Use of Polymerase Chain Reaction to Detect<i>Gaeumannomyces graminis</i>DNA in Plants Grown in Artificially and Naturally Infested Soil

Henson, Joan M.; Goins, Tresa; Grey, W. E.; Mathre, D. E.; Elliott, Monica L.

doi:10.1094/phyto-83-283

Cited by 51 publications

(23 citation statements)

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“…For example, sequence data from a cloned mitochondrial (mt) DNA fragment specific to Gaeumannomyces graminis facilitated primer design for fungal DNA amplification in infected wheat or bermudagrass (63). Primers have also been based on pathogen-specific plasmid sequences, such as those used to develop PCR diagnostic assays for Agrobacterium tumefaciens (40), Erwinia amylovora (12), and Xanthomonas campestris pv.…”

Section: < Zmentioning

confidence: 99%

“…Rollo et al (118) used boiled mycelium collected from infected lemon trees to detect the fungal pathogen Phoma tracheiphila, and boiling infected barley leaf tissue was sufficient to detect specific DNA sequences of Pyrenophora teres (B. Baltazar, A. Scharen, V. Raboy, unpublished data). Boiled wheat roots or crowns infected with G. graminis, infested oat seeds used as field inoculum, or even a single, boiled ascospore (containing 100-1200 copies of mitochondrial DNA target molecules) apparently released enough target mtDNA to produce visible amplified products on agarose gels (63).…”

Section: Sample Preparationmentioning

confidence: 99%

“…Amplified products are not produced from samples containing up to 40 ascospores when 1 gm of soil is added. Yet samples with entire perithecia containing up to 100 ascospores are positive, indicating that even soil inhibition is overcome if large numbers of target molecules are present in the sample (63).…”

Section: Sample Preparationmentioning

confidence: 99%

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The Polymerase Chain Reaction and Plant Disease Diagnosis

Henson¹,

French²

1993

Annu. Rev. Phytopathol.

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387

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Section: < Zmentioning

confidence: 99%

Section: Sample Preparationmentioning

confidence: 99%

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The Polymerase Chain Reaction and Plant Disease Diagnosis

Henson¹,

French²

1993

Annu. Rev. Phytopathol.

Self Cite

387

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“…However, identification of isolates of the G-P complex by conventional methods is time consuming, and on occasions, inconclusive. As a result of these problems, different molecular approaches have been developed for identification (Henson, 1989 ;Schesser et al, 1991 ;Bateman et al, 1992 ;OhDell et al, 1992 ;Ward & Gray, 1992 ;Henson et al, 1993 ;Tan et al, 1994 ;Ward & Akrofi, 1994 ;Fouly et al, 1997). In recent years, the use of random amplified polymorphic DNA (RAPD) has provided a fast and reproducible tool to distinguish species and varieties of Gaeumannomyces, and were shown to be in complete correspondence with the differentiation by other molecular methods (Fouly et al, 1996 ;Wetzel et al, 1996 ;Augustin et al, 1999 ;Bryan et al, 1999).…”

Section: mentioning

confidence: 99%

Identification and characterization of a new group of root‐colonizing fungi within the Gaeumannomyces–Phialophora complex

2000

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A new group of darkly pigmented root-infecting fungi was isolated from cereal roots obtained from six different locations in northeastern Germany. Similar random amplified polymorphic DNA (RAPD) patterns and restriction profiles of amplified rDNA were used as a basis for classifying the isolates in a separate group. The isolates demonstrating mycelial and infection characteristics typical of Gaeumannomyces graminis could be differentiated from the varieties of G. graminis as well as from Gaeumannomyces cylindrosporus\Phialophora graminicola using RAPD Polymerase chain reaction (PCR) and rDNA Restriction-fragment length polymorphism (RFLP) analysis. Phylogenetic analysis of the Internal transcribed spacer (ITS) regions suggests that the isolates form a distinct group (named group ' E') situated within the Gaeumannomyces-Phialophora complex between the branch of the G. graminis varieties and Gaeumannomyces incrustans\Magnaporthe poae. Isolates of group E produced lobed hyphopodia and were shown in biotests to be non-pathogenic to wheat, oats, Italian Ryegrass and Chewings Fescue, suggesting it is a benign parasite which colonizes cereals or grasses without destroying vascular tissue. Furthermore, curved phialospores could be found. Summarizing the results presented, this new group could be classified as a new species of Phialophora. Although isolates of group E were found at only six of the 32 investigated locations, they composed up to 50% of total isolates of the Gaeumannomyces-Phialophora complex at these sites. Because of the non-pathogenic behaviour, the new group may be of value as biological control agents for pathogenic fungi.

