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1.  Extracellular Localization of the Diterpene Sclareol in Clary Sage (Salvia sclarea L., Lamiaceae) 
PLoS ONE  2012;7(10):e48253.
Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L.) inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction). Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy). According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces.
doi:10.1371/journal.pone.0048253
PMCID: PMC3484996  PMID: 23133579
2.  Production and Emission of Volatile Compounds by Petal Cells 
Plant Signaling & Behavior  2007;2(6):525-526.
We localized the tissues and cells that contribute to scent biosynthesis in scented and non-scented Rosa × hybrida cultivars as part of a detailed cytological analysis of the rose petal. Adaxial petal epidermal cells have a typical conical, papillate shape whereas abaxial petal epidermal cells are flat. Using two different techniques, solid/liquid phase extraction and headspace collection of volatiles, we showed that, in roses, both epidermal layers are capable of producing and emitting scent volatiles, despite the different morphologies of the cells of these two tissues. Moreover, OOMT, an enzyme involved in scent molecule biosynthesis, was localized in both epidermal layers. These results are discussed in view of results found in others species such as Antirrhinum majus, where it has been shown that the adaxial epidermis is the preferential site of scent production and emission.
PMCID: PMC2634358  PMID: 19704548
floral scent; petal epidermis; Rosa; terpenes; volatiles
3.  Chemical and Histochemical Analysis of ‘Quatre Saisons Blanc Mousseux’, a Moss Rose of the Rosa × damascena Group 
Annals of Botany  2006;97(2):231-238.
• Background and Aims Moss roses are old garden roses covered with a mossy growth on flower pedicel and calyx. This moss releases a pine-scented oleoresin that is very sticky and odoriferous. Rosa × centifolia ‘muscosa’ was the first moss rose to be obtained by bud-mutation but, interestingly, R. × damascena ‘Quatre Saisons Blanc Mousseux’ was the first repeat-blooming cultivar, thus interesting breeders. In the present study, the anatomy of these sports (i.e. bud-mutations) is characterized and the volatile organic compounds (VOCs) produced by the moss versus the petals are identified. They are compared between the two lines and their respective parents.
• Methods Anatomy of the moss is studied by environmental scanning electron microscopy and histochemical light microscopy. Sudan Red IV and Fluorol Yellow 088 are used to detect lipids, and 1-naphthol reaction with N,N-dimethyl-p-phenylenediamine to detect terpenes (Nadi reaction). Head-space or solid/liquid extraction followed by gas chromatography and mass spectrometry are used to identify VOCs in moss, trichomes and petals.
• Key Results Moss of the two cultivars has the same structure with trichomes on other trichomes but not exactly the same VOCs. These VOCs are specific to the moss, with lots of terpenes. An identical VOC composition is found in leaves but not in petals. They are nearly the same in the moss mutants and in the respective wild types.
• Conclusions Sepals of moss roses and their parents have a specific VOC pattern, different from that of the petals. The moss corresponds to a heterochronic mutation with trichomes developing on other trichomes. Such a mutation has probably appeared twice and independently in the two lines.
doi:10.1093/aob/mcj034
PMCID: PMC2803356  PMID: 16344264
Rosa × damascena ‘bifera’; Rosa × damascena ‘Quatre Saisons Blanc Mousseux’; Rosa × centifolia; Rosa × centifolia ‘muscosa’; moss roses; glandular trichomes; histochemistry; volatile organic compounds; gas chromatography; sport; terpenoids; benzenoids

Results 1-3 (3)