Linking Colorimetric Variation with Non-Volatile and Volatile Components of Carob Flour.

Chromatic variation was examined for its association with flour composition and quality. Carob samples from variable altitudes and genetic backgrounds were milled and assessed for colorimetric parameter L* (lightness) and analyzed for phenols, tannins, antioxidant capacity, soluble carbohydrates (HPLC-RID), organic acids and protein (IC-CD), and volatile organic compounds (VOCs; HS-SPME/GC-MS). Higher altitudes and grafted genotypes yielded lighter-colored flours of higher antioxidant potential, phenols, tannins, sucrose, and malic acid concentrations. VOCs were mainly acids, esters, aldehydes, ketones, and alcohols. Acids were the most abundant and correlated negatively with L*, though correlation for many individual acids was non-significant, including 2-methyl-propanoic acid, widely considered the carob signature aroma (cheesy acidic buttery). The compositional and quality indexing potential of L* is more robust for grafted than non-grafted material, owing putatively to a narrower genetic basis. Antioxidant capacity and concentrations of phenolics, tannins and sucrose correlated positively with L*, indicating increased levels in carob flours sourced from grafted trees at higher altitudes. These flours also have a lower content of reducing sugars, the implication of which in the darkening of carob flour warrants further investigation. Overall, L* constitutes a reliable index for ranking carob flours for key compositional attributes and may be further reinforced by multiple-year data.

Genome Size Variation across a Cypriot Fabeae Tribe Germplasm Collection.

DNA content is an important trait linked to the evolutionary routes of taxa and often connected to speciation. In the present study, we studied C-values variation across the Cypriot Fabeae gene pool. Several hundred plants (Vicia spp., Lens spp., Pisum spp.) were sampled across Cyprus. Accurate estimates were established by flow cytometry and propidium iodine staining for 155 discrete populations/accessions. A ten-fold variation was detected across lineages with 1C DNA content varying from 1.584 pg for V. cretica (ARI02420) to 13.983 pg for V. faba (ARI00187). In general, flow cytometry was precise for the characterization of species, even though there were instances of genome overlapping across taxa. Most analyses in the current work refer to species that have not been characterized before by flow cytometry (or any other DNA content estimation method). Still, a correlation to C-values previously reported in Kew Plant DNA C-values database was attempted. A high degree of correlation except for V. dalmatica was established. The evaluation of genome size trait in relation with the Fabeae phylogeny, revealed that Pisum and Lens genera were rather homogenous, but an astonishing fluctuation was shown for Vicia spp. Moreover, it was established that genome up- or down-scaling was not directly linked to speciation drivers. The genomic size measurements presented here could deliver extra quality control for the identification and characterization of taxa in germplasm collections, particularly in cases where species share morphological characters.

Adapting Agriculture to Climate Change: A Synopsis of Coordinated National Crop Wild Relative Seed Collecting Programs across Five Continents.

The Adapting Agriculture to Climate Change Project set out to improve the diversity, quantity, and accessibility of germplasm collections of crop wild relatives (CWR). Between 2013 and 2018, partners in 25 countries, heirs to the globetrotting legacy of Nikolai Vavilov, undertook seed collecting expeditions targeting CWR of 28 crops of global significance for agriculture. Here, we describe the implementation of the 25 national collecting programs and present the key results. A total of 4587 unique seed samples from at least 355 CWR taxa were collected, conserved ex situ, safety duplicated in national and international genebanks, and made available through the Multilateral System (MLS) of the International Treaty on Plant Genetic Resources for Food and Agriculture (Plant Treaty). Collections of CWR were made for all 28 targeted crops. Potato and eggplant were the most collected genepools, although the greatest number of primary genepool collections were made for rice. Overall, alfalfa, Bambara groundnut, grass pea and wheat were the genepools for which targets were best achieved. Several of the newly collected samples have already been used in pre-breeding programs to adapt crops to future challenges.

Morphological Diversity, Genetic Characterization, and Phytochemical Assessment of the Cypriot Tomato Germplasm.

Tomato (Solanum lycopersicum L.) is considered one of the most valuable and versatile vegetable crops globally and also serves as a significant model species for fruit developmental biology. Despite its significance, a severe genetic bottleneck and intense selection of genotypes with specific qualitative traits have resulted in the prevalence of a restricted number of (geno)types, also causing a lack of diversity across widespread cultivated types. As a result, the re-emergence of landraces as well as traditional and heirloom varieties is largely acknowledged as a countermeasure to restore phenotypic, phytochemical and genetic diversity while enriching the aroma/taste tomato palette. On those grounds, the Cypriot tomato germplasm was assessed and characterized. Ten landrace accessions were evaluated under greenhouse conditions and data were collected for 24 IPGRI discrete phenotypic traits. Grouping of accessions largely reflected the fruit shape and size; four different fruit types were recorded across accessions (flattened, heart-shaped, rounded and highly rounded). Moreover, a single run panel consisting of ten SSRs was developed and applied in order to genetically characterize 190 Cypriot genotypes and foreign heirloom varieties. Based on genetic indexes it was established that tomato landraces have a rather low level of heterogeneity and genetic variation. Finally, mineral and phytochemical analyses were conducted in order to estimate biochemical attributes (total phenolics, ascorbic acid, lycopene, ß-carotene, total soluble content, titratable acidity) across genotypes; thus, ascertaining that the Cypriot panel has a high nutritional value. Due to the thermo-drought adaptation and tolerance of these genotypes, the current study serves as a roadmap for future breeding efforts in order to incorporate desirable traits or develop novel tomato lines combining resilience and alimentary value.

