Molecular Analysis of Freeze-Tolerance Enhanced by Treatment of Trinexapac-Ethyl in Kentucky Bluegrass

Trinexapac-ethyl[ 4-(cyclopropyl- $\alpha$ -hydroxy-methylene)-3,5-dioxocyclohexane carboxylic acid ethylester] is a growth-retardant for plants by inhibiting a key step in biosynthesis of GA. A treatment of trinexapacethyl generally induces a reduction in vegetative growth and also inhibits heading. In addition, the trinexapacethyl was known to enhance the freeze-tolerance in annual bluegrass, however, the mechanism is not known yet. One possible reason for the enhanced freeze-tolerance may be the antifreeze protein known to be accumulated in intercellular space of the leaf during cold acclimation. In order to see the possible in-duction of the synthesis of antifreeze proteins by trinexacpacethyl, the apoplastic proteins extracted from Kentucky bluegrass treated with trinexapacethyl were analyzed by SDS-PAGE and the presence of the antifreeze protein was observed. In addition, western analysis showed the identity of the protein induced by both a cold acclimation and a trinexapacethyl treatment. It appears that an enhanced freeze-tolerance of the turf grass by trinexapacethyl is due to the synthesis and/or accumulation of the antifreeze protein similar to the enhanced freeze tolerance induced by cold acclimation.