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The flavonoid and organic acid profiles of one cultivated tetraploid and six wild diploid blueberry species (Vaccinium spp.) were systematically investigated using high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC-ESI-MS-MS). Eighteen individual anthocyanins from five aglycone classes were characterized among species, with malvidin and delphinidin glycosides accounting for 31.4% and 29.1% of total anthocyanins. Twenty-three flavonol glycosides from six aglycone classes were identified, among which quercetin and myricetin glycosides accounted for more than 80% of total flavonols in most species. Both inter- and intraspecies differences in anthocyanin and flavonol composition were observed, as described by principal component analysis. Only B-type proanthocyanidins were found in blueberry species, and highly polymerized molecules with degree of polymerization greater than 10 appeared to be the most abundant fraction. Although overall proanthocyanidin levels varied from 27.7 to 146.3 mg/100 g fruit, all species exhibited similar proanthocyanidin composition. Citric, quinic, and shikimic acid were the major identified blueberry organic acids. However, their relative abundance varied across species. In certain species either citric acid (e.g., Vaccinium darrowii) or quinic acid (e.g., Vaccinium corymbosum) was lacking.

Cranberry (Vaccinium macrocarpon) is a member of the Heath family (Ericaceae) and is a temperate low-growing woody perennial native to North America that is both economically important and has significant health benefits. While some native varieties are still grown today, breeding programs over the past 50 years have made significant contributions to improving disease resistance, fruit quality and yield. An initial genome sequence of an inbred line of the wild selection ‘Ben Lear,’ which is parent to multiple breeding programs, provided insight into the gene repertoire as well as a platform for molecular breeding. Recent breeding efforts have focused on leveraging the circumboreal V. oxycoccos, which forms interspecific hybrids with V. macrocarpon, offering to bring in novel fruit chemistry and other desirable traits. Here we present an updated, chromosome-resolved V. macrocarpon reference genome, and compare it to a high-quality draft genome of V. oxycoccos. Leveraging the chromosome resolved cranberry reference genome, we confirmed that the Ericaceae has undergone two whole genome duplications that are shared with blueberry and rhododendron. Leveraging resequencing data for ‘Ben Lear’ inbred lines, as well as several wild and elite selections, we identified common regions that are targets of improvement. These same syntenic regions in V. oxycoccos, were identified and represent environmental response and plant architecture genes. These data provide insight into early genomic selection in the domestication of a native North American berry crop.

Blueberries (Vaccinium spp.) are highly vulnerable to changing climatic conditions, especially increasing temperatures. To gain insight into mechanisms underpinning the response to heat stress, two blueberry species were subjected to heat stress for 6 and 9 h at 45 °C, and leaf samples were used to study the morpho-physiological and transcriptomic changes. As compared with Vaccinium corymbosum, Vaccinium darrowii exhibited thermal stress adaptation features such as small leaf size, parallel leaf orientation, waxy leaf coating, increased stomatal surface area, and stomatal closure. RNAseq analysis yielded ~135 million reads and identified 8305 differentially expressed genes (DEGs) during heat stress against the control samples. In V. corymbosum, 2861 and 4565 genes were differentially expressed at 6 and 9 h of heat stress, whereas in V. darrowii, 2516 and 3072 DEGs were differentially expressed at 6 and 9 h, respectively. Among the pathways, the protein processing in the endoplasmic reticulum (ER) was the highly enriched pathway in both the species: however, certain metabolic, fatty acid, photosynthesis-related, peroxisomal, and circadian rhythm pathways were enriched differently among the species. KEGG enrichment analysis of the DEGs revealed important biosynthesis and metabolic pathways crucial in response to heat stress. The GO terms enriched in both the species under heat stress were similar, but more DEGs were enriched for GO terms in V. darrowii than the V. corymbosum. Together, these results elucidate the differential response of morpho-physiological and molecular mechanisms used by both the blueberry species under heat stress, andhelp in understanding the complex mechanisms involved in heat stress tolerance.

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 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

The metabolomic profiles of American cranberry ( Vaccinium macrocarpon) fruits and their variation among 10 diverse cultivars were investigated by ultraperformance liquid chromatography ion-mobility high-resolution mass spectrometry (UPLC-IM - HRMS). Over 80 metabolites, belonging to various phenolic compound groups, were putatively characterized. An HRMS data matrix consisting of 4778 unique ions across the 10 cultivars was built and analyzed by orthogonal projections to latent structures discriminant analysis (OPLS-DA). The 10 cultivars segregated into 4 clusters on the basis of their metabolomic similarities, which largely reflected their genetic backgrounds. Anthocyanins exhibited the most extensive variations among all the cultivars, reflecting the effects of cranberry breeding selection on fruit color. Flavonols, phenolic acid derivatives, and proanthocyanidins also varied among the different cultivars. The nontargeted metabolomic comparison using multivariate analysis proved to be efficient and robust for determining specific metabolite differences among the cultivars.

