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Downy mildew of hops represents a serious disease affecting hops production in all growing regions. Disease management is primarily based on the application of fungicides at regular intervals based on a short-term forecasting methodology that is essential for evaluating the occurrence of theoretical infections. To enable a more reliable assessment of the pathogen’s presence in a given area, spore traps capturing airborne Pseudoperonospora humuli sporangia can be utilized. The use of quantitative real-time PCR (qRT-PCR) for the detection of sporangia collected by these traps allows for the elimination of laborious and time-consuming microscopic counting. Among four tested P. humuli-specific nuclear DNA sequences, an effective qRT-PCR detection method was developed based on the c127233.5e3 sequence. This detection approach was used for the quantification of sporangia from volumetric spore trap samples collected in situ under field conditions at three selected localities in Bohemia and Moravia during the 2021–2022 period. The obtained results were compared with the short-term forecasting method of the downy mildew (HDM) weather index (I) based on meteorological data. The overall course of the HDM weather index (I) closely correlated with the occurrence of sporangia: after reaching the maximum HDM weather index (I) value, the highest sporangium detection was observed with a time delay of 1–2 weeks at all the monitored sites. The results corresponded well with data obtained from volumetric spore traps in Germany, and the qRT-PCR method proved to be fully comparable to light microscopy. The combination of volumetric spore traps and qRT-PCR can significantly improve the precision of short-term forecasting systems for P. humuli infection, thereby enabling more efficient fungicide application programs in hops protection and contributing to a better understanding of the pathogen’s dispersal dynamics.

Background Hop ( Humulus lupulus L.) bitter acids are valuable metabolites for the brewing industry. They are biosynthesized and accumulate in glandular trichomes of the female inflorescence (hop cone). The content of alpha bitter acids, such as humulones, in hop cones can differentiate aromatic from bitter hop cultivars. These contents are subject to genetic and environmental control but significantly correlate with the number and size of glandular trichomes (lupulin glands). Results We evaluated the expression levels of 37 genes involved in bitter acid biosynthesis and morphological and developmental differentiation of glandular trichomes to identify key regulatory factors involved in bitter acid content differences. For bitter acid biosynthesis genes, upregulation of humulone synthase genes, which are important for the biosynthesis of alpha bitter acids in lupulin glands, could explain the higher accumulation of alpha bitter acids in bitter hops. Several transcription factors, including HlETC1, HlMYB61 and HlMYB5 from the MYB family, as well as HlGLABRA2, HlCYCB2–4, HlZFP8 and HlYABBY1, were also more highly expressed in the bitter hop cultivars; therefore, these factors may be important for the higher density of lupulin glands also seen in the bitter hop cultivars. Conclusions Gene expression analyses enabled us to investigate the differences between aromatic and bitter hops. This study confirmed that the bitter acid content in glandular trichomes (lupulin glands) is dependent on the last step of alpha bitter acid biosynthesis and glandular trichome density.

Traditional hop (Humulus lupulus L.) cultivars have been used in the brewing industry for a long time. Globally, about ten new breeding lines were released to the market in each decade from ~1970 to 1999. Since 2006, the rate of release of new cultivars has increased tenfold. It is, therefore, important to identify their genotype and origin. Molecular genetic methods based on DNA are the most appropriate technology for this purpose. Recently, we developed an efficient marker system for the authenticity control of hop genotypes based on expressed sequence tag-simple sequence repeats (EST-SSR). In the present study, we enlarged the previously established EST-SSR set with 27 new polymorphic markers and evaluated molecular genetic variability within 135 traditional and new world hop cultivars. Two sets of 10 markers effectively differentiated all used cultivars, with the exception of cultivars derived from the same original genotype such as Saaz, Spalt, Tettnang and Nadwislawsky. Results of molecular genetic variability analyses corresponded with the genealogical and geographical origin of the key cultivars.

