摘要
We used callus of Populus euphratica Olive to isolate protoplasts, and IT fluxes across plasma membrane were investigated. The concentration of enzymes for protoplast isolation, e.g. cellulase, pectolyase, macerozyme, hemicellulase, and sorbitol content, incubation time were systemically studied. High yield and viability of protoplast was achieved after 6-8 hours incubation of P. euphratica callus in enzyme solution containing 1.5% （w：v） cellulase R-10, 0.1% （w：v） pectolyase Y-23, 0.2% （w：v） macerozyme R-10, 0.05% （w：v） hemicellulase and 0.75M）.80 mol·L^-1 sorbitol. Non-invasively ion selective microelectrode technique was used to access proton fluxes in the absence and presence of NaCl （20 mmol.L-1）. Salt-induced transient net IT effiux was observed in the plasma membrane ofP. euphratica cells. The shift of IT flux response to NaC1 shock and the relevance to salt tolerance were discussed.
We used callus of Populus euphratica Olive to isolate protoplasts, and IT fluxes across plasma membrane were investigated. The concentration of enzymes for protoplast isolation, e.g. cellulase, pectolyase, macerozyme, hemicellulase, and sorbitol content, incubation time were systemically studied. High yield and viability of protoplast was achieved after 6-8 hours incubation of P. euphratica callus in enzyme solution containing 1.5% （w：v） cellulase R-10, 0.1% （w：v） pectolyase Y-23, 0.2% （w：v） macerozyme R-10, 0.05% （w：v） hemicellulase and 0.75M）.80 mol·L^-1 sorbitol. Non-invasively ion selective microelectrode technique was used to access proton fluxes in the absence and presence of NaCl （20 mmol.L-1）. Salt-induced transient net IT effiux was observed in the plasma membrane ofP. euphratica cells. The shift of IT flux response to NaC1 shock and the relevance to salt tolerance were discussed.
基金
the key project of National Natural Science Foundation of China （30430430）
the HI-TECH Research and Development Program of China （863 Program, 2006AA10Z131）
a Foundation for the Author of National Excellent Doctoral Dissertation of PR China （200152）
the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education Institution of MOE, PRC （2002-323）.
