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a Data provided by Polysciences, Inc, Warrington, PA.

a Data provided by Polysciences, Inc, Warrington, PA.

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In contrast to general DNA dyes, immunostaining with fluorescent-labeled antibodies [11] or fluorescence in situ hybridization (FISH) with 16S rRNA probes [12-16] may provide high specificity to detect a given bacterial species or strain among the plant microflora. Also, fluorescently labeled lectins have proven useful to probe bacteria associated with the exopolysaccharide matrix of aggregates on plants and in biofilms in general [17,18]. While fluorescein and rhodamine bioconjugates have been popular fluorochromes for the visualization of bacteria on plant tissue, newer products for labeling of bioconjugates, such as Alexa Fluor® (Molecular Probes) and CyTM dyes (Amersham Biosciences Corp., Piscataway, NJ), exhibit brighter fluorescence and enhanced photostability.

General stains and bioconjugates can be used alone or in combination with fluorescent proteins to probe complex microenvironments, processes, or biological parameters governing bacterial behavior on plants. In the sections below, we discuss various applications of fluorescence microscopy to the study of the ecology of plant-associated bacteria and human enteric pathogens in the plant environment. Table 26.1 lists the excitation and emission peaks, as well as specific applications, of a variety of fluorescent proteins and stains, some of which are mentioned in this chapter.

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