A convenient sample preparation protocol for scanning electron microscope examination of xylem-occluding bacterial biofilm on cut flowers and foliage

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dc.contributor.author Ratnayake, Kamani
dc.contributor.author Joyce, D.C.
dc.contributor.author Webb, R.I.
dc.date.accessioned 2021-04-22T04:48:38Z
dc.date.available 2021-04-22T04:48:38Z
dc.date.issued 2012
dc.identifier.citation Scientia Horticulturae 140 (2012) 12–18 en_US
dc.identifier.issn 0304-4238
dc.identifier.uri http://repository.wyb.ac.lk/handle/1/3118
dc.description.abstract Microbes and their exopolysaccharides (EPS) can block xylem vessels, thereby increasing the hydraulic resistance and decreasing the vase life of cut flowers and foliage. Scanning electron microscopy (SEM) provides a powerful tool for investigation of bacteria-induced xylem occlusion. However, conventional preparation protocols for SEM involving chemicals can cause loss of hydrated EPS material, and thereby damage the bacterial biofilms during dehydration. A modified chemical fixation protocol involving prefixation with 75 mM lysine plus 2.5% glutaraldehyde followed by the normal fixation in 3% glutaraldehyde was, therefore, tested for improved preservation of bacterial biofilm at the stem-ends of cut Acacia holosericea foliage stems. Stem-end segments with different stages of bacterial growth were obtained from stems stood into water. The lysine-based protocol was compared with four other processing protocols of critical point drying (CPD) without fixation (control), freeze-drying (FD), conventional chemical fixation followed by drying with hexamethyldisilazane (HMDS), and conventional chemical fixation with CPD. The non-fixed control, FD and the glutaraldehyde fixation with HMDS drying gave poor preservation of hydrated material, including bacterial EPS. Conventional glutaraldehyde fixation followed by CPD was superior to these three methods in terms of better preserving the EPS. However, this fourth method gave condensation of biofilms during dehydration. In contrast, the modified lysine-based protocol resulted in superior preservation of EPS and biofilm structure. Thus, this fifth method was the most appropriate for examination of bacterial stem-end blockage in cut ornamentals. en_US
dc.language.iso en en_US
dc.publisher Elsevier en_US
dc.subject Acacia en_US
dc.subject Bacteria en_US
dc.subject Dehydration en_US
dc.subject Exopolysaccharides en_US
dc.subject Fixative en_US
dc.subject Lysine en_US
dc.subject SEM en_US
dc.title A convenient sample preparation protocol for scanning electron microscope examination of xylem-occluding bacterial biofilm on cut flowers and foliage en_US
dc.type Article en_US


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