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| 咖啡叶片原生质体分离条件的研究 | |
| 引用本文： | 范睿,闫林,谭乐和,黄丽芳,董云萍,郝朝运. 咖啡叶片原生质体分离条件的研究[J]. 热带作物学报, 2012, 33(5): 860-865 |
| 作者姓名： | 范睿 闫林 谭乐和 黄丽芳 董云萍 郝朝运 |
| 作者单位： | 1. 中国热带农业科学院香料饮料研究所,海南万宁571533;海南省热带香辛饮料作物遗传改良与品质调控重点实验室,海南万宁571533 2. 中国热带农业科学院香料饮料研究所,海南万宁571533;农业部香辛饮料作物遗传资源利用重点实验室,海南万宁 571533 3. 中国热带农业科学院香料饮料研究所,海南万宁571533;海南省热带香辛饮料作物遗传改良与品质调控重点实验室,海南万宁571533;农业部香辛饮料作物遗传资源利用重点实验室,海南万宁 571533 |
| 基金项目： | 公益性行业（农业）科研专项经费（No. 200903024）；农业部热带作物种质资源保护项目（No. 12RZZY-20）；中国热带农业科学院香料饮料研究所科技基金项目（No. XYS1102）。 |
| 摘 要： | 以中粒种咖啡（Coffea. canephora）叶片组织为材料，研究酶液组合、酶解时间、酶液渗透压及预处理措施对其原生质体分离效果的影响，以期确定能够酶解出高数量且高活力的原生质体的酶解条件。结果表明：获得有活力原生质体的最佳酶液组合为2％纤维素酶+0.5％果胶酶+0.3％离析酶，同时酶解时间为20 h、酶液渗透压即甘露醇浓度为0.7 mol/L，预处理措施为黑暗24 h的咖啡叶片组织得到8.1×105个/g、活力达到73％的原生质体。 |
| 关 键 词： | 咖啡；酶解条件；原生质体 |
Optimization Conditions of Coffea Mesophyll Protoplast Isolation |
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| FAN Rui,YAN Lin,TAN Lehe,HUANG Lifang,DONG Yunping and HAO Chaoyun. Optimization Conditions of Coffea Mesophyll Protoplast Isolation[J]. Chinese Journal of Tropical Crops, 2012, 33(5): 860-865 | |
| Authors: | FAN Rui YAN Lin TAN Lehe HUANG Lifang DONG Yunping HAO Chaoyun |
| Affiliation: | 1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences2 Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops;1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences 3 Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture;1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences,2 Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops3 Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture;1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences,2 Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops;1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences 3 Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture;1 Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences 3 Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture |
| Abstract: | Hydrolysis parameters of protoplast with high yield and activity were determined for C. canephora. The effects of enzyme combinations, mannitol concentration in the mixed enzyme solution, enzymolysis time and pretreatment time on protoplast isolations were studied. The results are as follow: Leaves grown for 10~15 days were cut and then treated with enzymes consisting of 2% cellulase, 0.5% pectinase, 0.3% macerozyme and 0.7 mol/L mannitol and are sustainied for 20 hours after 24-hour dark pretreatment, its protoplast yield and activity was the highest. |
| Keywords: | Coffee Enzymolysis conditions Protoplast |
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