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《Northern Horticulture》 2009-10
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The Proplast Isolation and Purification of Orange-heading Chinese Cabbage

WU Yun-xia1,ZHANG Lu-gang1,ZHANG Hua-ming1,FU Wen-ting1,XU Yu-jian1,GUAN Qing-hua2(1.College of Horticulture,Northwest Agricultural and Forestry University,Yangling,Shanxi 712100,China;2.Park River system of Taiyuan,Taiyuan,Shanxi 030002,China)  
Protoplasts of orange-heading chinese cabbage were isolated from cotyledons and hypocotyls.The different factors affecting the protoplasts yield and different methods of purification were investigated.The best enzyme solution of cotyledons contained CPW salts,DPD medium,0.8% cellulase R-10,0.1% macerozyme R-10,0.5 mol/L mannitol incubated at 27℃ for 2 hours.The protoplasts yield was 3.52 ×106/(g FW) and the viability was 60.3% under the condition.The best enzyme solution of hypocotyls contained CPW salts,DPD medium,1.5% cellulase R-10,0.3% macerozyme R-10,0.5 mol/L mannitol incubated at 27℃ for 18 hours.The protoplasts yield was 2.20×106/(g FW) and the viability was 63.7% under the condition.Moreover,the best protoplasts purification method was rising method with sucrose isodensity centrifugation at concentration of 30%,with the.centrifugal speed of 800 rpm,and the centrifugal time of 5 min.
【Fund】: 农业部公益性行业科研专项资金资助项目(nyhyzx07-007);; 陕西省13115工程资助项目(2007ZDKG-05)
【CateGory Index】: S634.1
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