Early Diagnosis of Productivity Through a Clock Gene Promoter Activity Using a Luciferase Bioluminescence Assay in Arabidopsis thaliana

• Published in: Environment control in biology 2011, Vol.49(2), pp.51-60
• Main Authors: FUKUDA, Hirokazu, ICHINO, Takatoshi, KONDO, Takao, MURASE, Haruhiko
• Format: Article
• Language: English
• Subjects: Arabidopsis thaliana; circadian clock; circadian resonance; medical molecular farming; plant factory; selection of seedling
• ISSN: 1880-554X; 1883-0986
• DOI: 10.2525/ecb.49.51

A method for early diagnosis of plant productivity using a luciferase bioluminescence assay at the early stage of cultivation was developed for transgenic Arabidopsis thaliana carrying the CCA1::LUC construct, in which the promoter of the CCA1 clock gene had been fused to a modified firefly luciferase (LUC) gene. Because the intensity of luciferase bioluminescence is proportional to the CCA1 promoter activity and the expression level of LUC, the dynamics of both can be investigated by measuring the bioluminescence. In this paper, plant biomass was considered a measure of plant production andLUC protein a measure of foreign protein production. The leaf area and bioluminescence of seedlings at the early stage of cultivation were investigated as indexes for early diagnosis of productivity; leaf area was a reflection of morphological information, and bioluminescence, molecular information. Bioluminescence showed higher correlation to productivity based on biomass andLUC protein than the leaf area under various growth conditions, indicating that bioluminescence is a better index for early diagnosis. In addition, we developed a general statistical method for selecting superior seedlings based on this early diagnosis and a general theory for determining an optimal parameter set for maximizing profit. This early diagnostic methodology appears suited to enhance the quality of the products in plant factories.