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Figure 1 | Plant Methods

Figure 1

From: Isolation of intact sub-dermal secretory cavities from Eucalyptus

Figure 1

Flow chart of the successful secretory cavity isolation protocol. (a) Leaf strip dissected from a Eucalyptus polybractea leaf. (b) Leaf strip after 12 h digestion with Aspergillus P-4716 pectinase (Sigma) and peeling away of the epidermis (E) with attached cuticle. (c) Digested leaf strip with removed epidermis showing freed cavities (arrow) and vasculature (V). (d) Mixture of isolated cavities (arrow) and mesophyll tissue (arrowhead) after leaf strip was teased apart with forceps. (e) Secretory cavities freed from the mesophyll tissue in which they were embedded. Isolated cavities can be collected via pipette or for larger numbers of cavities, after sieving and density separation steps. (f) Autofluorescence of isolated cavities imaged under UV illumination with GFP2 filter set show no evidence of characteristic red chlorophyll fluorescence. (g) Isolation of the epidermis is an added advantage of the protocol that may be of use in examinations of cuticular or epidermal properties such as stomatal (arrow) arrangement. (h) An incompletely isolated cavity using Rhizopus pectinase (Sigma). (i) An over-digested cavity using Trichoderma cellulose (Yakult). Scale bars represent 1 mm for panels a to d, and 200 μm for panels e to i.

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