花生原生质体分离方法探究

doi:10.3969/j.issn.1009-7791.2023.03.007

亚热带植物科学 ›› 2023, Vol. 52 ›› Issue (3): 220-227.DOI: 10.3969/j.issn.1009-7791.2023.03.007

• 植物生理生化与分子生物学 • 上一篇下一篇

花生原生质体分离方法探究

伍顺达，梁绮雯，孙敏，王思远，苏益，肖浪涛^*

(湖南农业大学生物科学技术学院，湖南长沙 410128)

收稿日期:2023-03-24 接受日期:2023-05-08 出版日期:2023-06-20 发布日期:2023-08-24
通讯作者: 肖浪涛
基金资助:
国家自然科学基金(32261143733)；湖南省自然科学基金(2022JJ30294)

Study on Protoplasts Isolation of Arachis hypogaea

WU Shun-da, LIANG Qi-wen, SUN Min, WANG Si-yuan, SU Yi, XIAO Lang-tao^*

(College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, Hunan China)

Received:2023-03-24 Accepted:2023-05-08 Online:2023-06-20 Published:2023-08-24
Contact: XIAO Lang-tao

摘要/Abstract

摘要： 以花生多粒型四粒红Arachis hypogaea var. fastigiate ‘Silihong’为材料，调节不同酶的浓度，优化原生质体分离条件。结果显示，混合酶解液的分离效果显著优于崩溃酶。制备花生60 d龄植株叶片原生质体的适宜条件为：2%纤维素酶+ 3%果胶酶+1%离析酶+ 0.02 mol·L^–1 KCl + 0.02 mol·L^–1 MES + 0.01 mol·L^–1 CaCl₂ + 0.6 mol·L^–1甘露醇+ 0.1% BSA，28 ℃黑暗酶解3 h。在此条件下，花生叶片原生质体产量为 2.965×10⁵个·g^–1，花瓣原生质体产量为2.55×10⁵ 个·g^–1。此外，花瓣与黄化叶均能成功分离得到大量原生质体，而花生苗的下胚轴以及果针未分离出原生质体。

关键词: 花生, 原生质体, 分离, 混合酶

Abstract: Using leaves and petal of Arachis hypogaea var. fastigiate ‘Silihong’ as materials, protoplast isolation conditions were optimized through modifying the concentrations of different enzymes. The results showed that the protoplast outcome by using the mixed enzymes was significantly better than that by using driselase. The optimum conditions for the preparation of leaves of peanut at 60 days old protoplasts were 2% cellulose + 3% pectinase + 1% diastase + 0.02 mol·L^–1 KCl + 0.02 mol·L^–1 MES + 0.01 mol·L^–1 CaCl₂ + 0.6 mol·L^–1 mannitol + 0.1% BSA, 28 ℃ for 3 hours in dark. Under this condition, the protoplast yield of peanut leaves was 2.965×10⁵ cells·g^–1, and peanut petal protoplast yield was 2.55×10⁵cells·g^–1. In addition, A large number of protoplasts were successfully isolated from peanut petals and yellow leaves, but protoplasts were not isolated from hypocotyls of peanut seedlings and peanut pegs.

Key words: Arachis hypogaea, protoplast, isolation, mixed enzymes

中图分类号:

S565.2

伍顺达, 梁绮雯, 孙敏, 王思远, 苏益, 肖浪涛. 花生原生质体分离方法探究[J]. 亚热带植物科学, 2023, 52(3): 220-227.

WU Shun-da, LIANG Qi-wen, SUN Min, WANG Si-yuan, SU Yi, XIAO Lang-tao. Study on Protoplasts Isolation of Arachis hypogaea[J]. Subtropical Plant Science, 2023, 52(3): 220-227.

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花生原生质体分离方法探究

Study on Protoplasts Isolation of Arachis hypogaea

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