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Same as above, the association between wPip groups and Cx.
To investigate the association between wPip groups and mtDNA variation, we sequenced the cytb gene (that have been shown polymorphic in the Cx. pipiens complex, see [ 14]) in a subsample of 184 specimens from 101 Cx. pipiens natural populations and 44 isofemale laboratory lines.
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Aside from the wPip-II/ wPip-III mixed populations found in Western Europe, at least five other geographic contact zones between wPip groups exist: in the North (wPip-II/ wPip-IV) and in the South of Italy (wPip-I/ wPip-IV), in North Africa (wPip-I/ wPip-IV), in South America (wPip-I/ wPip-III) and Eastern Asia (wPip-I/ wPip-V).
For instance, pipiens harbours infections of all five wPip groups and quinquefasciatus of three wPip groups (wPip-I, wPip-III and wPip-V).
Pairwise tests revealed significant LD for wPip groups and cytb haplotypes (P = 10-5) with a clear pattern of cytb haplotype specificity to wPip groups (Table 2).
However, we did not observe this pattern: similar wPip groups and mtDNA haplotypes are found in different taxa and no cytoplasm type is specific to a given Cx.
Some specificity in the cidA-cidB repertoire was also detected, with none of the variants shared between strains from different wPip groups (Figs. 3b and 4b).
When analysing the congruence between the five mtDNA and wPip groups, very little incongruence were observed.
The association between Cx. pipiens taxa and wPip groups was highly significant (Fisher exact test, P < 10-4), ashownwithith quinquefasciatus that appeared mainly infected with the wPip-I group (133 of 201 individuals), pipiens with wPip-II (55 of 118) and pallens with wPip-V (19 of 20) (Table 3).
The presence of shared wPip groups between Cx. pipiens members thus suggests that wPip infections undergo repeated transfers between quinquefasciatus, pipiens, molestus and pallens.
This suggests that the wPip ancestor may have initially spread in European populations where it evolved in five divergent groups, and that newly emerged wPip groups have only secondarily expanded outside Europe.
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