广东石油化工学院学报2026，Vol.36Issue(3)：32-36,5.DOI:10.26962/j.cnki.1991.2026.0047
菜心茎尖原生质体酶解体系构建
Construction of Enzymatic Hydrolysis System for Protoplasts of Flowering Chinese Cabbage Stem Tip
摘要
Abstract
To provide a reliable cell source for single-cell transcriptome sequencing of flowering Chinese cabbage stem tips,we optimized the concentrations of pectinase,cellulase,and macerozyme,as well as the enzymatic digestion time,to establish a suitable method for protoplast preparation from stem tips.Experimental results demonstrated that the combination of 1.50%cellulase,0.50%pectinase,and 0.6%macerozyme under 2-hour enzymatic hydrolysis yielded optimal protoplast yield and viability from flowering Chinese cabbage stem tip.Further validation confirmed the system's broad applicability across different developmental stages of flowering Chinese cabbage,achieving protoplast yields of 2.90×107,2.62×107,2.47×107 pieces/g,respectively,with viability rates consistently exceeding 92%.Both the quantity and viability of obtained protoplasts met stringent criteria for single-cell transcriptome sequencing,indicating that this enzymatic system can provide high-quality cellular samples for single-cell studies on flowering Chinese cabbage stem tip.关键词
菜心/原生质体/酶解体系Key words
flowering Chinese cabbage/protoplast/enzymatic hydrolysis system分类
农业科技引用本文复制引用
林思敏,王锶珩,宴子巧,关雪怡,刘梓怡,何勤勤,黄新敏..菜心茎尖原生质体酶解体系构建[J].广东石油化工学院学报,2026,36(3):32-36,5.基金项目
国家自然科学基金(32002028) （32002028）
广东省自然科学基金(2022A1515010647) （2022A1515010647）
广东石油化工学院大学生创新训练项目(S20221.1656104) （S20221.1656104）
