Suppression of wheat defences by Fusarium graminearum

将进行两个实验，向学生介绍烟草和小麦的病理系统、两个禾谷镰刀菌菌株、BSMV-Vox载体以及通过Western blotting检测病毒和使用荧光定量MUG、组织培养和/或光学显微镜检测真菌。实验1.烟草叶片中FG效应功能的分析。采用完全随机和重复的实验设计，将两个已有的BSMV：FG效应体和两个对照构建体(BSMV：MCS和BSMV：Ilov)一起接种到烟草上，以探索FG效应子的过度表达如何影响这种非寄主物种的镰刀菌感染。试验2.Fg-Gus和Fg-tri5突变体对小麦穗部侵染的分析。采用重复配对试验设计，以小麦品种为材料。用对照BSMV：Ilov载体或模拟接种BSMV：Ilov，7天后，用FG-GUS株系或FG-tri5突变体接种整个小麦穗。将每天对FG症状的发展进行目测评估，并在第7天对小麦小穗进行解剖，以探索和相互比较每个FG菌株在有无病毒的情况下的定殖程度。

Two experiments will be done to introduce the student to the tobacco and wheat pathosystems, the two F. graminearum strains, the BSMV-VOX vector as well as virus detection by Western blotting and fungus detection using a fluorescent quantitative MUG assay, tissue plating and/or light microscopy.Experiment 1. Analysis of Fg effector function in tobacco leaves. Using a fully randomised and replicated experimental design two already available BSMV:Fg effector constructs will be inoculated onto tobacco plants along with two control constructs (BSMV:MCS and BSMV:iLov) to explore how the over-expression of Fg effectors affects Fusarium infection of this non-host species. Experiment 2. Analysis of wheat spike infection by the Fg GUS and Fg tri5 mutant. Using a replicated pair wise experimental design, the wheat cv. Bobwhite will either be inoculated with the control BSMV:iLov construct or mock inoculated and then 7 days later the entire wheat spike will be inoculated with either the Fg GUS strain or the Fg tri5 mutant. Fg symptom development will be visually assessed on a daily basis and the at day 7 the wheat spikelets will be dissected to explore and inter-compare the extent of colonisation of each Fg strain with and without the virus.