Molecular and cytogenetic evidence of three sibling species of the Anopheles barbirostris Form A (Diptera:Culicidae) in Thailand

Abstract

Nine isoline colonies of Anopheles barbirostris Form A, derived from individual isofemale lines from Chiang Mai, Phetchaburi, and Kanchanaburi, were established in our insectary at Chiang Mai University. All isolines shared the same mitotic karyotype (X(1), X(2), Y(1)). Molecular analysis of deoxyribonucleic acid (DNA) sequences and polymerase chain reaction (PCR) products of ITS2, COI, and COII regions revealed three distinct groups: A1 (Chiang Mai), A2 (Phetchaburi), and A3 (Kanchanaburi). Crossing experiments among the three groups exhibited strong reproductive isolation, producing low and/or non-hatched eggs, and inviable and/or abnormal development of the reproductive system of F(1)-progenies. Asynaptic regions along the five polytene chromosome arms of F(1)-hybrid larvae clearly supported the existence of three sibling species within A. barbirostris Form A, provisionally named species A1, A2, and A3.

Karyotypic variation and geographic distribution of Anopheles campestris-like (Diptera: Culicidae) in Thailand

“…campestris-like were arbitrarily selected from the 30 isoline colonies as representatives of the three karyotypic forms, i.e., Form B (AKpB1), Form E (HCE6, AKkE4, AMkE1, AMsE3, HSkE3, ACpE6) and Form F (AUdF5, ACiF1, AAyF2, HCtF4, APkF1) (Table II). These isolines were used for crossing experiments, in order to determine post-mating reproductive isolation by employing the techniques previously reported by Saeung et al (2007Saeung et al ( , 2008.…”

“…Metaphase karyotype -Metaphase chromosomes were prepared from 10 samples of the early fourth-instar larval brains of F 1 and/or F 2 -progenies of each isoline using the techniques previously described by Saeung et al (2007Saeung et al ( , 2008. Identification of karyotypic forms followed the cytotaxonomic key of Baimai et al (1995).…”

“…barbirostris s.l. is a cryptic species consisting of at least four sibling species, i.e., A1, A2, A3 (Saeung et al 2008) and A4 (Suwannamit et al 2009). The An.…”

Susceptibility of Anopheles campestris-like and Anopheles barbirostris species complexes to Plasmodium falciparum and Plasmodium vivax in Thailand

“…campestris-like Forms B (Chiang Mai), E (Chiang Mai & Sa Kaeo) and F (Ayuttaya & Udon Thani) and An. barbirostris species A1 (Chiang Mai), A2 (Phetchaburi), A3 (Kanchanaburi) and A4 (Chiang Mai) were established based on morphological, cytogenetic and molecular investigations and crossing experiments, as previously reported by Saeung et al (2007Saeung et al ( , 2008, Suwannamit et al (2009) and Thongsahuan et al (2009). These colonies were successfully reared using the methods of Choochote et al (1983) and Kim et al (2003) in an insectary room at 27 ± 2ºC and 70-80% relative humidrelative humidity (RH).…”

“…campestris-like and An. barbirostris species A1, A2, A3 and A4 (Saeung et al 2007, 2008). Little is known about the potential vector for P. falciparum and/or P. vivax in these five sibling species members; thus, intensive comparison of their vector-competence is needed to elucidate their potential vector status.…”