Mycosystema 菌 物 學 報 15 November 2008, 27(6): 873-883
[email protected] ISSN1672-6472 CN11-5180Q
2008 Institute of Microbiology, CAS, all rights reserved.
青藏高原黃綠蜜環(huán)菌純培養(yǎng)菌種的分離培養(yǎng)及分子鑒定
李海波1 吳學謙1,2* 王立武3 付立忠1,2 魏海龍1,2 吳慶其1,2
1浙江省林業(yè)科學研究院 浙江省森林資源生物與化學利用重點實驗室 杭州 310023
2 麗水市食用菌研究開發(fā)中心 麗水 323000
3西藏那曲地區(qū)林業(yè)局 那曲 852000
摘 要：首次從采自青藏高原,、與高原牧草嵩草屬Kobresia草本植物形成外生菌根的黃綠蜜環(huán)菌Armillaria luteo-virens子實體中分離獲得一組織分離菌株，運用rDNA-ITS和rDNA-IGS-1測序技術(shù)對該組織分離菌株是否為黃綠蜜環(huán)菌的純培養(yǎng)菌種進行分子鑒定,，并基于黃綠蜜環(huán)菌的5.8S/ITS和IGS-1序列進行核酸序列數(shù)據(jù)庫GenBank同源性檢索比對,、構(gòu)建系統(tǒng)發(fā)育樹。結(jié)果表明,，本研究獲得的黃綠蜜環(huán)菌子實體組織分離菌株即為其純培養(yǎng)菌種,。基于ITS的系統(tǒng)發(fā)育分析表明黃綠蜜環(huán)菌與口蘑科內(nèi)其它屬間物種的系統(tǒng)發(fā)育關(guān)系較遠,；基于IGS-1的系統(tǒng)發(fā)育分析表明黃綠蜜環(huán)菌與蜜環(huán)菌屬內(nèi)的其它種序列差異較大,，系統(tǒng)發(fā)育關(guān)系較遠，而與Lepiota屬內(nèi)的部分種具有較近的系統(tǒng)發(fā)育關(guān)系,。本研究首次基于分子手段對我國青藏高原的黃綠蜜環(huán)菌種進行了分離培養(yǎng),、分子鑒定和系統(tǒng)發(fā)育分析，為黃綠蜜環(huán)菌的科學分類提供了分子依據(jù),。
關(guān)鍵詞：外生菌根,，分離培養(yǎng)，rDNA-ITS序列分析,，rDNA-IGS序列分析,，系統(tǒng)發(fā)育
Pure culture isolation, cultivation and molecular identification of Armillaria luteo-virens from Tibet Plateau
LI Hai-Bo1 WU Xue-Qian1,2 WANG Li-Wu3 FU Li-Zhong1,2 WEI Hai-Long1,2 WU Qing-Qi1,2
1Zhejiang Forestry Academy; Zhejiang Provincial Key Laboratory of Biological and Chemical Utilization of Forest Resource, Hangzhou 310023, China
2Lishui Edible Fungal Research and Development Center, Lishui 323000, China
3Nagqu Forestry Bureau of Tibet, Nagqu 852000, China
Abstract: Armillaria luteo-virens is a ectomycorrhizal fungus symbiotic with some species of Kobresia in Tibet Plateau, China. A strain was recently isolated from fruitbody tissue of the fungus. To determine whether the strain is the pure culture of the fungus, the internal transcribed spacer (ITS) region and the first intergenic spacer region (IGS-1) of the nuclear ribosomal DNA of fruiting body and the isolated strain were sequenced and compared. Further, the 5.8S/ITS and IGS-1 sequences of the species were BLAST matched with GenBank database on homology and a phylogenetic tree was constructed based on 5.8S/ITS sequence and two phylogenetic trees were constructed separately based on two IGS-1 sequences. The results demonstrated that the strain isolated from fruitbody tissue of Armillaria luteo-virens was the true pure culture of the fungus. Phylogenetic analysis based on 5.8S/ITS sequence indicated Armillaria luteo-virens has a distant phylogenetic relationship with other genera of Tricholomataceae. Phylogenetic analysis based on two IGS-1 sequences indicated there were significant differences between the IGS-1 sequence of Armillaria luteo-virens and that of other species of Armillaria. Armillaria luteo-virens has a relatively close phylogenetic relationship with some species of Lepiota.
Key words: ectomycorrhizal fungi, rDNA-ITS sequence analysis, IGS sequence analysis, phylogeny
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