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An efficient transient expression system for gene function analysis in rose

Author:
Lu, Jun, Bai, Mengjuan, Ren, Haoran, Liu, Jinyi, Wang, Changquan
Source:
Plant methods 2017 v.13 no.1 pp. 116
ISSN:
1746-4811
Subject:
Agrobacterium, DNA, Rosa chinensis, binding capacity, cut flowers, flowering, gardens, genetic transformation, genotype, luciferase, models, odors, ornamental plants, plant growth, protein-protein interactions, reporter genes, roots, seedlings, shoots, signal transduction, transcription (genetics), transcription factors
Abstract:
BACKGROUND: Roses are widely used as garden ornamental plants and cut flowers. Rosa chinensis cv ‘Old Blush’ has been used as a model genotype in rose studies due to its contribution to recurrent flowering and tea scent traits of modern roses. The deficiency of efficient genetic transformation systems is a handicap limiting functional genetics studies of roses. Agrobacterium-mediated transient transformation offers a powerful tool for the characterization of gene function in plants. RESULTS: A convenient and highly efficient Agrobacterium mediated genetic transformation protocol using R. chinensis cv ‘Old Blush’ seedlings in vitro as an expression system is described in this paper. The most important factor affecting transformation efficiency in this system is seedling age; 3/4-week-old rose shoots with or without roots from sub-culturing are optimal for transformation, requiring no complicated inoculation media, supplements, or carefully tuned plant growth conditions. This transient expression system was successfully applied to analysis of the gene promoter activities, DNA binding capacity of transcription factors, protein–protein interaction in physiological contexts using luciferase as a reporter gene. CONCLUSIONS: This transient transformation system was validated as a robust and efficient platform, thus providing a new option for gene function and signaling pathway investigation in roses and further extending the utility of R. chinensis cv ‘Old Blush’ as a model plant to study diverse gene function and signaling pathways in Rosaceae.