A cassava protoplast system for screening genes associated with the response to South African cassava mosaic virus.

ABSTRACT:

Background

The study of transient gene expression in cassava plants during virus infection using existing protocols is laborious and may take approximately fifteen weeks due to cassava's recalcitrance to transformation. The combination of a protoplast system with CRISPR-mediated gene editing promises to shorten the turnaround time from plant tissue culture to high-throughput gene expression screening for candidate genes. Here, we detail a protocol for screening genes associated with the response to South African cassava mosaic virus (SACMV) in cassava protoplasts, with reference to the ubiquitin E3 ligase gene, MeE3L.

Methods

Cassava protoplasts of model, and SACMV-susceptible and -tolerant genotypes, were transformed with SACMV infectious clones and/or a CRISPR-editing cons

SUBMITTER: Chatukuta P 

PROVIDER: S-EPMC7685591 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature