DNA barcodes evidence the contact zone of eastern and western caddisfly lineages in the Western Carpathians

The region of the Western Carpathians is, among other aspects, very important for survival and diversity of European freshwater fauna due to the presence of a large number of (sub)mountain springs and streams. However, these ecologically and faunistically diversified habitats are still understudied in the context of genetic diversity and population structure of their inhabitants. This study focuses on genetic diversity and distribution patterns of the caddisfly Rhyacophila tristis, common and widespread representative of mountain freshwater fauna. Analysis of the COI mitochondrial marker revealed presence of the western and eastern lineages, with samples from both lineages being grouped in BOLD (Barcode of Life Data System) into separate BINs (Barcode Index Numbers). Our data indicates that eastern lineage (BIN_E) is more closely related to the Balkan populations than to co-occurring western lineage (BIN_W), and that the contact zone of the lineages passes through the W Carpathians. The study revealed phylogeographic and demographic differences between lineages, supporting hypothesis of their evolutionary independence and specific ecological preferences. The obtained genetic data of the R. tristis population from W Carpathians improved our knowledge about population genetics of this aquatic species and can contribute to understanding the state and evolution of biodiversity of freshwater ecosystems in Europe.

Clinging to survival: Critically Endangered Chapman's pygmy chameleon Rhampholeon chapmanorum persists in shrinking forest patches

The Critically Endangered Chapman's pygmy chameleon Rhampholeon chapmanorum is endemic to the low elevation rainforest of the Malawi Hills in southern Malawi. Much of this forest has been converted to agriculture and it was uncertain whether chameleon populations have persisted. We used current and historical satellite imagery to identify remaining forest patches and assess deforestation. We then surveyed forest patches for the presence of this chameleon, and assessed its genetic diversity and structure. We estimated that 80% of the forest has been destroyed since 1984, although we found extant populations of the chameleon in each of the patches surveyed. Differentiation of genetic structure was strong between populations, suggesting that gene flow has been impaired. Genetic diversity was not low, but this could be the result of a temporal lag as well as lack of sensitivity in the mitochondrial marker used. Overall, the impact of forest loss is assumed to have led to a large demographic decline, with forest fragmentation preventing gene flow.

Quick Identification of Genetic Provenances and Best Practice Woodland Management of Chinese Pine (Pinus tabuliformis)

Chinese pine (Pinus tabuliformis) is one of the most widespread, ecologically and economically important tree species in North China. In this study, we analyzed and compared the genetic diversity and population structure of 158 individuals from 17 populations of P. tabuliformis, by group III, a new mitochondrial marker system (nad1-2, nad4-3, nad5-1 and nad7-1) with two other groups of marker systems. In contrast to the conservation in the evolution of the mitochondrial sequence, the new mitochondrial marker system of P. tabuliformis shows as extremely high polymorphism as 25, whose haplotypes are more than four times of the group I marker system (nad1-2, nad4-3, and nad5-1) as 8 haplotypes. Although the group II, nad7-1 (19 haplotypes), showed high resolution in the provenance identification of P. tabuliformis, the new mitochondrial marker system is more accurate for detection of specific groups like HL, WT and NS and powerful to differentiate populations between GD and SS. The results suggested that the new mitochondrial marker system is as high as the resolution of GBS (genotype by seqencing). It is much more available and will be of great help to provenance identification and molecular assisted breeding of P. tabuliformis. This study will make theoretical foundation for the following studies on the evaluation, cultivation and germplasm management of P. tabuliformis populations and aid the breeding, biodiversity and conservation programs of forest species.

Phylogeography of a Darevskia (caucasica) complex (Lacertidae: Sauria) based on the cytochrome b mitochondrial gene analysis

The range of superspecific complex of rock lizards Darevskia (caucasica) (D. alpina, D. c. caucasica, D. c. vedenica, D. daghestanica) includes the Main Caucasian ridge and its spurs on the southern and northern macroslopes within heights from 500 m to 3200 m above sea level. A fragment of the mitochondrial cytochrome b gene (1096 bp) was analyzed in 48 specimens of these lizards. Specimens were collected in 23 localities. We have found 39 unique haplotypes. The minimum genetic diversity was found in D. c. vedenica, maximum in D. c. caucasica. All taxa of the complex, except for D. c. vedenica, demonstrate genetic heterogeneity. Four clades can be distinguished on the dendrogram of phylogenetic relationships. They correspond to D. alpina, D. c. caucasica, D. c. vedenica, D. daghestanica. D. c. caucasica forms two distinct haplogroups. On the phylogenetic tree, the sample of D. c. vedenica formed an independent clade close to D. c. caucasica. The Darevskia (caucasica) complex is polyphyletic according to the analysis of the mitochondrial marker. Nucleotide sequences of D. alpina formed a single clade with Darevskia (saxicola) complex representatives and combined with the sequences of D. brauneri into subclade. On the example of the population of D. daghestanica, found in a forest biotope without rocky outcrops and genetically not differing from individuals from the adjacent territory, the ability of rocky lizards to master biotopes unusual for them in a short period of time was demonstrated. This example demonstrates the ability of rock lizards to adapt to unusual biotopes in a short period of time. Genetic distance between D. c. vedenica and D. c. caucasica is comparable to the distance between D. c. caucasica and D. daghestanica. We assume the existence of an independent form of D. caucasica ssp. inhabiting South Ossetia and adjacent territories.

