Junkum Anuluck, Komalamisra Narumon, Jitpakdi Atchariya, Jariyapan Narissara, Min Gi-Sik, Park Mi-Hyun, Cho Kang-Hyun, Somboon Pradya, Bates Paul A, Choochote Wej
Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
J Vector Ecol. 2005 Dec;30(2):213-24.
Iso-female lines (isolines) of Anopheles aconitus collected from Mae Hong Son, Phet Buri, and Chiang Mai Provinces were successfully identified to karyotypic forms. The results of identification revealed that An. aconitus Form B (X1, X2, Y2) was obtained from four and 48 isolines in Phet Buri and Chiang Mai Provinces, respectively, and Form C (X1, X2, Y3) was recovered from three and 41 isolines in Mae Hong Son and Chiang Mai Provinces, respectively. When comparing band to band on the same arm of ovarian nurse cell polytene chromosomes of An. aconitus Form B (Phet Buri: four isolines) and C (Mae Hong Son: three isolines, Chiang Mai: 20 isolines) to the standard chromosome mapping of An. aconitus Form B (Chiang Mai: 20 isolines), no major chromosomal rearrangements that related to the karyotype variations were demonstrated. The investigations on allelic frequencies of 4th stage larvae and adult females of three (Form C: Mae Hong Son), four (Form B: Phet Buri), 41 (Form C: Chiang Mai) and 48 (Form B: Chiang Mai) isolines suggested that An. aconitus Form B and C of all strains have similar allelic frequencies. This was observed at 10 isoenzymes 16 loci in 4th stage larvae, and 11 isoenzymes 13 loci in adult females. Hybridization tests among the four laboratory-raised isolines of An. aconitus Form B (Chiang Mai and Phet Buri) and C (Chiang Mai and Mae Hong Son) were employed by induced copulation. The results of crosses indicated that they were genetically compatible, providing viable progeny and completely synaptic salivary gland polytene chromosomes. The complete sequences ofrDNA internal-transcribed spacer two (ITS2) and partial sequences of mitochondrial cytochrome c oxidase subunit I and II (COI and COII) from genomic DNA of 12 isolines of An. aconitus Form B and C were identified. Total sequence lengths (ITS2+COI+COII) of An. aconitus isolines varied from 1550bp to 1556bp. Conspecific relationships between the two An. aconitus forms were well supported by low values of intraspecific distances (ranged from 0.1% to 1.0%) and genetic differentiation (d(xy): 0.01322) between the two forms. Based on evidence of no pre- and post-mating isolations, and nearly identical of DNA sequences of ITS2, COI and COII regions between An. aconitus Form B and C, we conclude that they are conspecific cytological races in the Thai population.
从夜丰颂府、碧武里府和清迈府采集的乌头按蚊同雌品系（纯系）已成功鉴定出核型形式。鉴定结果显示，分别从碧武里府的4个和清迈府的48个纯系中获得了乌头按蚊B型（X1、X2、Y2），分别从夜丰颂府的3个和清迈府的41个纯系中获得了C型（X1、X2、Y3）。将乌头按蚊B型（碧武里府：4个纯系）和C型（夜丰颂府：3个纯系，清迈府：20个纯系）卵巢滋养细胞多线染色体同一臂上的带与乌头按蚊B型（清迈府：20个纯系）的标准染色体图谱进行逐带比较，未发现与核型变异相关的主要染色体重排。对3个（C型：夜丰颂府）、4个（B型：碧武里府）、41个（C型：清迈府）和48个（B型：清迈府）纯系的四龄幼虫和成年雌蚊的等位基因频率进行调查，结果表明，所有菌株的乌头按蚊B型和C型具有相似的等位基因频率。在四龄幼虫的10种同工酶16个位点以及成年雌蚊的11种同工酶13个位点上均观察到这一点。通过诱导交配，对实验室饲养的4个乌头按蚊B型（清迈府和碧武里府）和C型（清迈府和夜丰颂府）纯系进行了杂交试验。杂交结果表明它们在遗传上是相容的，能产生有活力的后代和完全联会的唾液腺多线染色体。鉴定了12个乌头按蚊B型和C型纯系基因组DNA的核糖体DNA内转录间隔区2（ITS2）的完整序列以及线粒体细胞色素c氧化酶亚基I和II（COI和COII）的部分序列。乌头按蚊纯系的总序列长度（ITS2 + COI + COII）在1550bp至1556bp之间变化。两种乌头按蚊形式之间的种内距离值较低（范围为0.1%至1.0%）以及两种形式之间的遗传分化（d(xy)：0.01322），有力地支持了它们之间的同种关系。基于没有交配前和交配后的隔离证据，以及乌头按蚊B型和C型之间ITS2、COI和COII区域的DNA序列几乎相同，我们得出结论，它们是泰国种群中的同种细胞学族。
