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The fruit of commercial cranberry (Vaccinium macrocarpon Ait.) cultivars have relatively high concentrations of malic acid (MA) and citric acid (CA), and, to a lesser extent, quinic acid. These acids contribute to the high titratable acidity (TA), a measure of tartness, of cranberry fruit, which typically ranges from 2.3 to 2.5% citric acid equivalents in commercial cultivars. Thus, considerable amounts of sugar are added (“added sugar”) in products such as sweetened-dried cranberries and juices. Within our cranberry germplasm collection, a unique accession was identified with fruit having a TA ≈ 1.5%, where MA concentration was reduced to ~ 4 mg/g fresh weight (FW), compared to ~ 8 mg/g FW in current cultivars. Inbred crosses derived from this accession yielded progeny with a lower MA phenotype (~ 2 mg/g FW). Observed segregation indicated the accession was heterozygous (Mala/mala) for a low MA allele (mala) and very low MA progeny were homozygous (mala/mala). MA was reduced approximately 75% in these populations relative to standard cultivars. The homozygous mala/mala locus also depressed fruit CA and quinic acid concentrations. Quantitative trait loci mapping identified a region on chromosome 4 associated with low MA. The combined segregation of three half-sib populations derived from the low MA accession generated effective (within < 1 cM) Kompetitive allele-specific PCR (KASP) markers for use in breeding of cranberry cultivars with lower MA. Dihybrid populations were developed having a previously described low CA allele, cita, with the mala alleles of this study to explore the interaction of alleles at both loci.

The cranberry (Vaccinium macrocarpon Ait.) is a North American fruit crop domesticated less than 200 years ago. The USDA began the first cranberry breeding program in response to false-blossom disease in 1929, but after the first generation of cultivars were released in the 1950s, the program was discontinued. Decades later, renewed efforts for breeding cranberry cultivars at Rutgers University and the University of Wisconsin yielded the first modern cultivars in the 2000’s. Phenotypic data suggests that current cultivars have changed significantly in terms of fruiting habits compared to original selections from endemic populations. However, due to the few breeding and selection cycles and short domestication period of the crop, it is unclear how much cultivated germplasm differs genetically from wild selections. Moreover, the extent to which selection for agricultural superior traits has shaped the genetic and phenotypic variation of cranberry remains mostly obscure. Here, a historical collection composed of 362 accessions, spanning wild germplasm, first-, second-, and third-generation selection cycles was studied to provide a window into the breeding and domestication history of cranberry. Genome-wide sequence variation of more than 20,000 loci showed directional selection across the stages of cranberry domestication and breeding. Diversity analysis and population structure revealed a partially defined progressive bottleneck when transitioning from early domestication stages to current cranberry forms. Additionally, breeding cycles correlated with phenotypic variation for yield-related traits and anthocyanin accumulation, but not for other fruit metabolites. Particularly, average fruit weight, yield, and anthocyanin content, which were common target traits during early selection attempts, increased dramatically in second- and third-generation cycle cultivars, whereas other fruit quality traits such as Brix and acids showed comparable variation among all breeding stages. Genome-wide association mapping in this diversity panel allowed us to identify marker-trait associations for average fruit weight and fruit rot, which are two traits of great agronomic relevance today and could be further exploited to accelerate cranberry genetic improvement. This study constitutes the first genome-wide analysis of cranberry genetic diversity, which explored how the recurrent use of wild germplasm and first-generation selections into cultivar development have shaped the evolutionary history of this crop species.

The fruit of American cranberry (Vaccinium macrocarpon Ait.) is known for its tart, acidic taste. Although some acidity is required for expressing fruit flavor, the high acidity of cranberry requires considerable amounts of “added-sugars” to make cranberry products palatable. Reducing acidity of cranberry would allow for less sugar to be added to cranberry products. In fruit, total acidity is quantified as titratable acidity (TA) in citric acid equivalents and typically ranges from 2.3 to 2.5% in cranberry. This study characterized the genetics of a low acid trait (TA≈1.5%), due to lower citric acid (~ 2 mg/g fresh weight), identified in an undomesticated cranberry germplasm accession. To determine the inheritance of this trait, this accession was used in a series of crosses. The progeny of these crosses exhibited phenotypic segregation consistent with single-gene Mendelian inheritance, with the low citric acid trait being largely recessive. Bulk segregant analysis using simple sequence repeat (SSR) markers identified two SSR markers within

Informed assessment of priority genetic traits in plant breeding programs is important to improve the efficiency of developing cultivars suited to current climate and industry needs. The efficiency of genetic improvement is critical for perennial crops such as cranberries, as they usually involve more resources, time, and funding compared with other crops. This study investigated the relative importance of cranberry producers’ preferences for breeding traits related to fruit quality, productivity, plant physiology, and resistance to biotic and abiotic stresses. Industry responses revealed that fruit characteristics affecting fruit quality, including firmness, fruit size and anthocyanin content, and resistance to fruit rot, were the most desired traits in new cranberry cultivar release. These traits have the potential to increase the quality standards needed to process high-value sweetened dried cranberry products, positively affecting price premiums received by producers, which is critical for the economic viability of the cranberry industry. Our findings will be useful to breeders and allied scientists seeking to develop an advanced DNA-based selection strategy that would impact the global cranberry industry.

Developing new blueberry cultivars requires plant breeders to be aware of current and emerging needs throughout the supply chain, from producer to consumer. Because breeding perennial crop plants (such as blueberry) is time- and resource-intensive, understanding and targeting priority traits is critical to enhancing the efficiency of breeding programs. This study assesses blueberry industry breeding priorities for fruit and plant quality traits based on a survey conducted at commodity group meetings across nine U.S. states and in British Columbia (Canada) between Nov. 2016 and Mar. 2017. In general, industry responses signaled that the most important trait cluster was fruit quality including the firmness, flavor, and shelf life. Fruit quality traits affect price premiums received by producers; influence consumer’s preferences; and have the potential to increase the feasibility of mechanical harvesting, all critical to the economic viability of the industry. There were differences across regions in the relative importance assigned to traits for disease resistance, arthropod resistance, and tolerance to abiotic stresses. Our findings will be useful to researchers seeking solutions for challenges to the North American blueberry industry including development of new cultivars with improved traits using accelerated DNA-based selection strategies.

