Specific primers for the differentiation of Heterobasidion annosum (s.str.) and H. parviporum infected stumps in northern Europe

Hantula, Jarkko; Vainio, Eeva J.

doi:10.14214/sf.500

Cited by 27 publications

(28 citation statements)

References 14 publications

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“…The success of the infection and colonization of tree tissues by H. parviporum was determined by a combination of two standard methods, i.e., either by re-isolating the viable pathogen or by PCR amplification of ribosomal ITS region (Hantula and Vainio 2003) from the collected tissue samples, which had been stored at −20°C for 2 years. In addition, the hyphal colonization in tissue samples from clones V330 and V375 was observed microscopically (see "Microscopic documentation of the infection process" below).…”

Section: Validation Of Infection and Detection Of The Pathogen In Infmentioning

confidence: 99%

“…cases where re-isolation of the fungus was unsuccessful, specific primers designed for the recognition of H. parviporum based on the amplification of a 350 base pairs (bp) fragment in the ribosomal ITS region by PCR (Hantula and Vainio 2003) were used to detect the presence of the fungus in the spruce tissues. To confirm that the isolated fungal strains were identical to the original strain, pairing tests were performed (Korhonen 1978).…”

Section: B) A)mentioning

confidence: 99%

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Infection of Picea abies clones with a homokaryotic isolate of Heterobasidion parviporum under field conditions

et al. 2015

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Heterobasidion parviporum Niemelä & Korhonen is responsible for the majority of decay in conifers in northern Europe, which causes severe economic losses. In nature, heterokaryotic isolates of H. parviporum cause infection in Norway spruce (Picea abies (L.) Karst.). However, little is known on whether homokaryons of H. parviporum can infect trees under field conditions. In this study, 40-year-old clonal Norway spruce stems and roots were inoculated with a homokaryotic isolate of H. parviporum under field conditions. After four months, the infection frequency and necrotic lesion lengths were recorded. The homokaryon caused infection and provoked the development of necrotic lesions. Necrotic lesions were larger in roots than in stems. Among the studied Norway spruce genotypes, a Russian clone had the smallest necrotic lesions, whereas a Finnish clone developed the largest necrotic lesions. Clones with higher growth rates were more sensitive to fungal infection and wound damage. Under microscopic observation, H. parviporum grew adpressed to lumen cell walls, colonized tracheids next to rays, and induced lignification in cell walls close to the point of inoculation. This study provides a starting point for further studies on the ability of homokaryons to cause infection under field conditions and for discussions on factors affecting the resistance of Norway spruce against H. parviporum.Résumé : Chez les conifères d'Europe du Nord, la majeure partie de la carie qui est responsable d'importantes pertes économiques est causée par Heterobasidion parviporum Niemelä & Korhonen. En nature, des isolats hétérocaryotes de H. parviporum infectent l'épicéa commun (Picea abies (L.) Karst.). Cependant, on ne sait pas si des homocaryons de H. parviporum peuvent infecter les arbres sur le terrain. Dans cette étude, les racines et la tige d'individus clonaux d'épicéa commun âgés de 40 ans ont été inoculés avec un isolat homocaryote de H. parviporum sur le terrain. Après quatre mois, la fréquence des infections et la longueur des lésions nécrotiques ont été notées. L'homocaryon a causé une infection et provoqué le développement de lésions nécrotiques. Les lésions nécrotiques étaient plus grandes sur les racines que sur la tige. Parmi les génotypes d'épicéa commun étudiés, un clone russe avait les lésions nécrotiques les plus petites tandis que les plus grandes lésions se retrouvaient sur un clone finlandais. Les clones qui avaient le taux de croissance le plus élevé étaient plus sensibles aux infections fongiques et aux dommages causés par les blessures. Sous observation microscopique, H. parviporum croissait adossé à la paroi cellulaire à l'intérieur des cellules, colonisait les trachéides adjacentes aux rayons et induisait la lignification dans la paroi des cellules situées près du point d'inoculation. Cette étude constitue un point de départ pour d'autres études portant sur la capacité des homocaryons à causer des infections sur le terrain et pour alimenter la discussion concernant les facteurs qui influencent la résistance de l'ép...

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Section: Validation Of Infection and Detection Of The Pathogen In Infmentioning

confidence: 99%

Section: B) A)mentioning

confidence: 99%

Infection of Picea abies clones with a homokaryotic isolate of Heterobasidion parviporum under field conditions

et al. 2015

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“…Specific PCR tests have been developed for M. graminicola (Septoria tritici) in wheat (27) and to distinguish M. fijiensis and M. musicola from banana leaves (40). PCR detection may target specific genes or anonymous products, such as microsatellites, random amplified polymorphic DNA, or amplified fragment length polymorphism fragments (35,67,68,72). In order to provide simultaneous discrimination for several Mycosphaerella spp.…”

