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Ubiquitous datafication surrounds agriculture stakeholders with data as the raw material for additional value creation, resilience and innovation. Technical and soft competencies of data literacy (DL) are not comprehensively addressed in formal secondary education as an interdisciplinary and general concept. Aiming to assess DL structure set for the agricultural sector and the presence of open data (OD) in formal secondary education, we offer the methodological approach for landscaping 27 DL competencies over six areas of DL adapting the ODI Data Skills Framework for qualitative thematic analysis and quantifying DL structure. Three sources of data: undergraduate (BS) and graduate (MS) course descriptions (561), interviews (32) and course materials (8) offered to students of the University of Zagreb were used to contextualize the approach. At the MS level 47% of courses show DL with prevalent technical DL competencies. Limited DL was found at the BS level (17%) where soft and technical DL competence areas were balanced. Preliminary education guidelines regarding DL in the agricultural sector are given. Further development and use of our methodological approach can ensure core DL competencies and balance soft and entrepreneurship DL in formal secondary education for agricultural sector, but can also be extended to other domains.

This study investigates genetic diversity among five morphotypes and five chemotypes of Ocimum basilicum L. (sweet basil) using amplified fragment length polymorphism (AFLP) markers. Conducted on 80 basil accessions from the Collection of Medicinal and Aromatic Plants of the National Plant Gene Bank of the Republic of Croatia, this research aims to enhance the conservation and utilization of sweet basil’s genetic resources. AFLP analysis using extracted genomic DNA revealed high levels of polymorphism, particularly within the True basil morphotype, which displayed 95.6% polymorphic markers. The results showed genetic differentiation between the morphotypes, particularly between the ’green’ and ’purple’ groups, and within certain chemotypes, such as the High-linalool chemotype (Chemotype A). Principal coordinate analysis (PCoA) and Bayesian clustering further highlighted the genetic structures, with some admixture observed, particularly in the Purple basil B morphotype. Analysis of molecular variance (AMOVA) indicated that most of the genetic diversity was between accessions, emphasizing the value of individual variability. These findings underscore the genetic potential within sweet basil accessions, offering valuable insights for future breeding programs aimed at selecting basil cultivars with tailored biochemical and morphological traits suited for pharmaceutical, culinary, and ornamental applications. The study provides an important basis for the conservation and improvement of basil genetic resources.

The insecticidal compound pyrethrin is synthesized in Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevis.) Sch.Bip.; Asteraceae), a plant species endemic to the eastern Mediterranean. Pyrethrin is a mixture of six compounds, pyrethrin I and II, cinerin I and II, and jasmolin I and II. For this study we sampled 15 natural Dalmatian pyrethrum populations covering the entire natural distribution range of the species; Croatian coastal regions and the islands, inland Bosnia and Herzegovina and Montenegro. The plants were grown in a field experiment under uniform growing conditions to exclude a short-term response to environmental factors and instead observe variation in pyrethrin content and composition among and within populations due to genetic adaptation to the native environment. The drivers of local adaptation were explored by examining the role of bioclimatic factors as a cause of population differentiation. Pyrethrins were extracted by ultrasound-assisted extraction, and the extracts were analyzed by HPLC-UV-DAD. The populations differed significantly in the content and composition of pyrethrins. The highest levels of total pyrethrins (1.27% flower DW), were found in population P14 Budva and the significantly highest levels of pyrethrin I in population P14 Vranjske Njive, Podgorica (66.47% of total pyrethrin). Based on bioclimatic conditions of the sampling sites, populations were grouped into five bioclimatic groups (A, B, C, D, and E), which showed qualitative and quantitative variability in pyrethrin content. The most abundant bioclimatic group was bioclimatic group E, which was characterized by the highest average values for pyrethrin I (53.87% of total pyrethrin), total pyrethrin content (1.06% flower DW) and the ratio of pyrethrin I and II (1.85). The correlation analysis between the pyrethrin compounds and some of the bioclimatic variables (e. g., BIO03 Isothermality and BIO04 Temperature seasonality) showed their significant contribution in explaining the variation of pyrethrins in T. cinerariifolium. The differences in pyrethrin content and composition may be partly due to genetic adaptation to the ecological conditions of the native environment. The obtained data would enable the selection of source populations for breeding programs aimed at producing cultivars with desirable biochemical properties and adaptation to different bioclimatic conditions.

