The first two chromosome-scale genome assemblies of American hazelnut enable comparative genomic analysis of the genus Corylus.

The native, perennial shrub American hazelnut (Corylus americana) is cultivated in the Midwestern United States for its significant ecological benefits, as well as its high-value nut crop. Implementation of modern breeding methods and quantitative genetic analyses of C. americana requires high-quality reference genomes, a resource that is currently lacking. We therefore developed the first chromosome-scale assemblies for this species using the accessions 'Rush' and 'Winkler'. Genomes were assembled using HiFi PacBio reads and Arima Hi-C data, and Oxford Nanopore reads and a high-density genetic map were used to perform error correction. N50 scores are 31.9 Mb and 35.3 Mb, with 90.2% and 97.1% of the total genome assembled into the 11 pseudomolecules, for 'Rush' and 'Winkler', respectively. Gene prediction was performed using custom RNAseq libraries and protein homology data. 'Rush' has a BUSCO score of 99.0 for its assembly and 99.0 for its annotation, while 'Winkler' had corresponding scores of 96.9 and 96.5, indicating high-quality assemblies. These two independent assemblies enable unbiased assessment of structural variation within C. americana, as well as patterns of syntenic relationships across the Corylus genus. Furthermore, we identified high-density SNP marker sets from genotyping-by-sequencing data using 1343 C. americana, C. avellana and C. americana × C. avellana hybrids, in order to assess population structure in natural and breeding populations. Finally, the transcriptomes of these assemblies, as well as several other recently published Corylus genomes, were utilized to perform phylogenetic analysis of sporophytic self-incompatibility (SSI) in hazelnut, providing evidence of unique molecular pathways governing self-incompatibility in Corylus.

The Severity and Frequency of Systemic Reactions to Hazelnut Are Significantly Higher in Hazelnut Allergic Patients Monosensitized to Cor a 8 than in Patients Polysensitized to Cor a 1, Cor a 8, and Cor a 9.

INTRODUCTION: Hazelnuts are a leading trigger of food allergy. To date, several molecular components of hazelnut are available for component-resolved diagnosis. However, little is known about how simultaneous sensitization to multiple allergens affects the severity of the hazelnut-induced reaction. In a previous study, our group demonstrated a lower risk of systemic reactions to peach in patients sensitized to both Pru p 3 and Pru p 1 than in the patient monosensitized to peach LTP. We aimed to assess whether this was also true in hazelnut allergy in a cohort of adult patients. METHODS: Patients were selected based on a history of symptoms such as urticaria, vomiting, diarrhea, asthma, and anaphylaxis indicative of hazelnut IgE-mediated food allergy and graded according to a clinical severity scale. For all patients, specific IgE was determined for Cor a 1 and Cor a 8 and, for most patients, also Cor a 9. Patients were offered an oral food challenge in open format (OFC) with a cocoa-based roasted hazelnut spread on a voluntary basis in order to prescribe an appropriate diet. RESULTS: A total of two hundred and fourteen patients were recruited. Among these, 43 patients were monosensitized to Cor a 8. One hundred and seventy-one patients were sensitized to Cor a 1 (79.9%), and, among them, 48/171 (28.1%) were also Cor a 8 positive. Cor a 9 was evaluated in 124/214 patients, testing positive in 21/124 (16.9%). Patients monosensitized to Cor a 8 experienced systemic reactions more frequently than those sensitized to Cor a 1 ± Cor a 8 (p < 0.00001), with significantly more severe reactions (p < 0.0005) and testing more frequently positive at OFC (p < 0.0001). Regarding Cor a 9, the sensitized patients were significantly younger (p = 0.0013) and showed reactions of similar severity to patients who tested Cor a 9 negative, and these reactions were milder than in patients monosensitized only to Cor a 8. DISCUSSION/CONCLUSION: Sensitization to Cor a 1 seems to protect from the development of the severe systemic reactions induced by Cor a 8 sensitization, Cor a 9 does not influence the severity of symptoms in adult patients. The OFC with roasted hazelnut may help in dietary guidance.

Tolerance of peanuts and tree nuts in Spanish children with exclusive sensitization to lipid transfer proteins.

