Verticillium wilt of hop (Humulus lupulus L.), caused by the soil-borne pathogen Verticillium nonalfalfae, is a devastating disease with no effective chemical control. In European hop-growing regions, breeding resistant cultivars is the most effective strategy. The lack of response differences in earlier studies suggests constitutive resistance. We therefore conducted a genome-wide association study (GWAS) using a phased hop genome assembly to improve detection of Verticillium resistance loci. A bi-parental population of 142 genotypes, derived from a cross between resistant Wye Target and susceptible BL2/1, was phenotyped for Verticillium wilt resistance and genotyped by sequencing. Association analyses with five statistical models (MLM in TASSEL 5, MLM, MLMM, FarmCPU and BLINK in GAPIT) did not identify any significant SNPs; however, several candidate loci were identified using exploratory threshold, particularly in the phase 2 genome assembly, including a wall-associated kinase (WAK) consistently detected across both genome phases and all models. GWAS results were further assessed with a Random Forest model, which identified SNPs of high feature importance and showed adequate predictive power (accuracy ≈ 0.4, correlation ≈ 0.8) for preliminary breeding screening. These findings provide an initial set of candidate markers and exploratory prediction models for Verticillium wilt resistance in hop, representing a valuable genomic resource for future marker-assisted selection and breeding strategies.

This study evaluates the susceptibility of various hop varieties to three viroids: Hop Latent Viroid (HLVd), Hop Stunt Viroid (HSVd), and Citrus Bark Cracking Viroid (CBCVd), all of which pose significant threats to Slovenian hop production. The experiment was conducted under in vitro conditions, involving the cultivation of 13 different hop varieties in tissue cultures, alongside the in vitro synthesis of viroid transcripts and subsequent inoculation of the plants. The plants were then monitored for growth, development, and their responses to viroid infection. The findings reveal the complex interactions between hops and viroids, with successful inoculation observed in all varieties. However, infection rates varied, with HSVd showing the highest rates of infection, followed by CBCVd and HLVd. This research enhances the understanding of viroid dynamics in hop plants, which is crucial for developing effective disease management strategies. Despite the insights gained, no resistance or tolerance to the viroids was identified in any of the tested hop varieties.

Konoplja vsebuje zanjo specifične sekundarne metabolite kanabinoide, ki imajo veliko pozitivnih učinkov na zdravje ljudi in živali. Zaradi dvodomnosti in tujeprašnosti vrste ter populacijskih sort pogosto prihaja do pojava različnih fenotipov znotraj sorte. V naši raziskavi smo glede na vizualne lastnosti znotraj sorte Carmagnola selected (CS) in selekcije Finola odbrali različne fenotipe ter pri odbranih vzorcih analizirali kanabinoidni profil, kjer smo določili vsebnost 10 različnih kanabinoidov s tehniko HPLC. Podatke smo statistično obdelali ter ugotovili, da je bila najvišja vsebnost kanabinoida CBD-A pri fenotipih selekcije Finola (6,36–6,59 %), najvišja vsebnost CBG-A je bila pri fenotipu CI (1,62 %) najvišja vsebnost THC-A pa prav tako pri fenotipu CI (0,91 %). Z analizo PCA smo ugotovili, da se sorta CS in selekcija Finole med sabo ločita, medtem ko se na podlagi povprečij fenotipov in analiziranih kanabinoidov znotraj sorte ločita le fenotipa sorte CS. Z izračunom celokupnih vsebnosti kanabinoidov in razmerja med kanabinoidoma THC in CBD smo ugotovili, da lahko fenotip CI uvrstimo v kemotip II, vse ostale kemotipe pa v kemotip III. Fenotipi selekcije Finola so imeli daleč najvišjo celokupno vsebnost kanabinoida CBD (6,18–6,42 %).

This study evaluated the expression of two candidate transcripts (1902 and 94443) in response to Citrus Bark Cracking Viroid (CBCVd) infection in three hop cultivars: Celeia, Styrian Cardinal, and Styrian Wolf. A root grafting technique was used for plant inoculation, and leaf samples were collected before symptoms appeared. The transcripts were selected based on bioinformatic analysis of RNA-seq data, that suggested high transcript levels in CBCVd-infected plants. qPCR results did not fully confirm this in the early stages of infection, the highest transcript expression was observed in CBCVd-infected Styrian Wolf at 4 months past inoculation, but surprisingly, expression dropped by 5 months past inoculation. This initial analysis offers insights into transcript expression during CBCVd infection, but further research is needed to confirm these findings and explore the potential of these transcripts as early infection markers.