State-owned agricultural land is an asset that the state must manage in a responsible and transparent manner. Agricultural land is extremely important for farmers as it enables them to carry out agricultural activities. Due to its importance to farmers, it is often the subject of debate as stakeholders are often dissatisfied with the allocation and management of state-owned agricultural land. Qualitative research of the process of state agricultural land lease and the associated legislation in the Republic of Croatia enabled the analysis of the existing business model, with the results pointing to shortcomings in the Initial and Evaluation phases of the process. A steady rise in the number of tenders published in 2015–2022 was recorded. Local administrative units in the Continental region scored higher than those in the Adriatic region (both cities and municipalities) in terms of transparency. Unfortunately, the response rate from the local authorities was below 50% across both region and unit, further indicating low transparency. Based on the findings, a proposal of changes in the tendering process was made utilizing a digital platform as an environment for all stakeholders, which provides functionalities essential for the transparent implementation of tenders for the agricultural land lease in Croatia.

Teucrium montanum L. and T. capitatum L. are two plant species with overlapping distribution in the Balkan Peninsula, especially in Croatia, where several occurrences of their putative hybrid species T. × rohlenae have been recorded. Next-generation sequencing of both species and de novo assembly was carried out resulting in 120 contigs for T. montanum and 1685 contigs for T. capitatum assembled. The Dig-up primers pipeline was used for SSR mining of both assemblies, applying different criteria that resulted in 112 SSR candidates for testing. A subset of 41 SSRs was selected and after two rounds of testing, twelve SSRs were developed and characterized. A total of 232 alleles were detected with 5 to 29 alleles per locus, based on the test sample. The genetic diversity analysis of three Teucrium taxa from a single location revealed a higher level of diversity in T. montanum than in T. capitatum with intermediate values for the hybrid species. The NeighborNet diagram and genetic structure analysis grouped the taxa into two separate clusters, the first of which consisted exclusively of T. montanum, while the other was composed of intermixed T. capitatum and T. × rohlenae individuals. The availability of SSR markers for two Teucrium taxa will allow in-depth analysis of genetic diversity and structure, as well as molecular identification of their putative hybrids in the future.

Abstract Saprolegnia parasitica causes saprolegniosis, a disease responsible for significant economic losses in aquaculture and declines of fish populations in the wild, but the knowledge of its distribution and prevalence in the environment is limited. We developed a fast, sensitive and specific S. parasitica droplet digital PCR (ddPCR) assay and demonstrated its applicability for the detection and quantification of the pathogen in environmental samples: swab DNA collected from the host (trout skin, surface of eggs) and environmental DNA extracted from water. The developed assay was used to assess how abiotic (i.e. physico-chemical parameters of the water) and biotic (health status of the host) factors influence the S. parasitica load in the environment. The pathogen load in water samples was positively correlated with some site-specific abiotic parameters such as electrical conductivity (EC) and calcium, while fluorides were negatively correlated, suggesting that physico-chemical parameters are important for determining S. parasitica load in natural waters. Furthermore, skin swabs of injured trout had significantly higher pathogen load than swabs collected from healthy fish, confirming that S. parasitica is a widespread opportunistic pathogen. Our results provide new insights into various environmental factors that influence the distribution and abundance of S. parasitica.

Abstract Studying the population-genetic and phylogeographic structures of a representative species of a particular geographical region can not only provide us with information regarding its evolutionary history, but also improve our understanding of the evolutionary processes underlying the patterns of species diversity in that area. By analysing eight highly polymorphic microsatellite loci and two chloroplast DNA regions, we have investigated the influence of Pleistocene climate fluctuations on the evolutionary history of Salvia officinalis L. (common sage). The populations with the highest genetic diversity were located in the central parts of the Balkan distribution range. A large group of closely related haplotypes was distributed throughout the Balkans and the central Apennines, while the private lineage occupied the southern Apennines. In addition, two highly differentiated lineages were scattered only over the Balkans. The results suggest that a single refugium of the studied species from the last glacial period was located in the central part of the range in the Balkans. Numerous microrefugia, probably spanning several glaciation cycles, were scattered across the Balkans, while colonisation of the Apennines from the Balkans occurred at least on two occasions.

