Identification of Armillaria species from Hokkaido by analysis of the intergenic spacer (IGS) region of ribosomal DNA using PCR-RFLP

Abstract

The intergenic spacer (IGS) region, which is located between the 3' end of 26S ribosomal DNA (rDNA) and the 5' end of 5S rDNA, of six Armillaria species from Hokkaido was investigated using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). Restriction with only Alu I could distinguish A. mellea subsp, nipponica from the other species. With Alu I and Dde I, A. ostoyae and A. gallica could be distinguished from the other species. Digestion with Alu I resulted in two patterns (types A and B) of A. singula and three patterns (types A, B, and C) of A.[ezoensis. One pattern (type B) of the former species and two patterns (types B and C) of the latter species were each different from those of the other species. Armillaria sinapina gave only one Alu I digestion pattern, which was identical to that of A. jezoensis (type A) and A. singula (type A). However, by digestion with Dde I, A. singula (type A) could be distinguished from A. jezoensis (type A) and A. sinapina.

Identification of Armillaria nabsnona in Gastrodia Tubers

“…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][16][17] PCR-RFLP analysis profiles of intergenic spacer 1 (IGS-1) have been successfully applied to the Armillaria species identification.…”

“…12,[15][16][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.…”

“…13,14) Methods based on molecular techniques are nowadays routinely used and provided a new perspective for phylogenetic and taxonomic studies.…”

Genetic variation in Armillaria mellea subsp. nipponica estimated using IGS-RFLP and AFLP analyses

“…nipponica (Terashima et al 1998a;Coetzee et al 2000), and restriction fragment length polymorphism (RFLP) analyses of IGS have been applied to identify A. mellea subsp. nipponica (Harrington and Wingfield 1995;Terashima et al 1998b;Fukuda et al 2003).…”

“…IGS-RFLP analysis was carried out using the technique of Harrington and Wingfield (1995) with small modification (Terashima et al 1998b). The IGS region of the ribosomal RNA gene was amplified with the LR12R and O-1 primers (Anderson and Stasovski 1992), yielding an amplified fragment of approximately 830 bp in all isolates examined in this study.…”