植物研究2025，Vol.45Issue(2)：202-210,9.DOI:10.7525/j.issn.1673-5102.2025.02.006
马尾松原生质体制备及瞬时转化体系的建立
Establishment of a System for Preparation and Transient Transformation of Protoplasts of Pinus massoniana
摘要
Abstract
This study aimed to establish a system of the protoplast preparation in Pinus massoniana for the transient transformation of genes,so as to promote the study of gene function and the development of genetic transformation system in P.massoniana.The explants were selected from different developmental periods of P.massoniana,including needle leaf,hypocotyls and callus tissues.The effects of different enzymatic conditions on the isolation efficiency of protoplasts were explored.The protoplasts preparation process involving different types of tissues,different osmotic pressures and enzymatic hydrolysis time were optimized.The results showed that pine needle tissue in 0.5 mol⋅L-1 mannitol osmotic solution for 6 h was the optimal condition for protoplasts extraction.Based on this,a transient transformation system of exogenous genes expression in P.massoniana protoplasts was constructed by comparing different transformation conditions.The optimized protoplast preparation and transformation system could significantly improve the quality and transformation efficiency of protoplasts,and the transformation efficiency of protoplasts was as high as 47.83%when using a polyethylene glycol(PEG)-mediated transformation method in P.massoniana.In summary,this study successfully constructed an effective protoplast isolation system for P.massoniana,which provides a reliable transient expression platform for analyzing gene functions in coniferous species.关键词
马尾松/原生质体/瞬时转化/基因功能Key words
Pinus massoniana/protoplast/transient expression/gene function分类
生物工程引用本文复制引用
李岩硕,王民炎,刘殿宽,李伟,殷恒福..马尾松原生质体制备及瞬时转化体系的建立[J].植物研究,2025,45(2):202-210,9.基金项目
国家科技创新2030重大项目(2022ZD04016). （2022ZD04016）
