[Effects of cold-hardening on freezing tolerance and antioxidant enzyme activities in plantlets of Saussurea laniceps Hand.-Mazz].

Living conditions for plants in the mountains become increasingly less favorable with increasing altitude. In the alpine region, the plants are commonly exposed to daily rather than seasonal temperature fluctuations and by frequent freezing temperature. To elucidate the freezing tolerance mechanism of alpine plants, Saussurea laniceps Hand.-Mazz. was used as a model plant. It is a perennial herbal plant distributed in alpine regions of Yunnan and Tibet of China. It can survive on mountains with elevations over 4000 m. Wild S. laniceps plants are propagated only by seeds in the alpine areas. Micropropagation of S. laniceps through seed was a desirable method to get enough seedlings for freezing research. Micropropagation through plantlets derived from germinated S. laniceps seeds collected from Tibet was achieved successfully. Activities of antioxidant enzyme and solute contents were investigated in plantlets of S. laniceps. Freezing tolerance in plantlets increased after 7 or 15 d of cold-hardening (Table 1). Cold-hardening (2 degrees C) increased the activities of SOD, peroxidase, and catalase (Fig.1) in plantlets. A similar increase was also observed in the protein and proline content (Fig.1), whereas soluble carbohydrates changed little (Fig.1). These results obtained suggest that the higher activities of SOD, peroxidase, and catalase, as well as the higher protein and proline content may be biochemical adaptation for freezing toleranc in cold-hardened S. laniceps plantlets. Interestingly, deacclimation was slow; even after the plants were placed again under a temperature of 21-23 degrees C for 5 d, the higher freezing hardiness, enzyme activities, protein and proline content acquired after cold acclimation remained. In conclusion, our plantlet cultures have proved to be good materials for experimentation on freezing resistance in study of freezing-resistance mechanism in the alpine plant S. laniceps.