Morphological contrasting
We obtained mature sample peels was able on STRoentgenIBC (Letter =67), brand new American Museum from Sheer Records (AMNH, Letter =25), in addition to Us Federal Museum bird collection (USNM, Letter = 10) out-of Costa Rica, Panama, and Colombia for every from about three phenotypic qualities: i) dorsal plumage, ii) ventral plumage, and you will iii) facial secure ornamentation. Adopting the , MJM, Sel-, or over to 3 other observers (see More document step 1: Dining table S1) scored for every single bird alone into a level off 0 – 1, in which 0 depicted a great phenotypically sheer Jacana spinosa, and 1 represented a beneficial phenotypically natural Jacana jacana (centered on comparisons having peels out-of beyond your contact area), therefore women italian averaged the fresh new scores for every attribute since the phenotypic hybrid index for each specimen . Specimens with phenotypic hybrid indices less than 0.ten have been categorized as „pure“ spinosa, if you are an index out of 0.ninety or higher is obtained because the „pure“ jacana. A get ranging from 0.10 and you can 0.twenty-five try categorized just like the „spinosa-like“, while a directory between 0.75 and you will 0.90 are classified due to the fact „jacana-like“. Specimens having indices anywhere between 0.25 and you can 0.75 had been categorized since the „hybrids“.
Genetic investigation
We sequenced an excellent 652 legs pair fragment of the mitochondrial cytochrome oxidase I gene (COI: this new DNA barcode fragment) out-of 59 Panamanian jacanas, together with two specimens away from east Honduras provided for our analysis because of the J. Klicka together with Marjorie Barrick Museum. I selected it gene when planning on taking benefit of present series analysis to the Genbank to have jacanas out of varied geographical places, which were together with put into the fresh dataset (N = 10). Laboratory methods follow practical tips which have been explained in other places . We experimented with but were not able generate succession data away from toe mat extractions on the Smith and Emlen group of specimens.
The latest COI research were utilized to help you imagine this new genetic distinctiveness off both jacana species and you can infer enough time from speciation between J. spinosa and you can J. jacana, and measure the degree to which topography restrictions gene flow in jacanas (i.age. isolation-by-distance). I generated median-joining system during the PopART . We made use of Super so you’re able to assess the typical length utilizing the HKY make of unit progression to examine divergence into the jacanas some other avian kinds whereby hereditary point has been calculated making use of the HKY model .
I tested into the effectation of geographical range towards the genetic variations (i.elizabeth., isolation by point) thru an excellent Mantel attempt adopted in the program Alleles in space , and this measures up hereditary and you will geographic distances one of people in place of communities. For both J. spinosa and you can J. jacana mtDNA succession establishes, i synchronised uncorrected pairwise sequence divergence with pairwise geographical point to own all of the you are able to pairs. Importance of the brand new correlation was determined by evaluating the latest noticed relationship in order to haphazard correlations predicated on ten,100 permutations.
In order to fit new mtDNA-oriented analyses off genetic introgression across the jacana crossbreed region, i sequenced a few autosomal introns (gapdh 11: , and you may 10551: ) to own STRIBC specimens obtained around the Panama. Based on an initial assessment regarding series type from a number of samples away from for each geographic endpoint (away from crossbreed region) of our own testing construction, we known just one nucleotide polymorphism for every single intron: gapdh 3- anti-sense reputation 136 (cytosine-thymine: C-T changeover), and you may 10551 updates 153 (adenine-guanine: A-G change). Positions was relative to lined up sequences of each intron toward J. spinosa test MJM8238 (Genbank accessions: gapdh: KM891734, 10551: KM891735).
Cline figure analyses
We estimated the shape of the phenotypic and genotypic clines using ClineFit , which applies the cline fitting formulas developed by Szymura and Barton . The shape of a cline is determined by three formulas. The first formula (1) defines a symmetric S-shaped curve around the center of the hybrid zone, and relies on two parameters: c – for the center of the curve, and w – which is 1/maximum slope of the cline, and represents the cline width. Two additional formulae define the cline tails (2,3) and add four additional parameters: z L and z R , which represent the distance from c to a vertical asymptote for the exponential decay of allele frequencies to the left and right sides of the hybrid zone, while ? L and ? R define the rates of this exponential decay relative to the shape of the central cline (1).