Investigating resistance to Barley yellow dwarf virus in wheat

研究小麦对大麦黄矮病毒的抗性

• 批准号: 2594275
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2594275
• 关键词: Investigating; resistance; Barley; yellow; dwarf

Barley yellow dwarf virus (BYDV) threatens sustainable UK wheat production. Control of BYDV has historically relied upon vector-targeted mitigation strategies underpinned by pesticides, removing aphid species which acquire and spread the virus during feeding. Prior to late 2018 when a ban came into force, neonicotinoid seed treatments were widely used to protect winter wheat. Growers retain the option of alternative insecticide use, such as pyrethroids, but these require foliar sprays which are neither always possible in autumnal conditions nor specific, killing beneficial insects. Increasing levels of pyrethroid resistance by key aphid species are also being recorded. Combined with the demand for pyrethroids, a markedly high risk for rapid spread of insecticide resistance exists, further reducing the chemical arsenal available for controlling BYDV. Sources of genetic resistance are consequently of value and have become a renewed focus by the wheat breeders. This includes RAGT Seeds who have recently developed the BYDV resistant cultivar RGT Wolverine. This PhD project would involve screening the Wolverine source of BYDV resistance (and/or tolerance) against informative BYDV isolates, qualifying the level of resistance to different virus strains and determining whether strain complementation could overcome resistance. Through a series of experiments including antibody-based whole plant tissue printing combined with western blotting, the spatial and temporal movement of BYDV within resistant and susceptible plant lines will also be explored. Using the 'MutChromSeq' approach, or transcriptomics combined with bulk segregant analysis, the causal genomic loci underpinning the BYDV resistance will be elucidated. In concert to this work, an additional focus will be the development of a mid/high-throughput RT-qPCR assay for BYDV, as well as new approaches and technologies to help identify and/or validate other resistances. Limited reliable in-field BYDV resistance screening methodologies currently exist with most testing relying upon natural BYDV presence. During this PhD, prevalent strains of BYDV will be identified (via the Rothamsted UK insect suction trap network) and used to develop infectious clones (ICs). The use of these ICs with biolistic inoculation strategies will be explored and an IC 'toolkit' for screening wheat germplasm potentially developed. This would allow simplified maintenance of pure BYDV strains in bacterial cultures, rapid generation of a desired inoculum and robust investigations into BYDV strain complementation. This PhD represents an opportunity to generate valuable applied outputs with training provided (where required) on molecular biology/virology techniques, bioinformatics, entomology, critical thinking and conceptualising, data interpretation, and glasshouse and field work.

大麦黄矮病毒（BYDV）威胁着英国小麦的可持续生产。BYDV的控制历来依赖于以杀虫剂为基础的媒介靶向缓解策略，清除在进食过程中获得和传播病毒的蚜虫物种。在2018年底禁令生效之前，类烟碱种子处理剂被广泛用于保护冬小麦。种植者保留使用替代杀虫剂的选择，如拟除虫菊酯，但这些需要叶面喷雾，这在秋季条件下既不总是可能的，也不特定，杀死有益昆虫。据记录，主要蚜虫物种对拟除虫菊酯的抗药性水平也在不断提高。再加上对拟除虫菊酯的需求，杀虫剂耐药性迅速传播的风险显着增加，进一步减少了可用于控制黄矮病的化学武器库。因此，遗传抗性的来源是有价值的，并已成为小麦育种家的新焦点。这包括RAGT种子公司，他们最近开发了抗BYDV品种RGT金刚狼。这个博士项目将涉及筛选金刚狼来源的BYDV抗性（和/或耐受性）对信息BYDV分离物，鉴定不同病毒株的抗性水平，并确定菌株互补是否可以克服抗性。通过一系列实验，包括基于抗体的全植物组织打印结合蛋白质印迹，也将探索抗和感植物系中BYDV的空间和时间移动。使用“MutChromSeq”方法，或转录组学结合批量分离分析，将阐明支持BYDV抗性的致病基因组座位。与这项工作相一致，另一个重点将是开发BYDV的中/高通量RT-qPCR检测方法，以及帮助识别和/或验证其他抗性的新方法和技术。目前存在有限的可靠的田间BYDV抗性筛选方法，大多数测试依赖于天然BYDV的存在。在本博士期间，将鉴定BYDV的流行株（通过英国Rothamsted昆虫抽吸诱捕网络）并用于开发感染性克隆（IC）。将探索这些IC与生物射弹接种策略的使用，并开发用于筛选小麦种质的IC“工具包”。这将允许在细菌培养物中简化纯BYDV菌株的维持，快速产生所需的接种物和对BYDV菌株互补的稳健研究。该博士学位提供了一个机会，可以通过提供分子生物学/病毒学技术，生物信息学，昆虫学，批判性思维和概念化，数据解释以及温室和实地工作的培训（如需要）产生有价值的应用成果。