Bilateral BBSRC-Embrapa: Using disease risk forecasting, NGS and HIGS to explore and control Fusarium Head Blight disease in wheat fields

双边 BBSRC-Embrapa：利用疾病风险预测、NGS 和 HIGS 探索和控制麦田赤霉病

基本信息

批准号：
BB/N018095/1
负责人：
Kim Hammond-Kosack
金额：
$ 49.84万
依托单位：
Rothamsted Research
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2016
资助国家：
英国
起止时间：
2016 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FN018095%2F1
关键词：
Bilateral BBSRC Embrapa Using disease

项目摘要

This is a highly innovative and truly ground breaking research project devised by scientists based at Embrapa Wheat, Soybean, Genetic Resources and Biotechnology and Bioinformatics in Brazil and Rothamsted Research in the UK. The problem to be addressed is the control of a fungal disease called Fusarium head blight (FHB) which is one of the most serious and hazardous crop diseases worldwide. The main consequence of FHB is that trichothecene mycotoxins, such as deoxynivalenol (DON), accumulate in the grain, presenting a health risk to humans and animals. In Southern Brazil, where 90% of Brazilian wheat is grown, severe FHB epidemic years occur at a minimum of every 4 or 5 years. Legal limits have been set on the DON levels permitted in harvested grain used for different purposes. However, even moderate FHB years are highly problematic causing the lack of available safe grain for use either on farms or for sale into the market. For low income Brazilian farmers, FHB disease reduces the standards of living of farmer's families and that of their local communities. There is a pressing need to develop novel and effective FHB control options.In this project, we intend to take a novel whole fungal genome and disease modelling guided approach to develop a pipeline of genetically modified wheat genotypes harbouring T-DNA constructs, which can silence Fusarium genes critical for wheat infection via host-induced gene silencing (HIGS). We also intend to determine the plant and fungal mechanisms that control the HIGS phenomenon. HIGS could be used to control multiple pathogens.This project has six main research steps.1. To explore using next generation sequencing the genomes of the five FHB causing species in Southern Brazil. Define the core and species-specific proteome of the FHB species complex (FGSC) for the development of molecular diagnostic tools and the selection of HIGS targets. Monitor for possible genome alterations over 3 years.2. To enhance FHB disease risk forecasting, by establishing a spore sampler network that can detect and quantify Fusarium species. Devise and use diagnostic assays to identify and monitor FGSC diversity. Sample atmospheric Fusarium spores to identify potential inoculum sources and population structure. Incorporate data on the dynamics of airborne Fusarium spore populations into the existing regional FHB risk model. 3. To develop various T-DNA based constructs to silence Fusarium gene expression by HIGS, thereby controlling Fusarium infections. Evaluate single gene HIGS constructs using a novel cut wheat tillers (transient assay) and via stable transformation into Arabidopsis or lettuce. Two lead HIGS constructs targeting multiple Fusarium genes will be transform into a moderately FHB resistance Brazilian wheat cultivar. In the resulting transgenic plant populations, FHB severity and DON levels will be quantified. 4. To explore the underlying mechanisms of HIGS three cutting edge experiments will be completed. We will establish if long or short RNA molecules move between fungal and wheat cells, determine if the gene silencing phenomena once initiated operates systemically and investigate if transported RNA molecules are cargoed using vesicular transport to the plant cell surface for delivery into fungal cells.5. To perform two years of GM wheat trial in two locations in Brazil using the four best HIGS lines and three appropriate control lines. Assess in-field FHB symptoms, airborne inoculum, grain quality, DON contamination and fungal spore production. To collect Fusarium isolates able to cause any disease on the HIGS lines and complete a full genome analyse.6. The research team will engage with farmers, farmer co-operatives, grain purchasers, plant breeders in Brazil and the UK and academics globally to explain the project, the research findings and discuss ways to implement the new modelling/forecasting technologies, to use the novel GM trait and further understand the HIGS phenomenon.

这是一个高度创新和真正突破性的研究项目，由巴西 Embrapa 小麦、大豆、遗传资源和生物技术和生物信息学以及英国洛桑研究中心的科学家设计。要解决的问题是控制一种名为赤霉病（FHB）的真菌病害，它是全世界最严重和最危险的作物病害之一。 FHB 的主要后果是单端孢菌素真菌毒素，如脱氧雪腐镰刀菌烯醇 (DON)，在谷物中积累，对人类和动物造成健康风险。在巴西南部，巴西90%的小麦都产自这里，至少每4年或5年就会发生一次严重的FHB流行年。法律对用于不同用途的收获谷物中允许的呕吐毒素水平设定了限制。然而，即使是中等的 FHB 年份也存在很大问题，导致缺乏可用的安全谷物用于农场或销售到市场。对于低收入的巴西农民来说，FHB 疾病降低了农民家庭及其当地社区的生活水平。迫切需要开发新颖有效的 FHB 控制方案。在这个项目中，我们打算采用一种新颖的全真菌基因组和疾病建模指导方法来开发一系列含有 T-DNA 构建体的转基因小麦基因型，该基因型可以沉默镰刀菌基因通过宿主诱导的基因沉默（HIGS）对小麦感染至关重要。我们还打算确定控制 HIGS 现象的植物和真菌机制。 HIGS可用于控制多种病原体。本项目有六个主要研究步骤： 1．利用新一代测序技术探索巴西南部五种 FHB 致病物种的基因组。定义 FHB 物种复合体 (FGSC) 的核心和物种特异性蛋白质组，用于开发分子诊断工具和选择 HIGS 靶点。监测 3 年内可能发生的基因组改变。2．通过建立可检测和量化镰刀菌属物种的孢子采样器网络，加强 FHB 疾病风险预测。设计并使用诊断分析来识别和监测 FGSC 多样性。对大气镰刀菌孢子进行采样，以确定潜在的接种源和种群结构。将空气中镰刀菌孢子种群动态数据纳入现有的区域 FHB 风险模型。 3.开发各种基于T-DNA的构建体，通过HIGS沉默镰刀菌基因表达，从而控制镰刀菌感染。使用新型切小麦分蘖（瞬时测定）并通过稳定转化到拟南芥或生菜中来评估单基因 HIGS 构建体。两个针对多个镰刀菌基因的领先 HIGS 构建体将被转化为中度 FHB 抗性巴西小麦品种。在所得的转基因植物群体中，FHB 严重程度和 DON 水平将被量化。 4. 为了探索HIGS的潜在机制，将完成三个前沿实验。我们将确定长RNA分子还是短RNA分子在真菌和小麦细胞之间移动，确定基因沉默现象一旦启动是否会系统性地起作用，并研究运输的RNA分子是否通过囊泡运输被运送到植物细胞表面以递送到真菌细胞中。5．使用四个最好的 HIGS 品系和三个适当的对照品系在巴西的两个地点进行为期两年的转基因小麦试验。评估田间 FHB 症状、空气接种菌、谷物质量、DON 污染和真菌孢子产生。收集能够在HIGS品系上引起任何疾病的镰刀菌分离株并完成全基因组分析。6．研究团队将与巴西和英国的农民、农民合作社、粮食购买者、植物育种者以及全球学者合作，解释该项目、研究结果，并讨论实施新建模/预测技术的方法，以使用新颖的方法GM特质并进一步了解HIGS现象。