Cranberry flavonoids, including anthocyanins, flavonol glycosides and proanthocyanidins, and organic acids were characterized and quantified by HPLC and LC-MS/MS during fruit development and ripening in eight cranberry cultivars. Anthocyanin biosynthesis initiated at early fruit development and reached highest level in mature fruit, with significant differences between cultivars. Major flavonol glycosides, including the most abundant quercetin-3-galactoside and myricetin-3-galactoside, showed consistent concentrations during the season with moderate fluctuation, and were at similar levels in mature fruits of the eight cultivars. Proanthocyanidins declined during fruit development and then increased slightly in later maturation stages. Levels of various proanthocyanidin oligomers/polymers with different degree-of-polymerization were highly correlated within a cultivar during fruit development. Cultivars with coancestry exhibited similar levels (high/low) of anthocyanins or proanthocyanidins, indicating genetic effects on biosynthesis of such flavonoids. All cultivars showed similar levels of malic and citric acids, and declining levels of quinic acid during fruit development. Benzoic acid was extremely low early in the season and increased sharply during fruit ripening. Levels of quinic and citric acids were significantly different among cultivars in the mature fruit. Concentrations of proanthocyanidins, anthocyanins, quinic acid and benzoic acid have a strong developmental association in developing ovaries.

Eggplant fruits are known to contain different classes of phenolic phytochemicals (flavonols, phenolic acids, and anthocyanins) that can exert beneficial effects on human health. This study developed methods for the qualitative and quantitative composition analysis of phenolic compounds in the skin of eggplant fruits harvested following conventional and certified organic farming conditions. Eggplant skin was extracted using aqueous methanol prior to phenolic profiling with UHPLC-ESI-MS-MS. Eggplant skin extracts yielded a profile of 16 phenolic acids, 4 anthocyanins, and 11 flavonols, the first report of quercetin-3-diglucoside, myricetin-3-neohesperidoside, myricetin-3-galactoside, kaempferol-3,7-diglucoside, kaempferol-diglucoside and quercetin-3-rhamnoside. Polyphenolic extracts from all sources potently delayed the cupric ion-mediated lag-time for LDL lipid oxidation and protected Apo-B100 proteins against oxidative modification. Organic growing environment positively influences eggplant skin extract phenolic profile but not antioxidant capacity. In conclusion, eggplant skin has a robust profile of phenolic phytochemicals with excellent antioxidant properties.

Blueberries (Vaccinium section Cyanococcus) are perennial shrubs widely cultivated for their edible fruits. In this study, we performed admixture and genetic relatedness analysis of northern highbush (NHB, primarily V. corymbosum) and southern highbush (SHB, V. corymbosum introgressed with V. darrowii, V. virgatum, or V. tenellum) blueberry genotypes, and progenies of the BNJ16-5 cross (V. corymbosum &times, V. darrowii). Using genotyping-by-sequencing (GBS), we generated more than 334 million reads (75 bp). The GBS reads were aligned to the V. corymbosum cv. Draper v1.0 reference genome sequence, and ~2.8 million reads were successfully mapped. From the alignments, we identified 2,244,039 single-nucleotide polymorphisms, which were used for principal component, haplotype, and admixture analysis. Principal component analysis revealed three main groups: (1) NHB cultivars, (2) SHB cultivars, and (3) BNJ16-5 progenies. The overall fixation index (FST) and nucleotide diversity for NHB and SHB cultivars indicated wide genetic differentiation, and haplotype analysis revealed that SHB cultivars are more genetically diverse than NHB cultivars. The admixture analysis identified a mixture of various lineages of parental genomic introgression. This study demonstrated the effectiveness of GBS-derived single-nucleotide polymorphism markers in genetic and admixture analyses to reveal genetic relatedness and to examine parental lineages in blueberry, which may be useful for future breeding plans.

Background Highbush blueberry (Vaccinium corymbosum) has long been consumed for its unique flavor and composition of health-promoting phytonutrients. However, breeding efforts to improve fruit quality in blueberry have been greatly hampered by the lack of adequate genomic resources and a limited understanding of the underlying genetics encoding key traits. The genome of highbush blueberry has been particularly challenging to assemble due, in large part, to its polyploid nature and genome size. Findings Here, we present a chromosome-scale and haplotype-phased genome assembly of the cultivar “Draper,” which has the highest antioxidant levels among a diversity panel of 71 cultivars and 13 wild Vaccinium species. We leveraged this genome, combined with gene expression and metabolite data measured across fruit development, to identify candidate genes involved in the biosynthesis of important phytonutrients among other metabolites associated with superior fruit quality. Genome-wide analyses revealed that both polyploidy and tandem gene duplications modified various pathways involved in the biosynthesis of key phytonutrients. Furthermore, gene expression analyses hint at the presence of a spatial-temporal specific dominantly expressed subgenome including during fruit development. Conclusions These findings and the reference genome will serve as a valuable resource to guide future genome-enabled breeding of important agronomic traits in highbush blueberry.

Cranberry plants bearing disfigured, scarred fruit were reported by growers in the major cranberry-growing region of central Wisconsin in July 2012. Plants bearing scarred fruit have since been observed in Massachusetts and New Jersey. Three complementary methods provided evidence of Tobacco streak virus (TSV) in symptomatic plants: (i) leaves and scarred berries tested positive for TSV by double-antibody sandwich enzyme-linked immunosorbent assay; (ii) quasi-isometric particles approximately 33 nm in diameter were extracted from leaves of symptomatic plants and visualized using transmission electron microscopy; and (iii) coat protein gene sequence analysis revealed 94 to 99% nucleotide similarity with reference TSV sequences. In newer cultivars, 99% of uprights with scarred berries tested positive for TSV. In older cultivars, 31% of uprights with scarred berries tested positive for TSV and the remaining 69% of uprights with scarred berries tested positive for Blueberry shock virus. TSV overwintered in cranberry plants, and leaves, pollen, and fruit tested positive for TSV the year following symptom occurrence. Attempts to inoculate cranberry using infected pollen or sap as inoculum failed, but several herbaceous hosts tested TSV positive following mechanical inoculation. Phylogenetic analysis of the coat protein gene of 26 TSV isolates from various cultivars of cranberry in Wisconsin, New Jersey, and Massachusetts revealed diversity. This work provides information that will be useful in understanding the epidemiology of TSV in cranberry and in the development of management strategies.