BACKGROUND: Allergy to peanuts and tree nuts is a common cause of food allergy in Spain, with lipid transfer proteins (LTP) being the most frequently recognized panallergen. LTP sensitization often leads to multiple food group sensitivities, resulting in overly restrictive diets that hinder patient's quality of life. This study aimed to assess the tolerance of peanuts and tree nuts (hazelnuts and walnuts) in children sensitized to LTP, potentially mitigating the need for such diets. METHODS: This prospective study enrolled individuals diagnosed with allergy to peanuts, hazelnuts, or walnuts. Data were collected from medical records, including demographics and clinical history. Allergological assessment comprised skin prick tests using commercial extracts and the nuts in question, alongside measurements of total and specific IgE to nuts and their primary molecular components. Participants showing positive LTP sensitization without sensitization to seed storage proteins underwent open oral nut challenges. RESULTS: A total of 75 individuals labeled as allergic to peanuts, 44 to hazelnuts, and 51 to walnuts were included. All of them underwent an open oral provocation test with the incriminated nut, showing a high tolerance rate. Peanut was tolerated by 98.6% of patients, 97.72% tolerated hazelnut, and 84.3% tolerated walnut. CONCLUSION: The findings suggest that the majority of patients allergic to peanuts, hazelnuts, or walnuts, due to LTP sensitization and lacking IgE reactivity to seed storage proteins, can tolerate these nuts. This supports the need for personalized nut tolerance assessments to avoid unnecessary dietary restrictions.

First report of silverleaf caused by Chondrostereum purpureum in hazelnut (Corylus avellana) in Chile.

Hazelnut is among the most important nut crops in Chile, currently covering 46,000 ha. In 2023, the country exported 30,000-ton. In recent years the incidence of plants with internal discoloration, cankers and dieback has been increasing. In some cases, the trees died and had to be removed and, after a year, purple resupinate fruiting bodies were observed growing from the stumps. To determine the etiology of the symptoms and signs, wood samples (n=318) were collected since 2020, from 38 symptomatic orchards from Maule to La Araucanía Regions, primarily from the cvs. Tonda di Giffoni and Lewis. Wood sections 0.5 cm diameter were cut from the symptomatic tissues, disinfected using a sodium hypochlorite (10%) solution, and plated on a quarter-strength acidified potato dextrose agar (aPDA1/4). The plates were incubated and purified on PDA. Subsequently, isolates were identified by morphological and molecular means. Almost half of the isolates (47%) were preliminarily identified as basidiomycetes, based on mycelial features such as the presence of clamp connections, with 45% of them exhibiting abundant whitish cottony fast-growth mycelia, resembling Chondrostereum purpureum (Grinbergs et al., 2020). DNA was extracted and the 500-bp fragment, located between 5S and 18S ribosomal regions, was amplified using APN1 specific primers (Becker et al. 1999), identifying the isolates as C. purpureum. In addition, 5.8S gene of RGM1 (35°13'40.9"S 71°25'14.1"W), RGM2 (36°31'27.95"S 71°46'58.31"W), RGM3 (37°10'54.8"S 72°03'39.6"W), RGM4 (35°19'25.2"S 71°19'54.7"W) and RGM5 (36°35'30.8"S 72°05'18.8"W) isolates, representing different locations within the hazelnut growing area, was amplified using ITS1/ITS4 primers (White et al., 1990). The PCR product was sequenced, and the analysis showed 100% homology among isolates (Genebank codes: PP839283, PP839284, PP839285, PP839286 and PP839287, respectively). To determine the pathogenicity of the isolates, 30-cm healthy cuttings cv. Lewis were inoculated with mycelial plugs, while control shoots were inoculated with sterile agar plugs. Cuttings were vertically arranged in pots with 3-cm water and incubated for 60-d at 22°C. In addition, fresh cuts of 3-y potted plants cv. Lewis were inoculated with mycelial plugs and incubated for 137-d in a shadehouse. After incubation, bark was removed from inoculated cuttings and the length of necrotic lesions was measured. Although discoloration was reproduced by all the isolates in both pathogenicity tests, RGM1 isolate was the most aggressive, causing the complete discoloration of the cuttings and the death of the inoculated plants. To our knowledge this is the first report of C. purpureum causing wood disease in hazelnut. These findings are significant because the disease may not only reduce orchard longevity but also decrease fruit yield and quality, as observed in other fruit crops (Grinbergs et al., 2021).

Genetic Diversity Analysis of Anisogramma anomala in the Pacific Northwest and New Jersey.

Anisogramma anomala, a biotrophic ascomycete, causes eastern filbert blight (EFB) of hazelnuts (Corylus spp.). EFB is endemic in eastern North America, preventing the commercial production of European hazelnut (C. avellana L.). In contrast, the historic absence of A. anomala in the Pacific Northwest (PNW) supported the development of a robust hazelnut industry. Circa 1960, A. anomala was inadvertently introduced into southwestern Washington, causing orchard devastation. Distribution of the pathogen in the PNW has been hypothesized to be the result of a single-point introduction. This study aimed to investigate the single-point introduction hypothesis of A. anomala by comparing the genetic diversity of A. anomala samples from the PNW and New Jersey (NJ). Specimens from the main PNW production region (n = 60) and an area within the pathogen's native range, NJ (n = 151), were genotyped using 15 simple sequence repeat (SSR) markers. The following were used to assess genetic diversity and population structure: allelic summary statistics, discriminant analysis of principal components, network median-joining tree, analysis of multilocus genotypes, and allelic population diversity analysis. Analyses separated the samples into one cluster containing all the PNW isolates, and five clusters of NJ isolates. The PNW samples were nearly genetically uniform, and the NJ isolates were diverse. These findings support the hypothesis that A. anomala in the PNW was derived from a single-point introduction and corroborate previous studies that have shown A. anomala is very diverse in NJ. This indicates that maintaining restrictions on the movement of Corylus into the PNW is important to prevent the introduction of new populations of A. anomala, thus protecting the PNW hazelnut industry.

