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Proposal 2 Identification and Functional Analysis of Small RNAs in Systemic Acquired Resistance in Arabidopsis thaliana Summary This is an important topic to study because systemic acquired resistance (SAR) can lead to faster and strong defense responses in systemic tissue, to a broader spectrum of pathogens. The gaps in knowledge being addressed in this proposal are the unknown signal(s) that initiates systemic acquired resistance (SAR) and molecular mechanism underlying SAR. There appears to be a connection between sRNAs and pathogen resistance response because sRNAs have been identified in playing a role in PAMP-triggered immunity and effector-triggered immunity. There are also four major families of proteins involved in sRNA biogenesis and function in Arabidopsis. Based on these connections established by the authors, they hypothesize that sRNAs and sRNA pathway components function in SAR. The first objective is to perform a genetic evaluation of the function of the four major families involved in sRNA biogenesis and function (DCL1-4, RDR 1-6, AGO1-10, polIV, and polV). Their plan is to use loss-of-function mutants in a bacterial infection assay and gene expression study. The second objective is to perform high-throughput sequencing of the sRNAs involved in SAR. Their plan is to produce HA-tagged AGO overexpression plants and perform CoImmunoprecipitation followed by sRNA sequencing. The third objective is to identify and characterize the sRNAs and their target genes. Their plan is to collaborate with Xiu-Jie Wang’s lab from the State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences for the identification of the sRNAs and their target genes. Then to determine the functions of the sRNA and target genes in SAR they plan to use a combination of qt-PCR, western blotting, and loss-offunction mutants. Strengths/Weaknesses Strengths o The introduction and literature supporting the proposal was well researched, strengthening their hypothesis and basis of the proposal. o Experiments are detailed showing excellent preparation and the ability to begin the experiments in a timely manner. o They clearly outline expected results and identify problems they might come across demonstrating familiarity with the techniques and procedures. Weaknesses o The future directions section does not seem to be well related to the proposal; the projects in the future directions do not seem to stem from the results of this proposal. Broader Impacts Identifying new components in SAR and having a better understanding of the mechanism will be integrated into crop plants as an additional layer of protection against pathogens as well as generating new cultivars of disease-resistant crop plants.