Identification of the biological species of Armillaria associated with Wynnea and Entoloma abortivum using PCR-RFLP analysis of the intergenic region (IGR) of ribosomal DNA

Fukuda, Masaki; Nakashima, Etsuko; Hayashi, Kyoko; Nagasawa, Eiji

doi:10.1017/s0953756203008633

Cited by 17 publications

(17 citation statements)

References 18 publications

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“…Those results matched with those found by Fukuda et al 29) The sequencing of the IGS-1 region demonstrated a perfect match between them and with the Japanese A. nabsnona tester strain. Comparison of our isolates with other A. nabsnona deposited in the GenBank indicated a deletion event consisted of 22 bp, between the 430th and 451st position from the 3Ј end of the LR12R primer sequence.…”

Section: Discussionsupporting

confidence: 90%

Identification of Armillaria nabsnona in Gastrodia Tubers

Sekizaki

Kuninaga

Yamamoto

et al. 2008

Biological & Pharmaceutical Bulletin

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Armillaria species (Basidiomycota, Agaricales, Tricholomataceae) are well known to be the cause of root rot and death to woody plants in boreal, temperate and tropical regions worldwide.1) Generally this species has a broad variety of hosts, presenting pathogenic or symbiotic properties depending on the available substrate. 1,2)Armillaria has been identified as an associate in certain achlorophyllous taxa of Orchidaceae including Gastrodia elata BL. 3,4) and Galeola septentrionalis REICHB. 5) Although many reports refer to Armillaria mellea as the main fungus symbiont of G. elata, 3,6) 4)In eastern traditional medicine, the tubers of this orchid have been used for the treatment of many kinds of diseases including headache, rheumatism, seizure, dizziness and circulation problems. 7,8) G. elata is considered as a difficult species to cultivate due to the need for different kinds of fungi colonization to achieve its full development.6) To increase the knowledge of the cultivation process and understand its relationship with the Armillaria, it is important to first identify which Armillaria species is involved in the association.In past decades, the identification of Armillaria spp. was based on mating tests and morphological characteristics. 2,9,10) However, those methods are very limited due to the great number of biological species and similar morphological characteristics among them.11,12) Furthermore, mating pairings require fresh haploid cultures that sometimes are not available and a long period is required to obtain results. 13,14) Methods based on molecular techniques are nowadays routinely used and provided a new perspective for phylogenetic and taxonomic studies.12,15-17) PCR-RFLP analysis profiles of intergenic spacer 1 (IGS-1) have been successfully applied to the Armillaria species identification. 12,13,18,19) Sequencing of the internal transcribed spacer (ITS) and intergenic spacer 1 (IGS-1) regions 14,20,21) has also been used for identification. The ITS and IGS sequences available in the GenBank and the development of efficient software have provided a valuable tool for differentiation as well as for the detection of polymorphism and heterogeneity among species. The ITS and IGS-1 of the ribosomal DNA are known to be highly polymorphic enabling the accurate species differentiation of the Armillaria.11) In some cases, however, ITS1 region sequences were proven to be nearly uniform among the Armillaria of northern hemisphere species. 11)The aim of this study was to apply molecular biological tools to identify which Armillaria species are involved in the association with G. elata. The ITS and IGS-1 regions of rDNA were amplified and sequenced for further comparison with other Armillaria species. PCR-RFLP of the IGS-1 region was performed with three different endonucleases. Morphological characteristics and compatibility tests were also used in the analysis. MATERIALS AND METHODS Fungal Isolation and CultivationFungal samples were harvested in July 2005, found in different root parts of G. elata, which ...

