Meeting Abstract
Alternative reproductive systems, wherein individuals do not exclusively reproduce sexually, may provide unique insight regarding sexual conflict. Leiobunum manubriatum and L. globosum are facultatively parthenogenetic Japanese harvestman which vary in sex ratio across their geographic distribution. Males of both species possess morphology which suggest coercion may be common during mating events. In previous efforts, we used genotyping-by-sequencing and a SNP array panel to determine the reproductive mode of females. We found that despite the availability of males, females in populations with high male frequency reproduced primarily through parthenogenesis. Additionally, females from populations with few males were not significantly more fecund than females in equal sex ratio populations. These results also identified that a more accurate, cost-effective method to rapidly genotype thousands of samples was necessary. 3RAD Capture combines the low input DNA requirements and decreased PCR error rate of 3-enzyme RADseq with the high coverage of bait capture sequencing. First, specialized capture baits were designed from previous RAD sequencing of L. manubriatum. These baits target loci suitable for genotyping, maximizing usable reads from minimal template DNA. Samples were digested using three enzymes, and custom oligonucleotides were ligated to cut ends. Following this preparation, the library is hybridized with bait probes which are targeted with streptavidin beads. The captured DNA can then be isolated and sequenced with high efficacy. Rapid and accurate genotyping facilitates sire assignment of thousands of egg specimens, providing insight on the factors which maintain sex in these facultative parthenogens.
