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

• 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

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开创性的研究项目，由巴西Embrapa小麦，大豆，遗传资源和生物技术以及生物信息学和英国Rothamsted研究所的科学家设计。要解决的问题是一种称为镰刀菌头枯病（FHB）的真菌病害的控制，这是世界上最严重和最危险的作物病害之一。FHB的主要后果是，诸如脱氧雪腐镰刀菌烯醇（DON）的多孢霉烯真菌毒素在谷物中积累，对人类和动物造成健康风险。在巴西南部，90%的巴西小麦种植在那里，严重的FHB流行年至少每4或5年发生一次。法律的限制已经设定了用于不同目的的收获谷物中允许的DON水平。然而，即使是中等的FHB年份也存在很大的问题，导致缺乏可用于农场或销售到市场的安全谷物。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。本项目拟采用一种新的真菌全基因组和病害模型指导的方法，开发一系列含有T-DNA构建体的转基因小麦，通过宿主诱导基因沉默（HIGS）的方式，使小麦感染镰刀菌的关键基因沉默。我们还打算确定控制HIGS现象的植物和真菌机制。HIGS可用于多种病原菌的控制，本项目主要研究步骤有六个.探索使用下一代测序巴西南部五种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，用于分子诊断工具的开发和HIGS靶点的选择。监测可能的基因组改变超过3年。2.通过建立一个能够检测和量化镰刀菌种类的孢子采样器网络，加强对FHB疾病风险的预测。设计和使用诊断分析来识别和监测FGSC多样性。采样大气镰刀菌孢子，以确定潜在的接种源和人口结构。将空气中镰刀菌孢子种群的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。使用新的切割小麦分蘖（瞬时测定）并通过稳定转化到拟南芥或莴苣中来评估单基因HIGS构建体。将靶向多个镰刀菌属基因的两个前导HIGS构建体转化到中等FHB抗性巴西小麦栽培品种中。在所得转基因植物群体中，将定量FHB严重性和DON水平。4.为了探索HIGS的潜在机制，将完成三个前沿实验。我们将确定长或短RNA分子是否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系统性地运作，并调查是否使用囊泡运输将运输的RNA分子运送到植物细胞表面以递送到真菌细胞中。在巴西的两个地点使用四个最好的HIGS系和三个适当的对照系进行两年的转基因小麦试验。评估田间赤霉病症状，空气中的接种物，粮食质量，DON污染和真菌孢子生产。收集能在HIGS系上引起任何疾病的镰刀菌分离物并完成全基因组分析。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的学者进行交流，解释该项目、研究结果，并讨论如何实施新的建模/预测技术，使用新的转基因性状，进一步了解HIGS现象。

项目成果

Achieving durable disease resistance in cereals

实现谷物的持久抗病性

• DOI: 10.19103/as.2021.0092.07
• 发表时间: 2021
• 作者: Kanja C

Annotation of Fusarium graminearum (PH-1) Version 5.0.

禾谷镰刀菌 (PH-1) 5.0 版注释。

• DOI: 10.1128/genomea.01479-16
• 发表时间: 2017
• 期刊: Genome announcements
• 作者: King R

Analysis of small RNA silencing in the interactions between wheat and an exclusively apoplastic fungal pathogen Zymoseptoria tritici

小麦与纯质外体真菌病原体小麦发酵锈病菌之间相互作用的小 RNA 沉默分析

• 发表时间: 2019
• 期刊: MOLECULAR PLANT-MICROBE INTERACTIONS
• 影响因子: 3.5
• 作者: Kettles G. J.

Inter-genome comparison of the Quorn fungus Fusarium venenatum and the closely related plant infecting pathogen Fusarium graminearum.

• DOI: 10.1186/s12864-018-4612-2
• 发表时间: 2018-04-19
• 期刊: BMC genomics
• 影响因子: 4.4
• 作者: King R;Brown NA;Urban M;Hammond-Kosack KE
• 通讯作者: Hammond-Kosack KE

Identification and functional characterisation of a locus for target site integration in Fusarium graminearum.

禾谷镰刀菌靶位点整合位点的鉴定和功能表征。

• DOI: 10.1186/s40694-024-00171-8
• 发表时间: 2024
• 期刊: Fungal biology and biotechnology
• 作者: Darino M