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Section: Discussionsupporting

confidence: 90%

Identification of Armillaria nabsnona in Gastrodia Tubers

Sekizaki

Kuninaga

Yamamoto

et al. 2008

Biological & Pharmaceutical Bulletin

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“…et Curt.) in Japan were identified using PCR-restriction fragment length polymorphism (RFLP) analysis of the intergenic region (IGR) of ribosomal DNA (Fukuda et al 2003). In the Armillaria-E. abortivum and ArmillariaWynnea association, it is uncertain what kind of relationship exists between them.…”

Section: Discussionmentioning

confidence: 99%

Identification of Armillaria species associated with Polyporus umbellatus using ITS sequences of nuclear ribosomal DNA

Kikuchi

Yamaji

2010

Mycoscience

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The sclerotia of Polyporus umbellatus were collected from three locations in Japan and three locations in China. All the collected sclerotia were adhered to by rhizomorphs of the symbionts. When the sclerotium of P. umbellatus was cross sectioned, the internal part of the sclerotium was cream colored, and many black regions surrounding the invading rhizomorphs were observed. The surrounding zone contained string-like, gelatinous masses composed of hyphae, and its outside was brown in color. All isolates were similar in colony morphology and grew well on PDA medium with well-developed rhizomorphs. All the isolates showed typical morphology of Armillaria. The isolated fungi were identified via the ITS region of the nuclear ribosomal DNA sequence. Phylogenetic analysis based on the neighbor-joining method showed that all the isolates clustered with fungi belonging to Armillaria species. Among them, five species (A. sinapina, A. calvescens, A. gallica, A. cepistipes, and A. nabsnona) and the symbiont formed a highly supported clade. We report on the case where Armillaria has a relationship in the sclerotium of Polyporus umbellatus.

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“…the internal transcribed spacer (ITS) and intergenic spacer (IGS) (Anderson and Stasovski 1992;Harrington and Wingfield 1995;Banik et al 1996;Terashima et al 1998a,b;Pérez-Sierra et al 1999;Fukuda et al 2003;Lochman et al 2004a;Matsushita and Suzuki 2005;Schnabel et al 2005;Keča et al 2006;Sekizaki et al 2008). More recently the translation elongation factor-1 alpha (EF-1 alpha) gene, which encodes an essential part of the protein translation machinery, was shown to have high phylogenetic utility and has been used in studies on fungi.…”

Section: Introductionmentioning

confidence: 99%

Populations of Armillaria species in pine plantations in west-central Poland

2015

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Two diploid isolates of Armillaria gallica and 143 diploid isolates of A. solidipes were obtained from 145 samples of rhizomorphs, fruit bodies and infected wood of sessile oak and Scots pines from 5-10-year-old Scots pine plantations in three Forest Districts located 100-350 km apart in west-central Poland. Based on pairings among the 145 isolates, 18 somatic compatibility groups (genets) of A. solidipes were distinguished in the three plantations. Sequencing of ITS1/2 rDNA of 18 isolates representing the 18 genets delineated four nuclear haplotypes. This suggests that many of the A. solidipes genets are closely related and were possibly established by sib-related basidiospores. With a few exceptions there was general geographical specialization of genets but not haplotypes. Geographical specialization of genets and the dominance of single haplotypes suggest that the A. solidipes population results from clonal rather than sexual reproduction. Sequencing of the ITS1/2 and IGS-1 rDNA showed small nucleotide diversity in ITS1/2 rDNA and much more diversity in IGS-1 rDNA of the isolates of A. solidipes studied. However, none of these regions has sufficient resolution for the clear differentiation of A. solidipes from A. borealis. The sequences of the EF 1-alpha gene showed high interspecific variability in Armillaria species and very low intraspecific variability in A. solidipes. This gene is the most appropriate for reliable identification of biological species and subgroups of Armillaria. Its application is the most useful in ecological and epidemiological studies of Armillaria.

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Identification of the biological species of Armillaria associated with Wynnea and Entoloma abortivum using PCR-RFLP analysis of the intergenic region (IGR) of ribosomal DNA

Cited by 17 publications

References 18 publications

Identification of Armillaria nabsnona in Gastrodia Tubers

Identification of Armillaria nabsnona in Gastrodia Tubers

Identification of Armillaria species associated with Polyporus umbellatus using ITS sequences of nuclear ribosomal DNA

Populations of Armillaria species in pine plantations in west-central Poland

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