Dogs and dog owners are increasingly present in modern urban spaces, and the construction and maintenance of urban infrastructure that includes places for them has become standard in most major cities. We wanted to investigate the extent to which the City of Zagreb is adhering to open data principles when it comes to dog-related data it makes available to citizens.The openness and quality of dog-related data was analysed in three steps. First, dog-related data was searched on various official websites and portals of the city and the data formats were ranked according to the five-star system for open data. In the second step, based on the available data, a field survey was conducted in 2020 to verify the found datasets and geocode them using a GPS device. In addition, the locations obtained from the local community of dog owners through social media were reviewed. Finally, data obtained from the survey was cross-checked with the government data to assess their quality. Government data on the locations of 300 dog waste bins and 72 green areas where dogs can be walked off-leash were available in Croatian from one or more government sources. All data sets found received the lowest score in terms of open data formats. Field survey revealed differences between the data and reality. The location of 40 dog waste bins could not be confirmed, and additional 53 bins were found that were not mentioned in the data. As for green areas, there were reportedly 10 dog parks in the city of Zagreb. The survey confirmed all locations and discovered 12 more, five of which were mentioned in the data but not designated as dog parks. The results suggest that the municipality needs to update the already open datasets more frequently. Improved implementation of these datasets into existing city data portals or the creation of a separate hub for dog owners would greatly improve the availability and reuse of this data by citizens.

The successful adoption of digital innovations in agricultural production systems is based on the proactive participation of all stakeholders and represents an important step in establishing resilient agri-food chains and creating sustainable value. The key tool for the creation of sustainable value is integrating the nine aspects of the business process (cooperation; inclusion; financing; diversification; communication; policies; knowledge with entrepreneurship; and production) by re-using of open governmental and public endeavours data as well as by the contractual sharing. The objectives of this research are to identify stakeholders in the Croatian agricultural system, and to explore their roles and their potential for data supply and needs for data uptake. Open access repositories were queried to identify stakeholders. Direct observation methods and semi-structured conservational qualitative interviews were used for stakeholder characterisation and data flow detection. Stakeholder importance with respect to current data supply was analysed. Underdeveloped data flow relationships in the agricultural data ecosystem in Croatia could be built in a spontaneous process following the data opening of the Research and Consumer group of stakeholders and promoting data sharing initiatives of the early adopters in the Supplier group. In that way, data opening would be the driver of the effective cooperation creation required for sustainable value creation but also the adoption of the best management practices, sustainable solutions and digital development.

Fireflies (Coleoptera: Lampyridae), with more than 2 000 species in 100 genera worldwide, are a charismatic nocturnal species. Although popular in different cultures because of their association with warm summer evenings in childhood, fireflies are an under-researched insect. Like numerous other insects worldwide, fireflies have experienced declines in their distribution and abundance. Anthropogenic impacts and climate change are likely to influence their development, reproduction, and survival. A project called "Krešo Krijesnica" (eng. "Krešo the Firefly"), used a Citizen Science model of data collection, to determine where are the fireflies located and how abundant are they throughout Croatia. Citizen Science involves the participation of the general or non-scientific public in data collection so determining the basic demographic profile of the citizen scientists involved was also one of the project goals. During the first phase of the project (2019-2021), data on fireflies were provided by citizen scientists through a formal survey on social media (Facebook, Instagram). Phase two aims to open the fireflies' datasets to the public through various open data portals. In the three years of the project, more than 16 000 records of fireflies were collected and analysed from over 1800 sightings. Descriptive statistics showed that the highest firefly population density was found in central Croatia, which is consistent with the greater number of people living in this area and thus a greater chance of firefly detection. Higher number pf female reporters were noted during the project. The dataset collected in this Citizen Science project presents a valuable source of information to the scientific community, especially in the field of entomology, conservation biology and ecology.