The American cranberry, Vaccinium macrocarpon Ait., is an economically important North American fruit crop that is consumed because of its unique flavor and potential health benefits. However, a lack of abundant, genome-wide molecular markers has limited the adoption of modern molecular assisted selection approaches in cranberry breeding programs. To increase the number of available markers in the species, this study identified, tested, and validated microsatellite markers from existing nuclear and transcriptome sequencing data. In total, new primers were designed, synthesized, and tested for 979 SSR loci, 697 of the markers amplified allele patterns consistent with single locus segregation in a diploid organism and were considered polymorphic. Of the 697 polymorphic loci, 507 were selected for additional genetic diversity and segregation analyses in 29 cranberry genotypes. More than 95% of the 507 loci did not display segregation distortion at the p &lt, 0.05 level, and contained moderate to high levels of polymorphism with a polymorphic information content &gt, 0.25. This comprehensive collection of developed and validated microsatellite loci represents a substantial addition to the molecular tools available for geneticists, genomicists, and breeders in cranberry and Vaccinium.

Cranberry and blueberry are closely related and recently domesticated fruit crops in the genus Vaccinium. Both have a presumed American origin and likely evolved from a common ancestor; however, details of their adaptive radiation and the extent of their genomic divergence remains little understood. To better understand their evolutionary and genomic relationships, a set of 323 cross-transferable simple sequence repeat (SSR) markers were identified, added to existing marker datasets, and used to construct linkage maps for cranberry (582 SSRs) and an interspecific diploid blueberry population (V. darrowii x V. corymbosum) x V. corymbosum (409 markers, densest blueberry SSR map currently available). The maps allowed for the first comparative genetic mapping study in Vaccinium, and revealed a surprisingly high degree of macro-synteny and collinearity between the cranberry and blueberry genomes. Approximately 93% of the blueberry linkage map was collinear with cranberry, while the remaining 7% (66.3 cM) was spread across 15 non-collinear regions detected in eight of the 12 linkage groups. These observations suggest that large-scale genome differentiation between the cranberry and blueberry genomes has not occurred during their evolution, and that sequence information will be highly transferable between the species in future genetic research and breeding. Finally, the set of 323 cross-transferable SSRs and linkage maps they were used to construct can serve as a shared resource for the Vaccinium research community, enabling additional comparative mapping studies, the identification and transfer of quantitative trait loci and candidate genes between species, and future exploration of evolutionary relationships in Vaccinium.

Sustainability of the cranberry industry is threatened by widespread and increasing losses due to fruit rot in the field as well as increasing restrictions on fungicide inputs. Breeding for resistance offers a partial solution but is challenging because fruit rot is caused by a complex of pathogenic fungi that can vary by location and from year to year. We identified four genetically diverse germplasm accessions that exhibit broad-spectrum fruit rot resistance under field conditions. Three of these accessions were used in biparental crosses to develop four populations segregating for resistance. Genotyping by sequencing was used to generate single-nucleotide polymorphism (SNP) markers for development of high-density genetic maps and quantitative trait locus (QTL) analyses. Nineteen QTL associated with fruit rot resistance, distributed on nine linkage groups, were discovered in our populations. Three of these QTL matched previously reported fruit rot resistance QTL. Four newly reported QTL found on linkage group 8 (Vm8), which explain between 21 and 33% of the phenotypic variance for fruit rot, are of particular interest to our breeding program. The populations described herein were also phenotyped for other horticulturally important traits, and QTL associated with yield and berry weight were identified. These QTL provide markers for candidate gene discovery and for future breeding efforts to enhance and pyramid disease resistance and other traits into elite horticultural backgrounds.

This is the first de novo assembly and annotation of a complete mitochondrial genome in the Ericales order from the American cranberry (Vaccinium macrocarpon Ait.). Moreover, only four complete Asterid mitochondrial genomes have been made publicly available. The cranberry mitochondrial genome was assembled and reconstructed from whole genome 454 Roche GS-FLX and Illumina shotgun sequences. Compared with other Asterids, the reconstruction of the genome revealed an average size mitochondrion (459,678 nt) with relatively little repetitive sequences and DNA of plastid origin. The complete mitochondrial genome of cranberry was annotated obtaining a total of 34 genes classified based on their putative function, plus three ribosomal RNAs, and 17 transfer RNAs. Maternal organellar cranberry inheritance was inferred by analyzing gene variation in the cranberry mitochondria and plastid genomes. The annotation of cranberry mitochondrial genome revealed the presence of two copies of tRNA-Sec and a selenocysteine insertion sequence (SECIS) element which were lost in plants during evolution. This is the first report of a land plant possessing selenocysteine insertion machinery at the sequence level.

The complete plastid genome sequence of the American cranberry (Vaccinium macrocarpon Ait.) was reconstructed using next-generation sequencing data by in silico procedures. We used Roche 454 shotgun sequence data to isolate cranberry plastid-specific sequences of"HyRed" via homologycomparisonswithcompletesequencesfromseveral species available at the National Center for Biotechnology Information database. Eleven cranberry plastid contigs were selected for the construction of the plastid genome-based homologies and on raw reads flowing through contigs and connection information. We assembled and annotated a cran- berry plastid genome (82,284 reads; 185x coverage) with a length of 176 kb and the typical structure found in plants, but with several structural rearrangements in the large single-copy region when compared to other plastid asterid genomes. To evaluate the reliability of the sequence data, phylogenetic analysis of 30 species outside the order Ericales (with 54 genes) showed Vaccinium inside the clade Asteridae ,a s reported in other studies using single genes. The cranberry plastid genome sequence will allow the accumulation of crit- ical data useful for breeding and a suite of other genetic studies.

