A cellophane-supported Arabidopsis culture for seamless transfer between different media is useful for studying various nitrogen responses

Abstract

ABSTRACT Transferring plants between different nutritional media is a common technique in plant nutrition research. Solidified sterile media are often used for plant culture and subsequent transfer experiments. Transplanting is generally performed one plant at a time using sterile tweezers. This method has several challenges: it is time-consuming, which makes analyzing short-term responses to nutritional changes difficult; plants can be mechanically damaged, which can confound results; transferring mature plants is accompanied by root wounding due to root penetration into the solidified medium; and root shapes are altered, which precludes a precise evaluation on nutritional responses of root system architecture. To solve these problems, this study tried to adopt a system for Arabidopsis culture using solidified media covered with cellophane disks for the transfer experiments to analyze nitrogen responses as a case study. The cellophane disk prevented the penetration of roots into the solidified medium and fixed the roots loosely; thereby, facilitating sterile, rapid and noninvasive transfer of young and mature Arabidopsis plants between media without changing root shape. The plants-on-cellophane grown on the solidified media were directly transferred onto hydroponics, where plants vegetate and propagate seeds under strict nutritional control. The root proliferation on the cellophane disk allowed direct observations of root architecture and green fluorescent protein (GFP) fluorescence in the roots. Collectively, we conclude that the introduction of a cellophane sheet to the existing culture system would accelerate experiments in plant nutritional research.