Overexpression of BrAFP1 gene from winter rapeseed (Brassica rapa) confers cold tolerance in Arabidopsis

Antifreeze proteins (AFPs) can bind to ice crystals and restrain the formation of larger crystals, a strategy vital to the survival of plants in freezing environments. The BrAFP1 from winter rapeseed cultivars 'Longyou 7' with high cold tolerance was cloned and overexpressed in Arabidopsis. BrAFP1 was localized in the cytoplasm and nucleus. Under cold stress, SOD activity and free proline content were higher, MDA content and relative conductivity were lower in transgenic lines than those in wide-type Arabidopsis. Frostbite of transgenic plants was minimized, whereas frostbite of the Arabidopsis afp1 mutant was severe. Transition of the amino acid at position 17 of BrAFP1 was related to the increased winter survival of the rapeseed cultivar. Cultivars with higher survival rates had a predilection for tyrosine, not tryptophan, at the 17th site in the amino sequence of BrAFP1. Transcription of BrAFP1 was induced more rapidly, and the expression of the gene was also higher, in Longyou 7 than that in Tianyou 4 under cold stress. Overall, the high expression of BrAPF1 confers more cold-tolerance in Longyou 7.

Keywords: Antifreeze protein; Brassica rapa; Cold tolerance; Winter rapeseed.