Anti-Inflammatory, Antidiabetic Properties and In Silico Modeling of Cucurbitane-Type Triterpene Glycosides from Fruits of an Indian Cultivar of Momordica charantia L.

Diabetes mellitus is a chronic disease and one of the fastest-growing health challenges of the last decades. Studies have shown that chronic low-grade inflammation and activation of the innate immune system are intimately involved in type 2 diabetes pathogenesis. Momordica charantia L. fruits are used in traditional medicine to manage diabetes. Herein, we report the purification of a new 23-O-β-d-allopyranosyl-5β,19-epoxycucurbitane-6,24-diene triterpene (charantoside XV, 6) along with 25ξ-isopropenylchole-5(6)-ene-3-O-β-d-glucopyranoside (1), karaviloside VI (2), karaviloside VIII (3), momordicoside L (4), momordicoside A (5) and kuguaglycoside C (7) from an Indian cultivar of Momordica charantia. At 50 µM compounds, 2–6 differentially affected the expression of pro-inflammatory markers IL-6, TNF-α, and iNOS, and mitochondrial marker COX-2. Compounds tested for the inhibition of α-amylase and α-glucosidase enzymes at 0.87 mM and 1.33 mM, respectively. Compounds showed similar α-amylase inhibitory activity than acarbose (0.13 mM) of control (68.0–76.6%). Karaviloside VIII (56.5%) was the most active compound in the α-glucosidase assay, followed by karaviloside VI (40.3%), while momordicoside L (23.7%), A (33.5%), and charantoside XV (23.9%) were the least active compounds. To better understand the mode of binding of cucurbitane-triterpenes to these enzymes, in silico docking of the isolated compounds was evaluated with α-amylase and α-glucosidase.

Distribution and Genetic Diversity of Aedes aegypti Subspecies across the Sahelian Belt in Sudan

Aedes aegypti is the most important arboviral disease vector worldwide. In Africa, it exists as two morphologically distinct forms, often referred to as subspecies, Aaa and Aaf. There is a dearth of information on the distribution and genetic diversity of these two forms in Sudan and other African Sahelian region countries. This study aimed to explore the distribution and genetic diversity of Aedes aegypti subspecies using morphology and Cytochrome oxidase-1 mitochondrial marker in a large Sahelian zone in Sudan. An extensive cross-sectional survey of Aedes aegypti in Sudan was performed. Samples collected from eight locations were morphologically identified, subjected to DNA extraction, amplification, sequencing, and analyses. We classified four populations as Aaa and the other four as Aaf. Out of 140 sequence samples, forty-six distinct haplotypes were characterized. The haplotype and nucleotide diversity of the collected samples were 0.377–0.947 and 0.002–0.01, respectively. Isolation by distance was significantly evident (r = 0.586, p = 0.005). The SAMOVA test indicated that all Aaf populations are structured in one group, while the Aaa clustered into two groups. AMOVA showed 53.53% genetic differences within populations and 39.22% among groups. Phylogenetic relationships indicated two clusters in which the two subspecies were structured. Thus, the haplotype network consisted of three clusters.

A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging

Despite the urgent need to image living specimens for cutting-edge biological research, most existing fluorescent labeling methods suffer from either poor optical properties or complicated operations required to realize cell-permeability and specificity. In this study, we introduce a method to overcome these limits—taking advantage of the intrinsic affinity of bright and photostable fluorophores, no matter if they are supposed to be live-cell incompatible or not. Incubated with living cells and tissues in particular conditions (concentration and temperature), some Atto and BODIPY dyes show live-cell labeling capability for specific organelles without physical cell-penetration or chemical modifications. Notably, by using Atto 647N as a live-cell mitochondrial marker, we obtain 2.5-time enhancement of brightness and photostability compared with the most commonly used SiR dye in long-term imaging. Our strategy has expanded the scientist's toolbox for understanding the dynamics and interactions of subcellular structures in living specimens.

First record of albinism in spiny rats of genus Proechimys (Rodentia: Echimyidae) from Western Amazon

Albinism is a rare condition in natural populations. One of the factors that would lead to higher allele frequency for the albino phenotype is bottleneck events, causing small population sizes and loss of genetic diversity. Here, we report the first record of albinism in one specimen of Proechimys gardneri from Porto Velho, Rondônia, Brazil. This is also the first record of albinism for the genus and for the family Echimyidae. We identified the specimen through morphology and confirmed the identification by 801 base pairs (bp) of mitochondrial marker cytochrome b (Cyt b). We also discuss the selection against albino phenotype in natural populations.