Isolation of intact sub-dermal secretory cavities from Eucalyptus
AuthorGoodger, JQD; Heskes, AM; Mitchell, MC; King, DJ; Neilson, EH; Woodrow, IE
Source TitlePlant Methods
PublisherBIOMED CENTRAL LTD
University of Melbourne Author/sGoodger, Jason; HESKES, ALLISON; NEILSON, ELIZABETH; Woodrow, Ian; KING, DREW; MITCHELL, MADELINE CLAIRE
AffiliationSchool of BioSciences
School of Ecosystem and Forest Sciences
School of Botany
Document TypeJournal Article
CitationsGoodger, J. Q. D., Heskes, A. M., Mitchell, M. C., King, D. J., Neilson, E. H. & Woodrow, I. E. (2010). Isolation of intact sub-dermal secretory cavities from Eucalyptus. PLANT METHODS, 6 (1), https://doi.org/10.1186/1746-4811-6-20.
Access StatusOpen Access
ARC Grant codeARC/DP1094530
BACKGROUND: The biosynthesis of plant natural products in sub-dermal secretory cavities is poorly understood at the molecular level, largely due to the difficulty of physically isolating these structures for study. Our aim was to develop a protocol for isolating live and intact sub-dermal secretory cavities, and to do this, we used leaves from three species of Eucalyptus with cavities that are relatively large and rich in essential oils. RESULTS: Leaves were digested using a variety of commercially available enzymes. A pectinase from Aspergillus niger was found to allow isolation of intact cavities after a relatively short incubation (12 h), with no visible artifacts from digestion and no loss of cellular integrity or cavity contents. Several measurements indicated the potential of the isolated cavities for further functional studies. First, the cavities were found to consume oxygen at a rate that is comparable to that estimated from leaf respiratory rates. Second, mRNA was extracted from cavities, and it was used to amplify a cDNA fragment with high similarity to that of a monoterpene synthase. Third, the contents of the cavity lumen were extracted, showing an unexpectedly low abundance of volatile essential oils and a sizeable amount of non-volatile material, which is contrary to the widely accepted role of secretory cavities as predominantly essential oil repositories. CONCLUSIONS: The protocol described herein is likely to be adaptable to a range of Eucalyptus species with sub-dermal secretory cavities, and should find wide application in studies of the developmental and functional biology of these structures, and the biosynthesis of the plant natural products they contain.
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