Isolation of intact sub-dermal secretory cavities from Eucalyptus
Jason QD Goodger, Allison M Heskes, Madeline C Mitchell, Drew J King, Elizabeth H Neilson, Ian E Woodrow
PLANT METHODS | BMC | Published : 2010
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 lo..View full abstract
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Awarded by Australian Research Council
This research was funded by an Australian Research Council Discovery grant to JQDG and IEW (Project DP1094530). AH and EN were supported, in part, by the Holsworth Wildlife Research Endowment (managed by ANZ Trustees). We thank Assoc. Prof. Ed Newbigin (University of Melbourne) for supplying the initial enzymes, Assoc. Prof. Andrew Drinnan (University of Melbourne) for microscope usage and advice, and Dr. Janet Newman (CSIRO Molecular and Health Technologies) for use of the NanoDrop spectrophotometer.