G Protein Signaling in UV Protection: Methods for Understanding the Signals in Young Etiolated Seedlings

A seed is competent to respond to light soon after imbibition. A new developmental program begins in or on the ground where the young seedling may be exposed to heat, cold, drought, flooding (anoxia), salts, varying levels of visible light, and the topic of this paper, ultraviolet radiation. Herein what is described is a method for growing and maintaining seedlings, then methods of UV irradiation in order to measure discrete effects of UV wavelengths in signal transduction, very early in seedling development. The physiological response to an abiotic signal is partly dependent on the developmental state of the plant. Dark-grown seedlings of plant species possess young leaves or leaf primordia in a “suspended” state of development whereby exposure to sunlight, visible and UV, is required to initiate the leaf developmental program, including development of etioplasts or proplastids into fully functioning chloroplasts. In order for us to understand the initial and persisting effects of UV in seedlings, we “delay” light-induced development by carrying out all experiments in complete darkness between days 0 (seed) and day 7 (Arabidopsis). In this case, the UV regulation of a simple signaling pathway in Arabidopsis, G protein signaling in UV protection and acclimation early in development, is investigated with the use of several mutants and easily score-able phenotypes.