Cranberries (Vaccinium macrocarpon Ait.) are an economically important fruit crop derived from a North American native species. We report the application of 12 simple sequence repeats (SSR) or microsatellite markers to assess the genetic diversity of cranberry cultivars. We studied 164samplesof21differentcranberry cultivars,11 experimen- tal hybrids, and 6 representative accessions of wild species. Genetic cluster analysis, based on 117 SSR alleles, differenti- ated the major cranberry cultivars. However, some cranberry cultivar subclone variants and mislabeled samples were ob- served. Consensus genetic profiles identified the most likely clonal representatives of several important cranberry cultivars (e.g., "Ben Lear, "" Howes," and "Stevens"). The markers werefurther usedtoconfirm putativeparentsofseveral hybrid progenies. The long-term goal of our studies is to identify, preserve, and utilize unique genetic materials to breed im- proved cranberries. Attaining this goal will help growers maintain sustainability under changing economic and envi- ronmental conditions.

Culturable fungal population diversity and succession was investigated in developing cranberry ovaries of fruit rot-resistant and rot-susceptible cranberry selections, from flower through mature fruit. Fungi were recovered in culture from 1185 of 1338 ovary tissues collected from June to September, yielding 2255 isolates that represented 42 morphotaxa. During the season, species richness varied from 2 to 17 and 2 to 18 in rot-resistant and rot-susceptible selections, respectively, increasing from wk1 to wk10 and then gradually declining to wk14. Shannon-Wiener diversity index varied from 0.27 to 2.32 in rot-resistant and 0.18 to 2.38 in rot-susceptible, and Pielou’s evenness index varied from 0.11 to 0.63 and 0.06 to 0.64 in rot-resistant and rot-susceptible selections, respectively, confirming that diversity of fungi in developing ovaries was similar among rot-resistant and rot-susceptible selections, but varied among sampling time points. Principal component analysis grouped samples collected at the same sampling time point together regardless of rot susceptibility of the selections, and detected the predominant fungal species associated with each stage of development. Successional changes were observed in populations of endophytic, pathogenic and saprophytic fungi throughout the season as ovaries matured.

Consumer awareness, pesticide and fertiliser contaminations, and environmental concerns have resulted in significant demand for organically grown farm products. The present study evaluates the influence that organic and conventional farming practices exert on the total phenolic content in eggplant samples from two cultivars, Blackbell (American eggplant) and Millionaire (Japanese eggplant), grown by conventional and organic farming practices with similar climatic conditions. In addition, plant-to-plant variation in phenolic content was determined from eight eggplant samples collected from different plants grown at various field positions. Samples were separately assayed for phenolic content by the two independent procedures, HPLC/LC–MS and Folin–Ciocalteu assay. The results of the HPLC analysis showed significant plant-to-plant variation (% RSD ranged from 22% to 48%) in 5-caffeoylqunic acid content (the most abundant phenolic acid) in eight eggplant samples collected from eight different plants belonging to the same cultivar and grown under similar conditions. The cv. Millionaire showed a higher total phenolic content compared to the Blackbell cultivar. We did not observe significant consistent trend in the phenolic content of eggplant samples grown with organic and conventional farming practices with both eggplant cultivars. We, however, observed a strong correlation (R 2 = 0.87) between total phenolic content as assayed by Folin–Ciocalteu assay and 5-caffeoylqunic acid, a major phenolic acid in eggplant extract. This study concludes that multiple repetitive analyses of plant products collected from different plants grown over various time periods (seasons) at different locations should always be carried to unambiguously prove the impact of growing conditions on phenolic content or antioxidant activity. Published by Elsevier Ltd.

The influence of organic and conventional farming practices on the phenolic content in eggplant samples belonging to two cultivars, Blackbell (American eggplant) and Millionaire (Japanese eggplant) grown under similar environmental conditions was evaluated. Phytochemical investigation of the eggplant extracts showed that N -caffeoylputrescine, 5-caffeoylquinic acid, and 3-acetyl-5-caffeoylquinic acid made up the bulk of total eggplant phenolics. In addition, this is the first report on identification of trace quantities of three additional flavonols, namely, quercetin-3-glucoside, quercetin-3-rhamnoside, and myricetin-3-galactoside in freeze-dried eggplant pulp. The phenolic content of the conventionally grown Blackbell variety was marginally higher or equal to those grown organically. However, the phenolic content of the Millionaire organic variety was much greater than the conventionally grown samples. Eggplant extract inhibition of cupric-ion mediated oxidation of low-density lipoproteins was highly correlated with the content of 5-caffeolquinic acid ( R 2 = 0.9124), the most abundant phenolic acid identified.

Forty-one half-sib families (>3000 individuals) segregating for parthenocarpic fruit production were evaluated under field conditions. Within these families, ≈280 parthenocarpic individuals were identified. In general, three categories of segregants were observed: normal-seeded types, small/low-seeded types, and parthenocarpic types. Inheritance patterns suggested that the trait is recessive, but did not fit simple tetrasomic recessive or incomplete dominance models. Lack of fit may be attributable to environmental interactions or incomplete penetrance of this trait. Further evaluations are underway with small/low-seeded types with the expectation that offspring of these plants may express the trait at higher frequencies than in F1 × F1 crosses. The reduced vigor in many of the parthenocarpic segregants suggests that this germplasm will be most useful, initially, in improving fruit quality in the intermediate expression types, which have better fruit set and reduced seed development.

Cranberry fruit are a rich source of bioactive compounds that may function as constitutive or inducible barriers against rot-inducing fungi. The content and composition of these compounds change as the season progresses. Several necrotrophic fungi cause cranberry fruit rot disease complex. These fungi remain mostly asymptomatic until the fruit begins to mature in late August. Temporal fluctuations and quantitative differences in selected organic acid profiles between fruit of six cranberry genotypes during the growing season were observed. The concentration of benzoic acid in fruit increased while quinic acid decreased throughout fruit development. In general, more rot-resistant genotypes (RR) showed higher levels of benzoic acid early in fruit development and more gradual decline in quinic acid levels than that observed in the more rot-susceptible genotypes. We evaluated antifungal activities of selected cranberry constituents and found that most bioactive compounds either had no effects or stimulated growth or reactive oxygen species (ROS) secretion of four tested cranberry fruit rot fungi, while benzoic acid and quinic acid reduced growth and suppressed secretion of ROS by these fungi. We propose that variation in the levels of ROS suppressive compounds, such as benzoic and quinic acids, may influence virulence by the fruit rot fungi. Selection for crops that maintain high levels of virulence suppressive compounds could yield new disease resistant varieties. This could represent a new strategy for control of disease caused by necrotrophic pathogens that exhibit a latent or endophytic phase.