Viroids are small infectious pathogens, composed of a short single-stranded circular RNA. Hop (Humulus lupulus L.) plants are hosts to four viroids from the family Pospiviroidae. Hop latent viroid (HLVd) is spread worldwide in all hop-growing regions without any visible symptoms on infected hop plants. In this study, we evaluated the influence of HLVd infection on the content and the composition of secondary metabolites in maturated hop cones, together with gene expression analyses of involved biosynthesis and regulation genes for Saaz, Sládek, Premiant and Agnus cultivars. We confirmed that the contents of alpha bitter acids were significantly reduced in the range from 8.8% to 34% by viroid infection. New, we found that viroid infection significantly reduced the contents of xanthohumol in the range from 3.9% to 23.5%. In essential oils of Saaz cultivar, the contents of monoterpenes, terpene epoxides and terpene alcohols were increased, but the contents of sesquiterpenes and terpene ketones were decreased. Secondary metabolites changes were supported by gene expression analyses, except essential oils. Last-step biosynthesis enzyme genes, namely humulone synthase 1 (HS1) and 2 (HS2) for alpha bitter acids and O-methytransferase 1 (OMT1) for xanthohumol, were down-regulated by viroid infection. We found that the expression of ribosomal protein L5 (RPL5) RPL5 and the splicing of transcription factor IIIA-7ZF were affected by viroid infection and a disbalance in proteosynthesis can influence transcriptions of biosynthesis and regulatory genes involved in of secondary metabolites biosynthesis. We suppose that RPL5/TFIIIA-7ZF regulatory cascade can be involved in HLVd replication as for other viroids of the family Pospiviroidae.

Hop ( Humulus lupulus L.) is an emblematic industrial crop in the French North East region that developed at the same time as the brewing activity. Presently, this sector, especially microbreweries, are interested in endemic wild hops, which give beer production a local signature. In this study, we investigated the genetic and metabolic diversity of thirty-six wild hops sampled in various ecological environments. These wild accessions were propagated aeroponically and cultivated under uniform conditions (the same soil and the same environmental factors). Our phytochemical approach based on UHPLC-ESI-MS/MS analysis led to the identification of three metabolic clusters based on leaf content and characterized by variations in the contents of twelve specialized metabolites that were identified (including xanthohumol, bitter acids, and their oxidized derivatives). Furthermore, molecular characterization was carried out using sixteen EST-SSR microsatellites, allowing a genetic affiliation of our wild hops with hop varieties cultivated worldwide and wild hops genotyped to date using this method. Genetic proximity was observed for both European wild and hop varieties, especially for Strisselspalt, the historical variety of our region. Finally, our findings collectively assessed the impact of the hop genotype on the chemical phenotype through multivariate regression tree (MRT) analysis. Our results highlighted the ’WRKY 224’ allele as a key discriminator between high- and low-metabolite producers. Moreover, the model based on genetic information explained 40% of the variance in the metabolic data. However, despite this strong association, the model lacked predictive power, suggesting that its applicability may be confined to the datasets analyzed.

In vitro meristem cultures have been used for the production of hop (Humulus lupulus L.) virus-free rootstocks worldwide, because multipropagation is considered to preserve the genetic stability of the produced plantlet. Nevertheless, in vitro tissue cultures can cause genetic and epigenetic changes. Therefore, we studied the genetic and epigenetic variability of Saaz Osvald's clones, Sládek and Premiant cultivars on the DNA methylation level by methylation-sensitive amplification polymorphism (MSAP). In vitro propagated plants, acclimatised glasshouse rootstocks as well as derived mericlones and control plants under field conditions were used for the analyses. A total of 346 clearly and highly reproducible amplified products were detected in the MSAP analyses within the studied hop plants. We found 16 polymorphic products (4.6% of products) and 64 products with methylation changes (18.5% of products) in the analyses. The demethylation events were comparable to the de novo methylation events. Most demethylation changes were found in the in vitro plants, but only a few of them were found in the derived mericlones under field conditions. In contrast, the de novo methylation changes persisted in the acclimatised plants under glasshouse or field conditions. A hierarchical cluster analysis was used for the evaluation of the molecular genetic variability within the individual samples. The dendrogram showed that the individual samples of the same variety, more or less, clustered together. Because the methylation status varied during the virus-free rootstock production process, we suppose that de/methylation process is a natural tool of epigenetics and evolution in vegetatively propagated plants.

