Progress 12/01/22 to 11/30/23
Outputs
Target Audience:Some of the generated data were presented at the APS annual meeting with Plant Pathologists as the major target audiences. We have also presented/shared our generated data with researchers (Plant breeders and Genetisists) and industry partners at WGRC-IUCRC montly meetings. Changes/Problems:No major changes What opportunities for training and professional development has the project provided?The project has provided training opportunities for a postodc, a grdaute student and undergrads. A graduate student was recently recruited to conduct the experiments related to objective 3. Addiotionally,Nouri lab at KSU was the host of an undergraduate student for the Research and Extension Experience for Undergraduates (REEU) program for 8 weeks in summer 2023. The undergraduate was trained and performed some of the experiments related to objective 1 under the supervision of the PI Nouri and the assigned Postdoc. How have the results been disseminated to communities of interest?Presenting at meetings such as APS as well ascollaborative meetings between the Wheat Genomic Research Center scientists and wheat industry partners (WGRC-IUCRC). What do you plan to do during the next reporting period to accomplish the goals?Aim 1. Identify novel sources of genetic resistance and tolerance to WSMV inAe. taushii. We have completed this aim. Aim 2. Characterize genetic basis of resistance and tolerance genomic in a diverse panel ofAe. tauschiiaccessions through genome-wide association studies (GWAS). Curate final collection of complete accessions (n=250) with whole-genome sequencing data, generated k-mer matrix, and variant (SNP) genotyping. GWAS using both SNP variants and k-mers from preliminary completed set of WSMV screening (n~150 accessions) Evaluate population structure and geographic origin of accessions in relationship to WSMV resistance/tolerance profile. GWAS using both SNP variants and k-mers from preliminary completed set of mixed infection screening. Aim 3. Detailed evaluations of resistance and tolerance in selectedAe. tauschiito WSMV in single and mixed infections with TriMV. Our plan is to screen selectedAe. tauschiiaccessions from Aim 1 showed potential tolerance to WSMV single infections for the mixed-infections of WSMV+TriMV. Our criteria for the evaluation will be the same as Aim 1, symptom severity and the titers of both viruses in infected plants.
Impacts
What was accomplished under these goals?
Specific Aim 1:Identify novel sources of genetic resistance and tolerance to WSMV inAe. taushii. We screened extra 90 accessions ofAe. tauschiifor WSMV single infections under the controlled conditions to complete screening of the total of 250 accessions which was the goal of this aim. The seeds ofAe. tauschiigenotypes received from the Wheat Genomic Research Center (WGRC) at KSU. Our methods to perform experiments were explained in details in our last year report. Based on both symptom severity and the virus titers measured by the RT-quantitative PCR (RT-qPCR), we identified 52 accessions ofAe. tauschiias potential tolerant to WSMV single infections. We had previously identified 13 tolerant genotypes. Therefore, in total 65 accessions were determined as potential tolerant. We did not identify any complete resistant genotypes among our screenedAe. tauschiigenotypes. Specific Aim 2: Characterize genetic basis of resistance and tolerance genomic in a diverse panel ofAe. tauschiiaccessions through genome-wide association studies (GWAS). The phenotypic data including symptom severity scores and WSMV titers from all screenedAe. tauschiihave been shared with the Co-PI Poland`s research group at KAUST for conducting GWAS. Specific Aim 3: Detailed evaluations of resistance and tolerance in selectedAe. tauschiito WSMV in single and mixed infections with TriMV. "Tomahawk" wheat plants have been inoculated with both WSMV and TriMV to provide the source of mixed-infections. The standard curve has been created for TriMV which will be used to calculate the copy number of the genome.
