Bacteria in thin soil sections stained with the fluorescent brightener calcofluor white M2R

Postma, J.; Altemüller, Hans-Jürgen

doi:10.1016/0038-0717(90)90065-8

Cited by 51 publications

(16 citation statements)

References 10 publications

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“…Most suitable object for study is here the rhizoplane, thus excluding the loosely associated rhizosphere soil surrounding the root. Alternatively, the porous matrix of the rhizosphere soil (or bulk soil) may actually be accessible by SEM, provided the preparation includes imbedding in a resin, followed by thin-sectioning of the specimen (Postma and Altemüller 1990). Classical studies in the 1950´es were pioneering for direct observations of single microbial cells on root surfaces (Rovira 1956).…”

Section: Traditional and Environmental Semmentioning

confidence: 99%

“…Where this is a problem, a useful DNA stain is the green fluorescent SYBR Green II (Weinbauer et al 1998). Finally, a useful stain is also Fluorescent Brightener (FB) 28 (sometimes referred to as Calcofluor White M2R), applied for wall staining of fungi in root-soil microcosms (Thrane et al 1999) and imbedded thin-sections of soil (Postma and Altemüller 1990;Eickhorst and Tippkötter 2008). A more recent Fluorescent Brigthener Agent (FBA) 220 may actually improve staining of the soil thin-sections since crystallization problems can be avoided (Harris et al 2002).…”

Section: General Cell Stainsmentioning

confidence: 99%

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Molecular tools in rhizosphere microbiology—from single-cell to whole-community analysis

et al. 2009

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It is the aim of this chapter to present an overview of new, molecular tools that have been developed over recent years to study individual, single cells and composite, complex communities of microorganisms in the rhizosphere. We have carefully focused on culture-independent assays and selected methodologies that have already been or will soon be applicable for rhizosphere microbiology. Emphasis is placed on rhizosphere bacteria and the review first describes a number of the new methodologies developed for detection and localization of specific bacterial populations using modern electron and fluorescence microscopy combined with specific tagging techniques. First half of the chapter further comprises a thorough treatise of the recent development of reporter gene technology, i.e. using specific reporter bacteria to detect microscale distributions of rhizosphere compounds such as nutrients, metals and organic exudates or contaminants. Second half of the chapter devoted to microbial community analysis contains a thorough treatise of nucleotide-and PCRbased technologies to study composition and diversity of indigenous bacteria in the natural rhizosphere. Also included are the most recent developments of functional gene and gene expression analyses in the rhizosphere based on specific mRNA transcript or transcriptome analysis, proteome analysis and construction of metagenomic libraries.

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Section: Traditional and Environmental Semmentioning

confidence: 99%

Section: General Cell Stainsmentioning

confidence: 99%

Molecular tools in rhizosphere microbiology—from single-cell to whole-community analysis

et al. 2009

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“…The inoculated plates were sealed with Parafilm2, incubated at 20 mC and inspected regularly for bacterial and fungal colonies. Fungi were also quantified microscopically by measuring hyphal length after staining with Calcofluor White M2R2 (Sigma) which has been recommended as an excellent fluorochrome for the study of filamentous fungi in soil (Postma & Altemu$ ller, 1990). An aliquot of the shaken and sonicated soil suspensions (0n5 ml) was added to 0n5 ml of filter-sterile Calcofluor White solution in demineralized water (2n3 mg ml −" ) and incubated for 30 min at 20 mC.…”

Section: Microbiological Analyses Of Soil Samplesmentioning

confidence: 99%

Suppression of hyphal growth of soil‐borne fungi by dune soils from vigorous and declining stands of Ammophila arenaria

1998

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A study was carried out to determine whether expansion of marram-grass stands (Ammophila arenaria (L.) Link) on acidic inner Dutch coastal dunes was caused by suppressiveness of soils from these stands against three potential pathogenic fungi of marram grass, namely Fusarium culmorum (W. G. Sm.) Sacc., Phoma exigua Desm. and a Ulocladium sp. The suppressiveness of the acidic inner dune soils was compared with that of lime-rich dune soils from vigorous and declining marram-grass stands. Suppressiveness of the dune soils against the saprotrophic fungi Chaetomium globosum Kunze : Fr, Mucor hiemalis Wehmer and Trichoderma harzianum Rifai was also determined. All fungi had been isolated from marram-grass roots. Suppressiveness was determined by comparing the formation of hyphae from potato-dextrose agar discs into (layer method) or on top of (surface method) dune soils with that of controls consisting of sterile, acid-washed beach sand. The growth of the three root-infecting fungi was strongly inhibited in all soils regardless of the method used. Hence, there were no indications that the potential pathogenic fungi were selectively suppressed by the acidic dune soils and, consequently, the results did not give any indication for the involvement of a fungal component in the decline of marram grass. Growth of the saprotrophs C. globosum and M. hiemalis was much less inhibited than that of the root-infecting fungi. Growth of T. harzianum was strongly inhibited in alkaline soils but not in the acid ones. The suppression of fungal growth could be partly or completely eliminated by a microwave treatment, indicating that biological components of the soil were essential to suppressiveness. The suppression of the fungi by colonies of dune soil micro-organisms on water-agar differed considerably from soil alone. Yet, all methods indicated the occurrence of general suppressiveness against fungi by dune soils, irrespective of the origin of the soil samples. This suppressiveness was probably not due to direct competition with other soil micro-organisms for nutrients but to inhibiting compounds produced by the soil micro-organisms.

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“…CFW (fluorescent salt of 1-anilino-8-naphthalene sulfonic acid), and FM1- Altemüller, 1990;Weaver and Zibilske, 1975;Nadeau et al, 2008) for this purpose. The cell membranes or cell walls surrounding the "cells" will be detected with this type of dye, which indicates the boundary between the inside and outside of the "cell".…”

Section: Fluorescence Microscopic Detection Of Lifementioning

confidence: 99%

Japan Astrobiology Mars Project (JAMP): Search for Microbes on The Mars Surface with Special Interest in Methane-Oxidizing Bacteria

et al. 2010

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Bacteria in thin soil sections stained with the fluorescent brightener calcofluor white M2R

Cited by 51 publications

References 10 publications

Molecular tools in rhizosphere microbiology—from single-cell to whole-community analysis

Molecular tools in rhizosphere microbiology—from single-cell to whole-community analysis

Suppression of hyphal growth of soil‐borne fungi by dune soils from vigorous and declining stands of Ammophila arenaria

Japan Astrobiology Mars Project (JAMP): Search for Microbes on The Mars Surface with Special Interest in Methane-Oxidizing Bacteria

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