Skip to main content
ARS Home » Southeast Area » Stuttgart, Arkansas » Dale Bumpers National Rice Research Center » Research » Research Project #437664

Research Project: Identification of miRNA’s Involvement with the Presence of Blast Resistance Gene Ptr Under Water Stress Conditions

Location: Dale Bumpers National Rice Research Center

Project Number: 6028-21000-011-33-I
Project Type: Interagency Reimbursable Agreement

Start Date: Jan 6, 2020
End Date: Apr 16, 2020

Objective:
The fellow’s current research goal is to identify microRNA that are involved in regulating stress-responsive networks at the molecular level and to develop stress resilient crops.

Approach:
The project is expected to be accomplished in 12 weeks within the following specific time intervals to identify microRNA’s involvement with the presence of blast resistance gene Ptr under water stress conditions. Week 1: Laboratory orientation and safety training, data analysis software JMP genomics training, order chemicals including one step Reverse Transcriptase - Polymerase Chain Reaction and Qiagen kits for DNA and RNA extraction, and software Vector NTI ($1500) for DNA and RNA analysis, and WD Element (budgeted) for data storage. Week 2: Sow rice seeds in the greenhouse and grow fungal pathogens for RNA and DNA extractions. Week 3-4: Visit Riceland Foods and Producers Rice Mill, Stuttgart, Arkansas, University of Arkansas- Fayetteville, Little Rock, and Pine Bluff, and Arkansas State University, Jonesboro to gain understanding of rice industry and research in the USA. Week 5-6: Treat rice plants growing in greenhouse with blast fungal spores grown in fungal culture room (Week 2) and water stress (drought) and collect leaf tissue at different time points of disease and water stress with non-treated control plants. Send harvested tissue/DNA to service provider Novogene for genome, microRNA and degradome sequencing analysis and attend World Food Prize meeting, Des Moines, Iowa: https://www.worldfoodprize.org/, Event dates: 10/16-18/2019. Week 7: Analyze data received from Novogene. Design and order primers and kit for one step Reverse Transcriptase-Polymerase Chain Reaction method and get ready for validation of microRNA and their predicted targets. Week 8-11: Perform real time Polymerase Chain Reaction (PCR) with one step Reverse Transcriptase-Polymerase Chain Reaction method and verify expression patterns of 5 to 10 microRNA and their predicted target genes using blast- and water stress- treated samples collected in week 5. Perform genome analysis of Single Nucleotide Polymorphism (SNP) and Indel of Ptr containing rice plant Katy and its near isogenic line M2354 using JMP Genomics and open source bioinformatic software. Week 12: Make an exit presentation at DBNRRC sharing results of fellowship, prepare manuscript, project review and develop a follow-up plan.