Wild Vaccinium Species Research Articles

Contribution to the biosystematics of the wild Vaccinium species distributed in Turkey

The genus Vaccinium L. (Ericaceae) consists of shrubs or dwarf shrubs species widely distributed in Southeast Asia, America, and Europe. In the present paper, members of the genus (V. arctostaphylos, V. myrtillus, V. uliginosum, V. vitis-idaea) naturally grown in Northeast Anatolia were compared in terms of leaf venation, seed micromorphology, and palynological properties based on 16 populations in detail. Present findings revealed that number of areoles (the small area bounded by secondary veinlet) is 8–9 for V. arctostaphylos, 5–7 for V. myrtillus, 3–4 for V. uliginosum and V. vitis-idaea. Scanning electron microscope observation shows that the leaf adaxial surface is fiber-fluted in V. arctostaphylos, ruminate in V. myrtillus, V. uliginosum, and V. vitis-idaea due to epicuticular waxes. In addition, the seed shape of V. arctostaphylos in outline varies from triangular, tetragonal to oval, whereas they are lunate to reniform or falcate in the rest of the examined taxa. Epidermal cells of the seed are quadrilateral with straight anticlinal walls in V. arctostaphylos, however, they are most elongate rectangle with weakly undulate anticlinal walls in the rest species. The mean tetrad diameter (D) of pollen belonging to all examined species is 29.89 (21.47–43.22) μm while the lowest mean D was found in V. uliginosum (24.59 μm) and the highest mean D was found in V. arctostaphylos (36.74 μm). Principal component analysis based on 47 traits (flower: 9, leaf: 18, seed and fruit: 9 and pollen: 11) showed that the leaf length and width, teeth numbers, tetrad diameter, equatorial axis, and polar axis were the most valuable characters to distinguish the examined taxa in the present paper.

Development of a targeted genotyping platform for reproducible results within tetraploid and hexaploid blueberry

Blueberry (Vaccinium spp.) is one of the most economically important berry crops worldwide. Validation of genetic mapping studies is often hindered by asynchronous marker technology. The development of a standardized genotyping platform that targets a specific set of polymorphic loci can be a practical solution to unify the scientific and breeding community toward blueberry improvement. The objective of this study was to develop and evaluate a targeted genotyping platform for cultivated blueberries that is affordable, reproducible, and sufficiently high density to warrant large-scale adoption for genomic studies. The Flex-Seq platform was developed in a two-step procedure that resulted in 22,000 loci that yielded 194,365 single nucleotide polymorphisms when assessed in a diversity set of 192 samples including cultivated and other related wild Vaccinium species. Locus recovery averaged 89.4% in the cultivated polyploid blueberry (northern highbush [NHB], southern highbush [SHB], and rabbiteye [RE]) and on average 88.8% were polymorphic. While recovery of these loci was lower in the other Vaccinium species assayed, recovery remained high and ranged between 60.8% and 70.4% depending on the taxonomic distance to the cultivated blueberry targeted in this platform. NHB had the highest mean number of variants per locus at 9.7, followed by RE with 9.1, SHB with 8.5, and a range between 7.7 and 8.5 in other species. As expected, the total number of unique-in-state haplotypes exceeded the total number of variants in the domesticated blueberries. Phylogenetic analysis using a subset of the SNPs and haplotypes mostly conformed to known relationships. The platform also offers flexibility about the number of loci, depth of sequencing for accurate dosage calling, loci and haplotype reconstruction from increased fragment length. This genotyping platform will accelerate the development and improvement of blueberry cultivars through genomic-assisted breeding tools.

Polyphenol contents, antioxidant activities, and anti-cancer cell proliferation properties at each stage of fruit development in intersectional hybrids between highbush blueberry and shashanbo (Vaccinium bracteatum Thunb.)

BACKGROUND: In blueberries, the total polyphenol contents, total anthocyanin contents, and antioxidant activities of fruits in blueberries and their relatives are becoming important targets for breeders. Recently, intersectional hybrids between Japanese wild species Shashanbo and highbush blueberry were produced using polyploid breeding for the first time. However, the polyphenol contents, antioxidant activities, and anti-cancer cell proliferation properties of the fruits of these intersectional hybrids have not been studied sufficiently. OBJECTIVE: The objectives of this study were to determine the anthocyanin contents, polyphenol contents, and antioxidant activities at each stage of fruit development in intersectional hybrids, and to examine the effects of the fruit extracts on anti-cancer cell proliferation properties using human promyelocytic leukemia (HL-60) cells. METHODS: Freeze-dried fruit samples of three intersectional hybrids and their parents at five developmental stages were used to evaluate their polyphenol contents, anthocyanin contents, and levels of two types of antioxidant activities (DPPH and ORAC). The anti-cancer cell proliferation properties of each extract were also evaluated according to the survival rate of HL-60 cells. RESULTS: The total polyphenol contents in fruits of the intersectional hybrids gradually decreased as the fruits grew, but the contents increased slightly at the mature stage. Meanwhile, the total anthocyanin contents increased strongly to the mature stage. In addition, the fruit extract of intersectional hybrid JM1 was found to be more effective for inhibiting the growth of HL-60 human leukemia cells in vitro. Total polyphenol content correlated highly with antioxidant activities and anti-cancer cell proliferation properties. CONCLUSIONS: We clarified that the intersectional hybrids had produced have abundant bioactive compounds, high levels of antioxidant activities, and anti-cancer cell proliferation properties. The results proved the effectiveness of the use of Japanese wild Vaccinium species for the strategic breeding of blueberries with high levels of bioactive compounds.