The use of artificial neural network for modelling adsorption of Congo red onto activated hazelnut shell.

Activated hazelnut shell (HSAC), an organic waste, was utilized for the adsorptive removal of Congo red (CR) dye from aqueous solutions, and a modelling study was conducted using artificial neural networks (ANNs). The structure and characteristic functional groups of the material were examined by the FTIR method. The BET surface area of the synthesized material, named HSAC, was 812 m2/g. Conducted in a batch system, the adsorption experiments resulted in a notable removal efficiency of 87% under optimal conditions. The kinetic data for hazelnut shell activated carbon (HSAC) removal of CR were most accurately represented by the pseudo-second-order kinetic model (R2 = 0.998). Furthermore, the equilibrium data demonstrated a strong agreement with the Freundlich model. The maximum adsorption capacity of HSAC for CR was determined to be 34.8 mg/g. The optimum adsorption parameters were determined to be pH 6, contact time of 60 min, 10 g/L of HSAC, and a concentration of 400 mg/L for CR. Considering the various experimental parameters influencing CR adsorption, an artificial neural network (ANN) model was constructed. The analysis of the ANN model revealed a correlation of 98%, indicating that the output parameter could be reliably predicted. Thus, it was concluded that ANN could be employed for the removal of CR from water using HSAC.

Inducing Effect of Corylus avellana on Cytotoxic Activity in Lung and Breast Cancer Cells via Apoptosis.

Turkish hazelnut (Corylus avellana L. cv Tombul) is a widely used nut in the chocolate industry and is also rich in polyphenol content, which promises anticancer effects. The anti-cancer and apoptotic effects of hazelnut leaves extracts examined on lung and breast cancer cells. Sulforhodamine B (SRB) and Adenosine 5'- triphosphate (ATP) assays were carried out for cell viability measurement. The mode of cell death was shown morphologically by the double fluorescence staining. Apoptosis was determined by performing caspase-mediated cytokeratin 18 (M30 ELISA) and western blot analysis. PARP, caspase 3, caspase 8, DR4, and GAPHD (Glyceraldehyde-3-phosphate Dehydrogenase) protein bands were visualized as markers of apoptosis. A wound healing test was employed to measure cell migration. Methanol extract of hazelnut leaf exhibited inhibition of cell growth activities in a dose-dependent manner. IC50 values were determined as 32.17 µg/ml in MCF-7, 32.16 µg/ml in MDA-MB-231, 20.40 µg/ml in A549 and 12.04 µg/ml in H1299 cells for ethanol extract while it was determined as 21.08 µg/ml in MCF-7, 40.16 µg/ml in MDA-MB-231, 22.04 µg/ml in A549 and 5.91 µg/ml in H1299 cells in methanol extract. In comparison, methanol leaf extracts were more effective in H1299 cells (IC50 value was 5.91 µg/ml).In comparison, ethanol leaf extracts were more effective in H1299 cells (IC50 value was 9.722 µg/ml). Western blot analysis demonstrated that hazelnut leaf extract treatment of cancer cells led to cell death via apoptosis and inhibited cell migration in lung and breast cancer cell lines. The cytotoxic effects of hazelnut extract on breast and lung cancer cells might be valuable and promising in elucidating cell death mechanisms for the development of new methods in cancer treatment.

GRAS-Di SNP-based molecular characterization and fingerprinting of a Turkish Corylus avellana core set provide insights into the cultivation and breeding of hazelnut in Turkey.

Hazelnut (Corylus avellana L.) is an economically and socially important product for Turkey, the country that leads global production of this crop. The preservation of Turkish hazelnut genetic diversity and informed breeding of new cultivars are crucial for maintaining quality and crop yield stability. In this study, genotyping by random amplicon sequencing (GRAS-Di) was used to identify single-nucleotide polymorphisms (SNPs) in a panel of 96 individuals representing the Turkish national hazelnut collection. The resulting 7609 high-quality SNPs were physically mapped to the Tombul cultivar reference genome and used for population structure and diversity analyses. These analyses revealed that cultivars are not less diverse than wild accessions and that 44% of the panel had admixed ancestry. The results also indicated that recently released Turkish cultivars are highly similar to each other, suggesting that diversity analysis is an important tool that should be employed to prevent future genetic bottlenecks in this crop. A minimal marker algorithm was used to select a set of seven SNP markers that were capable of differentiating the panel accessions. These fingerprinting markers should be useful for the propagation of true-to-type elite cultivars that can be used to renew Turkey's aging hazelnut orchards.

