Production of monoclonal antibodies against surface antigens of spores from arbuscular mycorrhizal fungi by an improved immunization and screening procedure

Hahn, Alexander; Bonfante, Paola; Horn, Katrin; Pausch, Friederike; Hock, Bertold

doi:10.1007/bf00204061

Cited by 30 publications

(10 citation statements)

References 18 publications

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“…Growth stimulation of AMF by bacteria of the Bacillus group, e.g., by Bacillus mycoides (21) or Paenibacillus (10), has been described previously. The stimulation of mycorrhizal colonization observed by Budi et al (10) and the promotion of hyphal growth up to spore formation (this report) may be based on the same chemical factors produced by Paenibacillus.…”

Section: Vol 68 2002 Towards Growth Of Am Fungi Independent Of a Plmentioning

confidence: 84%

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Hildebrandt

Janetta

Bothe

2002

Appl Environ Microbiol

128

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When surface-sterilized spores of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices Sy167 were germinated on agar plates in the slightly modified minimum mineral medium described by G. Bécard and J. A. Fortin (New Phytol. 108:211-218, 1988), slime-forming bacteria, identified as Paenibacillus validus, frequently grew up. These bacteria were able to support growth of the fungus on the agar plates. In the presence of P. validus, hyphae branched profusely and formed coiled structures. These were much more densely packed than the so-called arbuscule-like structures which are formed by AMF grown in coculture with carrot roots transformed with T-DNA from Agrobacterium rhizogenes. The presence of P. validus alone also enabled G. intraradices to form new spores, mainly at the densely packed hyphal coils. The new spores were not as abundant as and were smaller than those formed by AMF in the monoxenic culture with carrot root tissues, but they also contained lipid droplets and a large number of nuclei. In these experiments P. validus could not be replaced by bacteria such as Escherichia coli K-12 or Azospirillum brasilense Sp7. Although no conditions under which the daughter spores regerminate and colonize plants have been found yet, and no factor(s) from P. validus which stimulates fungal growth has been identified, the present findings might be a significant step forward toward growth of AMF independent of any plant host.The roots of more than 80% of land plants are colonized by arbuscular mycorrhizal fungi (AMF). Under diverse stress conditions, the hyphae of the fungi exploit water and minerals from soils better than the roots and effectively transfer them to the plant macrosymbiont. Thus, arbuscular mycorrhiza is likely the most important symbiosis in nature. Its study is hampered by the fact that AMF are obligate biotrophs (19,20). However, as originally described by B. Mosse and C. M. Hepper (25,26) and subsequently elaborated in much more detail by others, some AMF, such as Gigaspora spp. and Glomus intraradices, can fulfil their life cycles up to spore formation when grown in a dual system with root tissues from either carrots (5), tomatoes (3), or clover (13). The root factors which enable the AMF to grow in this monoxenic culture have not yet been fully elucidated. When spores are placed on agar containing medium only, they germinate and form so-called runner hyphae. After some weeks, however, this growth stops, the hyphae septate, and the cytoplasm is retracted. In the monoxenic culture with carrot roots, completion of the life cycle of AMF requires the synthesis of lipids which are deposited into the newly formed spores (4) and CO 2 fixations for anabolism (6). These reactions are apparently turned on by unidentified factors produced and excreted into the medium by the plant tissues before any contact of the hyphae with a root tissue cell.Numerous investigations have indicated that rhizosphere bacteria have strong impacts on the growth of AMF (1, 2, 18). The so-called plant growth-promoting rhizoba...

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Section: Vol 68 2002 Towards Growth Of Am Fungi Independent Of a Plmentioning

confidence: 84%

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Hildebrandt

Janetta

Bothe

2002

Appl Environ Microbiol

128

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“…To address this limitation, a wide variety of tools have been examined as means for the reliable and accurate detection of the presence of multiple AMF isolates. Some of these include antibodies (21), isozyme patterns (26,46), lipid profiles (30), and PCR (13,34,38). Of these, conventional PCR-based methods have been acknowledged to be the best because they are sensitive, relatively easy to apply, invariant, and highly robust.…”

mentioning

confidence: 99%

Analysis of Quantitative Interactions between Two Species of Arbuscular Mycorrhizal Fungi,Glomus mosseaeandG. intraradices, by Real-Time PCR

Alkan

Gadkar

Yarden

et al. 2006

Appl Environ Microbiol

104

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Arbuscular mycorrhizal fungi (AMF) are obligate biotrophs, known to play an important role in ecological processes. Conventional light microscopy is the most common method used to detect their presence in planta, but this method fails to discern the presence of multiple AMF species and is not quantitative. These two factors are critically important in ecological studies, where the symbiotic contribution of each isolate needs to be defined. This paper describes the use of quantitative real-time PCR (qRT-PCR) as a detection system to address this issue. We used two Glomus spp., namely, G. intraradices and G. mosseae, to show that it is possible to study the interactions between these two isolates during the cocolonization of a single root system. Three different physiological studies were set up to assess how the interactions affected the occupancy of these fungi intraradically on a temporal basis. These treatments included saline and phosphorus stress, spatial distribution in the root zone, and preference for a particular host. qRT-PCR could prove a valuable tool in the area of AMF field ecology, where such data are critically important for defining the role of each species in the community structure.

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“…Approaches using isozymes, antibodies and DNA probes have been, or are being, explored but they have met with limited success, largely due to their low specificity (Hepper et al, 1988;Rosendahl, 1989;Rosendahl and Sen, 1992;Hahn et al, 1993;Cordier et al, 1994Cordier et al, , 1996Giovannetti and Gianinazzi-Pearson, 1994;Sanders et al, 1996). An alternative approach could be through the identification of taxon-specific proteins that, after purification, can be used for antibody production or sequencing to produce specific oligonucleotide probes.…”

Section: Discussionmentioning

confidence: 99%

Soluble proteins and polypeptide profiles of spores of arbuscular mycorrhizal fungi. Interspecific variability and effects of host (myc+) and non-host (myc^-) Pisum sativum root exudates

Samra

Dumas‐Gaudot²,

Gianinazzi‐Pearson³

et al. 1996

Agronomie

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Production of monoclonal antibodies against surface antigens of spores from arbuscular mycorrhizal fungi by an improved immunization and screening procedure

Cited by 30 publications

References 18 publications

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Analysis of Quantitative Interactions between Two Species of Arbuscular Mycorrhizal Fungi,Glomus mosseaeandG. intraradices, by Real-Time PCR

Soluble proteins and polypeptide profiles of spores of arbuscular mycorrhizal fungi. Interspecific variability and effects of host (myc+) and non-host (myc^-) Pisum sativum root exudates

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Production of monoclonal antibodies against surface antigens of spores from arbuscular mycorrhizal fungi by an improved immunization and screening procedure

Cited by 30 publications

References 18 publications

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Towards Growth of Arbuscular Mycorrhizal Fungi Independent of a Plant Host

Analysis of Quantitative Interactions between Two Species of Arbuscular Mycorrhizal Fungi,Glomus mosseaeandG. intraradices, by Real-Time PCR

Soluble proteins and polypeptide profiles of spores of arbuscular mycorrhizal fungi. Interspecific variability and effects of host (myc+) and non-host (myc-) Pisum sativum root exudates

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Soluble proteins and polypeptide profiles of spores of arbuscular mycorrhizal fungi. Interspecific variability and effects of host (myc+) and non-host (myc^-) Pisum sativum root exudates