Phenolic Antioxidants in Aerial Parts of Wild Vaccinium Species: Towards Pharmaceutical and Biological Properties.

Phenolic compounds are a widespread group of secondary metabolites found in all plants, representing the most desirable antioxidants due to their potential to be used as additives in the food industry (inhibition of lipid oxidation), and in cosmetology and medicine (protection against oxidative stress). In recent years, demand for the identification of edible sources rich in phenolic antioxidants, as well as the development of new natural plant products to be used as dietary supplements or pharmaceuticals, has been a great preoccupation. At present, from the “circular economy” perspective, there is an increased interest to use agricultural waste resources to produce high-value compounds. Vaccinium leaves and stems are considered essentially an agro-waste of the berry industry. Scientific studies have shown that phenolic compounds were found in a markedly higher content in the leaves and stems of Vaccinium plants than in the fruits, in agreement with the strongest biological and antioxidant activities displayed by these aerial parts compared to fruits. This paper aims to review the current state of the art regarding the phenolic antioxidants from leaves and stems of two wild Vaccinium species, bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.), as promising natural resources with pharmaceutical and biological activity.

Evaluation of some horticulturally important morphological traits of Turkish wild Vaccinium species

Evaluation of some horticulturally important morphological traits of Turkish wild <i>Vaccinium</i> species

Haplotype-phased genome and evolution of phytonutrient pathways of tetraploid blueberry.

BackgroundHighbush 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.FindingsHere, 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.ConclusionsThese findings and the reference genome will serve as a valuable resource to guide future genome-enabled breeding of important agronomic traits in highbush blueberry.

Short communication: Identification of cultivated and wild Vaccinium species grown in Portugal

Vaccinium crops offer a variety of benefits for human health due their high levels of antioxidants. Genetic diversity between two Vaccinium species (sixteen cultivars of Vaccinium corymbosum and three wild populations of Vaccinium myrtillus) were evaluated using Inter Simple Sequence Repeat (ISSR) markers. In V. corymbosum 74 polymorphic markers corresponding to 83.2% of polymorphism were obtained while in V. myrtillus only four polymorphic markers corresponding to 83.2% and 10.6% of polymorphism were observed. The dendrogram obtained showed a clear division into two distinct groups corresponding to the two analyzed species. V. corymbosum group is divided in different sub-clusters based on cultivars pedigree relationships. Twenty-eight specific bands were detected in total; 6 for V. corymbosum and 22 for V. myrtillus. Results allowed the selection of five primers due it potential to detect specific bands in the two species. These markers could be useful for identifying species and cultivars and consequently help in the management of germplasm collections and in breeding programs.

Recognition of candidate transcription factors related to bilberry fruit ripening by de novo transcriptome and qRT-PCR analyses

Bilberry (Vaccinium myrtillus L.) fruits are an excellent natural resource for human diet because of their special flavor, taste and nutritional value as well as medical properties. Bilberries are recognized for their high anthocyanin content and many of the genes involved in the anthocyanin biosynthesis have been characterized. So far, neither genomic nor RNA-seq data have been available for the species. In the present study, we de novo sequenced two bilberry fruit developmental stages, unripe green (G) and ripening (R). A total of 57,919 unigenes were assembled of which 80.2% were annotated against six public protein databases. The transcriptome served as exploratory data to identify putative transcription factors related to fruit ripening. Differentially expressed genes (DEGs) between G and R stages were prominently upregulated in R stage with the functional annotation indicating their main roles in active metabolism and catalysis. The unigenes encoding putative ripening-related regulatory genes, including members of NAC, WRKY, LOB, ERF, ARF and ABI families, were analysed by qRT-PCR at five bilberry developmental stages. Our de novo transcriptome database contributes to the understanding of the regulatory network associated with the fruit ripening in bilberry and provides the first dataset for wild Vaccinium species acquired by NGS technology.