Does hazelnut consumption affect brain health and function against neurodegenerative diseases?

INTRODUCTION: A healthy daily diet and consuming certain nutrients, such as polyphenols, vitamins, and unsaturated fatty acids, may help neuronal health maintenance. Polyphenolic chemicals, which have antioxidant and anti-inflammatory properties, are involved in the neuroprotective pathway. Because of their nutritional value, nuts have been shown in recent research to be helpful in neuroprotection. OBJECTIVE: Hazelnut is often consumed worldwide in various items, including processed foods, particularly in bakery, chocolate, and confectionery products. This nut is an excellent source of vitamins, amino acids, tocopherols, phytosterols, polyphenols, minerals, and unsaturated fatty acids. Consuming hazelnut may attenuate the risk of neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, and Huntington's disease due to its anti-inflammatory and anti-oxidant qualities. RESULTS: Many documents introduce hazelnut as an excellent choice to provide neuroprotection against neurodegenerative disorders and there is some direct proof of its neuroprotective effects. DISCUSSION: So hazelnut consumption in daily diet may reduce neurodegenerative disease risk and be advantageous in reducing the imposed costs of dealing with neurodegenerative diseases.

Characterization of phenolic constituents in hazelnut kernels.

Hazelnuts contain biologically active phenolic compounds and are widely used for their nutritional value. In this study, the phenolic compounds contained in hazelnuts were isolated from the kernels of Corylus avellana L. and investigated. Spectral analyses revealed 2 new acetophenone glycosides, characterized as 2',4',6'-trihydroxyacetophenone-4'-O-(2-O-ß-d-apiosyl)-ß-d-glucoside and 2',4',6'-trihydroxyacetophenone-4'-O-(2-O-ß-d-apiosyl-6-O-α-l-arabinosyl)-ß-d-glucoside, and 4 known compounds. Four high-molecular-mass condensed tannin fractions were detected in the water-soluble fraction of the extract, characterized as B-type procyanidin consisting of extension and terminal units. Gel permeation chromatography analyses revealed that the average molecular mass, based on the polystyrene standard, was approximately 15 000-113 000. These high-molecular-mass condensed tannin fractions were chemically characterized and exhibited different molecular weights. The fractions of high-molecular-mass condensed tannins were obtained from hazelnuts and tested for 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. The EC50 values indicated significant activity for all the fractions.

De novo Transcriptome Analysis and Gene Expression Profiling of Corylus Species.

Hazelnut (Corylus), which has high commercial and nutritional benefits, is an important tree for producing nuts and nut oil consumed as ingredient especially in chocolate. While Corylus avellana L. (Euro-pean hazelnut, Betulaceae) and Corylus colurna L. (Turkish hazelnut, Betulaceae) are the two common hazelnut species in Europe, C. avellana L. (Tombul hazelnut) is grown as the most widespread hazelnut species in Turkey, and C. colurna L., which is the most important genetic resource for hazelnut breeding, exists naturally in Anatolia. We generated the transcriptome data of these two Corylus species and used these data for gene discovery and gene expression profiling. Total RNA from young leaves, flowers (male and female), buds, and husk shoots of C. avellana and C. colurna were used for two different libraries and were sequenced using Illumina HiSeq4000 with 100 bp paired-end reads. The transcriptome data 10.48 and 10.30 Gb of C. avellana and C. colurna, respectively, were assembled into 70,265 and 88,343 unigenes, respectively. These unigenes were functionally annotated using the TRAPID platform. We identified 25,312 and 27,051 simple sequen-ce repeats (SSRs) for C. avellana and C. colurna, respectively. TL1, GMPM1, N, 2MMP, At1g29670, CHIB1 unigenes were selected for validation with qPCR. The first de novo transcriptome data of C. co-lurna were used to compare data of C. avellana of commercial importance. These data constitute a valuable extension of the publicly available transcriptomic resource aimed at breeding, medicinal, and industrial research studies.

Occurrence of powdery mildew caused by Erysiphe corylacearum on Hazelnuts in Israel.

