Description of Neoperla mindoroensis sp. nov., the first record of a stonefly from Mindoro, Philippines (Plecoptera, Perlidae), and identification of its life stages using COI barcodes

The new stonefly species, Neoperla mindoroensissp. nov. (Perlidae), from Mindoro island is described. The new species is assigned to the N. recta species complex of the N. montivaga group on account of its obvious T7 and T8 with pointed processes and the presence of basolateral lobes in the everted aedeagal sac. The male adult is distinguishable by its aedeagus with a slightly raised mediodorsal lobe, fully covered with fine spinules, while the female adult has comparably small eggs (240 × 220 μm) with a punctate, chorionic surface with punctae arranged in polygonal FCIs. The life stages and sexes were assigned using COI mtDNA barcodes (2.2% maximum intraspecific genetic distance), which were compared with available barcodes of congeners, which had interspecific genetic distances varying by at least 23.5%. Biogeographic aspects, ecological habitat requirements, and suitability as potential bioindicator of the species are also briefly discussed.

Shedding Light on a Secretive Tertiary Urodelean Relict: Hynobiid Salamanders (Paradactylodon persicus s.l.) from Iran, Illuminated by Phylogeographic, Developmental, and Transcriptomic Data

The Hyrcanian Forests present a unique Tertiary relict ecosystem, covering the northern Elburz and Talysh Ranges (Iran, Azerbaijan), a poorly investigated, unique biodiversity hotspot with many cryptic species. Since the 1970s, two nominal species of Urodela, Hynobiidae, Batrachuperus (later: Paradactylodon) have been described: Paradactylodon persicus from northwestern and P. gorganensis from northeastern Iran. Although P. gorganensis has been involved in studies on phylogeny and development, there is little data on the phylogeography, systematics, and development of the genus throughout the Hyrcanian Forests; genome-wide resources have been entirely missing. Given the huge genome size of hynobiids, making whole genome sequencing hardly affordable, we aimed to publish the first transcriptomic resources for Paradactylodon from an embryo and a larva (9.17 Gb RNA sequences; assembled to 78,918 unigenes). We also listed 32 genes involved in vertebrate sexual development and sex determination. Photographic documentation of the development from egg sacs across several embryonal and larval stages until metamorphosis enabled, for the first time, comparison of the ontogeny with that of other hynobiids and new histological and transcriptomic insights into early gonads and timing of their differentiation. Transcriptomes from central Elburz, next-generation sequencing (NGS) libraries of archival DNA of topotypic P. persicus, and GenBank-sequences of eastern P. gorganensis allowed phylogenetic analysis with three mitochondrial genomes, supplemented by PCR-amplified mtDNA-fragments from 17 museum specimens, documenting <2% uncorrected intraspecific genetic distance. Our data suggest that these rare salamanders belong to a single species P. persicus s.l. Humankind has a great responsibility to protect this species and the unique biodiversity of the Hyrcanian Forest ecosystems.

COI barcoding of plant bugs (Insecta: Hemiptera: Miridae)

The family Miridae is the most diverse and one of the most economically important groups in Heteroptera. However, identification of mirid species on the basis of morphology is difficult and time-consuming. In the present study, we evaluated the effectiveness of COI barcoding for 123 species of plant bugs in seven subfamilies. With the exception of threeApolygusspecies—A. lucorum,A. spinolae, andA. watajii(subfamily Mirinae)—each of the investigated species possessed a unique COI sequence. The average minimum interspecific genetic distance of congeners was approximately 37 times higher than the average maximum intraspecific genetic distance, indicating a significant barcoding gap. Despite having distinct morphological characters,A. lucorum,A. spinolae, andA. watajiimixed and clustered together, suggesting taxonomic revision. Our findings indicate that COI barcoding represents a valuable identification tool for Miridae and can be economically viable in a variety of scientific research fields.

Intrapopulational variation in color pattern of Trichomycterus davisi (Haseman, 1911) (Siluriformes: Trichomycteridae) corroborated by morphometrics and molecular analysis

Color patterns of the body are commonly used to distinguish and identify species of Trichomycterus. Therefore, variation in color pattern in a population can cause doubt concerning species identification. With the purpose to test the hypothesis of high variation in color pattern of Trichomycterus davisi (Haseman, 1911), 118 specimens were collected in a stream of a private Ecological Park in southern Brazil, of which 88 were used in the morphological analysis and 30 for DNA barcoding analysis. Three phenotypic classes were determined analyzing the distribution, size and shape of dark brown spots and blotches. The results of morphometric analysis indicate a tendency of association of those pigmentation patterns with the standard length, evidencing ontogenetic variation of color pattern in the species. The results of K2P intraspecific genetic distance (<0.72%), haplotypes network and Bayesian phylogenetic tree corroborate the existence of only one species with a high variable color pattern.

DNA profiles to identify Dillenia species (Dilleniaceae) in Thailand

Surveying of the species throughout Thailand revealed 13 identified and one unidentified species including D. aurea, D. excelsa, D. grandifolia, D. hookeri, D. indica, D. obovata, D. ovata, D. parviflora, D. pentagyna, D. philippinensis, D. pulchella, D. reticulata, D. suffruticosa and Dillenia sp. They were phylogenetically examined based on RAPD profiles of 2,290 discrete characters including a monomorphic and 226 polymorphic characteristics. The phylogenetic relationships calculated from theses banding data show that the intraspecific genetic similarity (S) values ranged from 0.990 to 1.000, and the interspecific S values ranged from 0.520 to 0.790. The RAPD method can be effectively used in Dillenia study. However, its quality of unreproducible method, it cannot be reasonable used in any study on specific genes or areas. In additions, sequences from two molecular regions, rbcL gene and psbA-trnH spacer, were analyzed and determined for genetic distances. The rbcL gene sequences were rather ineffective, as all of the paired species displayed no or low genetic distance values. The psbA-trnH spacer sequences were rather effective, with only one pair (D. reticulata and D. parviflora) showing low genetic distance values. The other species pairs indicated rather far genetic distances, ranging from 0.006 (D. parviflora and D. grandifolia) to 0.376 (Dillenia specie and D. indica). Intraspecific genetic distance values ranged from 0–0.003 and 0–0.013 for rbcL and psbA-trnH spacer regions, respectively. Based on the results from both nucleotide variations and DNA fingerprinting, D. grandifolia and D. ovata were distinct Dillenia species.