Analysis of major paralogs encoding the Fra a 1 allergen based on their organ-specificity in Fragaria × ananassa

Ishibashi, Misaki; Nabe, Takeshi; Nitta, Yoko; Tsuruta, Hiroki; Iduhara, Miho; Uno, Yuichi

doi:10.1007/s00299-017-2237-6

Cited by 15 publications

(35 citation statements)

References 38 publications

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“…The major allergen in F. × ananassa is a member of the PR-10 subfamily, Fra a 1 (Hjernø et al, 2006;Karlsson et al, 2004), and that in Fragaria vesca is Fra v 1 (Hyun and Kim, 2011). The number of paralogs in the genome differ among genotypes; there are over 30 paralogs in F. × ananassa (Ishibashi et al, 2018), and 13 paralogs in F. vesca (Hyun and Kim, 2011). The Fra a 1.01 protein has been targeted as one of the major strawberry allergens (Ishibashi et al, 2018;Musidlowska-Persson et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Seasonal Variation of the Major Allergen Fra a 1 in Strawberry Fruit

et al. 2019

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Strawberry (Fragaria × ananassa) contains a major allergen, Fra a 1, which causes oral allergic syndrome. Fra a 1 is a PR-10 homolog that is regulated by environmental conditions. The allergenicity of fruit caused by Fra a 1 may depend on the genotype or growing conditions. We analyzed the Fra a 1.01 transcript levels and Fra a 1.01 protein levels in strawberry fruits of several genotypes across all seasons. In the preliminary rough screening, we selected the line WH1 bearing white fruit and the red-fruited cultivar 'Akihime'. Under the same environmental conditions, there was no significant difference in Fra a 1.01 levels between the two cultivars over several months, suggesting that receptacle color was not indicative of allergenicity caused by Fra a 1.01. Fruits cultivated under the same environmental conditions should be used for comparisons of the allergenicity among genotypes. Both 'Akihime' and WH1 accumulated significantly higher levels of Fra a 1.01 protein in winter than in spring. We investigated the effects of irradiation and low temperature as environmental factors controlling the accumulation of Fra a 1.01 in winter. A shading treatment on fruit did not significantly affect Fra a 1.01 protein accumulation in strawberry fruits. Regarding variations over time, the Fra a 1.01 protein content was higher in fruits harvested at midnight in January than in those harvested at other times and in other months. These findings suggested that the Fra a 1.01 protein accumulates in response to environmental factors such as cold stress.

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Section: Introductionmentioning

confidence: 99%

“…The number of paralogs in the genome differ among genotypes; there are over 30 paralogs in F. × ananassa (Ishibashi et al, 2018), and 13 paralogs in F. vesca (Hyun and Kim, 2011). The Fra a 1.01 protein has been targeted as one of the major strawberry allergens (Ishibashi et al, 2018;Musidlowska-Persson et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Seasonal Variation of the Major Allergen Fra a 1 in Strawberry Fruit

et al. 2019

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“…To validate the quality of RNA for downstream assays, RNA samples were used in real-time PCR and NGS analyses. Target gene was Fra a 1, encoding major allergenic protein in F. ·ananassa because we have already established the evaluation system by real-time PCR (Ishibashi et al, 2018). In both Fra a 1.01 and Fra a 1.02 genes, no statistically significant difference was found in relative transcript levels between RNAs isolated by manual and automated methods (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The cDNA libraries were sequenced on the HiSeq4000 platform (Illumina, San Diego, CA) using paired-end chemistry with 100-base pair reads. Bioinformatic analysis was carried out using scientifically recognized algorithms according to Ishibashi et al (2018). Briefly, sequencing reads were filtered by SOAPnuke (Chen et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Efficient Isolation of High-quality Total RNA from Strawberry

Ishibashi¹,

Nabe²,

Nitta³

et al. 2019

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Sufficient yields of high-quality RNA are needed for next-generation sequencing and high-throughput real-time polymerase chain reaction analyses. In the case of strawberry (Fragaria 3ananassa) fruits, successful RNA isolation requires removal of abundant inhibitory substances (polysaccharides and polyphenols) that greatly reduce quality and yield. In this study, we applied various combinations of RNA isolation protocols directed at reproductive organs. The best manual isolation method involved nonionic polymer and modified acid guanidinium thiocyanate-phenol-chloroform treatments followed by phenol/chloroform/isoamyl alcohol extraction. Compared with other methods, this approach gave significantly higher yields [84.0 mg/g fresh weight (FW)] of RNA of greater purity (A260/A280 = 1.99; A260/230 = 1.51). Better-quality RNA (A260/ 230 = 2.11) was obtained using an automated method, but the yield was lower (18.1 mg/g FW) than that obtained manually. This automated method consisted of pretreatment with nonionic polymer followed by a silica-based system extraction. Although RNA of sufficient quality [RNA Integrity Number (RIN) ‡ 6.5 and 28S/18S ‡ 1.0] for RNA sequencing was obtained from receptacles using both automated and manual methods, the manual method yielded high-quality RNA from achenes and anthers. The automatic method features 6-fold faster high-throughput capacity, whereas the manual method has wider applicability to different tissues.

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“…Crystallization and Structural Characterization of Fra a 1.02. Since Fra a 1.02 is the most expressed allergen in the ripe strawberry fruit and has also been identified as highly allergenic, 5,9,16 it is considered as the main responsible allergen causing type I pollen-related allergies to strawberry. The…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Structural Bases for the Allergenicity of Fra a 1.02 in Strawberry Fruits

Orozco-Navarrete

Kaczmarska

Dupeux

et al. 2019

J. Agric. Food Chem.

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Although strawberries are highly appreciated fruits, their intake can induce allergic reactions in atopic patients. These reactions can be due to the patient's previous sensitization to the major birch pollen allergen Bet v 1, by which IgE generated in response to Bet v 1 cross-reacts with the structurally related strawberry Fra a 1 protein family. Fra a 1.02 is the most expressed paralog in ripe strawberries and is highly allergenic. To better understand the molecular mechanisms regulating this allergic response, we have determined the three-dimensional structure of Fra a 1.02 and four site-directed mutants that were designed based on their positions in potential epitopes. Fra a 1.02 and mutants conform to the START fold. We show that the cross-reactivity of all the mutant variants to IgE from patients allergic to Bet v 1 was significantly reduced without altering the conserved structural fold, so that they could potentially be used as hypoallergenic Fra a 1 variants for the generation of vaccines against strawberry allergy in atopic patients. Article pubs.acs.org/JAFC Cite This: J. Agric. Food Chem. XXXX, XXX, XXX−XXX

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Analysis of major paralogs encoding the Fra a 1 allergen based on their organ-specificity in Fragaria × ananassa

Cited by 15 publications

References 38 publications

Seasonal Variation of the Major Allergen Fra a 1 in Strawberry Fruit

Seasonal Variation of the Major Allergen Fra a 1 in Strawberry Fruit

Efficient Isolation of High-quality Total RNA from Strawberry

Structural Bases for the Allergenicity of Fra a 1.02 in Strawberry Fruits

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