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.

Despite increasing knowledge about the impacts of pathogens on the interactions between plants and insect vectors, relatively little is known about their effects on other, non-vector, organisms. In cranberries, phytoplasma infection causes false blossom disease, which is transmitted by leafhoppers. We hypothesized that changes in plant chemistry induced by phytoplasma infection might affect the performance and feeding behavior not only of vectors but also of other phytophagous insects. To test this, we measured growth, survival, and the number of leaves damaged by larvae of three common non-vector herbivores: spotted fireworm (Choristoneura parallela Robinson), Sparganothis fruitworm (Sparganothis sulfureana Clemens), and gypsy moth (Lymantria dispar L.) on phytoplasma-infected and uninfected cranberries (Vaccinium macrocarpon Ait.). We also assessed the effects of phytoplasma infection on nutrients and phytochemistry related to defenses. In general, larvae of all three herbivore species grew 2–3 times bigger, and damaged 1.5–3.5 times more leaves, while feeding on infected vs. uninfected plants. Survival of Sparganothis fruitworm larvae was also ~1.5 times higher on infected plants, while spotted fireworm and gypsy moth larval survival was not affected. In a long-term (5-week) assay, gypsy moth larval survival and mass were enhanced when feeding on phytoplasma-infected leaves. Levels of important plant nutrients (e.g., N, P, K, Ca, S, Mn, Fe, B, Al, and Na) were higher in infected plants, while levels of defensive proanthocyanidins were reduced by 20–40% compared to uninfected plants. In contrast, levels of Mg were lower in infected plants, while concentrations of Cu, Zn, and defensive flavonols were not affected. Taken together, these findings suggest that phytoplasma infection enhances plant nutritional quality, while reducing plant defenses in cranberries. These effects, in turn, may explain the observed enhancement of non-vector herbivore performance, as well as the higher number of damaged leaves, on infected plants. Improved understanding of the ecology of pathogen-plant-herbivore interactions could aid efforts to enhance plant resistance and suppress disease transmission in agricultural settings., Frontiers in Ecology and Evolution, 7, ISSN:2296-701X

Mummy berry disease caused by Monilinia vaccinii-corymbosi is the most widespread economically important problem of cultivated blueberry in North America. In an attempt to identify new sources of resistance to the fruit rot (mummification) phase of mummy berry, 140 accessions from a total of 21 populations from seven wild diploid species of blueberry were evaluated for resistance under greenhouse conditions. Six isolates of M. vaccinii-corymbosi from three states were used as inoculum. A highly resistant response to mummy berry fruit rot was exhibited by all accessions of Vaccinium boreale, V. myrtilloides, V. pallidum, and V. tenellum, and by most accessions of V. darrowi. Most of the V. corymbosum and V. elliottii accessions were moderately to highly susceptible. Introgression of the resistance found in the wild diploid species into horticulturally desirable cultivars could significantly improve available resistance.

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.

Cranberry is reported to have health benefits, including prevention of urinary tract infections and other chronic diseases, due to the high content of polyphenols, including flavonols and flavan-3-ols. The aim of this study was to determine the clearance of flavonol glycosides and flavan-3-ols and/or their metabolites in human urine. Ten healthy women volunteers ingested 240 mL of cranberry juice containing flavonol glycosides. Urine samples were collected at 0, 90, 225, and 360 min postingestion. While flavan-3-ols were not detected, five flavonol glycosides common in cranberry were identified. Quercetin-3-galactoside, the most abundant cranberry flavonol, exhibited the highest peak urine concentration (Cmax) of 1315 pg/mg creatinine, followed by quercetin-3-rhamnoside, quercetin-3-arabinoside, myricetin-3-arabinoside, and myricetin-3-galactoside. Quercetin-3-arabinoside showed delayed clearance, Cmax at 237 min (Tmax), relative to other flavonols (90–151 min). Both aglycone and the conjugated sugar moie...

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.

