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首页> 外文期刊>Medical mycology: official publication of the International Society for Human and Animal Mycology >Phylogenetic analysis of Pythium insidiosum Thai strains using cytochrome oxidase II (COX II) DNA coding sequences and internal transcribed spacer regions (ITS).
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Phylogenetic analysis of Pythium insidiosum Thai strains using cytochrome oxidase II (COX II) DNA coding sequences and internal transcribed spacer regions (ITS).

机译：利用细胞色素氧化酶II（COX II）DNA编码序列和内部转录间隔区（ITS）对泰国腐霉菌菌株进行系统发育分析。

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摘要

To investigate the phylogenetic relationship among Pythium insidiosum isolates in Thailand, we investigated the genomic DNA of 31 P. insidiosum strains isolated from humans and environmental sources from Thailand, and two from North and Central America. We used PCR to amplify the partial COX II DNA coding sequences and the ITS regions of these isolates. The nucleotide sequences of both amplicons were analyzed by the Bioedit program. Phylogenetic analysis using genetic distance method with Neighbor Joining (NJ) approach was performed using the MEGA4 software. Additional sequences of three other Pythium species, Phytophthora sojae and Lagenidium giganteum were employed as outgroups. The sizes of the COX II amplicons varied from 558-564 bp, whereas the ITS products varied from approximately 871-898 bp. Corrected sequence divergences with Kimura 2-parameter model calculated for the COX II and the ITS DNA sequences ranged between 0.0000-0.0608 and 0.0000-0.2832, respectively. Phylogenetic analysis using both the COX II and the ITS DNA sequences showed similar trees, where we found three sister groups (A(TH), B(TH), and C(TH)) among P. insidiosum strains. All Thai isolates from clinical cases and environmental sources were placed in two separated sister groups (B(TH) and C(TH)), whereas the Americas isolates were grouped into A(TH.) Although the phylogenetic tree based on both regions showed similar distribution, the COX II phylogenetic tree showed higher resolution than the one using the ITS sequences. Our study indicates that COX II gene is the better of the two alternatives to study the phylogenetic relationships among P. insidiosum strains.
机译：为了调查泰国腐霉菌分离株之间的系统发育关系，我们研究了从泰国的人类和环境来源以及从北美洲和中美洲的两种来源分离的31株P. Insidiosum菌株的基因组DNA。我们使用PCR扩增了这些分离株的部分COX II DNA编码序列和ITS区。通过Bioedit程序分析两个扩增子的核苷酸序列。使用MEGA4软件使用遗传距离方法和邻居加入（NJ）方法进行了系统发育分析。其他三个腐霉属物种，大豆疫霉菌和巨大紫花鱼的其他序列也被作为外群。 COX II扩增子的大小在558-564 bp之间变化，而ITS产物在大约871-898 bp之间变化。用针对木炭二代和ITS DNA序列计算的Kimura 2参数模型校正的序列差异分别在0.0000-0.0608和0.0000-0.2832之间。使用COX II和ITS DNA序列进行的系统发育分析显示出相似的树，在这里我们发现了P. insidiosum菌株中的三个姐妹组（A（TH），B（TH）和C（TH））。来自临床病例和环境来源的所有泰国分离株都被分为两个独立的姐妹组（B（TH）和C（TH）），而美洲分离株被分为A（TH）。尽管基于这两个区域的系统发育树显示出相似分布，COX II系统发育树显示出比使用ITS序列的树更高的分辨率。我们的研究表明，COX II基因是研究P. insidiosum菌株之间亲缘关系的两个替代方法中较好的一种。

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