Hazelnut (Corylus avellana L) is an emerging crop in Israel, primarily cultivated as a host plant to establish truffle plantations through symbiosis with ectomycorrhizal fungi. A significant damage and yield reduction is caused by the prevalent occurrence of powdery mildew in hazelnut trees (Sezer et al., 2017). Until recently, Phyllactinia guttata was considered the primary pathogen in Western Asia, the Caucasus region, and Europe (Abasova et al. 2018; Arzanlou et al. 2018; Mezzalama et al. 2021). However, in the last years, a new destructive species Erysiphe corylacearum has been identified as the pathogen of powdery mildew on hazelnuts in these regions (Meparishvili et al. 2019; Mezzalama et al. 2021; Kalmár et al. 2022; Zajc et al. 2023). In May 2022, powdery mildew symptoms were observed on hazelnut plants in the Ein-Zivan truffle plantation, gardens of Merom-Golan, and the adjacent garden of the packing house Pri-Beresheet in the northern Golan region of Israel. Symptoms were observed on the abaxial and adaxial leaf surfaces, fruits, and husks. Disease incidence and severity ranged between 30-70% and 5-90%, respectively. Disease severity was significantly greater on the leaves of the offshoots compared to those on the tree canopy. Morphological characterization of leaf samples from ten different trees showed the following characteristics: hyphal appressoria were lobed, solitary, 1-4 µm in diameter; mycelium was amphigenous, hyaline, and septate; conidiophores vertically elevated from the mycelium 50- 80 µm long. Conidia (n= 30) on conidiophores were hyaline, ellipsoid to ovoid, 24 - 34 µm long and 15.5 - 23 µm wide. Chasmothecia in several maturation degrees appeared in October on both sides of the leaves. They were spherical (n= 30) 86 - 125 µm in diameter, with 7 - 14 aseptate straight appendages, 67 - 96 µm long, 4.9 - 7.1 µm wide, dichotomous branched at the end 42 - 56 µm wide. In each chasmothecia, there were 3-5 asci (n=30) with a width of 38 - 43 µm and a length of 48-64 µm of oval-ellipsoid shape. Asci contained 4-8 ascospores (n=30), 18 - 26 µm long and 11 - 15.5 µm wide. A pathogenicity test was conducted to fulfill Koсh's postulates. Both detached leaves and plants of C. avellana were artificially infected by brushing conidia from infected leaves. Inoculated leaves in Petri dishes on 2% water agar (n= 5), and plants (n= 5) were incubated under 25°C and 12-h photoperiod/day. Untreated leaves and plants served as control. Typical symptoms appeared on the upper surface of the leaves within 7-10 days after inoculation. No symptoms were found on untreated control plants or detached leaves. The fungus isolated from the inoculated leaves was morphologically identical to the original isolates from natural diseased plants. DNA was extracted and the rDNA internal transcribed spacer region of five isolates originated from leaves of the tree canopy and offshoots was amplified using primers ITS1 and ITS4, and sequenced. BLAST analysis of 595 bp fragments (all identical and represented by isolate Cora1, GenBank Accession No. OR752437) showed 99% identity to ITS rDNA sequences of E. corylacearum from Georgia (MK157199) and 100% identity to isolates from Azerbaijan, Turkey and Italy (LC270863, KY082910 and MW045425, respectively) and only 83% similarity to P. guttata (accession number AB080558). To the best of my knowledge this is the first report on E. corylacearum causing powdery mildew in Israel. Future control measures to manage the disease on hazelnuts in truffle plantations in Israel should be considered.

Molecular Characterization and Pathogenicity of Diaporthe Species Causing Nut Rot of Hazelnut in Italy.

Hazelnut (Corylus avellana), a nut crop that is rapidly expanding worldwide, is endangered by a rot. Nut rot results in hazelnut defects. A survey was conducted in north-western Italy during 2020 and 2021 to identify the causal agents of hazelnut rots. Typical symptoms of black rot, mold, and necrotic spots were observed on hazelnut nuts. The prevalent fungi isolated from symptomatic hazelnut kernels were Diaporthe spp. (38%), Botryosphaeria dothidea (26%), Diplodia seriata (14%), and other fungal genera with less frequent occurrences. Among 161 isolated Diaporthe spp., 40 were selected for further analysis. Based on morphological characterization and multi-locus phylogenetic analysis of the ITS, tef1- α, and tub2, seven Diaporthe species were identified as D. eres, D. foeniculina, D. novem, D. oncostoma, D. ravennica, D. rudis, and D. sojae. D. eres was the main species isolated from hazelnut rots, in particular from moldy nuts. Pathogenicity test performed on hazelnut nuts 'Tonda Gentile del Piemonte' using a mycelium plug showed that all the Diaporthe isolates were pathogenic on their original host. To our knowledge, this work is the first report of D. novem, D. oncostoma and D. ravennica on hazelnut nuts worldwide. Diaporthe foeniculina, D. rudis, and D. sojae were reported for the first time as agents of hazelnut nut rot in Italy. Future studies should focus on the comprehension of epidemiology and climatic conditions favoring the development of Diaporthe spp. on hazelnut. Prevention and control measures should target D. eres, representing the main causal agents responsible for defects and nut rot of hazelnuts in Italy.

Erysiphe corylacearum as a new pathogen of hazelnut in the Czech Republic.

