Analysis of Methylesterase Gene Family in Fragaria vesca Unveils Novel Insights into the Role of FvMES2 in Methyl Salicylate-Mediated Resistance against Strawberry Gray Mold.

Methylesterases (MESs) hydrolyze carboxylic ester and are important for plant metabolism and defense. However, the understanding of MES' role in strawberries against pathogens remains limited. This study identified 15 FvMESs with a conserved catalytic triad from the Fragaria vesca genome. Spatiotemporal expression data demonstrated the upregulated expression of FvMESs in roots and developing fruits, suggesting growth involvement. The FvMES promoter regions harbored numerous stress-related cis-acting elements and transcription factors associated with plant defense mechanisms. Moreover, FvMES2 exhibited a significant response to Botrytis cinerea stress and showed a remarkable correlation with the salicylic acid (SA) signaling pathway. Molecular docking showed an efficient binding potential between FvMES2 and methyl salicylate (MeSA). The role of FvMES2 in MeSA demethylation to produce SA was further confirmed through in vitro and in vivo assays. After MeSA was applied, the transient overexpression of FvMES2 in strawberries enhanced their resistance to B. cinerea compared to wild-type plants.

Sulfur-Induced Resistance against Pseudomonas syringae pv. actinidiae via Triggering Salicylic Acid Signaling Pathway in Kiwifruit.

Sulfur has been previously reported to modulate plant growth and exhibit significant anti-microbial activities. However, the mechanism underlying its diverse effects on plant pathogens has not been elucidated completely. The present study conducted the two-year field experiment of sulfur application to control kiwifruit canker from 2017 to 2018. For the first time, our study uncovered activation of plant disease resistance by salicylic acid after sulfur application in kiwifruit. The results indicated that when the sulfur concentration was 1.5-2.0 kg m-3, the induced effect of kiwifruit canker reached more than 70%. Meanwhile, a salicylic acid high lever was accompanied by the decline of jasmonic acid. Further analysis revealed the high expression of the defense gene, especially AcPR-1, which is a marker of the salicylic acid signaling pathway. Additionally, AcICS1, another critical gene of salicylic acid synthesis, was also highly expressed. All contributed to the synthesis of increasing salicylic acid content in kiwifruit leaves. Moreover, the first key lignin biosynthetic AcPAL gene was marked up-regulated. Thereafter, accumulation of lignin content in the kiwifruit stem and the higher deposition of lignin were visible in histochemical analysis. Moreover, the activity of the endochitinase activity of kiwifruit leaves increased significantly. We suggest that the sulfur-induced resistance against Pseudomonas syringae pv. actinidiae via salicylic activates systemic acquired resistance to enhance plant immune response in kiwifruit.

Cloning and immunobiochemical analyses on recombinant chymopapain allergen Cari p 2 showing pollen-fruit cross-reaction.

Papaya is reported to trigger food and respiratory allergy. Here, we identified chymopapain Cari p 2 as an allergen that can sensitize atopic individuals through fruit consumption followed by respiratory hazards through pollen exposure. Recombinant Cari p 2 displayed IgE-reactivity with 78% of papaya allergic sera. rCari p 2 also displayed allergenic activity through basophil degranulation. rCari p 2 is correctly folded and showed irreversible denaturation in the melting curve. rCari p 2 displayed IgE-cross-reactivity with homologous cysteine proteases from kiwi and pineapple. Cari p 2 transcript was also detected in papaya pulps. rCari p 2 was resistant to pepsin digestion and retained IgE-reactivity after 60 minutes of pepsin digestion. In mouse model, rCari p 2 was found to elicit inflammatory responses in the lung and gastrointestinal epithelium. Hence, Cari p 2 is a newly characterized allergen with diagnostic and immunotherapeutic potential for managing allergic disorders in papaya sensitized individuals.

LTP Allergy Follow-Up Study: Development of Allergy to New Plant Foods 10 Years Later.

