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使用原生细胞转化系统,使Mangifera indica的功能研究成为可能

Mark Owusu Adjei^1,2, Huan Zhao¹, Xiaoguang Tao¹

¹College of Landscape Architecture, Sichuan Agricultural University, Chengdu 611130, China.

International journal of molecular sciences

|August 12, 2023

概括

研究人员优化了果原生细胞的隔离和转化,用于基因研究. 这种新方法有效地转化了果原塑,有助于在这个重要的果树中研究分子机制.

科学领域:

植物生物技术植物生物技术
分子生物学分子生物学
园艺园艺园艺园艺

背景情况:

果 (Mangifera indica L.) 的转化技术发展不足.
原生体技术为短暂表达和转化试验提供了一种快速的方法,特别是对于缺乏Agrobacterium介导系统的植物.

研究的目的:

为了优化果的原生质隔离和转化条件.
为果基因表达调节研究建立一个高效的系统.

主要方法:

优化酶度 (细胞酶R-10,麦克罗酶R-10) 和消化时间用于原生细胞分离.
确定了原生质细胞产量,生存能力和计数的最佳曼尼托度.
使用的聚乙烯甘醇 (PEG) -4000-CaCl2介导的转化隔离的果叶原生质.

主要成果:

确定了最佳条件:150毫克/毫升细胞酶R-10,180毫克/毫升麦克罗酶R-10,pH值5.6,消化12小时.
通过特定的曼尼托度 (0.16M在洗中,0.08M在MMG中) 实现高原生质产量.
使用PEG介导的转化证明了84.38%的转化效率,具有最佳的曼尼托尔可活性和成功的GFP表达.

结论:

开发了一种高效的果原生细胞分离和PEG介导转化协议.
该协议为果的快速分子机制研究提供了有效的工具.
优化的系统有助于在果中调节基因表达的研究.

关键词:

曼吉弗拉指南 L. 曼吉弗拉指南 L.聚乙烯糖介导的聚乙烯糖介导的原生质细胞隔离隔离.转化转化转化转化转化转化转化转化短暂的表达方式短暂的表达方式

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