In July 2021, leaves and shoot tops of the common hazel (Corylus avellana L.), with a whitish coating, were found in the Czech Republic (southern Moravia region). The infected hazel bushes were found along a road in a deciduous forest and in an urban garden. In most European countries, Phyllactinia guttata is found on the abaxial surface of the leaves in the form of a continuous whitish to light grey mycelium, possibly with large black chasmothecia. In our case, the mycelium was present on both sides of the leaves, but the symptoms and the incidence were much stronger on the adaxial side. The first symptoms usually appeared on the adaxial side of the leaves as small white radially expanding patches of mycelium. In the final stage, the spots merged and covered a substantial part of the leaf blade (50-85 % on the adaxial side, 5-25 % on the abaxial side). When the abaxial side of the leaves was infected, chlorotic spots were evident on the adaxial side. The spots of powdery mildew were small (3-15 mm), whitish, rounded to irregular, effuse eventually becoming confluent, and occurred primarily on the adaxial side of the leaves. Conidiophores (30-53×4-6 µm) grew on the amphigenic mycelium, were erect, consisted of 1-3 cells, i.e. cylindrical foot cell and followed 1-2 cells, from which hyaline ellipsoid to doliform-limoniform conidia (17-34 ×15-21) (n = 50) were individually detached. Single or in groups dark brown chasmothecia (77-116 µm in diameter) had up to hyaline 8-15 aseptate straight appendages (50-102 µm) with multiple (3-5×) dichotomously branched apexes and recurved tips. Chasmothecia contained 3-6 asci (42-62 × 34-55 µm) with 4-8 obovoid to broadly ellipsoidal hyaline ascospores (14-22 × 75 µm). Based on morphological characters, the powdery mildew was identified as Erysiphe corylacearum (2). Morphological identification was confirmed by molecular analysis of samples. DNA was extracted from symptomatic leaves tissue using the DNeasy Plant Mini Kit, (Quiagen, Hilden, Germany) and the rDNA internal transcribed spacer region of 2 isolates was amplified using primers PMITS1 and PMITS2 (Cunnington et al. 2003) and sequenced. BLAST analysis of our 720bp fragments (both identical and represented by GenBank accession no. OR432526) showed 100% sequence identity to ITS rDNA sequences of E. corylacearum other countries of Central Europe for example from Austria (MW031866), Italy (MW045428), Hungary (OQ411007), Germany (OP554268) or Slovakia (MT176105). Pathogenicity was verified on two-year-old plants of Corylus avellana. Healthy leaves were artificially infected by dusting conidia from infected leaves. Inoculated plants were incubated under controlled conditions (21-23 °C, 70-80 % relative humidity). Characteristic symptoms of powdery mildew appeared on the adaxial side of the leaves 9-12 days after inoculation. Control plants treated with distilled water remained symptomless. Powdery mildew isolated from inoculated leaves was morphologically identical to isolates from naturally infected leaves. The first record of E. corylacearum in Europe on cultivated hazelnut species was reported by Sezer et al. (2017) in Turkey in 2013. Within a few years, the E. corylacearum spread and was recorded on various species of Corylus in other European countries (for example Mezzalama et al., 2020; Rosati et al., 2021; Beenken et al., 2022; Boneva et al., 2023), East Asia (Arzanlou et al., 2018) and the USA (Meparishvili 2019). To our knowledge, this is the first report of Erysiphe corylacearum in the Czech Republic.

First Report of Xanthomonas arboricola pv. corylina Causing Bacterial Blight on Hazelnut Tree (Corylus avellana) in Montenegro.