INTRODUCTION: Allergy to nonspecific lipid transfer protein (nsLTP) is the main cause of plant-food allergy in Spain. nsLTPs are widely distributed in the plant kingdom and have high cross-reactivity but extremely variable clinical expression. Little is known about the natural evolution of this allergy, which complicates management. The objective of this study was to assess the development of allergy to new plant foods in nsLTP-sensitized patients 10 years after diagnosis. METHODS: One hundred fifty-one patients showing specific IgE to nsLTP determined by ISAC (Thermofisher) were included. After clinical workup (i.e., anamnesis, skin test, and challenge when needed), these patients were divided into two groups: 113 patients allergic to one or more plant food (74.5%) and 38 patients not allergic to any plant food (25.1%). Ten years later, a telephone interview was conducted to check whether patients had developed additional allergic reactions to plant foods. RESULTS: Ten years after diagnosis, 35 of the 113 (31%) plant-food-allergic patients sensitized to nsLTP reported reactions to new, previously tolerated plant foods, mainly Rosaceae/Prunoideae fruits and nuts followed by vegetables, Rosacea/Pomoideae fruits, legumes, and cereals. Five out of 38 (13.2%) patients previously sensitized to nsLTP but without allergy to any plant food had experienced allergic reactions to some plant food: two to Rosaceae/Prunoideae fruits, two to Rosaceae/Prunoideae fruit and nuts, and one to legumes. CONCLUSION: Patients sensitized to nsLTP developed allergic reactions to other plant foods, mainly Rosaceae-Prunoideae fruits and nuts. This was more frequent among plant-food-allergic patients than among those who had never had plant-food allergy.

Allergy to Gibberellin-Regulated Proteins (Peamaclein) in Children.

BACKGROUND: Gibberellin-regulated proteins (GRPs, Peamaclein) are allergens recently identified in plant-derived food allergy (FA), and little is known about the clinical manifestations of this allergic condition in the European population, especially in children. OBJECTIVE: Our study aimed to identify and characterize pediatric patients with pollen-FA due to GRP sensitization. METHODS: We retrospectively analyzed the charts of patients referred to the Allergy Unit of the Meyer Children's Hospital in Florence for suspected FA. Three main eligibility criteria based on the actual knowledge of GRP allergy were used to select patients deserving further investigations: (1) systemic reactions after consumption of fruit or an unknown culprit food, (2) positive skin prick tests to both cypress pollen and Pru p 3-enriched peach peel extracts, (3) negative in vitro test results for Pru p 3 serum-specific Immunoglobulin E (sIgE). We performed the in vitro test to determine the anti-rPru p 7 (Peamaclein) sIgE levels in the selected patients. RESULTS: We identified 10 pediatric patients with Pru p 7 allergy and described their characteristics. The use of our eligibility criteria showed a high accuracy in identifying these patients: 100% of the selected patients had positive in vitro results for Pru p 7. We therefore proposed a diagnostic algorithm for Pru p 7 allergy. CONCLUSION: This is the first case series of European pediatric patients with a demonstrated Peamaclein allergy. These findings broaden our knowledge on GRP allergy in pediatric populations and could help clinicians to suspect, diagnose, and manage this recently discovered plant-derived FA.

CsMYB96 enhances citrus fruit resistance against fungal pathogen by activating salicylic acid biosynthesis and facilitating defense metabolite accumulation.

Citrus fruit are generally confronted with various fungal diseases that cause fruit deterioration and economic loss. Salicylic acid (SA), a plant hormone, is an important signal molecule required for stimulating the disease resistance of plants. However, there has been limited information about the molecular mechanism of SA biosynthesis involving biotic stress response in citrus fruit. In the present study, an R2R3 MYB transcription factor (CsMYB96) was identified to mediate SA signaling in response to fungal diseases. The transient overexpression assay revealed that CsMYB96 contributed to the strong tolerance of citrus fruit to Penicillium italicum along with an increase in SA content; meanwhile, CsMYB96 conferred resistance to Botrytis cinerea in Arabidopsis plants. Further metabolomic profiling of stable transgenic Arabidopsis revealed that CsMYB96 participated in the regulation of various metabolism pathways and enhanced the accumulation of phenolic acids. RNA-seq analysis confirmed that overexpression of CsMYB96 activated the expression of genes involved in plant-pathogen interaction, phenylpropanoid biosynthesis, and SA signaling. Besides, CsMBY96 directly activated the transcription of calmodulin binding protein 60g (CsCBP60g), a predominant transcription factor required for the activation of SA signaling. In summary, our results reveal that CsMYB96 promotes SA biosynthesis and the accumulation of defense metabolites to enhance the fungal pathogen resistance of citrus fruit and Arabidopsis and provide new insights into the regulation of disease response.

Molecular and Immunological Identification of Low Allergenic Fruits among Old and New Apple Varieties.

