Project Number: 6028-21000-011-32-R
Project Type: Reimbursable Cooperative Agreement
Start Date: Dec 1, 2015
End Date: Nov 30, 2020
The objectives are to: (1) validate growth chamber determined cold tolerance traits in field experiments; (2) test various hypotheses regarding cold tolerance phenologies that might lead to gene discovery; and (3), validate and fine-map preliminarily mapped cold tolerance quantitative trait loci (QTL).
Our approach will be to: (1) validate those accessions identified as cold tolerant in the rice mini-core collection and Rice Diversity Panel 1 (RDP1) in a field environment; (2) test hypotheses for mechanisms of rice cold tolerance by examining the role of membrane damage (electrolyte leakage assays), antioxidant enzymes, and photoperiod/electrolyte leakage assays under cold stress; (3) fine-map strong-effect QTL using two bi-parental mapping populations derived from crosses between the most cold tolerant and least tolerant accessions in the mini-core; (4) identify putative candidate genes associated with cold tolerance QTL using publicly available online resources, including gene ontology (GO), the rice expression profile (RiceXPro), and mini-core single nucleotide polymorphism resequencing data, to search for candidate genes in genomic regions that logically associate with cold tolerance QTL to select putative candidate genes near QTL revealed in our studies, and (5) test selected candidate genes in transgenic plants using Agrobacterium-mediated transformation and CRISPR/Cas9 genome editing of rice.