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 ...

Development of a new preparation method of Lithospermi Radix enabled almost all the slender lateral roots (diameter smaller than 2 mm) to be preserved. The weight ratio of slender lateral roots was about 5.5% in the whole root. The amount of acetylshikonin in the slender lateral roots was two-fold greater than in its tap and thick lateral roots (diameter larger than 2 mm). The acid-insoluble ash content in slender lateral roots was 1.16%.