Isolation of protoplast from callus of Populus euphratica and H^+ fluxes across plasma membrane under NaCl stress  

作　　者：Gao Zhun Dai Song-xiang Chen Shao-liang Shen Xin Wang Rui-gang 

机构地区：[1]College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China [2]Key Laboratory of Biological Resources Protection and Utilization in Hubei Province, Hubei Institute for Nationalities, Enshi 445000, P. R. China

出　　处：《Forestry Studies in China》2007年第3期198-202,共5页中国林学（英文版）

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）.

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.

关 键 词：Populus euphratica protoplast isolation cellulase pectolyase macerozyme HEMICELLULASE SORBITOL non-invasivelyion-selective microelectrode technique 

分 类 号：S-3[农业科学]