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Isolation of protoplast from callus of Populus euphratica and H fluxes across plasma membrane under NaCl stress
作者姓名:Gao  Zhun  Dai  Song-xiang  Chen  Shao-liang  Shen  Xin  Wang  Rui-gang
作者单位:Gao Zhun1 Dai Song-xiang1 Chen Shao-liang1,2? Shen Xin1 Wang Rui-gang1 1College of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing 100083,P. R. China 2Key Laboratory of Biological Resources Protection and Utilization in Hubei Province,Hubei Institute for Nationalities,Enshi 445000,P. R. China
基金项目:国家自然科学基金;国家高技术研究发展计划(863计划);全国高等学校优秀博士学位论文作者专项基金;教育部高等学校优秀青年教师教学科研奖励计划
摘    要:We used callus of Populus euphratica Olive to isolate protoplasts, and H fluxes across plasma membrane were investi-gated. 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 0.75-0.80 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 H efflux was observed in the plasma membrane of P. euphratica cells. The shift of H flux response to NaCl shock and the relevance to salt tolerance were discussed.

关 键 词:原生质体离析  纤维素酶  半纤维素酶  山梨醇  微电极技术  质膜  橄榄树
收稿时间:6 April 2007
修稿时间:2007-04-06

Isolation of protoplast from callus of Populus euphratica and H+ fluxes across plasma membrane under NaCl stress
Gao Zhun Dai Song-xiang Chen Shao-liang Shen Xin Wang Rui-gang.Isolation of protoplast from callus of Populus euphratica and H fluxes across plasma membrane under NaCl stress[J].Forestry Studies in China,2007,9(3):198-202.
Authors:Zhun Gao  Song-xiang Dai  Shao-liang Chen  Xin Shen  Rui-gang Wang
Institution:1. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China
2. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China;Key Laboratory of Biological Resources Protection and Utilization in Hubei Province, Hubei Institute for Nationalities, Enshi 445000, P. R. China
Abstract:We used callus of Populus euphratica Olive to isolate protoplasts, and H+ 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 0.75–0.80 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 H+ efflux was observed in the plasma membrane of P. euphratica cells. The shift of H+ flux response to NaCl shock and the relevance to salt tolerance were discussed. Gao Zhun and Dai Song-xiang contributed equally to this work.
Keywords:Populus euphratica  protoplast isolation  cellulase  pectolyase  macerozyme  hemicellulase  sorbitol  non-invasively ion-selective microelectrode technique
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