Sweet cherries are self-incompatible, which is determined by a gametophytic self-incompatibility system (GSI). The self-incompatibility is controlled by a multi-allelic S-locus. Knowledge about the S-allele constitution of the cultivars is essential for fruit growers and breeders. Recently, molecular PCR-based methods have been developed to distinguish all S-alleles in sweet cherries. In our work, we analysed S-locus genotypes by 13 universal and allele-specific PCR primer combinations within 117 registered, old and local sweet cherry cultivars from the Czech genetic resources of the Research and Breeding Institute of Pomology in Holovousy, the Czech Republic. We confirmed the previous S-genotyping for 66 accessions except for Drogans Gelbe, Hedelfinger, Erika, Meckenheimer Frühe, Badeborner, Bing, Alfa, Gamma, Huldra, Rivan, Valerij Tschkalov, Viola and Winkler's Frühe. It could be due to either mislabelling or mistakes in the previous analyses. Newly, S-genotyping was determined for 51 accessions in which we found 4 new S-loci combinations. We detected the S-locus combinations in 19 incompatibility groups. The most frequent incompatibility groups were III (S3S4), II (S1S3), IV (S2S3), and VI (S3S6) with 22, 20, 12 and 12 genotypes, respectively.

The presence of genes for resistance to scab (Venturia inaequalis) and powdery mildew (Podosphaera leucotricha) was studied using molecular markers in a sample of 279 apple cultivars from the Czech collection of apple genetic resources. The sample comprised 37 cultivars supposed to have the Vf gene for scab resistance, 97 reference world cultivars and 145 old and local cultivars. Six PCR molecular markers for the scab resistance genes Vf, Vm, Vbj, Vr and Vh and three PCR molecular markers for the powdery mildew resistance genes Pl-w, Pl-1 and Pl-d were used. The marker for the major scab resistance gene Vf was detected in all cultivars supposed to have Vf, except in Romus 1, and in the three small-fruited cultivars Malus Evereste, Golden Gem and Hilleri. The markers of the Vr and Vh scab resistance genes were detected in 22 cultivars in combination with the marker for Vf, in 56 reference world cultivars and in 82 old and local apple cultivars. PCR molecular markers for one or two of the powdery mildew resistance genes were detected in the small-fruited cultivars Malus Evereste, Golden Gem, prof. Sprengeri and Hilleri; and in the larger fruited cultivars Hagloe Crab, Borovinka and Tita Zetei. We did not find markers for the scab resistance genes Vm and Vbj in any of the studied cultivars. They are absent also in the remaining part of the Czech collection of apple genetic resources.

Hop (Humulus lupulus L.) is an emblematic industrial crop in the French North East region that developed at the same time as the brewing activity. Presently, this sector, especially microbreweries, are interested in endemic wild hops, which give beer production a local signature. In this study, we investigated the genetic and metabolic diversity of thirty-six wild hops sampled in various ecological environments. These wild accessions were propagated aeroponically and cultivated under uniform conditions (the same soil and the same environmental factors). Our phytochemical approach based on UHPLC-ESI-MS/MS analysis led to the identification of three metabolic clusters based on leaf content and characterized by variations in the contents of twelve specialized metabolites that were identified (including xanthohumol, bitter acids, and their oxidized derivatives). Furthermore, molecular characterization was carried out using sixteen EST-SSR microsatellites, allowing a genetic affiliation of our wild hops with hop varieties cultivated worldwide and wild hops genotyped to date using this method. Genetic proximity was observed for both European wild and hop varieties, especially for Strisselspalt, the historical variety of our region. Finally, our findings collectively assessed the impact of the hop genotype on the chemical phenotype through multivariate regression tree (MRT) analysis. Our results highlighted the WRKY 224 allele as a key discriminator between high- and low-metabolite producers. Moreover, the model based on genetic information explained 40% of the variance in the metabolic data. However, despite this strong association, the model lacked predictive power, suggesting that its applicability may be confined to the datasets analyzed.Competing Interest StatementThe authors have declared no competing interest.