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刘艳丽, 金孝芳, 马林龙, 曹丹, 龚自明, 韦朝领. 茶树叶肉原生质体的分离与纯化[J]. 植物科学学报, 2017, 35(6): 908-911. DOI: 10.11913/PSJ.2095-0837.2017.60908
引用本文: 刘艳丽, 金孝芳, 马林龙, 曹丹, 龚自明, 韦朝领. 茶树叶肉原生质体的分离与纯化[J]. 植物科学学报, 2017, 35(6): 908-911. DOI: 10.11913/PSJ.2095-0837.2017.60908
Liu Yan-Li, Jin Xiao-Fang, Ma Lin-Long, Cao Dan, Gong Zi-Ming, Wei Chao-Ling. Isolation and purification of mesophyll protoplasts from the leaves of Camellia sinensis[J]. Plant Science Journal, 2017, 35(6): 908-911. DOI: 10.11913/PSJ.2095-0837.2017.60908
Citation: Liu Yan-Li, Jin Xiao-Fang, Ma Lin-Long, Cao Dan, Gong Zi-Ming, Wei Chao-Ling. Isolation and purification of mesophyll protoplasts from the leaves of Camellia sinensis[J]. Plant Science Journal, 2017, 35(6): 908-911. DOI: 10.11913/PSJ.2095-0837.2017.60908

茶树叶肉原生质体的分离与纯化

Isolation and purification of mesophyll protoplasts from the leaves of Camellia sinensis

  • 摘要: 以茶树幼苗叶片为材料,分析了甘露醇浓度、不同酶液组合、酶解时间等因素对叶肉原生质体分离及2种高密度分离液对原生质体纯化的影响。结果表明,叶片在含0.6 mol/L的甘露醇、1.5%的纤维素酶和2.5%的离析酶的酶解液中黑暗酶解16~18 h,叶肉原生质体的分离效果最好。此外,研究发现原生质体在10%的甘露醇与25%的碘克沙醇界面处聚集形成了一条带。

     

    Abstract: In the present study, the effect of mannitol concentration, combination of different enzyme (cellulase and macerozyme) solutions, and enzymolysis time on the separation of mesophyll protoplasts and purification of protoplasts from two high density separating solutions were studied in the leaves of tea seedlings. The results demonstrated that high quality mesophyll protoplasts were obtained from tea leaves subjected to enzymatic digestion in a solution containing 0.6 mol/L mannitol, 1.5% cellulase, and 2.5% macerozyme for 16-18 h in the dark. In addition, protoplasts gathered into a band on the layer between 10% mannitol and 25% iodixanol.

     

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