Djadid Navid Dinparast, Gholizadeh Saber, Aghajari Mehrnoush, Zehi Abdolghafar Hassan, Raeisi Ahmad, Zakeri Sedigheh
Malaria Research Group, Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.
Acta Trop. 2006 Jan;97(1):65-74. doi: 10.1016/j.actatropica.2005.08.003. Epub 2005 Sep 26.
Anopheles stephensi is one of the most important malaria vectors in the Middle-East, the Indian subcontinent, the Far-East and is the main malaria vector in south of Iran. This vector is thought to be a single but polytypic species, despite its enormous geographical range. To examine this hypothesis, we analyzed the rDNA-ITS2 and RAPD loci in different populations of An. stephensi from Iran. rDNA-ITS2 region in all sequenced specimens of An. stephensi contained a (CA)7 microsatellite sequence. Construction of phylogenetic tree based on rDNA-ITS2 sequences revealed that there only is a minor polymorphism between the different populations, despite their vast geographical distances. RAPD-PCR could differentiate rural and urban populations of An. stephensi, but it is unclear whether these two samples represent mysorensis and the type form. Further characterization of interested RAPD fragments by cloning; have shown the nature of inverted repeats and the presence of microsatellite region (GT) in both ends near to inverted repeat sequences of primers. These results showed that An. stephensi in Iran could be considered a single species with different biological and ecological forms in different zoogeographical zones. Further studies are needed to demonstrate the relation between RAPD and microsatellite sequences and the differences seen in the field for this species. This data will serve as first report on the sequence of rDNA-ITS2 and a microsatellite-containing RAPD region, which could be used for species-specific diagnosis and differentiation of urban and rural populations in An. stephensi.
斯氏按蚊是中东、印度次大陆、远东地区最重要的疟疾传播媒介之一,也是伊朗南部的主要疟疾传播媒介。尽管其地理分布范围极广,但该媒介被认为是一个单一的多型物种。为了验证这一假设,我们分析了来自伊朗的不同斯氏按蚊种群的核糖体DNA内转录间隔区2(rDNA-ITS2)和随机扩增多态性DNA(RAPD)位点。在所有测序的斯氏按蚊标本中,rDNA-ITS2区域都包含一个(CA)7微卫星序列。基于rDNA-ITS2序列构建的系统发育树显示,尽管不同种群之间地理距离遥远,但仅存在微小的多态性。RAPD-PCR可以区分斯氏按蚊的农村和城市种群,但尚不清楚这两个样本是否分别代表米氏按蚊和指名亚种。通过克隆对感兴趣的RAPD片段进行进一步表征,结果显示了反向重复序列的性质以及在引物反向重复序列附近两端存在微卫星区域(GT)。这些结果表明,伊朗的斯氏按蚊可被视为一个单一物种,在不同动物地理区域具有不同的生物学和生态学形式。需要进一步研究来证明RAPD与微卫星序列之间的关系以及该物种在野外观察到的差异。这些数据将作为关于rDNA-ITS2序列和含微卫星的RAPD区域的首次报告,可用于斯氏按蚊的物种特异性诊断以及农村和城市种群的区分。