About 50-70% of patients allergic to birch pollen suffer from sensitization after apple ingestion. Apple allergenicity was established in only few varieties. Studies were performed on apple fruits of 21 traditional and nine modern varieties organically, intensively, or integratively produced. The aim of the study was to assess whether the factors like cultivation method, maturity stage, genotype, or type of tissue place an impact on the allergenic potential of apples. To answer these questions, we used semiquantitative real-time PCR, ELISA, and immunoblotting. Apple allergen genes present divergent expression across apple cultivars. Expression of the Mal d 1.06A correlates with the Mal d 1 level and is affected by the cultivation method and maturity of the fruit. The content of the main allergen Mal d 1 varied widely across cultivars. Interestingly, in our study, the Gala variety presented a low Mal d 1 concentration regardless of the cultivation method. Based on the Mal d 1.06A expression, the Mal d 1 protein content, and the immunoreactivity assay, the Kandil Sinap, Kosztela, Rumianka from Alma-Ata, Kantówka Gdanska, Reinette Coulon, and Gala cultivars emerged as potentially hypoallergenic apple cultivars. Our study allowed distinguishing between potentially low, medium, and highly allergenic varieties.

Birch Pollen Related Pear Allergy: A Single-Blind Oral Challenge TRIAL with 2 Pear Cultivars.

Approximately 70% of birch pollen allergic patients in Europe experience hypersensitivity reactions to Immunoglobulin E (IgE) cross-reactive food sources. This so-called pollen-food syndrome (PFS) is defined by allergic symptoms elicited promptly by the ingestion of fruits, nuts, or vegetables in these patients. So far, in the literature, less attention has been given to Bet v 1 cross-reactive symptoms caused by pear (Pyrus communis). In the Netherlands, pears are widely consumed. The primary objective of this study was to measure the type and severity of allergic symptoms during pear challenges in birch pollen allergic patients, with a positive history of pear allergy, using two different pear varieties. Fifteen patients were included, skin prick test (SPT), prick-to-prick test (PTP), specific Immunoglobulin E (sIgE), and single-blind oral challenges were performed with two pear (Pyrus communis) varieties: the 'Cepuna' (brand name Migo®) and the 'Conference' pears. All patients were sensitized to one or both pear varieties. A total of 12 out of 15 participants developed symptoms during the 'Cepuna' food challenge and 14/15 reacted during the 'Conference' challenge. Challenges with the 'Cepuna' pears resulted in less objective symptoms (n = 2) in comparison with challenges with 'Conference' pears (n = 7). Although we did not find significance between both varieties in our study, we found a high likelihood of fewer and less severe symptoms during the 'Cepuna' challenges. Consequently selected pear sensitized patients can try to consume small doses of the 'Cepuna' pear outside the birch pollen season.

Pollen-food allergy syndrome and component sensitization in adolescents: A Japanese population-based study.

Allergic rhino-conjunctivitis with pollen allergy has been prevalent worldwide and Pollen-food allergy syndrome (PFAS) refers to individuals with pollen allergy who develop oral allergy syndrome (OAS) on consuming fruits and vegetables. The prevalence of PFAS varies by region and that in Japanese adolescents remains to be elucidated. In this cross-sectional study, we examined the epidemiological characteristics of PFAS in a general population of Japanese adolescents according to pollen allergy, OAS, and IgE component sensitization. Participants comprised adolescents, at age 13 years, from a prospective birth cohort study in Japan. We administered questionnaires to collect information from parents regarding pollen allergy, PFAS and OAS at each child's age 13 years. ImmunoCAP ISAC was used to assess IgE component sensitization. Among 506 participants with a complete questionnaire and ISAC measurement results, 56.5% had a history of hay fever, 16.0% had a history of OAS, 51.0% had pollen allergy, and 11.7% had a history of PFAS; additionally, 72.7% were sensitized to one or more tree, grass, and/or weed allergens. The most common sensitization (95.7%) among adolescents with pollen allergy was to Japanese cedar (Cry j 1). The most common causal foods were kiwi and pineapple (both 39.0%). Knowledge levels about PFAS were poor among affected adolescents. We found a high prevalence of PFAS among adolescents in Japan. Although it affects approximately 1/10 adolescents in the general population, public awareness regarding PFAS is poor. Interventional strategies are needed to increase knowledge and to prevent PFAS in the general population.

