Investigating fungal pathogen effector localisation within plant cells

为不断增长的世界人口确保全球粮食安全是一个主要问题。真菌病原体每年摧毁大量的粮食作物（约15%）。各种镰刀菌引起的小麦穗状疫病尤其严重（Dean et al., 2012）。这些病原体分泌效应蛋白，操纵宿主促进感染。一些效应器被预测进入宿主细胞器使用软件工具，搜索与亚细胞靶向相关的序列特征。然而，这些预测可能并不准确，因为只有少数真菌效应器的亚细胞定位得到了验证。需要可靠的预测来确定与效应宿主亚细胞定位差异相关的病原体群体的毒力变化。我们将使用多种软件工具对多种谷物镰刀菌菌株的分泌组进行计算机荟萃分析预测，并识别可能具有宿主亚细胞定位的效应物，然后在实验室中使用植物表达和共聚焦显微镜验证这些预测。经过验证的定位数据将用于提高预测工具的准确性。然后利用免疫共沉淀法或近距离标记法对某些效应物中的植物蛋白靶点进行鉴定。植物靶标鉴定将允许阐明效应函数。这一新知识将允许学生和顾问团队设计新的谷草镰孢菌控制策略。该项目是多学科的，具有很强的数学、计算生物学和成像成分。前六个月，学生将在洛桑研究所（Rothamsted Research, RRes）接受生物信息学、克隆技术、生物成像和特定教学课程的初步研究培训。随后，该学生将转到该大学进行进一步的研究训练，然后返回RRes进行剩余的博士项目。巴斯大学将参与并提供基因组方法和分析方面的培训。咨询团队包括Nutzmann博士（大学首席导师）和Brown博士（第二大学导师）以及RRes的Darino博士，smith和Urban博士和Hammond-Kosack教授（主要导师）。学生将有机会获得世界一流的研究设施，并将从他们的顾问团队接受优秀的跨学科培训。

Ensuring global food security for the ever-growing world's population is a major concern. Fungal pathogensdestroy a substantial amount of all food crops each year (~15%). Cereal head blight diseases of wheat caused byvarious Fusarium fungi are particularly serious (Dean et al., 2012). These pathogens secrete effectorproteins that manipulate the host to promote infection. Some effectors were predicted to enter host organelles using software tools that search for sequence signaturesassociated with subcellular targeting. However, these prediction might not be accurate as only for a few fungaleffectors has their subcellular localisation been validated. Robust predictions are required to identify virulencechanges in the pathogen population associated with differences in effector host subcellular localisation. We will use multiple software tools to perform an in-silico meta-analytic prediction of the secretome of multipleFusarium graminearum strains and identify effectors likely to have host subcellular localisation before validating thesepredictions in the laboratory using in planta expression and confocal microscopy. Validated localisation data will beused to improve the accuracy of prediction tools. Then the plant protein targets from some effectors will beidentified by co-immunoprecipitation or proximity labelling followed by mass spectrometry. Plant targetsidentification will allow the elucidation of effector function(s). This new knowledge will permit the student and theadvisory team to devise new F. graminearum control strategies. The project is multidisciplinary with strong mathematical, computational biology and imaging components. The studentwill spend the first six months at Rothamsted Research (RRes) undertaking initial research training inbioinformatics, cloning techniques, bioimaging and specific taught courses. Subsequently, the student will transfer to theUniversity for a further research training period, before returning to RRes for the remainder of the PhD project. TheUniversity of Bath will be involved and will provide training in genomic approaches and analyses. The advisory teamincludes Drs Nutzmann (lead University supervisor) and Brown (2nd University supervisor) and at RRes Drs Darino, Smithand Urban and Prof. Hammond-Kosack (main supervisor). The student will have access to world class research facilitiesand will receive outstanding interdisciplinary training from their advisory team.