One of the main breeding objectives for hemp (Cannabis sativa L.) cultivation is higher seed yield important also for the oil yields. However, there has been limited research on hemp breeding that focuses on these topical features. In this study, 17 crosses were created using 12 different hemp varieties. For each variety, five female inflorescences were selected for targeted crossing and one for self-pollination. Seeds from the crosses were sown in separate plots with a seeding density of 30 kg/ha. The number of male and female plants, plant height, internode length, steam thickness, inflorescence length, number of inflorescences per plant, and plant number per plot were measured in crossbred family plants. The absolute weight of seeds per plot, the degree of marbling and oil content were determined. The progeny of the cross between Helena and Tiborszallasi were found to have the highest 100 seed mass (1.546 g, upper half). The offspring of self-pollinated varieties Finola, Tiborszallasi, Fedora 17 and the cross between Helena and Tiborszallasi produced the highest oil content. The progeny of the cross between Futura 75 and KC Dora showed the most promising results in terms of high yield production. This study provided the first important data pertaining to hemp breeding with the goal of increasing oil content and seed yield.

The antimicrobial activity of hop extracts obtained from different hop genotypes were investigated against Staphylococcus aureus and Lactobacillus acidophilus. In this study the pure xanthohumol, purified β-acids rich fraction, as well as α-acids with β-acids rich fraction were used to test antimicrobial activity against Staphylococcus aureus and Lactobacillus acidophilus; whereby, the antimicrobial activity of different hop extracts against Lactobacillus acidophilus was studied for the first time. Microbial susceptibility to purified hydroacetonic extracts from different hop varieties was investigated by the broth microdilution assay to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The hop hydroacetonic extracts were more effective against Staphylococcus aureus than against Lactobacillus acidophilus. Strong inverse correlations of MIC and MBC values were obtained with xanthohumol, cohumulone, n+adhumulone, colupulone and n+adlupulone contents, suggesting that the identified chemical hop compounds are directly responsible for antimicrobial effects. Moreover, the effect of the growth medium strength on the MIC values of hop extracts against Staphylococcus aureus was systematically investigated for the first time. The current study also reveals the effect of different hop extracts on Staphylococcus aureus, which responds to their presence by lag phase extension and generation time prolongation.

Hemp (Cannabis sativa) contains a great variety of secondary plant constituents. Besides cannabinoids and terpenoids, polyphenols also play an important role in hemp. Besides their role in plant defense mechanisms, they exhibit many positive effects, e.g. antioxidant acitivity, which may be beneficial for human health. In our study, total polyphenol content (TPC) and antioxidant activity in female inflorescences in six different hemp phenotypes of three varieties (Carmagnola Selected [C], Tiborszallasi [T], and Finola Selection [F]) were determined. Two different extraction solvents were used: water and ethanol (99.8 %). TPC was measured using Folin-Ciocalteu's reagent, and antioxidant activity was determined using the DPPH method. TPC and antioxidant activity analysis both showed better results for ethanolic extracts. The highest TPC was found in ethanolic extracts of the Finola Selection phenotype FIV (31.8 mg gallic acid equivalent/g dry matter) and the highest antioxidant activity in the FII phenotype (75.3 %). For water extracts the highest TPC was evaluated in the FII phenotype (6.4 mg GAE/g dry matter) and the highest antioxidant activity in CII and TIII phenotypes (65 %).

