介孔硅纳米粒作为植物细胞转基因载体的研究  

Genetic Transformation of Plant Cells Mediated by Mesoporous Silica Nanoparticles

作　　者：陆新华[1] 孙德权[1] 张秀梅[1] LU Xin-hua;SUN De-quan;ZHANG Xiu-mei(South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Tropical Fruit’s Biology,Ministry of Agriculture and Rural Affairs,Zhanjiang 524091)

机构地区：[1]中国热带农业科学院南亚热带作物研究所农业农村部热带果树生物学重点实验室,湛江524091

出　　处：《生物技术通报》2022年第7期194-204,共11页Biotechnology Bulletin

基　　金：中央级公益性科研院所基本科研业务费项目(1630062017037)。

摘　　要：随着纳米生物技术的快速发展,以纳米材料作为基因载体在植物细胞转导研究中取得了初步进展。本研究制备了两种尺寸的介孔硅纳米颗粒,氨基修饰后对其进行表征,并负载含smGFP基因的质粒DNA对拟南芥原生质体进行细胞转导。结果表明,直径约20 nm的氨基化介孔硅纳米颗粒(Am-MSN-20)呈类球形,花形结构的氨基化介孔硅纳米颗粒(Am-MSN-50)直径约50 nm,两者均带正电荷。Am-MSN-50结合DNA的能力高于Am-MSN-20,两种纳米载体都表现出了良好的稳定性,能够保护负载的pDNA不被细胞核酸酶降解,并且对原生质体没有毒害作用。与Am-MSN-20相比较,Am-MSN-50具备更高的转导效率。本研究表明,氨基修饰的MSNs有望成为一种安全高效的新型植物基因载体。With the rapid development of nanobiotechnology,nanoparticles have been applied as vectors for gene transformation in plant cells.In this study,mesoporous silica nanoparticles(MSNs)with different particle sizes were synthesized and then amine-functionalized.The transformation efficiencies of amine-functionalized MSNs(Am-MSNs)for delivering plasmid DNA encoding green fluorescent protein(smGFP)were investigated in the protoplasts of Arabidopsis thaliana.As results,the Am-MSN-20 exhibited monodispersed spheroids with average particle size of 20 nm,while Am-MSN-50 showed a flower-like structure and had an average particle size of 50 nm.Both amine-functionalized MSNs were positively charged.Am-MSN-50 had higher binding capacity of pDNA than that of Am-MSN-20.The Am-MSNs/pDNA complex showed good stability,which suggested that both Am-MSNs protected bounded pDNA efficiently against degradation by cellular nucleases.The MSNs also exhibited no cytotoxic effects on the protoplasts of A.thaliana.The Am-MSN-50 enabled much higher transformation efficiency of the pDNA encoding smGFP compared to that of Am-MSN-20.The results suggest that Am-MSNs can be employed as promising carriers for safe and effective gene delivery.

关 键 词：介孔硅纳米粒 氨基化 遗传转化 载体 植物细胞 

分 类 号：Q943.2[生物学—植物学]