Suezawa Yasuhiko, Kimura Isao, Inoue Masako, Gohda Nana, Suzuki Motofumi
Kagawa Prefectural Industrial Center, Kagawa, Japan.
Biosci Biotechnol Biochem. 2006 Feb;70(2):348-54. doi: 10.1271/bbb.70.348.
We analyzed sequences of the D1D2 domain of the 26S ribosomal RNA gene (26S rDNA sequence), and the region of internal transcribed spacer 1, 5.8S ribosomal RNA gene and internal transcribed spacer 2 (ITS sequence) of the miso and soy sauce fermentation yeasts, Candida etchellsii and Candida versatilis, in order to evaluate the usefulness of this sequence analysis for identification and typing of these two species. In the 26S rDNA sequence method, the numbers of base substitutions among C. etchellsii strains were up to 2 in 482 bp (99.6% similarity), and they were divided into three types (types A, B, and C). Those of C. versatilis strains were also up to 2 in 521 bp (99.6% similarity) and they were divided into three types (types 1, 2, and 3). In the ITS sequence method, those of C. etchellsii strains were zero in 433 bp (type a, 100% similarity). Those of C. versatilis were 5 in 409 bp (98.8% similarity), divided into 4 types (types I, II, III and IV). It was found that molecular methods based on the sequences of the 26S rDNA D1D2 domain and the ITS region were rapid and precise compared with the physiological method for the identification and typing of these two species.
我们分析了味噌和酱油发酵酵母埃氏假丝酵母(Candida etchellsii)和多变假丝酵母(Candida versatilis)的26S核糖体RNA基因的D1D2结构域序列(26S rDNA序列),以及内部转录间隔区1、5.8S核糖体RNA基因和内部转录间隔区2的区域(ITS序列),以评估这种序列分析方法对这两个物种进行鉴定和分型的实用性。在26S rDNA序列分析方法中,埃氏假丝酵母菌株间在482 bp中的碱基替换数最多为2个(相似度99.6%),它们被分为三种类型(A、B和C型)。多变假丝酵母菌株在521 bp中的碱基替换数也最多为2个(相似度99.6%),它们被分为三种类型(1、2和3型)。在ITS序列分析方法中,埃氏假丝酵母菌株在433 bp中的碱基替换数为零(a型,相似度100%)。多变假丝酵母在409 bp中有5个碱基替换(相似度98.8%),分为4种类型(I、II、III和IV型)。结果发现,基于26S rDNA D1D2结构域和ITS区域序列的分子方法与这两个物种的生理鉴定方法相比,快速且准确。