In addition to delta-9-tetrahydrocannabinol (THC) a nd cannabidiol (CBD), other phytocannabinoids, such as can-nabigerol (CBG) and cannabichromene (CBC), also have beneficial effects on human health. A high content of CBG is found in plants with the B-0 genotype, whereas CBC is independent of the allelic chemotype locus B. In basic research models such as mice or rats, CBG has demonstrated anticancer properties, particularly against breast cancer. CBG has shown anti-inflammatory effects on murine colitis and on inflammatory bowel disease as well as stimulatory effects on the feeding behaviors of mice. It has also exhibited inhibition of aldose reductase, which is known to cause an accumulation of sorbitol and increase glucose levels in the blood, which may lead to diabetes. Cannabinoid CBC has also shown anti-inflammatory effects and reduced hypermobility in the gut and has displayed potential in vitro effect on adult neural stem progenitor cells. CBC also exerts modest analgesic properties in rodents, as well as anti-fungal, anti-bacterial, pro-apoptotic, and anti-proliferative effects in tumor cells.

SummaryWe recently described, inCannabis sativa, the oldest sex chromosome system documented so far in plants. Based on our estimate of its age, we predicted that it should be shared by its sister genusHumulus, which is known to also possess XY sex chromosomes.Here, we used transcriptome sequencing of a F1 family ofHumulus lupulusto identify and study the sex chromosomes in this species using the probabilistic method SEX-DETector.We identified 265 sex-linked genes inH. lupulus, located on the chromosome that is also theC. sativasex chromosome pair. Using phylogenies of sex-linked genes, we show that a region of these chromosomes had already stopped recombining in the common ancestor of the two species. Furthermore, as inC. sativa, Y gene expression was reduced in correlation to the position on the X chromosome, and strongly Y degenerated genes showed dosage compensation.Here we report, for the first time in the Angiosperms, a sex chromosome system that is shared by two different genera. Recombination suppression started at least 21-25 My ago, and then (either gradually or step-wise) spread to a large part of the sex chromosomes, leading to a strongly degenerated Y.

Eryngium amethystinum L. is a wild vegetable used in Croatia. Both E. amethystinum and E. alpinum L. are decorative plants that can also be used in horticulture. To find out the hidden qualities of these two species, the following biological activities were studied: essential oil (EO) composition, analyzed by gas chromatography and gas chromatography with mass spectrometry (GC, GC–MS); phenolic compound content (PC), analyzed by high performance liquid chromatography (HPLC); total phenols as well as total flavonoids, analyzed by ultraviolet–visible spectrophotometry (UV/Vis); antioxidants, analyzed by 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity (DPPH), β-carotene-linoleic acid assay, chelating activity and reducing power of the extracts; and antimicrobial evaluation by micro-dilution assay and inhibition of Candida albicans blastospore germination. The major constituents of EO were β-caryophyllene (15.2%), α-pinene (10.2%) and 2,3,6-trimethylbenzaldehyde (9.3%) for E. amethystinum and caryophyllene oxide (27.9%), bicyclogermacrene (13.2%) and germacrene D (8.2%) for E. alpinum. The methanol extracts of both species showed a broad spectrum of antibacterial and antifungal activity with minimum inhibitory concentrations (MIC) less than or equal to 1.944 and 1.11 mg/mL, respectively.

Cannabinoid content in different hemp varieties from the Common catalogue of varieties of agricultural plant species is not well known. Hemp (Cannabis saliva L., subsp. saliva) contains a wide range of cannabinoids, where cannabidiol (CBD) and ( - )-trans-Delta(9)-tetrahydrocannabinol (Delta(9) -THC) are the constituents with known therapeutic activity. Also, Delta(9)-THC is recognized as an illicit drug; therefore, cultivation of hemp is restricted to a 0.2 % limit of THC content in many European countries. In this study, the cannabinoid profiles of 15 hemp varieties, accessible to our research group, were analysed. The content of 13 cannabinoids was determined with HLPC (high-performance liquid chromatography) analysis. Large variations in cannabinoid content among varieties that grew in uniform conditions (ANOVA p < 0.05) and also within a single variety were found, which shows on tinuniform genetic profiles of the seed material. The varieties Fedora 17, USO 31, Tisza, Tiborszallasi, and Antal all displayed good response to growth conditions, related to cannabinoid content, in Slovenia.

