2010-3: Characterizing the Mimicry of Plant Peptides by Effectors from the Soybean Cyst Nematode, Heterodera glycines Score: Very Good Briefly summarize objectives and approaches 1. Retesting of HgPECs parasitism-specific activation and effector-like expression a. 29 HgPECs found to be differentially expressed during parasitic stages from whole-nematode microarray b. In-situ hybridisations of HgPECs on preparasitic and parasitic stages of H. glycines c. Candidates will be prioritised depending on: i. Presence of secretion signal peptide ii. Absence of transmembrane domain iii. Plant homologs with functions that could contribute to host cell reprogramming d. Looking for: i. Transcription confirmed to be upregulated at the onset of parasitism ii. Transcripts localised primarily in esophageal gland cells e. Localisation will be comfirmed by HgPEC:GFP fusion transiently expressed in onion epidermal cells. 2. Testing HgPECs contribution to parasitism a. Effect of stably-expressed HgPECs on Arabidopsis morphology and susceptibility to H. schachtii b. SiRNA lines will also be tested for susceptibility 3. Test whether HgPECs functionally mimic host proteins a. Stably-expressed HgPECs will be crossed to HgPEC-homolog mutant lines b. Progeny observed for rescue of mutant phenotype 4. Discovering HgPEC-interacting proteins a. Y2H and Y1H activator assays with HgPEC as bait and cDNA library as preys b. Results verified by immunoprecipitation Analyse strengths of the scientific merit of the proposal Logical progression of the scientific methods used. Firm foundations of prior research to base proposal on Analyse weaknesses of the scientific merit of the proposal Objective 3. The stably expressing HgPEC lines will have functional alleles of HgPEC homologs, therefore there will always be rescue of the mutant phenotype. o Would need to backcross to mutant parent perhaps multiple times and then use markers to identify F2 expressing the HgPEC but homozygous for mutant HgPEC-homologue. Objective 4: Instead of using a cDNA library, would it not be more efficient to identify interactors of the HgPEC homologues and use these instead? Analyse strengths and weaknesses of broader impacts Work could indeed be the basis for resistance against H. glycines in the future, though the broadness or durability is questionable until results from objective 3 are obtained. The two or three undergraduates do not seem to be included in the finding justification, but do seem to be in the budget. Research is to be presented at appropriate meetings, however no money has been allocated for travel. Provide a couple summary sentences about your overall thoughts on the proposal, why it was particularly strong, or if you didn’t score an excellent, a main reason or two that kept you from scoring it higher. A very good proposal. The background seems solid and there is a logical progression through the objectives to reach the end goal and question the hypothesis. Only major comment would be that I am unsure whether the amount of experiments that they wish to perform are feasible within their budget. Minor details are that perhaps objective 3 hasn’t been thought out fully and objective 4 is not quite optimised.