Allele frequency and gene expression differences under key winter stresses in temporal populations of two timothy...

Timothy is the most important perennial forage grass species in northern Norway, a region that is predicted to experience variable winter weather conditions due to climate change. Knowledge about how timothy cultivars respond to a changing climate is crucial for safeguarding forage production at higher latitudes. In the current study, we investigated changes in gene expression under freezing and ice encasement stresses and SNP allele frequencies between temporal populations (seed generations) of the two northern-adapted timothy cultivars Engmo and Noreng. In general, there was a decrease in freezing tolerance (defined as LT₅₀, the temperature lethal to 50% of the population) and an increase in ice encasement tolerance (defined as LD₅₀, the duration lethal to 50% of the population) over time. Comparative transcriptome analyses identified several genes known to be involved in stress responses, such as ethylene-responsive transcription factors, dehydration-responsive element binding transcription factors, reversion to ethylene sensitivity 1, and abscisic acid repressor 1, as differentially expressed between the temporal populations of Noreng under freezing stress. Several loci with large allele frequency changes were observed to be in close proximity to the genes displaying patterns resembling shifts over time in Noreng. Very few gene expression differences between populations of both cultivars under ice encasement stress could be due to weak selection pressure during seed multiplication. There was a gradual decline in genetic diversity in populations of both cultivars over time. The results indicate that phytohormone-mediated transcriptional regulation might be one of the key mechanisms for adaptation to changing winter weather conditions at higher latitudes. These findings underscore the importance of monitoring genetic shifts during seed multiplication to maintain cultivar stability and suggest that the identified stress-responsive genes could serve as valuable targets for breeding climate-resilient forage crops.