Since its domestication 200 years ago, breeding of the American cranberry (Vaccinium macrocarpon) has relied on phenotypic selection because applicable resources for molecular improvement strategies such as marker-assisted selection (MAS) remain limited. To enable MAS in cranberry, the first high-density SSR linkage map with 541 markers representing all 12 cranberry chromosomes was constructed for the CNJ02-1 progeny from a cross of elite cultivars, CNJ97-105-4 and NJ98-23. The population was phenotyped for a 3-year period for total yield (TY), mean fruit weight (MFW), and biennial bearing index (BBI), and data were analyzed using mixed models and best linear unbiased predictors (BLUPs). Significant differences between genotypes were observed for all traits. Quantitative trait loci (QTL) analyses using BLUPs identified four MFW QTL on three linkage groups (LGs), three TY QTL on three LGs, and one BBI QTL which co-localized with a TY QTL. Local BLAST of a cranberry nuclear genome assembly identified homologous sequences for the mapped SSRs which were then anchored to 12 pseudo-chromosomes using the linkage map information. Analyses comparing coding regions (CDS) anchored in the cranberry linkage map with grape, kiwifruit, and tomato genomes were performed. Moderate micro-synteny between the cranberry and kiwifruit genomes was detected, although none of the regions overlapped with the QTL identified in this study. The linkage map, QTL, and elite genotypic constitutions identified herein may be applied in subsequent cranberry MAS programs for the development of new cultivars, and potential marker transferability should allow for comparative genomic studies within economically important Vaccinium species.

Cranberry fruit is a rich source of polyphenols, and has shown biological activities against Streptococcus mutans. In the present study, we examined the influence of extracts of flavonols (FLAV), anthocyanins (A) and proanthocyanidins (PAC) from cranberry on virulence factors involved in Streptococcus mutans biofilm development and acidogenicity. PAC and FLAV, alone or in combination, inhibited the surface-adsorbed glucosyltransferases and F-ATPases activities, and the acid production by S. mutans cells. Furthermore, biofilm development and acidogenicity were significantly affected by topical applications of PAC and FLAV (P

The occurrence of cyclical translocation involving three non-homologous chromosomes and affecting pollen stainability has been observed in two cranberry cultivars: Howes, and Wilcox, a progeny of Howes. These cultivars were crossed with six normal cranberry cultivars to study the transmission of the cyclical translocation to their progeny. The translocational progeny were determined by pollen tetrad analysis. A total of 102 individuals (6 crosses) were analyzed in the progeny of Wilcox and 116 individuals (5 crosses) in the progeny of Howes. The ratios observed in the progeny of Wilcox and Howes were 71 translocated: 31 normal, and 79 translocated: 37 normal, respectively. The segregations deviated from the expected 1 translocated: 1 normal progeny ratio, but fit either a 3:1 or 2:1 ratio. The altered segregations may indicate the presence of a balanced lethal system located in the translocated segments of both Howes and Wilcox. Sterile individuals were found in the progeny of Wilcox×Howes, which could indicate that the two parents have non-identical translocations. The translocated progeny of both cultivars had a normal distribution for pollen stainability, which indicated that both the occurrence of crossing over in the interstitial region and the segregation of chromosomes are under polygenic control.

Genus Vaccinium,consisting of blueberries, cranberries, lingonberries and many related wild species, includes diploid, tetraploid, and hexaploid species. Most evidence indicates that the tetraploid species are autotetraploids, with non-preferential bivalent chromosome pairing. Although homoploid interspecific crosses usually produce numerous fertile hybrids if the parents are from the same section of the genus, inter sectional crosses at the diploid level normally produce no seedlings, weak seedlings, or seedlings that are very low in fertility. There is a strong but not complete triploid block within Vaccinium. Even with insections, tetraploid × diploid (and the reciprocal) crosses normally give only a few tetraploid and a few triploid hybrids. Hexaploid × diploid crosses within sections are very hard to make, and the few hybrids that have been obtained are pentaploid. The frequency of 2n gametes varies,both among genotypes within species and among species. Vaccinium pollen is normally shed in tetrads, and the frequency of large pollen grains shed in dyads can be used to estimate 2n gamete frequency. Cultivated blueberries occur at both the tetraploid and the hexaploid levels, and there are important genetic resources in the diploids. Unreduced gamete production has permitted transfer of genetic material from the diploid to the tetraploid level and from the diploids and tetraploids to the hexaploid level via triploid hybrids. Intersectional crosses can occasionally produce tetraploid Vaccinium hybrids that appear to behave as amphidiploids and have medium to high fertility. CommercialVaccinium cultivars are normally propagated by cuttings. Intersectional hybridization, chromosome doubling, and asexual propagation could permit the production of novel hybrid combinations with value as ornamentals or in fruit production.

A clone representing a fragment of the dihydroflavonol-4-reductase ( DFR ) gene from cranberry was isolated from a genomic DNA library using the tomato DFR gene as a probe. Sequence analysis of the clone confirmed homology to published DFR gene sequences. 3′ and 5′ RACE (rapid amplification of cDNA ends) reactions from cranberry leaf total RNA were used to obtain the entire cDNA sequence. The sequence information was used to amplify a full-length clone by RT-PCR. Sequencing analysis to confirm the identity of the full-length DFR cDNA identified a putative second allele. Segregation analysis suggested that the two sequences are not allelic, but multi-locus. Nucleotide sequence homology of the full-length clones was highest to published DFR sequence from Camellia sinensis (about 80% identity) followed by Forsythia x intermedia , Antirrhinum majus , Rosa hybrida and Petunia hybrida . When expressed using the CaMV 35S promoter, the corolla of flowers of transgenic tobacco plants were much darker pink than the controls. Some flower parts not normally highly pigmented, such as the filaments, were also dark pink. These data confirm the identity and function of the cranberry clones and further suggest that overexpression of the cranberry DFR could be used to increase anthocyanin production in transgenic plants.