Because of its known phytochemical activity and benefits for human health, American cranberry (Vaccinium macrocarpon L.) production and commercialization around the world has gained importance in recent years. Flavonoid compounds as well as the balance of sugars and acids are key quality characteristics of fresh and processed cranberry products. In this study, we identified novel QTL that influence total anthocyanin content (TAcy), titratable acidity (TA), proanthocyanidin content (PAC), Brix, and mean fruit weight (MFW) in cranberry fruits. Using repeated measurements over the fruit ripening period, different QTLs were identified at specific time points that coincide with known chemical changes during fruit development and maturation. Some genetic regions appear to be regulating more than one trait. In addition, we demonstrate the utility of digital imaging as a reliable, inexpensive and high-throughput strategy for the quantification of anthocyanin content in cranberry fruits. Using this imaging approach, we identified a set of QTLs across three different breeding populations which collocated with anthocyanin QTL identified using wet-lab approaches. We demonstrate the use of a high-throughput, reliable and highly accessible imaging strategy for predicting anthocyanin content based on cranberry fruit color, which could have a large impact for both industry and cranberry research.

Fruit rot is the primary threat to cranberry (Vaccinium macrocarpon) production in the northeastern United States, and increasingly in other growing regions. Efficacy of chemical control is variable because the disease is caused by a complex of pathogenic fungi. In addition, cranberries are often grown in environmentally sensitive areas, placing restrictions on chemical control measures. Thus, a major focus of the cranberry breeding program is to develop cultivars with improved fruit rot resistance (FRR). Several genetically diverse sources of FRR have been identified in our germplasm collection. However, the most resistant accessions lack one or more attributes; e.g., productivity, required for commercial acceptance. These resistant accessions were used in crosses with elite high-yielding selections and in 2009, 1624 progeny from 50 crosses were planted in 2.3-m2 field plots. During 2011–13, under field conditions with very limited fungicide management, disease pressure was severe, allowing evaluation for FRR. Plots were rated on a 1–5 scale for incidence of fruit rot (where 1 = 0% to 20% rot and 5 = 81% to 100% rotted fruit), and rotted fruit counts were made from selected plots to validate the ratings. There was a good correlation in the ratings between years (2011 vs. 2012: r = 0.59, P < 0.0001; 2011 vs. 2013: r = 0.50, P < 0.0001; 2012 vs. 2013: r = 0.62, P < 0.0001), and between rot ratings and percent rotted fruit (r = 0.90, P < 0.0001). Significant differences were found between and within families for FRR. High heritability estimates (h2 = 0.81) were obtained with midparent-offspring regression of mean fruit rot ratings, indicating additive genetic variance for FRR. Introgression of FRR into higher yielding genetic backgrounds was also accomplished, as some progeny exhibiting high FRR also had commercially viable yield (>300 g/0.09 m2), as well as good berry size and color. Selections are being further evaluated for potential cultivar release.

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.

BACKGROUND Cranberry fruit (Vaccinium macrocarpon) is rich in polyphenols, particularly oligomeric proanthocyanidins (PACs) possessing antimicrobial and antioxidant properties. PACs may play a role in resistance to fruit rot. Although many cranberry cultivars are grown for use in foods, beverages and nutraceuticals, data on PAC content among cultivars is limited. Eight cultivars were sampled from four growing regions during the 2010 season and analyzed for PAC content and composition. RESULTS MALDI-TOF MS showed that isolated PACs had similar oligomer profiles among cultivars. The major constituents were A-type (epi)catechin oligomers of two to eight degrees of polymerization. Total PAC content ranged between 18 and 92 g PAC kg−1 dried fruit, quantified as procyanidin A2 by the dimethylaminocinnamaldehyde method. Among the cultivars sampled, Howes had the highest total PACs (76–92 g kg−1), followed by Mullica Queen and Early Black (48–82 g kg−1). Ben Lear, a disease-susceptible variety, was significantly lower in PACs than the other cultivars (P < 0.001). CONCLUSIONS Several traditional and newer cultivars of cranberry from various growing regions in North America are excellent sources of PACs, particularly the Howes, Mullica Queen and Early Black cultivars. PAC content may play a role in keeping quality. © 2014 Society of Chemical Industry

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.

