Confocal Laser Scanning Microscopy Detection of Chlorophylls and Carotenoids in Chloroplasts and Chromoplasts of Tomato Fruit
Plant cells are unique among eukaryotic cells because of the presence of plastids, including chloroplasts and chromoplasts. Chloroplasts are found in green tissues and harbor the photosynthetic machinery (including chlorophyll molecules), while chromoplasts are present in non-photosynthetic tissues and accumulate large amounts of carotenoids. During tomato fruit development, chloroplasts are converted into chromoplasts that accumulate high levels of lycopene, a linear carotenoid responsible for the characteristic red color of ripe fruit. Here, we describe a simple and fast method to detect both types of fully differentiated plastids (chloroplasts and chromoplasts), as well as intermediate stages, in fresh tomato fruits. The method is based on the differential autofluorescence of chlorophylls and carotenoids (lycopene) detected by Confocal Laser Scanning Microscopy.
KeywordsChloroplast Chlorophylls Chromoplast Carotenoids Lycopene Confocal microscopy Tomato fruit Fluorescence
Our work is funded by grants from the Catalan AGAUR (2009SGR-26 and XRB), Spanish DGI (BIO2011-23680 and PIM2010IPO-00660), and European Union FP7 (TiMet, contract 245143). We are members of the IBERCAROT network funded by CYTED (112RT0445). L.D. received a predoctoral fellowship of the Spanish Ministerio de Educación FPU program.
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