高质量番茄根原生质体制备体系的建立

Establishment of a High-quality Tomato Root Protoplast Preparation System

原生质体因其缺乏细胞壁,在植物育种、基因转化及基础研究中具有广泛应用。酶解法是制备植物原生质体的关键方法。番茄根作为番茄的重要器官之一,建立番茄根尖原生质体制备体系对于研究番茄根的发育和功能具有重要意义。以番茄(Solanum lycopersicum cv.Micro-Tom)根尖为材料,优化了纤维素酶和离析酶的浓度配比,旨在建立一种高质量番茄根原生质体的制备方法。研究结果表明:在25℃条件下,采用1.5%纤维素酶R-10和1%离析酶R-10的组合进行6 h酶解,可使原生质体的浓度达到1 100个·μL^(-1),活力超过90%。

Protoplasts,which lack cell walls,hold significant potential in plant breeding,genetic transformation,and fundamental research.Enzymatic digestion is a crucial method for preparing plant protoplasts.Tomato roots are vital components of the tomato plant,and establishing a system for preparing tomato root apical meristem protoplasts preparation system is essential for understanding the intricacies of tomato root development and function.In this study,tomato(Solanum lycopersicum,cv Micro-Tom)root apices were used as the experimental material,and the concentration ratio of cellulase and pectinase was carefully optimized to develop a protocol for producing high-quality tomato root protoplasts.The results indicated that a combination of 1.5%cellulase R-10 and 1%pectinase R-10,followed by a 6 h enzymatic digestion at 25℃,yielded a protoplast concentration of 1100 cells/µL with a viability rate exceeding 90%.