Selection and characterization of salt-tolerant calli of Taraxacum officinale / 生物工程学报

ABSTRACT

In order to obtain salt-tolerant calli of Dandelion (Taraxacum officinale Weber), calli were induced from leaf explants of Dandelion on Murashige and Skoog's medium supplemented with 2.0 mg/L 6-benzyladenine and 0.5 mg/L 2,4-dichlorophen oxyacetic acid With 1.5% NaCl as selection pressure, most calli became brown and dead, whereas some new cell clusters appeared at the edge of the brown calli after 2 to 3 weeks. The survived cells were picked out and sub-cultured every 3 weeks onto the fresh selection medium and salt-tolerant calli were finally obtained through 4 continuous selections on the selection medium supplemented with 1.5% NaCl. Salt-tolerant calli increased steadily under a fixed NaCl stress though their relative growth rate decreased with increased NaCl concentration whereas the control calli which were sub-cultured by 4 continus selections on salt free medium ceased to grow under the same condition. This result indicated that the salt-tolerance of the selected calli is improved and this character is stable. Compared with the control, the SDS-PAGE pattern of the salt-tolerant calli had a unique 34 kD protein band. Its 30 kD and 18 kD protein bands were up-regulated. Further more, within the NaCl stress range up to 1.5%, the activities of antioxidant enzymes such as super oxidase dimutase, peroxidase and catalase, and the proline contents of the salt-tolerant calli were higher than those of the control. The results indicated that the selected calli with improved and stable salt tolerance were cell variants. The accumulation of the organic compatible solutes including proteins and the enhanced antioxidant capabilities in the salt tolerant calli are the two ways for them to regulate their osmotic homeostasis and alleviate the secondary reactive oxygen spexies damage respectively.