Polyploidy has been important in the evolution of angiosperms and may significantly affect population genetic diversity and structure. Nineteen isoenzyme loci were studied in diploid and tetraploid populations of Vaccinium oxycoccos (Ericaceae), and the results are compared with data previously reported for the related V. macrocarpon. Diploid V. oxycoccos and V. macrocarpon were readily discriminated based on their allozymic variation. No evidence for fixed heterozygosity was found in tetraploid V. oxycoccos. In contrast, all polymorphic loci exhibited both balanced and unbalanced heterozygotes, with some individuals exhibiting a pattern consistent with the presence of three alleles. These results support an autopolyploid origin for tetraploid V. oxycoccos. However, tetraploid V. oxycoccos possessed a suite of alleles not found in diploid V. oxycoccos; half of these alleles were shared with V. macrocarpon. This suggests that autotetraploid V. oxycoccos may have undergone hybridization with V. macrocarpon or that the autotetraploid retained the genetic variation present in an ancestral diploid species. Following theoretical expectations, proportion of polymorphic loci, mean number of alleles, and observed heterozygosity were significantly higher for the autotetraploid than for the diploid. Mean inbreeding (F:(IS)) was similar for diploid and tetraploid V. oxycoccos. The latter exhibited population differentiation (F:(ST)) exceeding both diploid species.

Since up to 2.4 m (8 ft) of water may be applied annually to cranberry beds for various production purposes, water quality can alter soil chemical properties and potentially affect plant health. Many cranberry plantings have recently been developed in nontraditional production regions and on atypical sites, wherechemical properties of the available water may differ from those in cranberry sites in the traditional production regions. Water currently being used for cranberry production was sampled from farms in most major production regions to characterize its chemical characteristics. High alkalinity in many samples was a concern, since alkalinity can increase soil pH above the desired level for cranberries. Total soluble salt concentrations and sodium adsorption ratios were seldom high enough to be of concern. Water samples from long-established plantings were lower in alkalinity, pH, and soluble salt concentrations than samples from newer plantings without production histories.

Cranberry (Vaccinium macrocarpon Ait.) has the opportunity to partition resources into sexual and/or asexual (stolons) modes of reproduction. Nitrogen status has been shown to impact the degree of stoloniferous growth. To determine whether there is a genotypic response to varying nitrogen levels, six hybrid and four native cultivars were treated with three annual rates of nitrogen fertilizer (17, 34, or 67 kg·ha-1) for 4 years. Fruit yield was determined each year and asexual vegetative growth (stolons) weight was removed and measured in all but the first year of the experiment. Cultivars exhibited different patterns of yield and stolon weight response over the three nitrogen rates. Not all cultivars exhibited significant yield decreases at the high N levels. Vegetative growth (stolon weight) generally increased with increasing N, however, not all cultivars responded similarly over three N rates. Partitioning between yield and stolon production favored fruit yield at the lower N rates in three of the four native cultivars studied (`Cropper', `Early Black', and `Howes'). Yield over N rates was more stable for four of the six hybrid cultivars, which may be the result of greater heterozygosity in hybrids than natives, and/or genetic gain from one breeding and selection cycle, offering increased tolerance to nitrogen stress. This study indicates that genetic variation exists for yield, yield stability, and stolon production relative to nitrogen level, and that genetic gain in cranberry is possible for these traits. Future studies involving cranberry physiology and nutrition should consider the genotypes used.

Cranberry A-type proanthocyanidins (PACs) have been recognized for their inhibitory activity against bacterial adhesion and biofilm-derived infections. However, the precise identification of the specific classes of degree-of-polymerization (DP) conferring PACs bioactivity remains a major challenge owing to the complex chemistry of these flavonoids. In this study, chemically characterized cranberries were used in a multistep separation and structure-determination technique to isolate A-type PAC oligomers of defined DP. The influences of PACs on the 3D architecture of biofilms and Streptococcus mutans-transcriptome responses within biofilms were investigated. Treatment regimens that simulated topical exposures experienced clinically (twice-daily, 60 s each) were used over a saliva-coated hydroxyapatite biofilm model. Biofilm accumulation was impaired, while specific genes involved in the adhesion of bacteria, acid stress tolerance, and glycolysis were affected by the topical treatments (vs the vehicle-control). Genes (rmpC, mepA, sdcBB, and gbpC) associated with sucrose-dependent binding of bacteria were repressed by PACs. PACs of DP 4 and particularly DP 8 to 13 were the most effective in disrupting bacterial adhesion to glucan-coated apatitic surface (85% inhibition vs vehicle control), and gene expression (eg rmpC). This study identified putative molecular targets of A-type cranberry PACs in S. mutans while demonstrating that PAC oligomers with a specific DP may be effective in disrupting the assembly of cariogenic biofilms.

Progress was made in identifying molecular markers linked to genes which control chilling requirement and cold hardiness in a woody perennial, blueberry (Vaccinium, section Cyanococcus). 40Construction of RAPD (randomly amplified polymorphic DNA)-based genetic linkage maps using two testcross populations derived from interspecific hybrids of wild diploid blueberry species, V. darrowi Camp X V elliottii Chapm. and V. darrowi X V caesariense Mackenz. was continued. Mapping populations were segregating for chilling requirement and cold hardiness. Screening plants of the V. darrowi × V. caesariense-derived mapping population for cold acclimation capacity using leaf tissue collected before and after cold treatment and an electrolyte leakage assay was begun. In addition, a tetraploid breeding population (predominantly V. corymbosum L.) was screened for chilling requirement and efforts were begun to identify markers linked to genes determining chilling requirement using this population and a bulked-segregant-type analysis. [Article copies available from The Haworth Document Delivery Service: 1-800-342-9678.]