Hemp (Cannabis sativa L.) cannabinoids and terpenoids have therapeutic effects on human and animal health. Cannabis plants can often have a relatively high heterogeneity, which leads to different phenotypes that have different chemical profiles despite being from the same variety. Little information exists about cannabinoid and terpenoid profiles in different hemp phenotypes within the same variety. For this study, 11 phenotypes from three different varieties (“Carmagnola” selected (CS), “Tiborszallasi” (TS), and “Finola” selection (FS)) were analyzed. The components of essential oil (29) were analyzed using gas chromatography with flame ionization detection (GC/FID), and 10 different cannabinoids of each phenotype were determined using high-performance liquid chromatography (HPLC). Principal component analysis (PCA) and analysis of variance (ANOVA) showed that according to the components of essential oil, FS and TS plants were more uniform than CS plants, where there were great differences between CI and CII phenotypes. The content of cannabinoid CBD-A was the highest in all four FS phenotypes. By comparing cannabinoid profiles, FS was clearly separated from TS and CS, while these two varieties were not clearly distinguishable. Phenotypes TV and CI had the highest total content of Δ-9-THC, while all phenotypes of FS had the highest total content of CBD. The highest total content of CBG was determined in phenotype CI. Obtained results are useful for the development of new supplementary ingredients, for different pharmacy treatments, and for further breeding purposes.

A set of seven SNP markers was used to evaluate the genetic diversity of wild Portuguese hops in comparison with commercial cultivars. A collection of 110 wild genotypes and 33 cultivars was characterized by a high-resolution melting analysis of short amplicons targeting SNP loci. Most of the 143 genotypes (75%) could be differentiated. Phylogenetic analysis showed three main clusters, one included almost exclusively (98%) wild accessions, being the others constituted of both wild and commercial genotypes. The study of population genetic structure placed the accessions into three genetic units, being one exclusively of Portuguese genotypes. The study showed the great biodiversity of the Portuguese hop germplasm and the notable differences (FST = 0.163, p<0.00001) from commercial hops. Results support the usefulness of the use of these seven markers for hop discrimination, with the fast and high-throughput HRMA technique for allele calling and contribute to the affirmation of the high richness breeding potential of Portuguese wild hops.

This study aims to do a morphological and biochemical characterization of selected wild hop populations from the Banja Luka region, as plant material with potential value for introduction into plant selection. Female inflorescences (cones) from seven populations (DKH3, DKH5, DKH6, DKH8, DKH9, DKH10, and DKH11) were sampled in 2018 and 2019. Hop cones were collected in October 2018 and 2019 for morphological characterization, when 50 cones per population were analyzed, and also the cones were collected in mid-September in 2019 for biochemical characterization. Morphological parameters were carried out for: cone length (cm), number of nodes per cone spindle, cone density, and number of seeds per cone, all important agronomic characters in hop production. Also, 27 components of essential oil and 6 components of hop resins were analyzed. Statistical analysis of morphological characteristics was performed by factorial analysis of variance (year × population), and the significance of individual differences was determined by LSD test. The grouping of examined hop populations according to the manifested morphological characteristics was performed by cluster analysis. Statistical analysis for biochemical characteristics was performed by analysis of variance, and the significance of the differences was tested by Tukey’s statistical test. In order to determine similarities between different population multivariant statistical method Principal component analysis (PCA) was applied. According to morphological and biochemical analysis, DKH6 hop population is singled out as the most valuable for hop breeding.

Male specific DNA sequences were selected from a Diversity Arrays Technology (DArT) mapping study to evaluate their suitability for determination of the sex phenotype among young seedlings in a hop (Humulus lupulus L.) breeding program. Ten male specific DArT markers showed complete linkage with male sex phenotype in three crossing families. Following optimization, four were successfully converted into PCR markers and a multiplex PCR approach for their use was developed. Among 197 plants (97 from the world collection; 100 from three segregating families), 94-100% positive correlation with sex phenotypic data was achieved for the single PCR amplification, whereas the multiplex approach showed 100% correlation. To develop a fast and low-cost method, crude sample multiplex PCR was evaluated in 253 progenies from 14 segregating populations without losing accuracy. The study describes, for the first time, the routine application of molecular markers linked to male sex in an intensive Slovenian hop breeding program. The methods described could be employed for screening of sex at the seedling stage in other hop programs worldwide, thereby saving resources for desirable female plants.