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Progress 12/01/21 to 11/30/22
Outputs
Target Audience:The goal of this research project is to screen the genotypes of a wheat wild relative for identifying new sources of virus resistance/tolerance. The research communities in Plant Breeding, Plant Virology, and Plant Genetics were reached asthe target audience of this research. The wheat industry also benefited from the results of this research. Changes/Problems:None What opportunities for training and professional development has the project provided?The project has provided research opportunity for a Postdoc in PI Nouri`s lab.A second postdoc at KAUST is now working on the project for genomics and association mapping.The postdoc will gain further train in these areas. How have the results been disseminated to communities of interest?Some results were disseminated through presentations at meetings such as a collaborative meeting between the Wheat Genomic Research Center scientists and wheat industry partners. What do you plan to do during the next reporting period to accomplish the goals?Aim 1. Identify novel sources of genetic resistance and tolerance to WSMV inAe. taushii. We will continue screening moreAe. tauschiiaccessions for WSMV single infections based on both symptoms and virus titer measured by the real-time PCR. We will update our database constantly. We will also share our results with the Co-PI Poland at KAUST. The PI Nouri lab has advertised the current research to recruit an REEU student for summer 2023. Nouri`s lab will also recruit a graduate student for this project in fall 2023. Aim 2. Characterize genetic basis of resistance and tolerance genomic in a diverse panel ofAe. tauschiiaccessions through genome-wide association studies (GWAS). We have organized the SNP variants and k-mers for association mapping and will complete preliminary analysis confiming the overall analysis pipeline.With the completed datasets for symptom severity and virus copy counts, we will run the association mapping for the complete panel. Aim 3. Detailed evaluations of resistance and tolerance in selectedAe. tauschiito WSMV in single and mixed infections with TriMV. Our plan is to complete the Aim 1 first. Then, promising accessions will be selected to get screened for double infections of WSMV and TriMV.
Impacts
What was accomplished under these goals?
Specific Aim 1:Identify novel sources of genetic resistance and tolerance to WSMV inAe. taushii. We have screened 70 accessions ofAe. tauschiiso far for WSMV infection under controlled conditions. The seeds have been received from the Wheat Genomic Research Center (WGRC) at KSU. For infecting wild wheat varieties, 100mg of WSMV infected Tomahawk (susceptible variety), which was stored at -80°C, was crushed in 1.5 ml of 0.2 M phosphate buffer pH 7 in ice cold and sterile mortar-pestle. Carborundum was added in to this sap followed by mechanical inoculation with cotton bud on two leaves (three leaf stage). For control, mixture of phosphate buffer-carborundum was used. Symptom of infection was monitored till 31 days post inoculation (dpi) and scored 1-4, where 1 was given for only yellowing, 2 for mild mosaic/streak; 3, sever mosaic/streak; and 4, stunting with sever mosaic/streak symptoms. To know the exact copy number of viral RNA in mechanically infected plants, absolute quantification of WSMV coat protein RNA was conducted from leaf samples collected at 14, 21 and 31 dpi. For that, Total RNA was isolated and converted in to cDNA followed by absolute quantification with coat protein primers using the real-time PCR machine. In our earlier preliminary screens, we found that line TA2431 is a susceptible accession based on symptom and high titer.Therefore, we used this line as positive control and compared symptoms and titer of WSMV tested genotypes with symptoms and titer of WSMV of TA2431. Ae. tauschiiaccessions which showed mild symptoms (scores 1 and 2) either with high or low titer of the virus have been identified as "Tolerant"; while the accessions that showed mild symptoms (scores 1 and 2) with high virus titer have been selected as "True Tolerant" accessions. We also consider "delayed symptoms" as the tolerance response. Based on this criteria, we have identified15Ae. tauschiigenotypesso far as "Tolerant" and "True Tolerant". Specific Aim 2: Characterize genetic basis of resistance and tolerance genomic in a diverse panel ofAe. tauschiiaccessions through genome-wide association studies (GWAS). The phenotypic data including symptom severity scores and WSMV titers from 70 screenedAe. tauschiihave been shared with the Co-PI, Dr. Jesse Poland.We have organized phenotypes for association mapping. The genotyes for association mapping have been organized for theAe. tauschiipanel.This includes SNP variants and k-mer matrix.We are initializing association mapping for both symptom severity and virus copy number. Specific Aim 3: Detailed evaluations of resistance and tolerance in selectedAe. tauschiito WSMV in single and mixed infections with TriMV. Experiments related to this Aim will be conducted after completing the first Aim.
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