Hazelnut is a minor but rapidly increasing commercially grown species in Montenegro. In June 2021, severe infection, affecting more than 80% of the trees, was observed on 6-year-old hazelnut plants (Corylus avellana) cultivar Hall's Giant, in a 0.3ha plantation near Cetinje, central Montenegro. Numerous, small, 2-3mm in diameter, irregular, brown, necrotic spots, sometimes surrounded by a weak chlorotic halo, were observed on leaves. As the disease progressed, the lesions coalesced and formed large necrotic areas. Necrotic leaves remained attached to the twigs. Longitudinal brown lesions developed on twigs and branches, causing their dieback. Necrotic, unopened buds were noticed as well. No fruits were observed in the orchard. From the diseased leaf, bud and twig bark tissue, yellow, convex, and mucoid bacterial colonies were isolated on yeast extract dextrose CaCO3 medium and 14 isolates were subcultured. The isolates induced hypersensitive reaction in pelargonium leaves (Pelargonium zonale), were Gram-negative, catalase positive, oxidase negative, obligate aerobic, hydrolyzed starch, gelatin and esculin, did not reduce nitrate and did not grow at 37°C and in the presence of 5% NaCl, showing so the same biochemical profile of the reference strain Xanthomonas arboricola pv. corylina (Xac) NCPPB 3037. Using primer pair XarbQ-F/XarbQ-R (Pothier et al., 2011), a 402 bp product was amplified in all 14 isolates and the reference strain, confirming their affiliation to X. arboricola species. Additionally, the isolates were further identified by PCR analysis, using primer pair XapY17-F/XapY17-R (Pagani 2004; Pothier et al., 2011), resulting in a single band of 943 bp characteristic for Xac. The amplification and sequencing of the partial rpoD gene sequence of two selected isolates RKFB 1375 and RKFB 1370, were performed using a set of primers described by Hajri et al., 2012. The obtained DNA sequences showed that the isolates (GenBank Nos. OQ271224 and OQ271225) share 99.47% to 99.92% rpoD sequence identity with Xac strains CP076619.1 and HG992342.1 isolated from hazelnut in France and HG992341.1 in USA. Pathogenicity of all isolates was confirmed by spraying young shoots (20 to 30 cm long, with 5 to 7 leaves) on 2-year-old potted hazelnut plants (cv. Hall's Giant) using a handheld sprayer with the bacterial suspension (108 CFU/mL of sterile tap water), in three replicates. Sterile distilled water (SDW) and NCPPB 3037 Xac strain were used as negative and positive control, respectively. The inoculated shoots were incubated under plastic bags, providing high humidity conditions, in an acclimatized greenhouse at 22-26°C, for 72 h. Lesions surrounded by a halo appeared on leaves of all inoculated shoots within 5 to 6 weeks after inoculation, while leaves sprayed with SDW remained symptomless. Koch's postulates were confirmed by the re-isolation of the pathogen from the necrotic test plant tissue and identity checked by PCR using the primer set of Pothier et al., 2011. Based on pathogenic, biochemical, and molecular characteristics, the isolates from hazelnut plants in Montenegro were identified as X. arboricola pv. corylina. This is the first report of Xac affecting hazelnut in this country. Considering favorable environmental conditions, the pathogen can cause significant economic losses in hazelnut production in Montenegro. Therefore, phytosanitary measures have to be implemented to prevent introduction and spread of the pathogen in other areas.

Optimization of hazelnut husk medium for pullulan production by a domestic A. pullulans strain.

Hazelnut husk is one of the most abundant agricultural residue in Turkey. Valorization of this lignocellulosic biomass would provide a promoting alternative for economical production of pullulan. In this study, dried hazelnut husk hydrolysate was used directly as fermentation medium for pullulan production by a domestic strain of Aureobasidium pullulans. The aim of this work was the optimization of some fermentation medium parameters by central composite design using response surface methodology (RSM). The effects of (NH4)2SO4 concentration, the volume of concentrated H2SO4 and the amount of ground hazelnut husk on pullulan production were optimized by RSM. The optimum levels of the fermentation parameters defined as 7.2 gL-1, 2.5 mL and 20 g, respectively. The maximum pullulan and exopolysaccharide concentrations were determined as 74.39 and 75.95 gL-1, respectively in the optimum conditions. Specific growth rate of the strain was found as 0.097 h-1. FTIR spectral attributes confirmed the structure of pullulan. Thermal decomposition temperature of synthesized pullulan was found as 247.15 °C. This study showed that hazelnut husk was one of the novel substrate for production of the pullulan by A. pullulans AZ-6. No previous work was found to utilize dried hazelnut husk as fermentation medium for pullulan production by A. pullulans.

Chemical Composition of Hazelnut Skin Food Waste and Protective Role against Advanced Glycation End-Products (AGEs) Damage in THP-1-Derived Macrophages.

Glycation and the accumulation of advanced glycation end-products (AGEs) are known to occur during aging, diabetes and neurodegenerative diseases. Increased glucose or methylglyoxal (MGO) levels in the blood of diabetic patients result in increased AGEs. A diet rich in bioactive food compounds, like polyphenols, has a protective effect. The aim of this work is to evaluate the capacity of hazelnut skin polyphenolic extract to protect THP-1-macrophages from damage induced by AGEs. The main polyphenolic subclass was identified and quantified by means of HPLC/MS and the Folin-Ciocalteu method. AGEs derived from incubation of bovine serum albumin (BSA) and MGO were characterized by fluorescence. Cell viability measurement was performed to evaluate the cytotoxic effect of the polyphenolic extract in macrophages. Reactive oxygen species' (ROS) production was assessed by the H2-DCF-DA assay, the inflammatory response by real-time PCR for gene expression, and the ELISA assay for protein quantification. We have shown that the polyphenolic extract protected cell viability from damage induced by AGEs. After treatment with AGEs, macrophages expressed high levels of pro-inflammatory cytokines and ROS, whereas in co-treatment with polyphenol extract there was a reduction in either case. Our study suggests that hazelnut skin polyphenol-rich extracts have positive effects and could be further investigated for nutraceutical applications.

Hazelnut (Corylus avellana L.) shells as a potential source of dietary fibre: impact of hydrothermal treatment temperature on fibre structure and degradation compounds.

