Multiphoton fluorescence lifetime imaging shows spatial segregation of secondary metabolites in Eucalyptus secretory cavities

Multiphoton fluorescence lifetime imaging shows spatial segregation of secondary metabolites in Eucalyptus secretory cavities

Summary Multiphoton fluorescence lifetime imaging provides an excellent tool for imaging deep within plant tissues while providing a means to distinguish between fluorophores with high spatial and temporal resolution. Ideal candidates for the application of multiphoton fluorescence lifetime imaging...

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Bibliographic Details
Published in:Journal of microscopy (Oxford) Vol. 247; no. 1; pp. 33 - 42
Main Authors: HESKES, A.M., LINCOLN, C.N., GOODGER, J.Q.D., WOODROW, I.E., SMITH, T.A.
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Publishing Ltd 01-07-2012
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Subjects:
Cavities
Chemical compounds
Cineole
Essential oil
Essential oils
Esters
eucalypt
Eucalyptus
Eucalyptus - chemistry
Eucalyptus - ultrastructure
FLIM
Fluorescence
Fluorophores
Holes
Image Processing, Computer-Assisted
Imaging
Metabolites
Microscopy, Fluorescence, Multiphoton
monoterpene
oil gland
Oils & fats
Oils, Volatile - metabolism
oleuropeic acid
Organelles - chemistry
Organelles - ultrastructure
Plant Leaves - chemistry
Plant Leaves - ultrastructure
Secondary metabolites
Temporal resolution
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Summary:Summary Multiphoton fluorescence lifetime imaging provides an excellent tool for imaging deep within plant tissues while providing a means to distinguish between fluorophores with high spatial and temporal resolution. Ideal candidates for the application of multiphoton fluorescence lifetime imaging to plants are the embedded secretory cavities found in numerous species because they house complex mixtures of secondary metabolites within extracellular lumina. Previous investigations of this type of structure have been restricted by the use of sectioned material resulting in the loss of lumen contents and often disorganization of the delicate secretory cells; thus it is not known if there is spatial segregation of secondary metabolites within these structures. In this paper, we apply multiphoton fluorescence lifetime imaging to investigate the spatial arrangement of metabolites within intact secretory cavities isolated from Eucalyptus polybractea R.T. Baker leaves. The secretory cavities of this species are abundant (up to 10 000 per leaf), large (up to 6 nL) and importantly house volatile essential oil rich in the monoterpene 1,8‐cineole, together with an immiscible, non‐volatile component comprised largely of autofluorescent oleuropeic acid glucose esters. We have been able to optically section into the lumina of secretory cavities to a depth of ∼80 μm, revealing a unique spatial organization of cavity metabolites whereby the non‐volatile component forms a layer between the secretory cells lining the lumen and the essential oil. This finding could be indicative of a functional role of the non‐volatile component in providing a protective region of low diffusivity between the secretory cells and potentially autotoxic essential oil.
Bibliography:ark:/67375/WNG-TRK85K0S-1
ArticleID:JMI3593
istex:C77FAC378F6CD180A32C1EBD30906DEAA7F5B7A8
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0022-2720
1365-2818
DOI:10.1111/j.1365-2818.2011.03593.x