A high-efficiency PEG-Ca²⁺-mediated transient transformation system for broccoli protoplasts.

ABSTRACT: Transient transformation of plant protoplasts is an important method for studying gene function, subcellular localization and plant morphological development. In this study, an efficient transient transformation system was established by optimizing the plasmid concentration, PEG4000 mass concentration and genotype selection, key factors that affect transformation efficiency. Meanwhile, an efficient and universal broccoli protoplast isolation system was established. Using 0.5% (w/v) cellulase R-10 and 0.1% (w/v) pectolyase Y-23 to hydrolyze broccoli cotyledons of three different genotypes for 3 h, the yield was more than 5×10⁶/mL/g, and the viability was more than 95%, sufficient to meet the high standards for protoplasts to be used in various experiments. The average transformat

SUBMITTER: Yang D 

PROVIDER: S-EPMC9790990 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature