Volatilomics of raspberry fruit germplasm by combining chromatographic and direct-injection mass spectrometric techniques.

The application of direct-injection mass spectrometric (DI-MS) techniques, like Proton Transfer Reaction Time of Flight Mass Spectrometry (PTR-ToF-MS) has been suggested as a reliable phenotyping tool for fruit volatilome assessment in both genetic and quality-related studies. In this study the complexity of raspberry aroma was investigated by a comprehensive untargeted VOC analysis, done by combining SPME-GC-MS and PTR-ToF-MS assessments with multi-block discriminant analysis using the DIABLO mixOmics framework. The aim was to acquire an exhaustive characterization of the raspberry volatilome according to different fruit ripening stages (pink, ripe, and overripe) and genetic variances (50 accessions), as well as to investigate the potential of PTR-ToF-MS as a rapid and high throughput VOC phenotyping tool to address issues related to raspberry fruit quality. Results of this study demonstrated the complementarity between SPME-GC-MS and PTR-ToF-MS techniques to evaluate the raspberry aroma composition. PTR-ToF-MS generates reliable raspberry VOC fingerprints mainly due to a reduced compound fragmentation and precise content estimation. In addition, the high collinearity between isomers of monoterpenes and norisoprenoids, discovered by GC analysis, reduces the main analytic limitation of PTR-ToF-MS of not being able to separate isomeric molecules. The high similarity between the VOC matrices obtained by applying PTR-ToF-MS and SPME-GC-MS confirmed the possibility of using PTR-ToF-MS as a reliable high throughput phenotyping tool for raspberry volatiolome assessment. In addition, results provided by the germplasm collection investigation enabled to distinguish the best performing accessions, based on VOCs composition, to be used as superior parental lines for future breeding programs.

Ethylene Production Affects Blueberry Fruit Texture and Storability.

Ethylene, produced endogenously by plants and their organs, can induce a wide array of physiological responses even at very low concentrations. Nevertheless, the role of ethylene in regulating blueberry (Vaccinium spp.) ripening and storability is still unclear although an increase in ethylene production has been observed in several studies during blueberry ripening. To overcome this issue, we evaluated the endogenous ethylene production of a Vaccinium germplasm selection at different fruit ripening stages and after cold storage, considering also textural modifications. Ethylene and texture were further assessed also on a bi-parental full-sib population of 124 accessions obtained by the crossing between "Draper" and "Biloxi", two cultivars characterized by a different chilling requirement and storability performances. Our results were compared with an extensive literature research, carried out to collect all accessible information on published works related to Vaccinium ethylene production and sensitivity. Results of this study illustrate a likely role of ethylene in regulating blueberry shelf life. However, a generalisation valid for all Vaccinium species is not attainable because of the high variability in ethylene production between genotypes, which is strictly genotype-specific. These differences in ethylene production are related with blueberry fruit storage performances based on textural alterations. Specifically, blueberry accessions characterized by the highest ethylene production had a more severe texture decay during storage. Our results support the possibility of tailoring ad hoc preharvest and postharvest strategies to extend blueberry shelf life and quality according with the endogenous ethylene production level of each cultivar.

Development of a Novel Phenotypic Roadmap to Improve Blueberry Quality and Storability.

Improved fruit quality and prolonged storage capability are key breeding traits for blueberry (Vaccinium spp.) fruit. Until now, breeding selection was mostly oriented on the amelioration of agronomic traits, such as flowering time, chilling requirement, or plant structure. Up until now, however, the storage effect on fruit quality has not been extensively studied, mostly because objective and handy phenotyping tools to evaluate quality traits were not available. In this study we are proposing a novel phenotyping protocol to support breeding selection and quality control within the entire blueberry production chain. Volatile organic compounds (VOCs) and texture traits, were measured by Proton Transfer Reaction- Time of Flight- Mass Spectrometry (PTR-ToF-MS) and a texture analyzer respectively, taking into consideration the influence of prolonged storage. The exploitation of the genetic variability existing within the investigated blueberry germplasm collection (including both southern and northern highbush, hybrids, and rabbiteyes) allowed the identification of the best performing cultivars, based on texture and VOCs variability, to be used as superior parental lines for future breeding programs. The comprehensive characterization of blueberry aroma allowed the identification of a wide array of spectrometric features, mostly related to aldehydes, alcohols, terpenoids, and esters, that can be used as putative biomarkers to rapidly evaluate the blueberry aroma variations related to genetic differences and storability. In addition, this study revealed a lack of straightforward relationship between harvest and postharvest quality features, that might be genotype-dependent.

