An autoradiographic method for determining nutrient competition between leaf epiphytes and plant pathogens

Edwards, M. C.; Blakeman, J.P.

doi:10.1111/j.1365-2818.1984.tb00486.x

Cited by 7 publications

(4 citation statements)

References 21 publications

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“…Microautoradiography has been successfully applied to investigate microbial growth and substrate competition in systems other than activated sludge (4,5,8). Carman (4) used microautoradiography to investigate the uptake of radioactive tracers by a natural assemblage of sedimentary microorganisms.…”

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“…He demonstrated that [ 3 H]acetate and [ 3 H]thymidine were taken up only by heterotrophic bacteria and that [ 14 C]bicarbonate was taken up only by microalgae. Microautoradiography has also been used in the study of nutrient competition between leaf epiphytes and plant pathogens (5), as well as in a comparative study of photoheterotrophy, chemoheterotrophy, and photolithotrophy in a eutrophic reservoir and an oligotrophic lake (8). In the present study, microautoradiographic methods were applied to study the uptake of labeled organic substrates by several filamentous microorganisms under different conditions of incubation.…”

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Application of microautoradiography to the study of substrate uptake by filamentous microorganisms in activated sludge

Andreasen¹,

Nielsen²

1997

Appl Environ Microbiol

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Excessive growth of filamentous microorganisms in activated-sludge treatment plants is a major operational problem which causes poor settlement of activated sludge. An enhanced understanding of the factors controlling growth of different filamentous microorganisms is necessary in order to establish more successful control strategies. In the present study, the in situ substrate uptake was investigated by means of microautoradiography. It was demonstrated that the uptake of labeled organic substrates by the filamentous microorganisms, during short-term incubation, could be detected by microautoradiography. Viability and respiratory activity of the filaments were also detected by reduction of CTC (5-cyano-2,3-ditolyl tetrazolium chloride) and by incorporation of [ 3 H]thymidine. Gram, Neisser, and fluorescence staining techniques were used for the localization and identification of the filaments. Activated-sludge samples from five wastewater treatment plants with bulking problems due to filamentous microorganisms were investigated. Microthrix parvicella, Nostocoida limicola, and Eikelboom's type 0041 and type 021N were investigated for their ability to take up organic substrates. A panel of six substrates, i.e., [ 14 C]acetate, [ 3 H]glucose, [ 14 C]ethanol, [ 3 H]glycine, [ 3 H]leucine, and [ 3 H]oleic acid, was tested. The uptake response was found to be very specific not only between the different filamentous types but also among filaments of the same type from different treatment plants. Interestingly, M. parvicella consistently took up only oleic acid among the tested substrates. It is concluded that microautoradiography is a useful method for investigation of in situ substrate uptake by filamentous microorganisms in activated sludge.

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Application of microautoradiography to the study of substrate uptake by filamentous microorganisms in activated sludge

Andreasen¹,

Nielsen²

1997

Appl Environ Microbiol

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“…Populations of the epiphytic bacterium Pseudomonas syringae were shown to be more limited by the availability of carbon than by the availability of nitrogen (56). Competition for limiting nutritional resources has been reported between bacteria and germinating fungal spores (9,10,18,42), between yeasts (8,22), and between yeasts and phytopathogenic fungi (17,23).…”

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Coexistence among Epiphytic Bacterial Populations Mediated through Nutritional Resource Partitioning

Wilson

Lindow

1994

Appl Environ Microbiol

249

117

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The levels of coexistence between Pseudomonas syringae and various nonpathogenic epiphytic species in the phyllosphere of beans (Phaseolus vulgaris) were assessed by using replacement series. The epiphytic species Pseudomonas fluorescens, Pantoea agglomerans, Stenotrophomonas maltophilia, and Methylobacterium organophilum were all capable of exhibiting higher levels of coexistence with P. syringae than was observed with a near-isogenic P. syringae strain pair. The ecological similarity of the epiphytes was estimated with niche overlap indices derived from in vitro carbon source utilization profiles. The level of coexistence of the epiphytes was inversely correlated with the ecological similarity of the strains. Hence, the level of coexistence between the epiphytes was proportional to the degree of niche differentiation, defined as the ability to utilize carbon sources not utilized by a competing strain. Comparisons of utilization profiles for groups of carbon sources (amino acids, organic acids, and carbohydrates) indicated the types of carbon sources for which the strains likely competed in the bean phyllosphere. P. fluorescens and P. syringae strains probably competed for most carbon sources. S. maltophilia and M. organophilum strains probably competed with P. syringae for most organic acids but few amino acids or carbohydrates. P. agglomerans strains probably competed with P. syringae for most amino acids and organic acids but few carbohydrates. A variable level of coexistence observed between P. agglomerans and P. syringae probably reflected the variability in abundance in the bean phyllosphere of the carbohydrates that P. agglomerans utilized exclusively.

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“…Although different methods have been developed for detection and determination of microorganisms on the leaf surface, so far, they have the disadvantage of being extremely time consuming. Moreover, they interfere with the microorganisms and may also be destructive or harmful to the leaf material (Langvad, 1980; Edwards and Blakeman, 1984). Thus, it would be advantageous to develop a non‐destructive method for the study of the trilateral interrelationship between plant, pathogen and protective microorganisms in situ .…”

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Non‐invasive determination of plant‐associated bacteria in the phyllosphere of plants

Gau¹,

Dietrich²,

Kloppstech³

2002

Environmental Microbiology

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Epiphytic living Pseudomonas strains isolated from different Malus domestica cultivars were transformed with two reporter genes [green fluorescent protein (gfp) and luciferase (luxAB)]. The establishment and distribution of these bacteria on sterile, in vitro-propagated, and thus genetically identical, Malus domestica plants were continuously analysed with a cooled, back-illuminated, charge-coupled-device (CCD) camera system. The combination of the assessment of bioluminescence and the use of a CCD camera offer an intriguing method to study, non-invasively and in real time, plant-microbe interactions as well as the colonization of the phyllosphere by microorganisms. Here we report on the applicability and sensitivity of the method with the goal to investigate quantitatively the interaction of symbiotic and pathogenic microorganisms with the corresponding host plant. It will be shown that the three bacterial isolates of the genus Pseudomonas studied, differ considerably with respect to their establishment on the host plants. It will also be shown that the chosen host apple variety has an impact on the activity of the bacterial cultivars. Analysis by a laser scanning fluorescence microscope provides the first evidence for the mode by which the epiphytic microorganisms interact with the plant.

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An autoradiographic method for determining nutrient competition between leaf epiphytes and plant pathogens

Cited by 7 publications

References 21 publications

Application of microautoradiography to the study of substrate uptake by filamentous microorganisms in activated sludge

Application of microautoradiography to the study of substrate uptake by filamentous microorganisms in activated sludge

Coexistence among Epiphytic Bacterial Populations Mediated through Nutritional Resource Partitioning

Non‐invasive determination of plant‐associated bacteria in the phyllosphere of plants

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