While comparatively few amphibian species have been described on the North East Asian mainland in the last decades, several species have been the subject of taxonomical debates in relation to the Yellow sea. Here, we sampled Dryophytes sp. treefrogs from the Republic of Korea, the Democratic People's Republic of Korea and the People's Republic of China to clarify the status of this clade around the Yellow sea and determine the impact of sea level change on treefrogs' phylogenetic relationships. Based on genetics, call properties, adult morphology, tadpole morphology and niche modelling, we determined the segregated status species of D. suweonensis and D. immaculatus. We then proceeded to describe a new treefrog species, D. flaviventris sp. nov., from the central lowlands of the Republic of Korea. The new species is geographically segregated from D. suweonensis by the Chilgap mountain range and known to occur only in the area of Buyeo, Nonsan and Iksan in the Republic of Korea. While the Yellow sea is the principal element to the current isolation of the three clades, the paleorivers of the Yellow sea basin are likely to have been the major factor for the divergences within this clade. We recommend conducting rapid conservation assessments as these species are present on very narrow and declining ranges.

In recent years, animal ethics issues have led researchers to explore nondestructive methods to access materials for genetic studies. Cicada exuviae are among those materials because they are cast skins that individuals left after molt and are easily collected. In this study, we aim to identify the most efficient extraction method to obtain high quantity and quality of DNA from cicada exuviae. We compared relative DNA yield and purity of six extraction protocols, including both manual protocols and available commercial kits, extracting from four different exoskeleton parts. Furthermore, amplification and sequencing of genomic DNA were evaluated in terms of availability of sequencing sequence at the expected genomic size. Both the choice of protocol and exuvia part significantly affected DNA yield and purity. Only samples that were extracted using the PowerSoil DNA Isolation kit generated gel bands of expected size as well as successful sequencing results. The failed attempts to extract DNA using other protocols could be partially explained by a low DNA yield from cicada exuviae and partly by contamination with humic acids that exist in the soil where cicada nymphs reside before emergence, as shown by spectroscopic measurements. Genomic DNA extracted from cicada exuviae could provide valuable information for species identification, allowing the investigation of genetic diversity across consecutive broods, or spatiotemporal variation among various populations. Consequently, we hope to provide a simple method to acquire pure genomic DNA applicable for multiple research purposes.

Rapid anthropogenic alterations caused by urbanization are increasing temperatures in urban cores, a phenomenon known as the urban heat island (UHI) effect. Two cicada species, Cryptotympana atrata and Hyalessa fuscata (Hemiptera: cicadidae), are abundant in metropolitan Seoul where their population densities correlate strongly with UHI intensities. Such a positive correlation between cicada density and UHI intensity may be possible if cicada abundance is linked to a certain thermal tolerance. We tested this hypothesis by investigating variation in morphology and thermal responses of two cicada species along a thermal gradient in Seoul and surrounding areas. The morphological responses were measured by metrics such as length, thorax width and depth, and mass. The thermal responses were measured in terms of minimum flight temperature, maximum voluntary temperature and heat torpor temperature. First, we observed a species-specific variation in thermal responses, in which C. atrata displayed a higher thermal threshold for maximum voluntary and heat torpor temperatures than H. fuscata. Second, a positive association between temperature conditions and body sizes were displayed in females H. fuscata, but not in either conspecific males or C. atrata individuals. Third, C. atrata exhibited similar thermal responses regardless of habitat temperature, while H. fuscata in warmer areas showed an increase in heat tolerance. In addition, H. fuscata individuals with bigger thorax sizes were more heat-tolerant than those with smaller thoraxes. Overall, our research is the first to detect a variation in thermal responses and body size of H. fuscata individuals at a local scale. More investigations would be needed to better understand the adaptation mechanisms of insects linked to UHI effects.Temperature is one of the most important abiotic factors affecting daily activities and the life history of cicadas 1-6 . As temperature directly dictates chemical rates and metabolic processes in vivo, a habitat with fluctuating temperatures could drive insects to manage the high energy costs of maintaining body temperature, T b , within a certain range to coordinate reproductive activity 4 . The surrounding conditions may also have an effect on reproductive success and mortality 7-10 . If so, cicadas should select favorable thermal conditions to obtain optimal body temperatures via behavioral-or physiological mechanisms 4,11 .The urban heat island (UHI) effect, in which increased temperatures are associated with urban areas, is a ubiquitous consequence of microclimatic perturbation 12 due to human activities and energy consumption, along with modification of landscapes and urban geometry [13][14][15][16][17] . Elevated temperatures in urban environments are regarded as detrimental as they reduce plant photosynthesis capability 18 and biological diversity 19,20 . Nevertheless, for urban herbivorous insects such as cicadas, such warm habitats may prove advantageous. First, post-morphogenesis development of cicada eggs require a large degree o...

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