Hypercalcemia remains a major impediment to the clinical use of vitamin D in cancer treatment. Approaches to remove hypercalcemia and development of nonhypercalcemic agents can lead to the development of vitamin D-based therapies for treatment of various cancers. In this report, in vitro and in vivo anticancer efficacy, safety, and details of vitamin D receptor (VDR) interactions of PT19c, a novel nonhypercalcemic vitamin D derived anticancer agent, are described. PT19c was synthesized by bromoacetylation of PTAD-ergocalciferol adduct. Broader growth inhibitory potential of PT19c was evaluated in a panel of chemoresistant breast, renal, ovarian, lung, colon, leukemia, prostate, melanoma, and central nervous system cancers cell line types of NCI60 cell line panel. Interactions of PT19c with VDR were determined by a VDR transactivation assay in a VDR overexpressing VDR-UAS-bla-HEK293 cells, in vitro VDR-coregulator binding, and molecular docking with VDR-ligand binding domain (VDR-LBD) in comparison with calcitriol. Acute toxicity of PT19c was determined in nontumored mice. In vivo antitumor efficacy of PT19c was determined via ovarian and endometrial cancer xenograft experiments. Effect of PT19c on actin filament organization and focal adhesion formation was examined by microscopy. PT19c treatment inhibited growth of chemoresistant NCI60 cell lines (log10GI50 ~ -4.05 to -6.73). PT19c (10 mg/kg, 35 days) reduced growth of ovarian and endometrial xenograft tumor without hypercalcemia. PT19c exerted no acute toxicity up to 400 mg/kg (QDx1) in animals. PT19c showed weak VDR antagonism, lack of VDR binding, and inverted spatial accommodation in VDR-LBD. PT19c caused actin filament dysfunction and inhibited focal adhesion in SKOV-3 cells. PT19c is a VDR independent nonhypercalcemic vitamin D-derived agent that showed noteworthy safety and efficacy in ovarian and endometrial cancer animal models and inhibited actin organization and focal adhesion in ovarian cancer cells.

The American cranberry (Vaccinium macrocarponAit.) is a recently domesticated, economically important, fruit crop with limited molecular resources. New genetic resources could accelerate genetic gain in cranberry through characterization of its genomic structure and by enabling molecular-assisted breeding strategies. To increase the availability of cranberry genomic resources, genotyping-by-sequencing (GBS) was used to discover and genotype thousands of single nucleotide polymorphisms (SNPs) within three inter-related cranberry full-sib populations. Additional simple sequence repeat (SSR) loci were added to the SNP datasets and used to construct bin maps for the parents of the populations, which were then merged to create the first high-density cranberry composite map containing 6073 markers (5437 SNPs and 636 SSRs) on 12 linkage groups (LGs) spanning 1124 cM. Interestingly, higher rates of recombination were observed in maternal than paternal gametes. The large number of markers in common (mean of 57.3) and the high degree of observed collinearity (mean Pair-wise Spearman Rank Correlations > 0.99) between the LGs of the parental maps demonstrates the utility of GBS in cranberry for identifying polymorphic SNP loci that are transferable between pedigrees and populations in future trait-association studies. Furthermore, the high-density of markers anchored within the component maps allowed identification of segregation distortion regions, placement of centromeres on each of the 12 LGs, and anchoring of genomic scaffolds. Collectively, the results represent an important contribution to the current understanding of cranberry genomic structure and to the availability of molecular tools for future genetic research and breeding efforts in cranberry.

Background The application of genotyping by sequencing (GBS) approaches, combined with data imputation methodologies, is narrowing the genetic knowledge gap between major and understudied, minor crops. GBS is an excellent tool to characterize the genomic structure of recently domesticated (~200 years) and understudied species, such as cranberry (Vaccinium macrocarpon Ait.), by generating large numbers of markers for genomic studies such as genetic mapping. Results We identified 10842 potentially mappable single nucleotide polymorphisms (SNPs) in a cranberry pseudo-testcross population wherein 5477 SNPs and 211 short sequence repeats (SSRs) were used to construct a high density linkage map in cranberry of which a total of 4849 markers were mapped. Recombination frequency, linkage disequilibrium (LD), and segregation distortion at the genomic level in the parental and integrated linkage maps were characterized for first time in cranberry. SSR markers, used as the backbone in the map, revealed high collinearity with previously published linkage maps. The 4849 point map consisted of twelve linkage groups spanning 1112 cM, which anchored 2381 nuclear scaffolds accounting for ~13 Mb of the estimated 470 Mb cranberry genome. Bin mapping identified 592 and 672 unique bins in the parentals and a total of 1676 unique marker positions in the integrated map. Synteny analyses comparing the order of anchored cranberry scaffolds to their homologous positions in kiwifruit, grape, and coffee genomes provided initial evidence of homology between cranberry and closely related species. Conclusions GBS data was used to rapidly saturate the cranberry genome with markers in a pseudo-testcross population. Collinearity between the present saturated genetic map and previous cranberry SSR maps suggests that the SNP locations represent accurate marker order and chromosome structure of the cranberry genome. SNPs greatly improved current marker genome coverage, which allowed for genome-wide structure investigations such as segregation distortion, recombination, linkage disequilibrium, and synteny analyses. In the future, GBS can be used to accelerate cranberry molecular breeding through QTL mapping and genome-wide association studies (GWAS). Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-2802-3) contains supplementary material, which is available to authorized users.

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.