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“…Specific primers for PCR have been used effectively to detect and differentiate plant pathogenic fungi (e.g. Henson et al 1993;Ouellet and Seifert 1993).…”

Section: Introductionmentioning

confidence: 99%

Differentiation of entomopathogenic fungus Beauveria bassiana (Ascomycetes: Hypocreales) isolates by PCR-RFLP

Sabbahi

Lavallée

Merzouki

et al. 2010

phyto

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The entomopathogenic fungus Beauveria bassiana is a promising biological control agent of several insect pests in agriculture. Molecular approaches (PCR, DNA sequence analysis and PCR-RFLP) were used in our research as tools for the identification of different B. bassiana isolates. Our work consisted in identifying the 18S, ITS1, 5.8S, ITS2 and 28S regions of B. bassiana ribosomal DNA. The DNA sequences of the amplified regions showed that the 18S rDNA is the most conserved unit, with a high homology (99.5%) between the isolates studied, while the 3’ end of the 28S rDNA has a great variability, which makes it possible to differentiate the isolates. The PCR-RFLP method was used to monitor isolates of B. bassiana and distinguish them in a target pest, Lygus lineolaris. This method involved two main steps. First, PCR was used to amplify a region of the 28S gene of B. bassiana. Second, this PCR product was digested using restriction endonucleases, and the fragments produced were compared using gel electrophoresis. Because of the high specificity and sensitivity of PCR-RFLP, it was possible to discriminate between B. bassiana isolates using spores scraped from the surface of an infected insect as samples.Le champignon entomopathogène Beauveria bassiana suscite de plus en plus d’intérêt en recherche et constitue une avenue intéressante en lutte biologique contre plusieurs insectes ravageurs en agriculture. Différentes approches (PCR, analyse des séquences d’ADN et PCR-RFLP) ont été utilisées lors de cette étude comme outils moléculaires d’identification de différents isolats de B. bassiana. Notre travail a consisté à identifier les régions 18S, ITS1, 5.8S, ITS2 et 28S de l’ADN ribosomal de B. bassiana. Les séquences d’ADN des régions amplifiées ont démontré que la région 18S de l’ADNr était la sous-unité la plus conservée, avec une homologie de 99,5 % entre les isolats étudiés, tandis que l’extrémité 3’ du gène 28S a accumulé beaucoup de variabilité et peut donc être utilisée pour différencier les isolats de B. bassiana. La technique PCR-RFLP a été utilisée pour réaliser le suivi d’isolats de B. bassiana chez un ravageur ciblé, Lygus lineolaris, et pour les distinguer. Cette méthode comprenait deux étapes. Premièrement, la PCR était utilisée pour amplifier une région du gène 28S de B. bassiana. Deuxièmement, ce produit de PCR était digéré à l’aide des endonucléases de restriction et les fragments produits ont été comparés en utilisant l’électrophorèse sur gel. En raison de la grande spécificité et sensibilité de la PCR-RFLP, il a été possible de différencier les isolats de B. bassiana en utilisant comme échantillons des spores prélevées à la surface d’un insecte infecté

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Use of Polymerase Chain Reaction to DetectGaeumannomyces graminisDNA in Plants Grown in Artificially and Naturally Infested Soil

Cited by 51 publications

References 0 publications

The Polymerase Chain Reaction and Plant Disease Diagnosis

The Polymerase Chain Reaction and Plant Disease Diagnosis

Identification and characterization of a new group of root‐colonizing fungi within the Gaeumannomyces–Phialophora complex

Differentiation of entomopathogenic fungus Beauveria bassiana (Ascomycetes: Hypocreales) isolates by PCR-RFLP

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