title: Phylogenetic study of six species of Anopheles mosquitoes in Peninsular Malaysia based on inter-transcribed spacer region 2 (ITS2) of ribosomal DNA.
abstract: BACKGROUND
  Molecular techniques are invaluable for investigation on the biodiversity of Anopheles mosquitoes. This study aimed at investigating the spatial-genetic variations among Anopheles mosquitoes from different areas of Peninsular Malaysia, as well as deciphering evolutionary relationships of the local Anopheles mosquitoes with the mosquitoes from neighbouring countries using the anopheline ITS2 rDNA gene.
  
  
  METHODS
  Mosquitoes were collected, identified, dissected to check infection status, and DNA extraction was performed for PCR with primers targeting the ITS2 rDNA region. Sequencing was done and phylogenetic tree was constructed to study the evolutionary relationship among Anopheles mosquitoes within Peninsular Malaysia, as well as across the Asian region.
  
  
  RESULTS
  A total of 133 Anopheles mosquitoes consisting of six different species were collected from eight different locations across Peninsular Malaysia. Of these, 65 ITS2 rDNA sequences were obtained. The ITS2 rDNA amplicons of the studied species were of different sizes. One collected species, Anopheles sinensis, shows two distinct pools of population in Peninsular Malaysia, suggesting evolvement of geographic race or allopatric speciation.
  
  
  CONCLUSIONS
  Anopheles mosquitoes from Peninsular Malaysia show close evolutionary relationship with the Asian anophelines. Nevertheless, genetic differences due to geographical segregation can be seen. Meanwhile, some Anopheles mosquitoes in Peninsular Malaysia show vicariance, exemplified by the emergence of distinct cluster of An. sinensis population.
authors.forename: Jia-Siang
authors.initials: JS
authors.lastname: Sum
authors.forename: Wenn-Chyau
authors.initials: WC
authors.lastname: Lee
authors.forename: Amirah
authors.initials: A
authors.lastname: Amir
authors.forename: Kamil A
authors.initials: KA
authors.lastname: Braima
authors.forename: John
authors.initials: J
authors.lastname: Jeffery
authors.forename: Noraishah M
authors.initials: NM
authors.lastname: Abdul-Aziz
authors.forename: Mun-Yik
authors.initials: MY
authors.lastname: Fong
authors.affiliation: Tropical Infectious Disease Research and Education Center (TIDREC), Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia. lauyeeling@um.edu.my.
authors.forename: Yee-Ling
authors.initials: YL
authors.lastname: Lau
mesh_terms: D063147:Animal Distribution; D000818:Animals; D000852:Anopheles / Q000235:genetics*; D044822:Biodiversity; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D008296:Malaysia; D010802:Phylogeny
chemical_list: D021903:DNA, Ribosomal Spacer