High-throughput sequencing has enabled genome skimming approaches to produce complete mitochondrial genomes (mitogenomes) for species identification and phylogenomics purposes. In particular, the portable sequencing device from Oxford Nanopore Technologies (ONT) has the potential to facilitate hands-on training from sampling to sequencing and interpretation of mitogenomes. In this study, we present the results from sampling and sequencing of six gastropod mitogenomes (Aplysia argus, Cellana orientalis, Cellana toreuma, Conus ebraeus, Conus miles and Tylothais aculeata) from a graduate level biodiversity course. The students were able to produce mitogenomes from sampling to annotation using existing protocols and programs. Approximately 4 Gb of sequence was produced from 16 Flongle and one MinION flow cells, averaging 235 Mb and N50 = 4.4 kb per flow cell. Five of the six 14.1–18 kb mitogenomes were circlised containing all 13 core protein coding genes. Additional Illumina sequencing revealed that the ONT assemblies spanned over highly AT rich sequences in the control region that were otherwise missing in Illumina-assembled mitogenomes, but still contained a base error of one every 70.8–346.7 bp under the fast mode basecalling with the majority occurring at homopolymer regions. Our findings suggest that the portable MinION device can be used to rapidly produce low-cost mitogenomes onsite and tailored to genomics-based training in biodiversity research.
Species delimitation can be difficult when the divergence between focal taxa is in the incipient stage of speciation, because conflicting results are expected among different data sets, and the species limits can differ depending on the species concept applied. We studied speciation history and investigated the impact on taxonomic decision-making when using different types of data in a Taiwanese endemic sister-species pair of stag beetles, Lucanus miwai and Lucanus yulaoensis, from sky island habitats. We showed that the two geographical taxa can be diagnosed by male mandibular shape. We found two mitochondrial co1 lineages with pairwise sequence divergence > 3%; however, L. miwai might not be monophyletic. The result of our multispecies coalescent-based species delimitation using five nuclear loci supported the evolutionary independence of the two sister species, but the calculated values of the genealogical divergence index (gdi) corresponded to the ambiguous zone of species delimitation. We also showed that post-divergence gene flow is unlikely. Our study demonstrates challenges in the delineation of incipient species, but shows the importance of understanding the speciation history and adopting integrative approaches to reconcile seemingly conflicting results before making evolutionarily relevant taxonomic decisions.
Species distribution models (SDMs) have conventionally been used for evaluating the distribution of individual species, but they can also be used, through comparing different SDMs, to evaluate the geographic similarity between taxa. In this study, we used a parasite and host system to infer the geographic overlaps between species with tight biological interaction, for example, parasites and their obligate host. Specifically, we used the horsehair worm Chordodes formosanus and its three mantis hosts to study the extent of niche overlap. We retrieved presence points for the host species and the parasite, and then we built SDMs with MaxEnt implemented in ENMeval using selected bioclim variables (based on variance inflation factor values) at 30s scale. The models showed that the hosts and parasite do not occur in the high elevation areas in Taiwan, which is expected based on their biology. Interestingly, the predicted parasite distribution included areas without collection records, implying local extinction or sampling bias. We subsequently evaluated niche overlap between hosts and the parasite according to five similarity indices (Schoener's D , I statistic, relative rank, Pearson correlation coefficient, and the rank correlation coefficient rho). Our models showed a high similarity of SDM predictions between hosts and the parasite. There were differences among metrics for which host shared the highest similarity with the parasite, but the majority of the results indicated that the Japanese boxing mantis had the highest niche similarity with the horsehair worm. The choice of the niche overlap metric to use can uncover information on the parasite's ecology, which can be important for endangered species. SDMs are reliable tools for host and parasite conservation management and could help improve our understanding of parasite biology and ecology.
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