A functional genomics toolkit for the wheat pathogen Mycosphaerella graminicola

小麦是欧洲的主要粮食作物，包括英国，小麦的年产量约为14亿英镑。预计到2050年，地球人口将增加到90亿以上，我们迫切需要找到提供更多粮食的方法来支持这一增长。真菌病是小麦生产的主要威胁之一。在英国，致病菌真菌是小麦的主要致病因子，也是仅在英国就花费超过5亿英镑用于保护谷类作物的杀菌剂的主要原因之一。尽管如此，仍有重大的产量损失。因此，迫切需要新的持久的疾病控制策略，包括新型杀菌剂。开发新型杀菌剂的主要困难是无法确定新分子如何杀死真菌。在酿酒酵母/面包酵母的模型真菌，酿酒酵母，强大的工具已经产生，如酵母菌株组；缺乏每种真菌基因的；每个蛋白质都被荧光蛋白标记，或者每个蛋白质都在高水平上表达。这些资源使我们能够大规模地确定各种化学物质是如何改变酵母的生物学特性的。我们打算构建一套类似的完整工具，使我们能够进行这些检测，从而确定新型杀菌剂如何在小麦病原菌中起作用。具体来说，我们将把它的大约10,000个基因中的每一个放入一个可移动的遗传元件（质粒）中，这将允许容易地转移到大范围的其他质粒（我们也将创建）中，这些质粒可以用来灭活，荧光标记和过表达每个基因，无论是单独的还是作为一个池。我们还将构建一系列禾本科真菌菌株，这将使我们能够确定天然真菌是如何入侵小麦的，我们将开发一个小麦感染的实验室模型来实现这一目标。这将显著增加当前小麦基因组测序投资的价值。然后，我们将利用这些资源来确定杀菌剂如何影响感染过程，从而绕过发展新控制措施的主要障碍。为了确保这些资源的广泛使用，我们将为这些质粒和菌株开发一个储存库，将它们放在国际储存库中，编写标准操作程序以最大限度地利用这些资源，并创建一个网站来宣传和允许访问这些资源。最后，我们将与西南地区（如BBSRC North Wyke的新综合农场平台）和全国（如洛桑研究所、塞恩斯伯里实验室和先正达）的利益相关者进行互动，以增加所产生工具的影响和传播。

Wheat is the principle food crop in Europe, including Britain where the annual value of wheat production is approximately £1.4 billion. With the expectation that planet's population will increase to over 9 billion by 2050, we urgently need to find ways to provide more food to support this growth. One of the major threats to wheat production is fungal disease. The pathogen Mycosphaerella graminicola is the major disease-causing agent of wheat in the UK and is one of the principle reasons why over £500 million of fungicides are applied to protect cereal crops in the UK alone. Despite this there are still major yield losses. Hence there is an urgent need for new durable disease control strategies, including novel fungicides. The main difficulty in developing novel fungicides is the inability to determine how new molecules work to kill the fungus. In the model fungus brewer's/baker's yeast, Saccharomyces cerevisiae, powerful tools have been generated, such as sets of yeast strains; lacking each fungal gene; that have each protein tagged with a fluorescent protein, or where each individual protein is expressed at a high level. These resources have allowed large-scale determination of how a wide range of chemicals function to alter the biology of yeast. We intend to construct a similar complete set of tools that will allow us to perform these assays, and hence determine how novel fungicides work, in the wheat pathogen M. graminicola. Specifically, we will place each of its ~10,000 genes into a mobile genetic element (plasmid) that will allow easy transfer into a large range of other plasmids (that we will also create) that can be used to inactivate, fluorescently tag and over-express each gene, both individually and as a pool. We will also construct a series of M. graminicola strains that will allow us to determine how the natural fungus invades wheat, and we will develop a laboratory model of wheat infection to achieve this. This will significantly add to the value of the current investment in sequencing the wheat genome. We will then use these resources to determine how fungicides impact on the infection process, and hence circumvent the principle impediment to the development of new control measures. To ensure that these resources are made widely available we will develop a repository for these plasmids and strains, place them in international depositories, write standard operating procedures to allow maximal exploitation and create a website to advertise and allow access to the resources. Finally, we will interact with stakeholders, both in the South-West (eg the new integrated farm platform at BBSRC North Wyke), and nationally (eg Rothamsted Research, Sainsbury Laboratory, and Syngenta) to increase the impact and dissemination of the tools generated.