Cranberry A-type proanthocyanidins (PACs) have been recognized for their inhibitory activity against bacterial adhesion and biofilm-derived infections. However, the precise identification of the specific classes of degree-of-polymerization (DP) conferring PACs bioactivity remains a major challenge owing to the complex chemistry of these flavonoids. In this study, chemically characterized cranberries were used in a multistep separation and structure-determination technique to isolate A-type PAC oligomers of defined DP. The influences of PACs on the 3D architecture of biofilms and Streptococcus mutans-transcriptome responses within biofilms were investigated. Treatment regimens that simulated topical exposures experienced clinically (twice-daily, 60 s each) were used over a saliva-coated hydroxyapatite biofilm model. Biofilm accumulation was impaired, while specific genes involved in the adhesion of bacteria, acid stress tolerance, and glycolysis were affected by the topical treatments (vs the vehicle-control). Genes (rmpC, mepA, sdcBB, and gbpC) associated with sucrose-dependent binding of bacteria were repressed by PACs. PACs of DP 4 and particularly DP 8 to 13 were the most effective in disrupting bacterial adhesion to glucan-coated apatitic surface (>85% inhibition vs vehicle control), and gene expression (eg rmpC). This study identified putative molecular targets of A-type cranberry PACs in S. mutans while demonstrating that PAC oligomers with a specific DP may be effective in disrupting the assembly of cariogenic biofilms.

The first genetic map of cranberry (Vaccinium macrocarpon) has been constructed, comprising 14 linkage groups totaling 879.9 cM with an estimated coverage of 82.2 %. This map, based on four mapping populations segregating for field fruit-rot resistance, contains 136 distinct loci. Mapped markers include blueberry-derived simple sequence repeat (SSR) and cranberry-derived sequence-characterized amplified region markers previously used for fingerprinting cranberry cultivars. In addition, SSR markers were developed near cranberry sequences resembling genes involved in flavonoid biosynthesis or defense against necrotrophic pathogens, or conserved orthologous set (COS) sequences. The cranberry SSRs were developed from next-generation cranberry genomic sequence assemblies; thus, the positions of these SSRs on the genomic map provide information about the genomic location of the sequence scaffold from which they were derived. The use of SSR markers near COS and other functional sequences, plus 33 SSR markers from blueberry, facilitates comparisons of this map with maps of other plant species. Regions of the cranberry map were identified that showed conservation of synteny with Vitis vinifera and Arabidopsis thaliana. Positioned on this map are quantitative trait loci (QTL) for field fruit-rot resistance (FFRR), fruit weight, titratable acidity, and sound fruit yield (SFY). The SFY QTL is adjacent to one of the fruit weight QTL and may reflect pleiotropy. Two of the FFRR QTL are in regions of conserved synteny with grape and span defense gene markers, and the third FFRR QTL spans a flavonoid biosynthetic gene.

Cranberry extracts may provide beneficial health effects in the treatment of various diseases, including cancer. However, the underlying molecular mechanisms of antineoplastic properties are not understood. We report the effect of a proanthocyanidin (PAC)-rich isolate from cranberry (PAC-1) as a therapeutic agent with dual activity to target both ovarian cancer viability and angiogenesis in vitro. PAC-1 treatment of chemotherapy-resistant SKOV-3 cells blocked cell cycle progression through the G2/M phase, increased the generation of reactive oxygen species (ROS), and induced apoptosis through activation of intrinsic and extrinsic pathway components. Cytotoxicity of PAC-1 was partially based on ROS generation and could be blocked by co-treatment with antioxidant glutathione. PAC-1 reduced the cell viability of both SKOV-3 ovarian cancer cells and HUVEC endothelial cells in a dose-dependent manner and blocked the activation of the pro-survival factor AKT. Furthermore, PAC-1 blocked vascular endothelial growth factor (VEGF)-stimulated receptor phosphorylation in endothelial cells, which correlated with the inhibition of endothelial tube formation in vitro. Our findings suggest that PAC-1 exerts potent anticancer and anti-angiogenic properties and that highly purified PAC from cranberry can be further developed to treat ovarian cancer in combinational or single-agent therapy.

Polyphenolic extracts of the principal flavonoid classes present in cranberry were screened in vitro for cytotoxicity against solid tumor cells lines, identifying two fractions composed principally of proanthocyanidins (PACs) with potential anticancer activity. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) analysis of the proanthocyanidins (PACs) fractions indicated the presence of A-type PACs with 1-4 linkages containing between 2-8 epicatechin units with a maximum of 1 epigallocatechin unit. PACs exhibited in vitro cytotoxicity against platinum-resistant human ovarian, neuroblastoma and prostate cancer cell lines (IC50 = 79-479 microg/mL) but were non-cytotoxic to lung fibroblast cells (IC50 > 1000 microg/ml). SKOV-3 ovarian cancer cells treated with PACs exhibited classic apoptotic changes. PACs acted synergistically with paraplatin in SKOV-3 cells. Pretreatment of SKOV-3 cells with PACs (106 microg/ml) resulted in a significant reduction of the paraplatin IC50 value. Similarly, in a BrdU incorporation assay, co-treatment of SKOV-3 cells with PACs and paraplatin revealed reduced cell proliferation at lower concentrations than with either individually. In SKOV-3 cell cultures co-treated with PAC-1 and paraplatin, an HPLC analysis indicated differential quantitative presence of various PAC oligomers such as DP-8, -9, -11 and -14 indicating either selective binding or uptake. Cranberry proanthocyanidins exhibit cell-line specific cytotoxicity, induce apoptotic markers and augment cytotoxicity of paraplatin in platinum-resistant SKOV-3 ovarian cancer cells.

