Development and utilization of a carrot protoplast-based gene transient expression system

Abstract

Carrots are the top ten most economically important vegetable crops worldwide by market value and production area. Protoplasts are generally isolated from Tobacco Bright Yellow 2 (BY2) cell lines, tobacco leaf cells and onion bulb cells. However, the complicated and expensive preparation method limits these protoplasts transient expression (TE). This study shows that carrot protoplasts have many advantages for TE, including easy access, low preparation cost, high repeatability, uniform size and strong vitality. We developed an efficient method for producing carrot protoplasts. Meanwhile, carrot protoplasts were successfully utilized for protein subcellular localization analysis and bimolecular fluorescence complementation (BiFC).