和锥虫染色表明,与转基因系相比,WT 受到的疾病影响更大。
我们的研究结果证实 GauERF105 是棉花抵御 Verticillium dahliae 入侵的防御机制中的一个关键候选基因,在这个过程中起着关键作用。这些结果有可能促进棉花种质资源的抗病性发展。