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《Acta Agrestia Sinica》 2010-01
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Protoplast Culture and Plant Regeneration of Midnight Ⅱ(Poa Pratensis L.)

MA Hui-ling,ZHAO Xiao-qiang,BAI Xiao-ming(Key laboratory of Grassland Ecosystem of Ministry of Education & Sino-US Centers for Grazingland Ecosystem Sustainability,Pratacultural College,Gansu Agricultural University,Lanzhou,Gansu Province 730070,China)  
The friable calli induced from mature seeds of Poa pratensis L.cv.Midnight II were used for protoplast preparation and the protoplasts were isolated using enzyme digestion.Calli appeared after sustained protoplast divisions in culture medium and the green shoot and albino plantlets were obtained from the protocalli on differentiated medium.The effects of different enzyme composition and enzyme digestion time on protoplast isolation were discussed in this paper.The results show that it was available to harvest high yield and viabilities of protoplasts from embryogenic calli with treatment of digestion in 1.0% Cellulase Onozuka R-10,1.0% Macerozyme R-10,0.3% Driselase,and 0.5% Pectolase Y-23 in 8 to 10 d.It was better to digest for 14-16 h in enzyme digestion solution mentioned above.Calli formed on KM8P medium supplemented with 1.0 mg/L 2,4-D,0.3 mg/L 6-BA,100 mg/L casein hydrolysate,100 mg/L lactablumin hydrolysate,1.0%(W/V) sucrose,and 0.4 mol/L mannitol at plating density of 2.0×105-5.0×105 mL-1.The protoplast-derived calli developed shoots and roots on differentiation media(MS+0.5 mg/L NAA+2.0 mg/L 6-BA) after further subculture,and then complete plants were obtained.
【Fund】: 甘肃农业大学草业学院草业科学国家级重点学科学术骨干科研项目暨草业生态系统教育部省部共建重点实验室资助项目(CY-GG-2006-10);; 甘肃省教育厅项目(0702-03)
【CateGory Index】: S688.4
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