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葡萄悬浮细胞系的建立

Establishment of suspension cell system of grape Kyoho

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【作者】 杜晓映张振文夏惠徐国前

【Author】 DU Xiao-ying,ZHANG Zhen-wen,XIA Hui,XU Guo-qian (College of Enology,Northwest A&F University,Yangling,Shaanxi 712100,China)

【机构】 西北农林科技大学葡萄酒学院

【摘要】 【目的】建立适合葡萄悬浮细胞系快速稳定生长的培养体系。【方法】以鲜食葡萄巨峰的果皮为材料,以B5为基本培养基,通过选择合适的激素配比及浓度、pH、蔗糖浓度等建立并优化葡萄愈伤组织培养体系及细胞悬浮培养体系。【结果】在B5+0.5 mg/L 6-BA+NAA条件下,NAA最适质量浓度为1 mg/L;在B5+0.5 mg/L 6-BA+2,4-D条件下,2,4-D最适质量浓度为1.0~2.0 mg/L;在B5+1 mg/L NAA+6-BA条件下,6-BA最适质量浓度为0.5~1.0 mg/L;在B5+2 mg/L 2,4-D+KT条件下,KT的最适质量浓度0.2 mg/L。在最佳蔗糖质量浓度30g/L、最适pH 5.5~5.8培养基条件下,建立了能快速生长的均一、稳定的巨峰葡萄细胞悬浮体系,优化了培养条件。【结论】适合葡萄愈伤组织生长的培养体系为:B5+250 mg/L水解酪蛋白+0.1 mg/L NAA+0.2 mg/L KT+30 g/L蔗糖+6 g/L琼脂;适合葡萄细胞悬浮培养的体系为:B5+1 mg/L NAA+0.5 mg/L 6-BA+30 g/L蔗糖和B5+250mg/L水解酪蛋白+0.1 mg/L NAA+0.2 mg/L KT+30 g/L蔗糖。

【Abstract】 【Objective】 The researth establishd a suitable method for the stable and rapid growth of grape suspension culture.【Method】 With the fruits of Grape Kyoho(V.vinifera L.) as materials,B5 medium as basic medium,at the condition of proper hormone concentration and ratio,pH and sucrose to induce callus,a suspension culture system was established and optimized.【Result】 The hormone concentration was determined in such a way:NAA 1 mg/L,at the condition of B5+0.5 mg/L 6-BA+NAA;2,4-D 1.0-2.0 mg/L,at the condition of B5+0.5 mg/L 6-BA +2,4-D;6-BA 0.5-1.0 mg/L,at the condition of B5+1 mg/L NAA +6-BA;KT 0.2 mg/L,at the condition of B5+2 mg/L 2,4-D+KT.Their effects compared the proper hormone ratio was decided,and the rapid growth,equal,and stabilized grape suspension culture was established at the condition of pH 5.8 and sucrose 30 g/L.【Conclusion】 The instance of callus induction and suspension celll system establishment of Grape Kyoho indicate: the suitable medium for inducing callus is B5+250 mg/L hydrolyzed casein+0.1 mg/L NAA +0.2 mg/L KT+30 g/L sucrose+6 g/L agar,while suitable liquid medium for suspension culture for Kyoho is B5+0.5 mg/L 6-BA+1 mg/L NAA+30 g/L sucrose and B5+250 mg/L hydrolyzed casein +0.1 mg/L NAA +0.2 mg/L KT+30 g/L sucrose.

【关键词】 巨峰葡萄愈伤组织悬浮细胞培养体系
【Key words】 grape Kyohocallussuspension cellculture system
【基金】 陕西省“13115”科技创新工程重大科技专项计划“葡萄与葡萄酒产业关键技术研究”(2007ZDKG-09)
  • 【文献出处】 西北农林科技大学学报(自然科学版) ,Journal of Northwest A & F University(Natural Science Edition) , 编辑部邮箱 ,2009年01期
  • 【分类号】S663.1
  • 【被引频次】16
  • 【下载频次】545
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