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The symbiosis with the arbuscular mycorrhiza (AM) was observed in the roots of Botrychium ternatum. The fungal partial 18S ribosomal DNA (rDNA:approximately 450/ 1800 bp) were amplified from the DNA e...xtracts of fern roots using fungal specific primer, and subcloned to cloning vector (pGEM-T) . According to the RFLP pattern of subclone, some clones were selected and their inserts were sequenced. One of them (GLA-006) was predominant in this analysis, however, four (or potentially five) different clone were found in the root of B. ternatum. To infer the phylogenetic relationship, sequences were donated on BLAST Search. According to the result of BLAST Search, neighbor joining analysis was performed. The partial sequencing data of 18S rDNA indicate that these isolates closely related to genus Glomus (AM fungi),especially G.proliferum and G.sinuosum. Three of them were 92-95 % identical to G.proliferum
Botrychium ternatum根茎の菌根菌について遺伝学的手法を用いて検討した.根茎より抽出したDNAを真菌類特異的18SrDNAプライマーにより増幅,クローニングし,得られたプラスミドのインサートの配列についてRFLP解析,シーケンシングを行った.塩基配列はBLAST Searchを用いてGenBankのデータと比較後,近縁種のデータを用いてNJ法で系統解析を行った.解析の結果,DNAフラグメントGLA-006,GLA-016,GLA-032はG.proliferumと92-95%の相同性を示した.RFLP解析より数種のAM菌根菌が見出されたが,B.ternatumの根茎ではGLA-006が優占種であることが明らかになった.show more
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