Cuticular wax biosynthesis in blueberries (Vaccinium corymbosum L.): Transcript and metabolite changes during ripening and storage affect key fruit quality traits.

In fruits, cuticular waxes affect fruit quality traits such as surface color at harvest and water loss during postharvest storage. This study investigated the transcriptional regulation of cuticular wax deposition in northern highbush blueberries (Vaccinium corymbosum L.) in relation to fruit water loss and surface color during ripening and postharvest storage, as well as the effects of abscisic acid (ABA)-mediated changes in cuticular wax deposition on these fruit quality traits. Total cuticular wax content (µg∙cm-2) decreased during fruit ripening and increased during postharvest storage. Transcriptome analysis revealed a transcript network for cuticular wax deposition in blueberries. Particularly, five OSC-Likes were identified as putative genes for triterpene alcohol production, with OSC-Like1 and OSC-Like2 encoding mixed amyrin synthases, OSC-Like3 encoding a lupeol synthase, and OSC-Like4 and OSC-Like5 encoding cycloartenol synthases. The expression of three CYP716A-like genes correlated to the accumulation of two triterpene acids oleanolic acid and ursolic acid, the major wax compounds in blueberries. Exogenous ABA application induced the expression of triterpenoid biosynthetic genes and the accumulation of ß-amyrin and oleanolic acid, as well as increased the ratio of oleanolic acid to ursolic acid. These changes were associated with reduced fruit water loss. The content of ß-diketones was also increased by ABA application, and this increase was associated with increased fruit lightness (measured as L* using CIELAB Color Space by a colorimeter). This study provided key insights on the molecular basis of cuticular wax deposition and its implications on fruit quality traits in blueberries.

Determination of bound volatiles in blueberries, raspberries, and grapes with an optimized protocol and a validated SPME-GC/MS method.

Bound volatiles are odorless aroma reservoirs that modify flavor when released during food processing, and their determination is important to understand the aroma of fruit beverages. However, the generation of oxidation/degradation artifacts during analyses of glycosidically-bound volatiles has not been compared across fruit species and their dependence on diverse acidic and enzymatic hydrolytic conditions remains unclear. This work aimed to optimize and compare different hydrolytic conditions for the analysis of glycosidically-bound volatiles in blueberries, raspberries, and grapes with a solid-phase microextraction - gas chromatography/mass spectrometry (SPME-GC/MS) methodology. Enzymatic hydrolyses using AR2000® at 100 mg.mL-1 and Pectinex Ultra SPL® at 25-100 µL.mL-1 showed profiles characterized by the expected alcohols, while using AR2000® at 200-400 mg.mL-1 and citric acid at 50-100 mM resulted in profiles defined by artifacts (hydrocarbons, norisoprenoids, and aldehydes). (Z)-3-hexen-1-ol, 3-methyl-1-butanol, linalool, citronellol, and geraniol presented Odor Activity Values (OAV) > 1 for most small fruit genotypes.

Investigation of Melissococcus plutonius isolates from 3 outbreaks of European foulbrood disease in commercial beekeeping operations in western Canada.

European foulbrood (EFB) disease is an economically important bacterial disease of honey bee larvae caused by enteric infection with Melissococcus plutonius. In this study, we investigated 3 clinical outbreaks of EFB disease in commercial beekeeping operations in western Canada in the summer of 2020 and characterized the Melissococcus plutonius isolates cultured from these outbreaks according to genetic multi-locus sequence type and i n vitro larval pathogenicity. We isolated M. plutonius sequence type 19 from EFB outbreaks in British Columbia and Alberta, and a novel M. plutonius sequence type 36 from an EFB outbreak in Saskatchewan. In vitro larval infection with each M. plutonius isolate was associated with decreased larval survival in vitro by 58.3 to 70.8% (P < 0.001) compared to non-infected controls. Further elucidation of mechanisms of virulence of M. plutonius, paired with epidemiologic investigation, is imperative to improve EFB management strategies and mitigate risks of EFB outbreaks in western Canada.

Enquête sur des isolats de Melissococcus plutonius provenant de trois éclosions de loque e uropéenne dans des exploitations apicoles commerciales de l'Ouest canadien. La loque européenne (EFB) est une maladie bactérienne économiquement importante des larves d'abeilles mellifères causée par une infection entérique par Melissococcus plutonius. Dans cette étude, nous avons enquêté sur trois éclosions cliniques de la maladie EFB dans des exploitations apicoles commerciales dans l'ouest du Canada à l'été 2020 et caractérisé les isolats de Melissococcus plutonius cultivés à partir de ces éclosions selon le typage génomique multilocus et la pathogénicité larvaire in vitro. Nous avons isolé le type de séquence 19 de M. plutonius des éclosions d'EFB en Colombie-Britannique et en Alberta, et une nouvelle séquence de type 36 de M. plutonius d'une éclosion d'EFB en Saskatchewan. L'infection larvaire in vitro avec chaque isolat de M. plutonius était associée à une diminution de la survie larvaire in vitro de 58,3 à 70,8 % (P < 0,001) par rapport aux témoins non infectés. Une élucidation plus poussée des mécanismes de virulence de M. plutonius, associée à une enquête épidémiologique, est impérative pour améliorer les stratégies de gestion de l'EFB et atténuer les risques d'épidémies d'EFB dans l'Ouest canadien.(Traduit par Dr Serge Messier).

Optimization and validation of a SPME-GC/MS method for the determination of volatile compounds, including enantiomeric analysis, in northern highbush blueberries (Vaccinium corymbosum L.).

Blueberry aroma is one of the most important quality traits that influences consumer purchasing decisions. This study aimed to optimize and validate a solid-phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) method for the quantification of 73 volatile compounds in northern highbush blueberries. A SPME extraction of blueberries with water and specific proportions of sodium chloride, citric acid, and ascorbic acid, for 60 min at 50 °C using a divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber was optimal. The method was validated for sensitivity, reproducibility, linearity, and accuracy, and used to quantify volatile compounds through matrix-matched calibration curves in six blueberry cultivars ('Duke', 'Draper', 'Bluecrop', 'Calypso', 'Elliott', and 'Last Call'). Terpenes represented the most abundant volatile fraction, followed by aldehydes and alcohols. Linalool and 2-(E)-hexenal were key compounds that differentiated blueberry cultivars via Principal Component Analysis (PCA). Enantiomeric analyses revealed an excess of (-)-limonene, (-)-α-pinene, and (+)-linalool for all cultivars with potential impacts on the blueberry aroma.