First assessment of the biodiversity of praying mantises (Insecta: Mantodea) in Cameroon with DNA barcoding

combination of morphological characters was not matched with any described species. DNA barcode of the sample has the distance of 14% with a sequence of typical S. lineola, while on phylogenetic tree the unidentified species formed a separate branch with same species. It might be suggested that femoral spots are an early developing character resulted from predator pressure, as the spots are a part of threat display of the mantis. Larger body size might also facilitate in repelling the offenders. One of the advantages of using DNA barcodes is identification of samples, otherwise impossible to determine due to life stage or sex. This is illustrated on obtained tree in Steno- pyga extera. One of the samples (VGPC23383) we obtained the sequence from was immature female mantis, belonging to genus Stenopyga. However, identification to species-level based on morphological characters was not possible, as the diagnostic characters for the genus include wing coloration (not developed at nymphal stage) and structure of male genitalia (obviously absent in a female) [20]. While building phylogenetic tree, the barcode of immature Stenopyga clustered with a sequence from same locality of an adult male Stenopyga extera, which was pos- sible to identify based on morphological characters. The distance between two samples is <0.2%, which confirms these two specimens are conspecific. DNA barcoding proved to be useful in identification of specimens with abnormal morpho- logical traits. One female of Polyspilota aeruginosa (VGPC23764) was found to possess black-

ened prosternum, atypical for this species. Given that P. aeruginosa is a taxon with high intraspecific variation [24], it is likely an individual aberration. Molecular analysis supports this hypothesis, as the maximum likelihood tree places this specimen close to sequences from CAR, moreover, the pairwise distance between a typical looking sample and the one with blackened prosternum both collected in Ebogo II is about 0.7%. Comparison of the dissected genitalia of this female with a typically looking female revealed no noticeable differences in copulatory organs. Cameroon is an often-disregarded region when it comes to biodiversity assessments, partic- ularly regarding Mantodea. Our research represents the first comprehensive study on praying mantises of Cameroon, enriching the catalogue of species and enhancing it through the use of DNA barcodes. The use of genetic information allowed us to compare existing phylogeny of the order and to complement existing efforts from neighboring countries. Phylogenetic tree built using sequences of Mantodea of Central African subregion is adequately congruent with current taxonomical consensus. However, several collected specimens likely belong to previ- ously unrecognized taxa, highlighting the importance of expanding the study area and increas- ing the frequency of sampling to reveal even more undescribed species. The tree, complemented by pairwise distances, demonstrate unresolved taxonomy of several genera, such as Negromantis, Plistospilota. To address these knowledge gaps, it is crucial to employ a

combination of morphological and molecular approaches to fill in the missing data on the biology and ecology of praying mantises in the Central African subregion and, by extension, the entire continent. Supporting information S1 Table. Primer pairs used for amplification of the samples and their molecular voucher and accession codes. (DOCX) S2 Table. Pairwise distances. (XLSX) PLOS ONE Diversity of praying mantises in Cameroon PLOS ONE | https://doi.org/10.1371/journal.pone.0304163 May 23, 2024 20 / 23