Phylogeny of Anophelinae using mitochondrial protein coding genes

Abstract

Malaria is a vector-borne disease that is a great burden on the poorest and most marginalized communities of the tropical and subtropical world. Approximately 41 species of Anopheline mosquitoes can effectively spread species of Plasmodium parasites that cause human malaria. Proposing a natural classification for the subfamily Anophelinae has been a continuous effort, addressed using both morphology and DNA sequence data. The monophyly of the genus Anopheles, and phylogenetic placement of the genus Bironella, subgenera Kerteszia, Lophopodomyia and Stethomyia within the subfamily Anophelinae, remain in question. To understand the classification of Anophelinae, we inferred the phylogeny of all three genera (Anopheles, Bironella, Chagasia) and major subgenera by analysing the amino acid sequences of the 13 protein coding genes of 150 newly sequenced mitochondrial genomes of Anophelinae and 18 newly sequenced Culex species as outgroup taxa, supplemented with 23 mitogenomes from GenBank. Our analyses generally place genus Bironella within the genus Anopheles, which implies that the latter as it is currently defined is not monophyletic. With some inconsistencies, Bironella was placed within the major clade that includes Anopheles, Cellia, Kerteszia, Lophopodomyia, Nyssorhynchus and Stethomyia, which were found to be monophyletic groups within Anophelinae. Our findings provided robust evidence for elevating the monophyletic groupings Kerteszia, Lophopodomyia, Nyssorhynchus and Stethomyia to genus level; genus Anopheles to include subgenera Anopheles, Baimaia, Cellia and Christya; Anopheles parvus to be placed into a new genus; Nyssorhynchus to be elevated to genus level; the genus Nyssorhynchus to include subgenera Myzorhynchella and Nyssorhynchus; Anopheles atacamensis and Anopheles pictipennis to be transferred from subgenus Nyssorhynchus to subgenus Myzorhynchella; and subgenus Nyssorhynchus to encompass the remaining species of Argyritarsis and Albimanus Sections.

Time and mode of Culicidae evolutionary history

“…The Bironella genus showed as an ancestral lineage in relationship to the Anopheles genus including all subgenera assessed in our analysis such as Kerteszia , Nyssorhynchus, Anopheles and Cellia . This contrasts with other findings that positioned this genus inside the Anopheles genus using mitochondrial protein sequences from mitogenomes 19 . Previous studies, using both nuclear ribosomal sequences and fragments of mitochondrial genes COI and COII of Bi.…”

“…Hence, there are still many non-studied species and unresolved phylogenetic relationships in genera such as Aedes, Armigeres, Coquillettidia, Culex, Mansonia, Mimomyia, Psorophora, Topomyia, Tripteroides, Toxorhynchites, Uranotaenia and Wyeomyia 61 . The Anophelinae subfamily phylogenetic relationship was recently investigated using the mitochondrial genomes from many species and the authors proposed a number of taxonomic status changes such as the elevation of some groups from the subgenus to genus level 19 . Some other studies also characterized the mitogenomes of some species to solve the phylogenetic positioning of culicids, however, most of them focused on a specific group ( Anopheles , Culex and Sabethes ) or few genera of the Culicidae family 10,11,[19][20][21]66 .…”

“…In addition, mitogenomes have a uniparental heritage, high copy number per cells and single-copy genes which facilitates DNA recovery and phylogenetic analysis [16][17][18] . Recently, some studies have sequenced a larger number of mitochondrial genomes from different mosquito species, but they are mostly focused on Anopheles species [19][20][21] . Mosquito mitogenomes are structurally conserved following the metazoa gene number and order, with few exceptions, showing 37 genes comprising 13 protein coding genes, 22 tRNAs and 2 rRNA genes [22][23][24] .…”

“…Other approaches available allow the recovery of mitogenomes by PCR amplification from environment samples and pooled DNA and mitogenome recovery from low-coverage sequencing [29][30][31] . Moreover, a number of bioinformatics tools were developed to specifically assembly and annotate mitogenomes [32][33][34][35][36] Most of the available mitogenomes belong to Anopheles species with much fewer genomes from Culex, Aedes and other genera such as Haemagogus, Bironella, Sabethes, and Lutzia [19][20][21]24,37 , but there is no available molecular data for the large majority of the species. Aiming to contribute with this basic and fundamental knowledge we performed low-coverage sequencing and data mining on already published Culicidae SRA data in order to characterize the mitogenomes from different species and genera and better understand the evolutionary history of mosquitoes focusing in Culicinae subfamily.…”

Ookinete-Specific Genes and 18S SSU rRNA Evidenced in Plasmodium vivax Selection and Adaptation by Sympatric Vectors

“…At least 10 species of the family Culicidae transmit malaria in this continent. The Subgenera Nyssorhynchus and Anopheles diverged 100 million years ago, and Nyssorhynchus was recently raised to genus status (Foster et al, 2017;Harbach, 2018). Nyssorhynchus albimanus and Anopheles pseudopunctipennis are abundant in Mexico, Central America, and the northern South America regions (Sinka et al, 2012).…”