Texture features explain the susceptibility of grapevine cultivars to Drosophila suzukii (Diptera: Drosophilidae) infestation in ripening and drying grapes.

Grapevine is a well-known host plant of the invasive pest Drosophila suzukii, but its susceptibility to pest oviposition and development greatly depends on the cultivar. To address environmental sustainability during viticultural zoning planning, new vineyard plantation and Integrated Pest Management programmes, it is essential to take pest pressure and cultivar susceptibility into account. To determine the different grapevine cultivars susceptibility to D. suzukii, we tested twelve widely spread cultivars during the ripening period. We also tested three cultivars during the drying period for raisin wine production. The infestation and emergence rates were consequently related to chemical and texture features of the berries to explain the role of skin and pulp characteristics in determining the nature of the susceptibility. Our results showed that susceptibility to D. suzukii infestation varies across cultivars. On ripening grapes, infestation is primarily influenced by skin and pulp firmness, elasticity and consistency. Suitability for egg development resulted mainly related to skin and pulp deformation. In a drying loft, infestation may also occur in relation to skin and pulp consistency. Lastly, we discuss the practical implication of the underestimated role of berry texture in D. suzukii oviposition and emergence success, in both ripening and drying grapes.

Exploring Blueberry Aroma Complexity by Chromatographic and Direct-Injection Spectrometric Techniques.

Blueberry (Vaccinium spp.) fruit consumption has increased over the last 5 years, becoming the second most important soft fruit species after strawberry. Despite the possible economic and sensory impact, the blueberry volatile organic compound (VOC) composition has been poorly investigated. Thus, the great impact of the aroma on fruit marketability stimulates the need to step forward in the understanding of this quality trait. Beside the strong effect of ripening, blueberry aroma profile also varies due to the broad genetic differences among Vaccinium species that have been differently introgressed in modern commercial cultivars through breeding activity. In the present study, divided into two different activities, the complexity of blueberry aroma was explored by an exhaustive untargeted VOC analysis, performed by two complementary methods: SPME-GC-MS (solid phase microextraction- gas chromatography-mass spectrometry) and PTR-ToF-MS (proton transfer reaction-time of flight-mass spectrometry). The first experiment was aimed at determining the VOC modifications during blueberry ripening for five commercially representative cultivars ("Biloxi," "Brigitta Blue," "Centurion," "Chandler," and "Ozark Blue") harvested at four ripening stages (green, pink, ripe, and over-ripe) to outline VOCs dynamic during fruit development. The objective of the second experiment was to confirm the analytical capability of PTR-ToF-MS to profile blueberry genotypes and to identify the most characterizing VOCs. In this case, 11 accessions belonging to different Vaccinium species were employed: V. corymbosum L. ("Brigitta," "Chandler," "Liberty," and "Ozark Blue"), V. virgatum Aiton ("Centurion," "Powder Blue," and "Sky Blue"), V. myrtillus L. (three wild genotypes of different mountain locations), and one accession of V. cylindraceum Smith. This comprehensive characterization of blueberry aroma allowed the identification of a wide pull of VOCs, for the most aldehydes, alcohols, terpenoids, and esters that can be used as putative biomarkers to rapidly evaluate the blueberry aroma variations related to ripening and/or senescence as well as to genetic background differences. Moreover, the obtained results demonstrated the complementarity between chromatographic and direct-injection mass spectrometric techniques to study the blueberry aroma.