The influence of organic and conventional farming practices on the phenolic content in eggplant samples belonging to two cultivars, Blackbell (American eggplant) and Millionaire (Japanese eggplant) grown under similar environmental conditions was evaluated. Phytochemical investigation of the eggplant extracts showed that N-caffeoylputrescine, 5-caffeoylquinic acid, and 3-acetyl-5-caffeoylquinic acid made up the bulk of total eggplant phenolics. In addition, this is the first report on identification of trace quantities of three additional flavonols, namely, quercetin-3-glucoside, quercetin-3-rhamnoside, and myricetin-3-galactoside in freeze-dried eggplant pulp. The phenolic content of the conventionally grown Blackbell variety was marginally higher or equal to those grown organically. However, the phenolic content of the Millionaire organic variety was much greater than the conventionally grown samples. Eggplant extract inhibition of cupric-ion mediated oxidation of low-density lipoproteins was highly correlated with the content of 5-caffeolquinic acid (R 2 = 0.9124), the most abundant phenolic acid

Most blueberry breeding activity is focused on northern highbush, southern highbush and rabbiteye types. The major objectives of blueberry breeders center on high plant vigor, improved disease resistance, flavor, longer storing fruit and expanded harvest dates. Cranberry breeders have concentrated on early maturing fruit, uniform large size, intense color, keeping quality, high productivity, disease resistance and plant vigor. Considerable variability exists in blueberry and cranberry for most of the horticulturally important traits, and while only a limited number of genetic studies have been performed, most inheritance patterns fit quantitative models. Several genes have been identified through molecular, genetic and genomic approaches that are associated with cold hardiness. Wide hybridization is commonly employed in blueberry breeding and southern highbush types were derived primarily by incorporating genes from the diploid species Vaccinium darrowii into the highbush background via unreduced gametes. A wide array of molecular markers has been used in blueberry for fingerprinting and linkage mapping, and a major QTL regulating the chilling requirement in diploids has been identified. Transgenic blueberries have been produced with herbicide resistance and the Bt gene (Bacillus thuringiensis) has been incorporated into cranberry. A large EST library of highbush blueberry has been produced.

Obesity is a major and independent risk factor for cardiovascular disease and it is strongly associated with the development of dyslipidemia, insulin resistance and type 2 diabetes. Flavonoids, a diverse group of polyphenol compounds of plant origin widely distributed in human diet, have been reported to have numerous health benefits, although the mechanisms underlying these effects have remained obscure. We analyzed the effects of chronic dietary supplementation with flavonoids extracted from cranberry (FLS) in normal and obese C57/BL6 mice compared to mice maintained on the same diets lacking FLS. Obese mice supplemented with flavonoids showed an amelioration of insulin resistance and plasma lipid profile, and a reduction of visceral fat mass. We provide evidence that the adiponectin-AMPK pathway is the main mediator of the improvement of these metabolic disorders. In contrast, the reduced plasma atherogenic cholesterol observed in normal mice under FLS seems to be due to a downregulation of the hepatic cholesterol synthesis pathway. Overall, we demonstrate for the first time that the molecular mechanisms underlying the beneficial effects of flavonoids are determined by the metabolic state.

A total of 1475 individuals belonging to 43 natural populations of seven diploid (2x) blueberry species (Vaccinium section Cyanococcus) and two natural interspecific 2x hybrid populations were evaluated for unreduced pollen production. Significant differences were found in the frequency of unreduced pollen producers between species and within and between populations of the same species. Individuals with 1% or more unreduced pollen were considered unreduced pollen producers. The average frequency of unreduced pollen producers in these diploid species was 13.5%, ranging from 7.4% (V. corymbosum) to 18.4% (V. darrowii). The frequency of unreduced pollen grains in individual clones varied from

Putative transgenic cranberry plants have been achieved via Agrobacterium-mediated transformation. Leaf explants were transformed with a supervirulent Agrobacterium tumefaciens strain EHA 105, harboring the binary vector P35SGUSint and nptII selectable marker genes. Inoculation of precultured explants (≈10 days on regeneration medium) coupled with sonicasion improved transformation efficiency significantly. Adventitious shoots were directly regenerated from explants. Putative transformed shoots were identified by being kanamycin-resistant and GUS-positive. Stable GUS gene expression (turning blue) could be detected within 1 h of incubation at 37 °C. Confirmation of transformation by molecular analysis is in progress. Eight putative transgenic cranberry plants were obtained. All appeared morphologically normal. This appears to be the first success in achieving cranberry transformed plants by Agrobacterium-mediated method. Optimizing the transformation system is ongoing.

We have established a very efficient cranberry regeneration (shoot organogenesis) system from leaf explants using a basal medium consisting of Anderson's salts and Murashige and Skoog (MS) organics supplemented with 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ) and N6-(-??-dimethyallylamino) purine) (2ip). Characteristics examined include combinations of varying levels of three plant growth regulators (TDZ, 2ip, and naphthaleneacetic acid (NAA), explant orientation (adaxial or abaxial side in contact with the media), and leaf position relative to the distal end of the shoot. Genotypes (`Early Black', `Pilgrim', `Stevens', `Ben Lear', and US#35) differed significantly in regeneration capacity, and there were no genotype by treatment interaction effects. Regeneration occurred on more than 95% of the explants with `Early Black' and `Pilgrim' producing as many as 100 shoot tips per explant with one particular treatment. Emerging adventitious shoots were always observed on the adaxial side of the leaves regardless of explant orientation. However, regeneration was much greater when the adaxial side was in contact with the media. Regeneration efficiency was not significantly affected by leaf position (10 leaves). Elongation of shoot tips began about 2 weeks after the regenerating explants were transferred to the basal medium without hormones and continued for several months. Elongated shoot cuttings rooted readily.

Four diploid (2x) clones of alfalfa, Medicago sativa L., which produced good seed set when used as male parents in 4x-2x crosses were selected for study. The 2x clones descended from 2x haploids of cultivated 4x alfalfa. Fertility in the 4x-2x cross was due to the production of pollen with the unreduced chromosome number (2n pollen) from the 2x parent. The cytological mechanism of 2n pollen formation was found to be disorientation of spindles at metaphase II in up to 38% of the pollen mother cells. Thus, both n and 2n pollen were produced by all four diploids examined. Normal spindles at metaphase II were oriented such that they defined the poles of a tetrahedron and resulted in normal tetrads in a tetrahedral arrangement. Disoriented spindles were basically parallel to each other and resulted in formation of dyads and occasionally a triad. Dyads developed into two 2n pollen grains; triads developed into one 2n and two n pollen grains. Since both n and 2n pollen grains are produced by the diploids, they can be maintained as diploids or they can be used as male parents in crosses to tetraploids. The genetic constitution of 2n pollen resulting from parallel spindles is similar to that expected after first division restitution of meiosis and much of the heterozygosity of the diploid parent is conserved in the gametes. The 2n gamete mechanism has potential application in germplasm transfer and in maximizing heterozygosity in tetraploid hybrids.