Oral allergy syndrome by fruit and vegetable PR-10 allergy: Accuracy of in vivo diagnosis

Routine diagnostic methods for allergies to plant-derived foods are based on skin prick test (SPT) with commercial extracts, prick-by-prick (PbP) with fresh food, serum-specific IgE measurement, and oral food challenge. We discuss the possibility and the advantages of performing, in patients with oral allergy syndrome (OAS) by fruit and vegetables (excluding nuts) PR-10 allergy, component-resolved diagnosis (CRD) by SPT and PbP with raw and cooked vegetables, rather than performing a CRD with in vitro tests by drawing blood. Based on our clinical experience and the studies published in the literature, we believe that, at least for the OAS by fruit and vegetables (excluding nuts) PR-10 allergy, the search for sensitizing allergens and related cross-reactive allergens with SPT and PbP can be performed routinely in clinical practice, even at the primary-care level (AU)

Oral allergy syndrome by fruit and vegetable PR-10 allergy: Accuracy of in vivo diagnosis.

Routine diagnostic methods for allergies to plant-derived foods are based on skin prick test (SPT) with commercial extracts, prick-by-prick (PbP) with fresh food, serum-specific IgE measurement, and oral food challenge.We discuss the possibility and the advantages of performing, in patients with oral allergy syndrome (OAS) by fruit and vegetables (excluding nuts) PR-10 allergy, component-resolved diagnosis (CRD) by SPT and PbP with raw and cooked vegetables, rather than performing a CRD with in vitro tests by drawing blood.Based on our clinical experience and the studies published in the literature, we believe that, at least for the OAS by fruit and vegetables (excluding nuts) PR-10 allergy, the search for sensitizing allergens and related cross-reactive allergens with SPT and PbP can be performed routinely in clinical practice, even at the primary-care level.

Host susceptibility factors render ripe tomato fruit vulnerable to fungal disease despite active immune responses.

The increased susceptibility of ripe fruit to fungal pathogens poses a substantial threat to crop production and marketability. Here, we coupled transcriptomic analyses with mutant studies to uncover critical processes associated with defense and susceptibility in tomato (Solanum lycopersicum) fruit. Using unripe and ripe fruit inoculated with three fungal pathogens, we identified common pathogen responses reliant on chitinases, WRKY transcription factors, and reactive oxygen species detoxification. We established that the magnitude and diversity of defense responses do not significantly impact the interaction outcome, as susceptible ripe fruit mounted a strong immune response to pathogen infection. Then, to distinguish features of ripening that may be responsible for susceptibility, we utilized non-ripening tomato mutants that displayed different susceptibility patterns to fungal infection. Based on transcriptional and hormone profiling, susceptible tomato genotypes had losses in the maintenance of cellular redox homeostasis, while jasmonic acid accumulation and signaling coincided with defense activation in resistant fruit. We identified and validated a susceptibility factor, pectate lyase (PL). CRISPR-based knockouts of PL, but not polygalacturonase (PG2a), reduced susceptibility of ripe fruit by >50%. This study suggests that targeting specific genes that promote susceptibility is a viable strategy to improve the resistance of tomato fruit against fungal disease.

Comprehensive polyphenolic profiling in promising resistant grapevine hybrids including 17 novel breeds in northern Italy.

BACKGROUND: A promising way to overcome the susceptibility of Vitis vinifera L. to fungal diseases is the integration of genetic resistance by the interspecific crossing between V. vinifera varieties and resistant species. However, the products of such hybrids are still not accepted by customers, particularly due to their organoleptic characteristics, not least influenced by their polyphenolic profile. RESULTS: A total of 58 resistant breeding lines, 41 from international programs and 17 new progeny individuals, were grown in one untreated vineyard to exclude any variances by climatic and pedologic conditions or vineyard practice. A total of 60 polyphenols (including acids, anthocyanins, flavonols, flavan-3-ols, and stilbenoids) were determined in grapevine berries by ultrahigh-performance liquid chromatography-mass spectrometry in two consecutive years. The overall profiles were rather consistent (variation P > 0.05) within the two harvests, with the exceptions of epicatechin and caftaric acid. Anthocyanin diglucosides were found in ten of the red breeding lines, malvidin-3,5-O-diglucoside being predominant in nine of them. Total polyphenol content of the unknown progeny individuals and international breeding lines was comparable, with the exception of significantly increased amounts of gallic acid and some flavonoids. CONCLUSION: The comprehensive study reported herein of the polyphenolic profile of hybrids from international breeding programs, but also of new breeds from private initiatives, all cultivated in the same vineyard, will support the selection of promising candidates for further breeding programs to overcome impairment due to undesired sensory characteristics of new highly resistant varieties.