BACKGROUND: Hemicellulose extraction from lignocellulosic biomasses has gained interest over the years, and hydrothermal treatment is one of the most common methods employed for this purpose. This work aimed to deeply study hazelnut (Corylus avellana L.) shells as a new source of dietary fibre, evaluating the effect of hydrothermal treatment temperatures on the type and structure of fibre extracted, but also on the formation of side-products derived from lignocellulose degradation. RESULTS: Different process temperatures led to diverse polysaccharides in the hydrothermal extract. Pectin was identified for the first time in hazelnut shells when experimenting with extraction at 125 °C, whereas at 150 °C a heterogeneous mixture of pectin, xylan, and xylo-oligosaccharides was present. The highest yield in terms of total fibre was gained at 150 and 175 °C, and then decreased again at 200 °C. Finally, more than 500 compounds from different chemical classes were putatively identified and they appeared to be present in the extracted fibre with a different distribution and relative amount, depending on the heat treatment severity. A generally high content of phenols, phenyls, oligosaccharides, dehydro-sugars, and furans was observed. CONCLUSIONS: Modulation of the hydrothermal treatment temperature allows fibre extracts with very different compositions, and therefore different potential end uses, to be obtained from hazelnut shells. A sequential temperature-based fractionation approach, as a function of the severity of the extraction parameters, can also be considered. Nevertheless, the study of the side-compounds formed from lignocellulosic matrix degradation, as a function of the applied temperature, needs to be fully addressed for a safe introduction of the fibre extract within the food chain. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Removal of cationic dye pollutants from wastewater with HS loaded semi-IPN composites: kinetic and thermodynamic studies.

In this study, methylene blue (MB) pollutant in water was removed using produced hazelnut shell loaded semi-interpenetrating polymer networks (HS loaded semi-IPN) adsorbent. The physical and chemical characterizations of the adsorbents were investigated using TGA, DSC, FT-IR, BET, FE-SEM, and EDX. Experimental parameters such as temperature, swelling, dye concentration, contact time, pH solution, and adsorbent dosage for MB adsorption were thoroughly investigated. It was determined that the HS loaded semi-IPN adsorbent removed 92.1% of MB dye. Subsequently, the adsorption properties between the adsorbent and dye were investigated in detail using several different kinetic, isotherm, and thermodynamic models. As a result of the obtained data, the interaction between adsorbent and dye molecules is discussed. Moreover, studies on the industrial usability of the adsorbent have been carried out, and it has been observed that the adsorbent can be employed even after four cycles.

Discrimination of ß-cyclodextrin/hazelnut (Corylus avellana L.) oil/flavonoid glycoside and flavonolignan ternary complexes by Fourier-transform infrared spectroscopy coupled with principal component analysis.

The goal of the study was the discrimination of ß-cyclodextrin (ß-CD)/hazelnut (Corylus avellana L.) oil/antioxidant ternary complexes through Fourier-transform infrared spectroscopy coupled with principal component analysis (FTIR-PCA). These innovative complexes combine the characteristics of the three components and improve the properties of the resulting material such as the onsite protection against oxidative degradation of hazelnut oil unsaturated fatty acid glycerides. Also, the apparent water solubility and bioaccessibility of the hazelnut oil components and antioxidants can be increased, as well as the controlled release of bioactive compounds (fatty acid glycerides and antioxidant flavonoids, namely hesperidin, naringin, rutin, and silymarin). The appropriate method for obtaining the ternary complexes was kneading the components at various molar ratios (1:1:1 and 3:1:1 for ß-CD hydrate:hazelnut oil (average molar mass of 900 g/mol):flavonoid). The recovering yields of the ternary complexes were in the range of 51.5-85.3% and were generally higher for the 3:1:1 samples. The thermal stability was evaluated by thermogravimetry and differential scanning calorimetry. Discrimination of the ternary complexes was easily performed through the FTIR-PCA coupled method, especially based on the stretching vibrations of CO groups in flavonoids and/or CO/CC groups in the ternary complexes at 1014.6 (± 3.8) and 1023.2 (± 1.1) cm-1 along the second PCA component (PC2), respectively. The wavenumbers were more appropriate for discrimination than the corresponding intensities of the specific FTIR bands. On the other hand, ternary complexes were clearly distinguishable from the starting ß-CD hydrate along the first component (PC1) by all FTIR band intensities and along PC2 by the wavenumber of the asymmetric stretching vibrations of the CH groups at 2922.9 (± 0.4) cm-1 for ternary complexes and 2924.8 (± 1.4) cm-1 for ß-CD hydrate. The first two PCA components explain 70.38% from the variance of the FTIR data (from a total number of 26 variables). Other valuable classifications were obtained for the antioxidant flavonoids, with a high similarity for hesperidin and naringin, according to FTIR-PCA, as well as for ternary complexes depending on molar ratios. The FTIR-PCA coupled technique is a fast, nondestructive and cheap method for the evaluation of quality and similarity/characteristics of these new types of cyclodextrin-based ternary complexes having enhanced properties and stability.