Changes in the primary and secondary metabolome of male green asparagus (Asparagus officinalis L.) as modulated by sequential harvesting.

Rising temperature and solar radiation drive the mobilization and depletion of crown-deposited metabolites harnessed for asparagus spear regeneration during the spring harvest season. We presently examined how successive same-season harvests impact the sensory, nutritive and bioactive composition of select green asparagus genotypes. Soluble carbohydrates were analyzed by HPLC-RI, organic acids and polyphenols by HPLC-DAD and metals by ion chromatography. Higher sugars and lower acids accentuated sweetness and lower polyphenols contributed to reduced astringency at the beginning of the harvest season. This trend was reversed as the season advanced and spear sensory quality was compromised by declining sugars and rising acids; however, functional quality improved as antioxidant capacity increased along with the concentrations of quercetin-3-O-rutinoside (rutin) and ascorbic acid. The compositional changes presently described were uniform across all genotypes examined and thus contribute toward our understanding of seasonal variation in the sensory and functional quality of this acclaimed health-promoting product.

Configuration of the Volatile Aromatic Profile of Carob Powder Milled From Pods of Genetic Variants Harvested at Progressive Stages of Ripening From High and Low Altitudes.

Carob powder is increasingly valued as a substitute for cocoa and as a flavor-enhancing component of processed foods. However, little is known about the impact of preharvest factors such as fruit maturity, genotype and altitude on its volatile organic compounds (VOCs) composition. The current study examined the VOCs composition of powder milled from pods of two genotypes cultivated at 15 and 510 m altitude and harvested at six progressive stages of maturity, ranging from fully developed immature green (RS1) to late ripe (RS6). Fifty-six VOCs categorized into acids, esters, aldehydes, ketones, alcohols, furans, and alkanes were identified through HS-SPME GC-MS analysis. Maturity was the most influential factor, followed by altitude and least by genotype. Aldehydes and alcohols correlated positively (r = 0.789; p < 0.001), both accumulated in immature carobs and decreased with progressive ripening, resulting in the attenuation of green grassy aroma. Conversely, acids increased with ripening and dominated the carob volatilome at full maturity, correlating negatively with aldehydes and alcohols (r = -0.835 and r = -0.950, respectively; p < 0.001). The most abundant VOC throughout ripening (17.3-57.7%) was isobutyric acid, responsible for the characteristic cheesy-acidic-buttery aroma of carob powder. The pleasurable aroma detected at the immature stages (RS2 and RS3) was traced to isobutyrate and methyl isobutyrate esters, rendering unripe green carob powder a potential admixture component for improving the aroma of novel food products. Lower altitude favored the accumulation of acids linked to less pleasant aroma, whereas isobutyric acid was more abundant at higher altitude. This constitutes a significant indication that higher altitude enhances the characteristic carob-like aroma and sensory quality of carob powder.

Pod Morphology, Primary and Secondary Metabolite Profiles in Non-grafted and Grafted Carob Germplasm Are Configured by Agro-Environmental Zone, Genotype, and Growing Season.

Carob is a predominantly rainfed tree crop of high nutritive value and a long history of adaptation to the edaphoclimatic stress conditions of the Mediterranean. However, declining attention to the carob tree in recent decades has aggravated genetic erosion. The extant in situ germplasm varies both in terms of pod morphology and composition, reflecting the genetic and physiological divide chiefly among grafted and non-grafted material, and possibly the impact of variable agro-environments. Accordingly, the present study aimed to establish a systematic categorization of the genetic and phenotypic diversity encountered across carob germplasm identified in situ throughout Cyprus, a historical center of production and genetic diversity for the species. Linking pod morphology, primary and secondary metabolite profiles with genotyped source material originating in different agro-environments and crop seasons would provide a framework for interpreting (a) the interaction of these factors in configuring carob pod physicochemical constitution, and (b) the relative stability of phenotypic traits against environmental and seasonal variation. Microsatellite analysis discriminated 36 genotypes out of the 124 trees located in nine traditional agro-environmental zones and revealed low genetic diversity within the grafted germplasm. Two landraces were identified: "Tillyria," which is widespread and predominant, and "Kountourka," which is mainly localized to the northeastern peninsula of Karpasia. Morphological traits, such as seeds-to-pod weight ratio, pod width and thickness were principally under genetic control. Contrarily, compositional traits, particularly total phenolic content-including condensed tannins, in vitro antioxidant capacity and to a lesser extent gallic acid, organic acids and minerals were under agro-environmental control. Agro-environmental zone also modulated principally fructose and glucose; sucrose was modulated equally by genotype and agro-environment, while total sugars were under genetic control. Statistically significant differences between seasons were detected for all traits except for the seeds-to-pod weight ratio, pod length and width. Hierarchical cluster analysis corroborates that Cyprus may be divided into two major agro-environmental zones modulating the compositional properties of the carob pulp. The present study provides a comprehensive insight into the extant carob genetic resources of Cyprus and advances our understanding of how genetic, agro-environmental and seasonal factors interact in shaping carob pod morphology and composition.