Potential of microbial endophytes to enhance the resistance to postharvest diseases of fruit and vegetables.

Food loss of fruit and vegetables caused by postharvest diseases is a major issue worldwide. The method used to prevent and control postharvest diseases is usually to use chemical fungicides, but long-term and large-scale use will make the pathogens resistant and potentially have a negative impact on human health and the ecological environment. Therefore, finding a safe and effective biological control method instead of chemical control is a hot research topic in recent years. Endophytes, colonizing plants asymptomatically, can promote the growth of the hosts and enhance their resistance. The use of endophytes as biological control agents for postharvest diseases of fruit and vegetables has attracted increasing attention in the last 20 years. Compared with chemical control, endophytes have the advantages of being more environmentally friendly, sustainable, and safer. However, there are relatively few relevant studies, so herein we summarize the available literature. This review focuses mainly on the recent progress of using endophytes to enhance the resistance of postharvest fruit and vegetables to diseases, with the emphasis on the possible mechanisms and the potential applications. Furthermore, this article suggests future areas for study using antagonistic endophytes to prevent and control fruit and vegetable postharvest diseases: (i) screening more potential broad-spectrum anti-pathogen endophytes and their metabolic active substances by the method of macrogenomics; (ii) elucidating the underlining molecular mechanism among endophytes, harvested vegetables and fruit, pathogens, and microbial communities; (iii) needing more application research to overcome the difficulties of commercialization practice. © 2020 Society of Chemical Industry.

Effects of Daily Raspberry Consumption on Immune-Metabolic Health in Subjects at Risk of Metabolic Syndrome: A Randomized Controlled Trial.

Consumption of red raspberries has been reported to exert acute beneficial effects on postprandial glycemia, insulinemia, triglyceridemia, and cytokine levels in metabolically disturbed subjects. In a two-arm parallel-group, randomized, controlled trial, 59 subjects with overweight or abdominal obesity and with slight hyperinsulinemia or hypertriglyceridemia were randomized to consume 280 g/day of frozen raspberries or to maintain their usual diet for 8 weeks. Primary analyses measured metabolic differences between the groups. Secondary analyses performed with omics tools in the intervention group assessed blood gene expression and plasma metabolomic changes following the raspberry supplementation. The intervention did not significantly affect plasma insulin, glucose, inflammatory marker concentrations, nor blood pressure. Following the supplementation, 43 genes were differentially expressed, and several functional pathways were enriched, a major portion of which were involved in the regulation of cytotoxicity, immune cell trafficking, protein signal transduction, and interleukin production. In addition, 10 serum metabolites were found significantly altered, among which ß-alanine, trimethylamine N-oxide, and bioactive lipids. Although the supplementation had no meaningful metabolic effects, these results highlight the impact of a diet rich in raspberry on the immune function and phospholipid metabolism, thus providing novel insights into potential immune-metabolic pathways influenced by regular raspberry consumption.

Nickel allergy in lipid transfer protein sensitized patients: Prevalence and clinical features.

Nickel (Ni), the main responsible for allergic contact dermatitis worldwide, is also involved in systemic condition called "Systemic Nickel Sulfate Allergy Syndrome (SNAS)." Likewise, IgE-mediated reactivity to Lipid Transfer Protein (LTP) represents the main cause of primary food allergy in adults of Mediterranean countries. We evaluated the prevalence of SNAS in LTP allergic patients and investigated patients' clinical features with double sensitization (LTP and Ni). A retrospective, single-center, observational study was conducted performing a complete allergological work-up including: (1) skin prick tests; (2) serum specific IgE for plant food allergens and rPru p3 (LTP); (3) patch test with 5% Ni sulfate in petrolatum. We enrolled 140 LTP allergic patients of which 36 patients (25.7% of sample) showed additional positivity to Ni patch test. Patients with double sensitization were more frequently females and reported fewer cutaneous symptoms. Higher values of sIgE for peach, apple, peanut, walnut, grain, corn, and garlic were found in LTP allergic patients, while higher values for hazelnut in the other subgroup. The prevalence of SNAS in the LTP allergic population is clinically relevant. Moreover, the clinical and immunological profiles of patients with double sensitization were different from patients monosensitized to LTP.