Abstract Immortelle (Helichrysum italicum (Roth) G. Don; Asteraceae) is a perennial plant species native to the Mediterranean region, known for many properties with wide application mainly in perfume and cosmetic industry. A total of 18 wild H. italicum populations systematically sampled along the eastern Adriatic environmental gradient were studied using AFLP markers to determine genetic diversity and structure and to identify loci potentially responsible for adaptive divergence. Results showed higher levels of intrapopulation diversity than interpopulation diversity. Genetic differentiation among populations was significant but low, indicating extensive gene flow between populations. Bayesian analysis of population structure revealed the existence of two genetic clusters. Combining the results of F ST - outlier analysis (Mcheza and BayeScan) and genome-environment association analysis (Samβada, LFMM) four AFLP loci strongly associated with the bioclimatic variables Bio03 Isothermality, Bio08 Mean temperature of the wettest quarter, Bio15 Precipitation seasonality, and Bio17 Precipitation of driest quarter were found to be the main variables driving potential adaptive genetic variation in H. italicum along the eastern Adriatic environmental gradient. Redundancy analysis revealed that the partitioning of genetic variation was mainly associated with the adaptation to temperature oscillations. The results of the research may contribute to a clearer understanding of the importance of local adaptations for the genetic differentiation of Mediterranean plants and allow the planning of appropriate conservation strategies. However, considering that the identified outlier loci may be linked to genes under selection rather than being the target of natural selection, future studies must aim at their additional analysis.

Tanacetum cinerariifolium is an endemic species of the eastern Adriatic coast that synthesizes the natural insecticide pyrethrin. We have characterized the complete chloroplast genome of the species and analyzed its phylogeny within the Asteraceae family. The complete chloroplast genome of T. cinerariifolium has a size of 150,136 bp, including a large single-copy (LSC) region of 82,717 bp, a small single-copy (SSC) region of 18,411bp, and a pair of inverted repeats (IRs) of 24,504 bp. The chloroplast genome of T. cinerariifolium encodes 108 genes, including 77 protein-coding genes (PCGs), 27 tRNA genes, and 4 rRNA genes. Phylogenetic analyses based on the complete chloroplast genomes placed T. cinerariifolium in a sister position to species of the genera Artemisia and Chrysanthemum.

The development of bioinformatic solutions is guided by biological knowledge of the subject. In some cases, we use unambiguous biological models, while in others we rely on assumptions. A commonly used assumption for genomes is that related species have similar genome sequences. This is even more obvious in the case of chloroplast genomes due to their slow evolution. We investigated whether the lengths of complete chloroplast sequences are closely related to the taxonomic proximity of the species. The study was performed using all available RefSeq sequences from the asterid and rosid clades. In general, chloroplast length distributions are narrow at both the family and genus levels. In addition, clear biological explanations have already been reported for families and genera that exhibit particularly wide distributions. The main factors responsible for the length variations are parasitic life forms, IR loss, IR expansions and contractions, and polyphyly. However, the presence of outliers in the distribution at the genus level is a strong indication of possible inaccuracies in sequence assembly.

Common bean (Phaseolus vulgaris L.) exhibits a wide range of seed coat colors and this morphological trait is widely used in cultivar identification and assessment of diversity within this species. With an advancement in technology and informatics, new methods of assessing seed color are emerging in addition to traditionally used visual observation. Due to a great variety of color measuring techniques, the evaluation of the agreement between methods is needed prior to using the methods interchangeably. Seed coat color in terms of CIE L*a*b* color coordinates of 100 common bean accessions belonging to five mono-colored landraces was assessed using two methods, colorimeter and Computer vision. The percentage difference between the two methods across all samples for L* color coordinate was 5.81%, for a* color coordinate 23.32% and for b* color coordinate 44.44%. According to Bland-Altman difference plot there is a considerable lack of agreement between the two methods. However, using stepwise discriminant analysis revealed that colorimeter method correctly classified 97% of accessions into their respective landrace, while the classification success of the Computer vision was 99%.

Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevis.) Sch. Bip.) is an outcrossing plant species (2n = 18) endemic to the eastern Adriatic coast and source of the natural insecticide pyrethrin. Due to the high repeatability and large genome (1C-value = 9.58 pg) our previous attempts to develop microsatellite markers using the traditional method were unsuccessful. Now we have used Illumina paired-end whole genome sequencing and developed a specific procedure to obtain useful microsatellite markers. A total of 796,130,142 high-quality reads (approx. 12.5× coverage) were assembled into 6,909,675 contigs using two approaches (de novo assembly and joining of overlapped pair-end reads). A total of 31,380 contigs contained one or more microsatellite sequences, of which di-(59.7%) and trinucleotide (25.9%) repeats were the most abundant. Contigs containing microsatellites were filtered according to various criteria to achieve better yield of functional markers. After two rounds of testing, 17 microsatellite markers were developed and characterized in one natural population. Twelve loci were selected for preliminary genetic diversity analysis of three natural populations. Neighbor-joining tree, based on the proportion of shared alleles distances, grouped individuals into clusters according to population affiliation. The availability of codominant SSR markers will allow analysis of genetic diversity and structure of natural Dalmatian pyrethrum populations as well as identification of breeding lines and cultivars.