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confidence: 99%

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

Glen¹,

Smith²,

Langrell³

et al. 2007

Phytopathology®

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Glen, M., Smith, A. H., Langrell, S. R. H., and Mohammed, C. L. 2007. Development of nested polymerase chain reaction detection of Mycosphaerella spp. and its application to the study of leaf disease in Eucalyptus plantations. Phytopathology 97:132-144.Mycosphaerella leaf disease (MLD) is a serious disease of two of the major eucalypt species grown in temperate regions worldwide, Eucalyptus globulus and E. nitens. More than 30 species of Mycosphaerella have been reported on eucalypts worldwide. Accurate, rapid, and early discrimination of Mycosphaerella spp. causing crown damage to E. globulus and E. nitens will assist the development of sustainable management strategies. This study describes the development, and incorporation in a nested polymerase chain reaction (PCR) approach, of specific primers for the detection and identification of Mycosphaerella spp. commonly reported from leaf lesions of E. globulus and E. nitens in Australia. Primer design was assisted by sequence alignment and phylogenetic analysis of 165 nonredundant sequences from the nuclear ribosomal DNA internal transcribed spacer regions of Mycosphaerella and related species. Phylogenetic analysis revealed very high sequence similarity for two taxon groups, Mycosphaerella grandis and M. parva, and M. vespa, M. ambiphylla, and M. molleriana, and primers were designed to differentiate each of the two groups. Three other species, M. cryptica, M. nubilosa, and M. tasmaniensis, were distinct and distinguished by species-specific primers. In double-blind trials, the detection test accurately and rapidly identified Mycosphaerella spp. in cultures and discriminated against other pathogens that co-occur in or on Eucalyptus leaves, thereby verifying its reliability. The detection test has an internal amplification control in the first-round PCR with fungal-specific primers to raise confidence in test results, particularly to highlight negative results due to PCR inhibition. When applied to DNA extracted from leaf or stem samples either as multiple or single lesions, it detected and identified up to five Mycosphaerella spp. or taxon groups in both positively identified and in young (putative) MLD lesions. The samples were 20 mm 2 or larger in surface area and were collected while undertaking disease rating assessments in an experimental investigation of Eucalyptus plantations and regrowth forest. Using nested PCR detection, Mycosphaerella spp. were positively identified in 2 days, 1 to 5 months earlier than by classical methods, demonstrating the potential application of this detection test to the early discrimination of MLD components in ecological, epidemiological, and genetic investigations.

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“…Although the use of DNA‐based specific probes was initially met with skepticism (Schulze and Bahnweg 1998), such technology is becoming an accepted tool in diagnostic laboratories (Garbelotto 2004). Hantula and Vainio (2003) developed specific molecular probes to differentiate Heterobasidion annosum from H. parviporum directly from infected stumps in northern Europe. Sicoli et al.…”

Section: Applications Of Dna Sequencing Technology Of Wood‐decay Fungimentioning

confidence: 99%

Use of fungal biosystematics and molecular genetics in detection and identification of wood‐decay fungi for improved forest management

Glaeser

Lindner

2010

Forest Pathology

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Advances in fungal biosystematics and molecular genetics have clarified relationships among the wood-decay fungi and are providing new tools for their detection and identification. Species complexes of forest pathogens, including those within Heterobasidion, Armillaria, Laetiporus, and Phellinus, are being resolved. The ability to isolate fungal DNA directly from wood without in intermediate culturing step will greatly facilitate sampling and disease detection and has applications in forest disease management, hazard tree assessment, invasive species detection, and carbon cycling, sequestration and climate change research. Recent changes in fungal nomenclature and their application to forest pathology are discussed.Forest pathologists have been interested in wood-decay fungi since Hartig published his first treatise on forest pathogens in the late 1800s (Hartig 1894). Heart-rot and root-rot fungi are among the most devastating of forest pathogens, while saprotrophic decay fungi play an essential role in forest health by recycling nutrients, providing animal habitat, and creating coarse woody debris that breaks down into humic substances, thus improving soil quality. Forestry schools have traditionally included the identification of decay fungi in their curriculum, so forest pathologists generally have a good working knowledge of the major decay fungi found in their region. Pathologists are often frustrated by what seems like the never-ending revision of fungal names by mycologists for what may seem like frivolous or arbitrary reasons. This is especially difficult when longestablished names of pathogens are revised, such as Phellinus pini or Fomes annosus. The objective of this review is to familiarize forest pathologists with recent advances in fungal biosystematics and show how the development of molecular tools and a better understanding of fungal phylogeny can help forest pathologists make better assessments and recommendations for overall improvement of forest health.

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Specific primers for the differentiation of Heterobasidion annosum (s.str.) and H. parviporum infected stumps in northern Europe

Cited by 27 publications

References 14 publications

Infection of Picea abies clones with a homokaryotic isolate of Heterobasidion parviporum under field conditions

Infection of Picea abies clones with a homokaryotic isolate of Heterobasidion parviporum under field conditions

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

Use of fungal biosystematics and molecular genetics in detection and identification of wood‐decay fungi for improved forest management

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