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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Sunflower and Plant Biology Research » Research » Research Project #441342

Research Project: Investigating Mutations in Arabidopsis That Increase Deacclimation Resistance

Location: Sunflower and Plant Biology Research

Project Number: 3060-21220-033-019-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Jan 15, 2022
End Date: Jan 14, 2023

The two objectives of this agreement proposal are as follows: 1) Characterization of arabidopsis lines carrying mutations in candidate genes identified in a GWAS study on deacclimation in canola and 2) Phenotypic evaluation of F3 RIL families from crosses between freezing tolerant and freezing susceptible lines of canola.

1. Perform experiments designed to analyze the phenotypes of Arabidopsis lines carrying mutations in VIP3 and PSK4 genes and statistically analyze the results and write the manuscript reporting the results. Mutant lines will be subjected to acclimation, deacclimation, and no acclimation followed by freezing at -14C for 4 hours and subsequent recovery and scoring for percent damage according to established protocols. Nine replicate pots of each line for each of the three treatments will be tested for freezing tolerance in a random complete block design for each experimental run. Two such runs have been completed. A third is planned. Additionally, initial phenotypic analysis as described above but with fewer (only three) technical replicates per line will be done for 20 additional mutant lines in other candidate genes. 2. We have produced seeds from crosses between a highly freezing tolerant and slow deacclimating canola line and a line with very poor freezing tolerance and rapid deacclimation. F2 seeds have been planted and F3 families are being generated to begin mapping the genes controlling freezing tolerance in canola. Once F3 seeds are available, twelve seeds from each F3 family will be grown to the 6-leaf stage and then acclimated at 5C for 6 weeks. Acclimated plants will be subjected to freezing at -13C for 4 hours according to established protocols and then scored for freezing damage. The resulting segregation of phenotypes will be used to predict the inheritance of freezing tolerance.