Genetic variability in landraces populations and the risk to lose genetic variation. The example of landrace 'Kyperounda' and its implications for ex situ conservation.

Genetic characterization enhances the development of rational conservation strategies and the utilization of germplasm to plant breeding programs. In the present study, 19 microsatellite markers were employed to evaluate the genetic diversity and the genetic affiliations across 20 Cypriot durum wheat (Triticum turgidum L. subsp. durum) landraces, 13 landraces from the broader Mediterranean basin and 22 modern varieties. Cluster analysis depicted a clear separation among modern varieties and landraces, regardless of their origin. Landraces presented the highest genetic variation (average discriminating power of 0.89) and a high number of private alleles (131) was detected; underlying the unique genetic mark-up of this genepool. AMOVA revealed that the highest variability was detected within the landraces originating from Cyprus and landraces from the broader Mediterranean basin. The Cypriot landrace 'Kyperounda' was selected for further evaluation of its' intra-genetic variation and it was determined that genetic diversity was higher in accessions conserved as sublines (He 0.643-0.731) than bulks (He 0.384-0.469). Bayesian analysis revealed substantial admixture within 'Kyperounda' accessions, depicted also by Principal Coordinate Analysis. The findings of the current manuscript emphasize that high intra-genetic diversity is retained when landraces are conserved as sublines in ex situ collections, while landraces that are conserved as bulks have a higher risk of bottleneck. Hence, a more exhausting diversity evaluation is needed in order to fully utilize landraces in breeding schemes and to prevent the loss of genetic variation.

Genetic diversity of Avena ventricosa populations along an ecogeographical transect in Cyprus is correlated to environmental variables.

Avena ventricosa Balansa ex Coss. is considered the C-genome donor of the cultivated hexaploid oat and is a 'priority' species for conservation, since it has limited geographic distribution and the only recorded populations in Europe are present in Cyprus. The current study attempts to characterize the genetic structure and fragmentation of the species via the application of genotypic markers. It was revealed that the genetic variety was mainly allocated among the populations collected, since clustering obtained was according to the geographic origin of the samples and the habitat. Species distribution modeling showed that the most important climatic variable defining A. ventricosa distribution is the mean diurnal temperature. Furthermore, significant association of the genetic structure to environmental variables was detected; overall, a negative association to precipitation was confirmed, while significant correlations of genetic structure and the temperature at the time of anthesis and germination were established. The safeguarding of this valuable genetic resource is discussed.

Assessment of Vegetation Indices Derived by UAV Imagery for Durum Wheat Phenotyping under a Water Limited and Heat Stressed Mediterranean Environment.

There is growing interest for using Spectral Vegetation Indices (SVI) derived by Unmanned Aerial Vehicle (UAV) imagery as a fast and cost-efficient tool for plant phenotyping. The development of such tools is of paramount importance to continue progress through plant breeding, especially in the Mediterranean basin, where climate change is expected to further increase yield uncertainty. In the present study, Normalized Difference Vegetation Index (NDVI), Simple Ratio (SR) and Green Normalized Difference Vegetation Index (GNDVI) derived from UAV imagery were calculated for two consecutive years in a set of twenty durum wheat varieties grown under a water limited and heat stressed environment. Statistically significant differences between genotypes were observed for SVIs. GNDVI explained more variability than NDVI and SR, when recorded at booting. GNDVI was significantly correlated with grain yield when recorded at booting and anthesis during the 1st and 2nd year, respectively, while NDVI was correlated to grain yield when recorded at booting, but only for the 1st year. These results suggest that GNDVI has a better discriminating efficiency and can be a better predictor of yield when recorded at early reproductive stages. The predictive ability of SVIs was affected by plant phenology. Correlations of grain yield with SVIs were stronger as the correlations of SVIs with heading were weaker or not significant. NDVIs recorded at the experimental site were significantly correlated with grain yield of the same set of genotypes grown in other environments. Both positive and negative correlations were observed indicating that the environmental conditions during grain filling can affect the sign of the correlations. These findings highlight the potential use of SVIs derived by UAV imagery for durum wheat phenotyping under low yielding Mediterranean conditions.