The family Thymelaeaceae Juss., includes numerous botanically and medicinally important species, yet its taxonomy, particularly within the genus Daphne Tourn. ex L., remains unresolved. In the present study, we sequenced and de novo assembled the chloroplast (cp) genome of Daphne mucronata Royle from Pakistan and assembled cp genomes of four additional Daphne species, one Dirca L. species and two Thymelaea Mill. species using public raw data. Comparative analyses across 14 genera, including 17 Daphne species, revealed substantial structural variation in cp genomes. The cp genomes contained 111-112 unique genes, including 77-78 protein-coding genes, 30 tRNA genes and four rRNA genes. When considering duplicated copies in the inverted repeat regions, the total gene count ranged from 128 to 142 (83-96 protein-coding genes, 37-38 tRNA genes and eight rRNA genes). Variability in total gene number was primarily driven by inverted repeat contraction and expansion and pseudogenization of some genes. ndhF was pseudogenized in seven Daphne species, whereas ndhI and ndhG were pseudogenized in one species each. Two large inversion events were detected, involving 19 and 40 genes, respectively; the first inversion was synapomorphic and provided significant phylogenetic signal. Phylogenetic analysis of shared protein-coding sequences from 74 species (87 individuals) recovered Daphne as a strongly supported monophyletic clade when taxa previously proposed for exclusion from the genus were omitted, whereas Wikstroemia Endl. and Gyrinops Gaertn. exhibited paraphyly. This study expands the cp genome resources for Thymelaeaceae and provides new insights into the structural evolution of cp genome.

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Due to the evident increase in tick-borne diseases worldwide, it is necessary to constantly update information on the distribution and zoonotic potential of hard ticks. We studied diversity, population structure, and seasonal dynamics of hard tick fauna, faunal similarity and the climate impact on tick occurrence in natural habitats (NHs) (forest communities) and anthropogenic habitats (AHs) (orchards, grasslands, degraded forests) in eastern and central parts of Continental Croatia. Host-seeking hard ticks were sampled by the flag-dragging method in lowland AHs (Bansko Hill (BH); 2023-2024 yr.) and in mountainous NHs (Medvednica Mountain (MM); 2019-2021, 2024 yr.). Overall, 2726 specimens belonging to eight hard tick species ( Ixodes ricinus , I. frontalis , I. hexagonus , I. kaiseri , Haemaphysalis inermis , H. concinna , Dermacentor marginatus , D. reticulatus ) were identified in AHs, while in NHs 1543 hard ticks, belonging to three species ( I. ricinus , I. frontalis , D. reticulatus ), were collected. The most abundant species in both habitat types (47.83% in AHs, 99.80% in NHs) was I. ricinus , showing unimodal seasonal activity within studied NHs and bimodal activity at AHs. Comparison of hard tick fauna in different habitats using the Sørenson index on BH and MM showed a high percentage of similarity (50.0-88.8). At AHs, a significant ( p N ) and the mean monthly air temperatures (°C) for H. inermis ( r = -0.5931; p = 0.0421) and D. reticulatus ( r = -0.6289; p = 0.0285), while their numbers positively correlated ( r = 0.5551; p = -0.2667; r = 0.4430; p = 0.1492) with air humidity (%). In contrast, the number of sampled host-seeking I. ricinus ticks at natural forest habitats on MM was positively associated with air temperature and negatively with air humidity at elevations from 200 to 1000 m a.s.l. ( r = -0.7684; p = 0.0259; at 200 m a.s.l.). Collected specimens of I. frontalis mark the first record for Osijek-Baranja County, while the sampled D. reticulatus on MM represents the first catch at 1000 m a.s.l. in Croatia. This new data on the distribution and seasonality of medically important hard tick species in Continental Croatia contributes to identifying tick-risk foci and high-risk periods.

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We investigated taxonomic and endemic richness, patterns of spatial distribution, cenotic and spatial diversification, and chorological and life form spectra of montane coniferous forests in the central part of the Balkan Peninsula. We collected information on 1435 taxa (1351 at the level of species and 84 subspecies) with 65,289 species-occurrence data, published in 1930 original plots with a total area of about 215 ha in the analysis. All statistical analyses (univariate and multivariate) were performed on binary matrices prepared for different levels of analysis. Our main results showed that the montane coniferous forests of the central Balkan Peninsula represent very species-rich vegetation. At the same time, the high proportion of endemics indicated that the montane coniferous forests of the central Balkan Peninsula differ significantly from Central European and boreal forests of a similar type. Furthermore, we found that there were regional differences in the species composition of the coniferous forests of the Balkan Peninsula, and that the primary centers of floristic richness are located in the area of the central and continental Dinarides. This latter finding suggested that the true centers of the richness of European coniferous forests are located south of the Limestone Alps-Western Dinarides-Carpathian Foothills line in Romania, which used to be considered the center of the richness of the coniferous forests in Europe.

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In order to well identify the 93 wild Cortex Daphnes samples from different species and habitats in western China and develop a standard operating procedure (SOP) for the authentication and quality of them in the future, a comprehensive and efficient identification system based on DNA barcoding and HPLC fingerprint technologies has been developed. The result showed that only 17 samples (18%) were Daphne giraldii Nitsche (DG), which is recorded in Chinese Pharmacopeia, while the others (82%) might have safety hazards. Additionally, the result of HPLC fingerprint analysis indicated that samples in the same species origins and wild distributions could be clustered together, which was consistent with DNA barcoding analysis. The study can provide a significant system for the authentication and quality of commercial Cortex Daphnes herbs. Undoubtedly, this study undoubtedly confirmed that the chemical compositions of Cortex Daphnes herbs were affected by both species origins and ecological environments, which is required more in-depth research.

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The tribe Aquilarieae of the family Thymelaeaceae consists of two genera, Aquilaria and Gyrinops , with a total of 30 species, distributed from northeast India, through southeast Asia and the south of China, to Papua New Guinea. They are an important botanical resource for fragrant agarwood, a prized product derived from injured or infected stems of these species. The aim of this study was to estimate the genome size of selected Aquilaria species and comprehend the evolutionary history of Aquilarieae speciation through molecular phylogeny. Five non-coding chloroplast DNA regions and a nuclear region were sequenced from 12 Aquilaria and three Gyrinops species. Phylogenetic trees constructed using combined chloroplast DNA sequences revealed relationships of the studied 15 members in Aquilarieae, while nuclear ribosomal DNA internal transcribed spacer (ITS) sequences showed a paraphyletic relationship between Aquilaria species from Indochina and Malesian. We exposed, for the first time, the estimated divergence time for Aquilarieae speciation, which was speculated to happen during the Miocene Epoch. The ancestral split and biogeographic pattern of studied species were discussed. Results showed no large variation in the 2C-values for the five Aquilaria species (1.35-2.23 pg). Further investigation into the genome size may provide additional information regarding ancestral traits and its evolution history.

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