Pyrethrin is a specialized metabolite of Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.), Asteraceae, known worldwide as an effective bioinsecticide. It consists of six active compounds: Pyrethrin I and II, cinerin I and II, and jasmolin I and II. Pyrethrin accumulates mainly in the flower heads and its content depends on numerous factors, such as the flower developmental stage. This study aims to investigate the accumulation patterns of six pyrethrin compounds in the flower heads of Dalmatian pyrethrum over the eight developmental stages (FS1 to FS8), and to make a comparison in six natural populations. Ultrasound assisted extraction was used to extract the pyrethrin, while qualitative and quantitative analysis was performed by High performance liquid chromatography. The accumulation patterns of different pyrethrin compounds were generally similar and also synchronous between different populations, while the pyrethrin I/pyrethrin II ratio showed irregular patterns. In all populations studied, the highest increase of all compounds was observed from FS1 to FS2. Their concentration continuously increased, reaching the highest values at FS4 stage (2–5 rows of open disc flowers), and generally decreased gradually towards FS6 or FS7 and then stagnated until the FS8 stage. Despite the very similar accumulation pattern of pyrethrin compounds in the different populations, the significant differences in their content suggest a different genetic background. Knowledge of the dynamics of pyrethrin compounds accumulation across flower development stages is valuable for determining the optimal harvest time of pyrethrum flower heads.

An ethnobotanical survey was carried out in the Knin area (northern Dalmatia, Croatia) with the aim of recording traditional plant use by the local (native) people and contributing to the knowledge of plant biodiversity in the investigated area. Ethnobotanical data were collected by interviewing 40 local people at 17 locations. Data on 123 plant taxa (on average 21 taxa per interview) and 122 unique medicinal and 18 food uses in the local community were recorded. Prevalent medicinal uses of the recorded taxa were the treatment of digestive, respiratory, cardiovascular disorders, and injuries. The most commonly used plant parts gathered were leaves (35%) and flowers (20%). The most commonly collected plants in the investigated area were: Urtica dioica, Thymus longicaulis, Sambucus nigra, and Hypericum perforatum.

Hop (Humulus lupulus L.) is used in the brewing industry as a source of compounds responsible for the bitterness, aroma, and preservative properties of beer. In this study, we used microsatellite markers to investigate genetic diversity and genetic differentiation of wild hop populations sampled in the northwestern part of Croatia. Analysis of 12 microsatellite loci revealed high diversity and weak population differentiation among wild hop populations. A total of 152 alleles were determined with an average of 12.67 alleles per locus. Observed heterozygosity ranged from 0.689 to 0.839 (average 0.767) and expected heterozygosity ranged from 0.725 to 0.789 (average 0.760). A total of 38 private alleles were detected. The data suggest that H. lupulus populations are not affected by recent bottlenecks. The degree of genetic differentiation among populations was low and not significant for most pairwise FST values, except for the pair of geographically most distant populations. The results did not indicate the existence of genetic structure among the sampled populations. The high genetic diversity and low differentiation among populations, combined with the absence of isolation by distance, indicate the existence of substantial gene flow among wild hop populations. Therefore, extensive sampling per population is clearly required to assess the genetic diversity of hop populations. Sampling strategies involving sampling across a large number of localities represented by only a few samples could lead to erroneous conclusions.

With the development of next-generation sequencing technology and bioinformatics tools, the process of assembling DNA sequences has become cheaper and easier, especially in the case of much shorter organelle genomes. The number of available DNA sequences of complete chloroplast genomes in public genetic databases is constantly increasing and the data are widely used in plant phylogenetic and biotechnological research. In this work, we investigated possible inconsistencies in the stored form of publicly available chloroplast genome sequence data. The impact of these inconsistencies on the results of the phylogenetic analysis was investigated and the bioinformatic solution to identify and correct inconsistencies was implemented. The whole procedure was demonstrated using five plant families (Apiaceae, Asteraceae, Campanulaceae, Lamiaceae and Rosaceae) as examples.