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Zhang Si,Liu Cui,Jin Yuemei,Chi Shan,Tang Xianming,Chen Fuxiao,Fang Xu,Liu Tao. 2014. Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii. Acta Oceanologica Sinica, 33(10):114-123
Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii
长心卡帕藻原生质体分离、培养研究
Received:April 16, 2013  Revised:November 12, 2013
DOI:10.1007/s13131-014-0546-y
Key words:Kappaphycus alvarezii  protoplast  regenerated plantlet  callus-like  young shoots
中文关键词:  长心卡帕藻  原生质体  再生植株  类愈伤组织  再生新枝
基金项目:The National Science Foundation Project under contract No. 2007FY210500; the National Department Public Benefit Research Foundation of China under contract No. 200805075; the Province Science and Technology in the Guangdong Project under contract Nos 2010B060200010 and 2010B020201015; the Science Expenditure in the Hainan Project under contract No. 11-20410-0015; the National Natural Science Foundation of China under contract Nos 41206106 and 41222038.
Author NameAffiliationE-mail
Zhang Si National Marine Environmental Forecasting Center, Beijing 100081, China
College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China 
 
Liu Cui College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China  
Jin Yuemei College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China  
Chi Shan College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China  
Tang Xianming Hainan Provincial Fisheries Research Institute, Haikou 570204, China  
Chen Fuxiao Hainan Provincial Fisheries Research Institute, Haikou 570204, China  
Fang Xu School of Life Science, Shandong University, Jinan 250100, China  
Liu Tao College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China liutao@ouc.edu.cn 
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Abstract:
      In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30℃ continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL-1, 65.20×104 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20℃, light intensity of 1 500-2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated: (1) callus-like cell mass and regenerated plantlet occurred on protoplast; (2) young shoots and calluslike cell mass occurred in tissue blocks after enzymolysis.
中文摘要:
      以长心卡帕藻为材料,利用海螺酶和鲍酶两种海藻工具酶与纤维素酶成功制备其原生质体。确定了最适酶比例为20%鲍酶与12%纤维素酶,渗透剂为2.0 mol/L葡萄糖,最适酶解条件为30℃黑暗酶解4.0 h,原生质体密度为32.60×104 个/mL,产量为65.20×104 个/g。20℃,光强1500-2000 lx,光周期12 h/d培养,原生质体出现类愈伤组织细胞团和再生植株两种分化途径,酶解后组织块培养出现了再生新枝和类愈伤组织细胞团两种分化途径。
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