An apoplastic chitinase CpCHT1 isolated from the corolla of wintersweet exhibits both antifreeze and antifungal activities

The shrub Chimonanthus praecox L. (wintersweet) which is native to Chinese montane forests produces its flowers in the midst of winter. This indicates that the floral organs of this species are adapted to growth and development under freezing temperatures. Here, we report the isolation and preliminary characterisation of a 33 kDa apoplastic antifreeze chitinase (CpCHT1) from the petals and its corresponding cDNA. The chitinase activity of CpCHT1 was confirmed by activity staining. Antifreeze activity was validated in terms of the formation of bipyramidal ice crystals and high thermal-hysteresis values. CpCHT1 was also found to affect the germination of fungal spores of four major plant pathogens. In addition, the gene and protein are expressed constitutively not only in flowers, but also in leaves, bark and root tissues. From these data we hypothesize that this protein is multifunctional and may protect wintersweet from freezing injury and provide nonspecific disease resistance. Additional key word: Chimonanthus praecox.