茶树叶肉原生质体的分离与纯化

刘艳丽; 金孝芳; 马林龙; 曹丹; 龚自明; 韦朝领

doi:10.11913/PSJ.2095-0837.2017.60908

茶树叶肉原生质体的分离与纯化

1. 湖北省农业科学院果树茶叶研究所, 武汉 430209;
2. 安徽农业大学茶树生物学与资源利用国家重点实验室, 合肥 230036

基金项目:

湖北省农业科学院果树茶叶研究所基金项目（GCJJ201601）；湖北省农业科学院基金项目（2017NKYJJ05）；茶树生物学与资源利用国家重点实验室开放基金课题（SKLTOF20160104）。

详细信息

作者简介:
刘艳丽(1977-),女,博士,助理研究员,研究方向为茶树资源与育种(E-mail:zkslyl@163.com)。

通讯作者:
金孝芳,E-mail:jxf1130@126.com

韦朝领,E-mail:weichl@ahau.edu.cn

中图分类号: Q945
计量
- 文章访问数: 915
- HTML全文浏览量: 4
- PDF下载量: 1227
出版历程
- 收稿日期: 2017-10-09
- 网络出版日期: 2022-10-31
- 发布日期: 2017-12-27

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

1. Institute of Fruit and Tea, Hubei Academy of Agricultural Science, Wuhan 430209, China;
2. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China

Funds:

This work was supported by grants from the Institute of Fruit and Tea, Hubei Academy of Agricultural Science (GCJJ201601), Fund of Hubei Academy of Agricultural Science (2017NKYJJ05), and Open Fund of State Key Laboratory of Tea Plant Biology and Utilization (SKLTOF20160104).

摘要

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。
- 茶树叶片 /
- 原生质体 /
- 分离 /
- 纯化
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.
- Tea leaves /
- Protoplast /
- Isolation /
- Purification

HTML全文

参考文献(9)

[1]	Guo JJ, Morrell-Falvey JL, Labbé JL, Muchero W, Kalluri UC, et al. Highly efficient isolation of Populus mesophyll protoplasts and its application in transient expression assays[J]. PLoS One, 2012, 7(9):e44908.
[2]	Confraria A, Baena-González E. Using Arabidopsis protoplasts to study cellular responses to environmental stress[J]. Meth Mol Biol, 2016, 1398:247.
[3]	Zhao FL, Li YJ, Hu Y, Gao YR, Zang XW, et al. A highly efficient grapevine mesophyll protoplast system for tran-sient gene expression and the study of disease resistance proteins[J]. Plant Cell Tiss Org, 2016, 125(1):43-57.
[4]	王璐, 钱文俊, 王新超, 李娜娜, 杨亚军. 一种茶树原生质体的提取方法:201610175155.8[P]. 2016-03-25.
[5]	杨子银, 梅鑫, 周瀛, 傅秀敏. 一种茶树花原生质体及其制备方法和应用:201610255637.4[P]. 2016-04-22.
[6]	刘艳丽, 姚家玲. 盾叶薯蓣原生质体分离[J]. 华中农业大学学报, 2008, 27(3):430-433. Liu YL, Yao JL. Protoplast isolation of Dioscorea zingiberensis Wright[J]. Journal of Huazhong Agricultural University, 2008, 27(3):430-433.
[7]	Duruflé H, Clemente HS, Balliau T, Zivy M, Dunand C, Jamet E. Cell wall proteome analysis of Arabidopsis tha-liana mature stems[J]. Proteomics, 2017, 17(8):1600449.
[8]	Gregory RP, Bendall DS. The purification and some pro-perties of the polyphenol oxidase from tea(Camellia sinensis L.)[J]. Biochem J, 1966, 101(3):569.
[9]	Ali HM, El-Gizawy AM, El-Bassiouny REI, Saleh MA. Inhibition of apple polyphenoloxidase(PPO) by ascorbic acid, citric acid and sodium chloride[J]. J Food Process Pres, 2010, 17(1):21-30.

施引文献(5)

期刊类型引用(4)

1.	李必聪，李慧英，肖遥，罗莎，周庆红，黄英金，朱强龙. 芋扩展蛋白基因家族的全基因组鉴定及其在球茎膨大中的表达分析. 浙江农业学报. 2023(07): 1604-1616 . 百度学术
2.	赵晓宇，苏二虎，王雪娇，刘坤雨，高圆丽，薛春雷，梁红伟，李强. 缺硼对大豆幼苗生长及保护性酶活的影响. 大豆科学. 2023(06): 718-725 . 百度学术
3.	罗萍，王晓萍，张昊楠，范春节，王玉娇，徐建民. 巨桉扩展蛋白EgrEXPA8和EgrEXPA10基因的克隆和表达特性分析. 热带亚热带植物学报. 2023(06): 827-834 . 百度学术
4.	侯佳玉，闫磊，程锦，曾紫君，张雅茹，鲁克嵩，姜存仓. L-天冬氨酸纳米钙促进油菜生长的机理机制. 农业环境科学学报. 2022(07): 1408-1416 . 百度学术