School of Botany - Research Publications
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ItemIsolation of intact sub-dermal secretory cavities from Eucalyptus(BMC, 2010-09-01)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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ItemCyanogenic polymorphism as an indicator of genetic diversity in the rare species Eucalyptus yarraensis (Myrtaceae)(CSIRO PUBLISHING, 2002)The rare Australian tree Eucalyptus yarraensis Maiden & Cambage is cyanogenic, a quantitative trait potentially indicative of genetic diversity. Cyanogenic plants are capable of releasing cyanide from endogenous cyanide-containing compounds. Cyanide is toxic or deterrent to generalist or non-adapted specialist herbivores. Consequently, cyanogenic plants are afforded an effective means of chemical defense. In this paper we characterize quantitative variation in cyanogenic capability, known as cyanogenic polymorphism, in E. yarraensis for the first time. We show that the cyanogenic glucoside prunasin (R-mandelonitrile-β-D-glucoside) is the only cyanogenic compound in E. yarraensis foliage. We also show that two natural populations of E. yarraensis display extensive intra- and inter-population variation in foliar prunasin concentration. The high prunasin concentrations reported in this paper represent the highest yet recorded for mature eucalypt leaves. The cyanogenic variation could not be attributed to measured physical and chemical parameters, supporting the hypothesis that the variation is genetically based. A preliminary progeny trial also supports this hypothesis, with narrow sense heritability estimated at 1.17 from three half-sibling families. The variation in cyanogenic capability may be a useful tool in the development of a conservation strategy for the species.
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ItemGrowth cost and ontogenetic expression patterns of defence in cyanogenic Eucalyptus spp.(SPRINGER HEIDELBERG, 2006-11)