Genome-wide association study for electrolyte leakage in rapeseed/canola (Brassica napus L.)

Freezing temperature/frosts can cause significant damage to plants by rupturing plant cells. Rapeseed/canola (Brassica napus L.) is susceptible to freezing temperature at early seedling stage. The degree of cell rupture or seedling damage can be evaluated through the measurement of electrolyte leakage. Here, we measured the electrolyte leakage of a diversity panel of B. napus germplasm accessions under simulated freezing conditions. Preliminary data for electrolyte leakage measurement indicated that cold acclimation of two-week-old seedlings for 7days at 4 degrees C followed by freezing treatment at -12 degrees C for 2h provided a reasonable diversity in response. With this protocol for electrolyte leakage, a genome-wide association study was conducted on 157 winter, semi-winter, and spring types of B. napus accessions that originated from 17 countries. A total of 37,454 single-nucleotide polymorphism (SNP) markers based upon genotyping-by-sequencing were used for the analysis. Ten QTL were identified as associated with electrolyte leakage of canola seedlings, which together explained 43% phenotypic variation. Five of the QTL were located on A-genome. We identified at least 33 orthologs of the functional candidate genes. Although no well-characterized cold regulatory genes were identified, there were some indications that genes involved in membrane structure, developmental processes, and extracellular transport may be involved in altering the electrolyte leakage following the short-term hard freeze and rapid defrosting suffered by the plants in our protocol.