Mitogen-Activated Protein Kinase Cascade and Reactive Oxygen Species Metabolism are Involved in Acibenzolar-S-Methyl-Induced Disease Resistance in Apples.

Apple fruits were subjected to dipping treatment to explore the effects of acibenzolar-S-methyl (ASM) and the mitogen-activated protein kinase (MAPK) inhibitor PD98059 on lesion growth in fruits inoculated with Penicillium expansum. We investigated the roles of the MAPK cascade and reactive oxygen species metabolism in disease resistance in apples. ASM treatment inhibited lesion growth; suppressed catalase (CAT) activity; increased H2O2 content; reduced glutathione and ascorbic acid contents; and increased glutathione reductase, ascorbate peroxidase, peroxidase, superoxide dismutase, and NADPH oxidase activities. Moreover, ASM upregulated MdSOD, MdPOD, MdGR, MdAPX, MdMAPK4, MdMAPK2, and MdMAPKK1 expressions and downregulated MdCAT and MdMAPK3 expressions. PD98059 + ASM treatment increased CAT activity and MdCAT and MdMAPK3 expressions; inhibited MdSOD, MdPOD, MdGR, MdAPX, MdMAPK4, MdMAPK2, and MdMAPKK1 expressions; reduced superoxide dismutase, peroxidase, ascorbate peroxidase, and glutathione reductase activities; and reduced glutathione content in apples. These findings indicate that ASM induces disease resistance in apples by regulating the expressions of key genes involved in reactive oxygen species metabolism and the MAPK cascade.

Can patients with oral allergy syndrome be at risk of anaphylaxis?

PURPOSE OF REVIEW: Oral allergy syndrome, also known as pollen-food syndrome (PFS), is a condition usually associated with adults and characterized by mild transient oropharyngeal symptoms. The purpose of this review is to determine whether systemic or anaphylactic reactions do occur and if so, who is affected and what are the triggers. RECENT FINDINGS: An increasing number of studies demonstrate that PFS occurs all age groups, and a significant number of affected adults do experience systemic and anaphylactic reactions. The upsurge in the adoption of vegan lifestyles, increase in consumption of fruits and vegetables including smoothies and juices, and use of plant foods in nutritional or body-building supplements, could exacerbate this. Changes in pollen and pollution levels, cofactors and sensitization to other plant food allergens may also be involved. SUMMARY: While the majority of those with PFS will continue to experience mild symptoms, all individuals should be properly advised regarding the dangers of concentrated or unusual forms of plant food allergens such as smoothies, juices, soy/nut milks and nutritional supplements. Further well characterized studies are needed to determine risk factors for severe reactions, and sensitization patterns to pollens and plant food allergens.

Evaluation of the phenolic profile and immunoreactivity of Mal d 3 allergen in ancient apple cultivars from Italy.

BACKGROUND: Since the second half of the 20th century, the cultivation of ancient and local apple cultivars has almost disappeared from orchards in Italy. Some of these ancient apple cultivars often possess high nutraceutical values and display lower allergenicity than the modern ones, supporting the so-called 'green revolution' theory. RESULTS: In this study, the phenolic composition and the antioxidant activity of five ancient apple cultivars ('Belfiore', 'Pomella Genovese', 'Gravenstein', 'Bella del Bosco', and 'Piatlin') were compared with a 'Golden Delicious' commercial cultivar. Additionally, apples were tested for their potential allergenicity by detecting the presence of Mal d 3, a non-specific lipid transfer protein that represents the main apples' allergen. All apples came from northern Italy (Trentino Region) and were organically produced. Results showed that, for all cultivars, the skins contained more polyphenols than the pulps. 'Bella del Bosco' had the highest amount of polyphenols and antioxidant activity, whereas 'Piatlin' had the lowest phenolic content. All ancient cultivars presented a higher amount of pulp phenolic compounds than 'Golden Delicious'. Immunoblotting techniques showed that 'Bella del Bosco' and 'Piatlin' had very low quantities of Mal d 3 allergen; hence, they can be considered hypoallergenic cultivars. CONCLUSIONS: The preservation of ancient apple cultivars would be of great importance, not only to maintain the biodiversity but also for their nutritional properties. The hypoallergenic activity of some of these cultivars could be of interest also for the preparation of different apple-based products. © 2020 Society of Chemical Industry.