The sensory profile of beer is influenced by the hop variety and hopping technology. This study examined pilot (50 L) pale lager beers, hopped in the kettle with (or without) single dry hopping. Seven Slovenian, four Czech and one American genotypes were used to determine the impact of dry hopping on the sensory and volatile profiles of beer. Descriptive and bitterness lingering assessment sensory methods were used. The essential oil composition of the hops and beers was accessed using GC/MS so as to clarify their correlation to sensory profiles of both brewing processes. Changes in essential oils and aroma profiles of beer related to storage were evaluated. Results showed differences among the genotypes in the hop oil profiles in the kettle-hopped beers. Based on the hop dose, dry hopping increased the monoterpene alcohol (r = 0.60) and monoterpene hydrocarbon (r = 0.74) content. Hoppy flavour correlated to the monoterpene alcohols and hydrocarbons. Hop-derived aroma profiles of the genotypes differed. Dry-hopped beers tended to have a more bitter aftertaste than solely kettle-hopped beers. Both kettle and dry hopped beers of all genotypes showed good sensory acceptance (3-4.5; nine-point scale). After storage of beer, the content of most essential oils and hoppy flavour significantly decreased. The sensory aging of the kettle and dry-hopped beers was comparable. Copyright (c) 2018 The Institute of Brewing & Distilling

Drought is one of the major environmental devastating stressors that impair the growth and productivity of crop plants. Despite the relevance of drought stress, changes in physiology and resistance mechanisms are not completely understood for certain crops, including hop (Humulus lupulus L.). In this research the drought response of hop was studied using a conventional physiological approach (gas exchange techniques, fluorescence, relative water content measurements) and proteomic analysis (2D-DIGE). Plants of two cultivars (Aurora and Savinjski golding) were exposed to progressive drought in a pot experiment and analysed at different stress stages (mild, moderate and severe). Measurements of relative water content revealed a hydrostable water balance of hop. Photosynthesis was decreased due to stomatal and non-stomatal limitation to the same extent in both cultivars. Of 28 identified differentially abundant proteins, the majority were down regulated and included in photosynthetic (41%) and sugar metabolism (33%). Fifteen % of identified proteins were classified into the nitrogen metabolism, 4% were related to a ROS related pathway and 7% to other functions.

Satureja montanaL. and S. subspicata Vis. (Lamiaceae) are used for centuries in traditional medicine of Balcanic people in the healing of the lymphatic nodule and respiratory system inflammation. In this paper the amount of total phenols and flavonoids (analyzed by UV/Vis spectrophotometry), phenolic compounds profile (analyzed by HPLC), antimicrobial and antioxidant activities were studied in samples collected in seven per species populations of S. montanaand S. subspicatain Croatia. Eight phenolic compounds (rutin, quercetin, caffeic, p-coumaric, ellagic, protocatehuic, rosmarinic, and syringic acid) were identified and quantified using HPLC in methanolic and ethanolic extracts. Results showed that both species contained polyphenolics and other antioxidant compounds with chelating and radical-scavenging properties. The extracts prepared from both species showed broad spectrum of antimicrobial activity on in vitrotested microbial species (Staphylococcus aureus, Escherichia coli, Candida albicans, C. dubliniensis, C. krusei, C. glabrata, C. parapsilosis, and Microsporum gypseum).

Although sources of resistance to major pathogens exist in cultivated hop germplasm, little effort has been invested to date in developing resistance-linked markers. The aim of this study was to design and evaluate resistance gene analogs (RGAs) potentially useful for marker-assisted selection towards novel resistant hop cultivars. A set of 34 putative hop RGAs was retrieved by screening publicly available hop expressed sequence tags (ESTs) for conserved sequence motifs of common resistance protein domains. Seventeen of these were identified as putative RGAs by BLAST analyses. Exon/intron boundary prediction enabled re-sequencing of 24 EST-RGAs, allowing the acquisition of approximately 5 kbp of novel intronic sequence and 8 kbp of re-sequenced exons. Fifteen EST-RGAs exhibited polymorphisms and were added to a framework linkage map of hop. In addition to providing EST-derived markers potentially useful for resistant hop cultivar development, this study provides valuable insights into the utility of targeting hop introns for marker development.