Input data needed for a risk model for the entry, establishment and spread of a pathogen (Phomopsis vaccinii) of blueberries and cranberries in the EU

AbstractInternational trade in live plant materials has increased worldwide over the last four decades. This has led to a dramatic increase in the introduction, establishment and spread of non‐native plant pathogens. Regulatory authorities need advice on measures that may mitigate these adverse consequences of trade. Risk models may be used to underpin such advice. In this review, we give a systematic overview of the data needed for a quantitative risk model for Phomopsis vaccinii, which causes stem and fruit infections on Vaccinium species, and sometimes death, potentially also on native wild Vaccinium species in the EU. P. vaccinii is a quarantine organism worldwide, except for North America, where it is endemic. Despite extensive knowledge of the aetiology of the diseases caused by this pathogen and its taxonomy, quantitative data on transportation and detection of infected plants for planting and berries are scarce, and quantitative assessment of the future introduction, establishment and spread of P. vaccinii is difficult. Estimation of the potential impact of this pathogen in production fields and wild Vaccinium stands is even more difficult. P. vaccinii is not unique in this respect, and this review indicates the need for more and better quantitative data for assessment of the risks posed by newly introduced plant pathogens in areas where they are not endemic.

Pest risk assessment of Diaporthe vaccinii for the EU territory.

As requested by the European Commission, the EFSA Panel on Plant Health (PLH) Panel assessed the risk of Diaporthe vaccinii in the EU, focusing on entry, establishment, spread and impacts on cultivated and wild Vaccinium species, the principal hosts being American and European cranberry and blueberry. Several outbreaks occurred in the EU since 1956, but most were eradicated except in Latvia. The Panel considered entry via fruits and plants for planting. The risk of establishment from discarded infected berries is much lower than from infected plants for planting, of which, potted plants and cuttings pose the greatest risk, while plug plants, derived from tissue culture and grown in pest free structures, pose a low risk. Nine per cent of the EU is highly suitable for establishment of the pathogen, mostly in the SE and NE. Following establishment, the pathogen could spread naturally over short range, and by human assistance over long range. Calculations with an integrated model for entry, establishment and spread, indicate that with current regulations, over a period of 5 years, a few hundred cultivated Vaccinium plants and several thousand Vaccinium plants in natural ecosystems would contract the disease. The associated loss of commercial production is small, less than one tonne of berries per year. On natural vegetation, the median impact after 5 years was estimated to be negligible affecting a negligible proportion of the natural Vaccinium population (2 × 10−8). However, the uncertainty of this estimate was high, due to uncertainty about the rate of spread; in a worst‐case scenario (99th percentile), almost 1% of plants in natural areas would become infected. Complete deregulation (scenario A1) was predicted to increase the impact substantially, especially in natural areas, while additional measures (scenario A2) would effectively eliminate the entry of infected plants for planting, further reducing the impacts below the current situation.

Differential iridoid production as revealed by a diversity panel of 84 cultivated and wild blueberry species.

Cultivated blueberry (Vaccinium corymbosum, Vaccinium angustifolium, Vaccinium darrowii, and Vaccinium virgatum) is an economically important fruit crop native to North America and a member of the Ericaceae family. Several species in the Ericaceae family including cranberry, lignonberry, bilberry, and neotropical blueberry species have been shown to produce iridoids, a class of pharmacologically important compounds present in over 15 plant families demonstrated to have a wide range of biological activities in humans including anti-cancer, anti-bacterial, and anti-inflammatory. While the antioxidant capacity of cultivated blueberry has been well studied, surveys of iridoid production in blueberry have been restricted to fruit of a very limited number of accessions of V. corymbosum, V. angustifolium and V. virgatum; none of these analyses have detected iridoids. To provide a broader survey of iridoid biosynthesis in cultivated blueberry, we constructed a panel of 84 accessions representing a wide range of cultivated market classes, as well as wild blueberry species, and surveyed these for the presence of iridoids. We identified the iridoid glycoside monotropein in fruits and leaves of all 13 wild Vaccinium species, yet only five of the 71 cultivars. Monotropein positive cultivars all had recent introgressions from wild species, suggesting that iridoid production can be targeted through breeding efforts that incorporate wild germplasm. A series of diverse developmental tissues was also surveyed in the diversity panel, demonstrating a wide range in iridoid content across tissues. Taken together, this data provides the foundation to dissect the molecular and genetic basis of iridoid production in blueberry.

Scientific Opinion on the pest categorisation of Diaporthe vaccinii Shear

The European Commission requested the EFSA Panel on Plant Health to perform a pest categorisation of Diaporthe vaccinii Shear, the fungal agent responsible for twig blight, canker, viscid rot, fruit rot and storage rot of several Vaccinium species. The pest is listed in Annex IIAI of Directive 2000/29EC. D. vaccinii is a single taxonomic entity and methods exist for its discriminative detection. The host is restricted to Vaccinium species, the main cultivated hosts being blueberries and cranberries. Hosts are cultivated throughout Europe, and wild Vaccinium species are common components of forests. Conditions are conducive to disease development in most areas of Europe, but not in all Member States (MSs). The disease is currently present in Latvia with restricted distribution and the pest is under surveillance in The Netherlands. In the one Latvian report, storage losses were observed on cranberry, but these losses were caused by a complex of pathogens, including D. vaccinii, which was isolated with a low incidence. Detection methods are available but cultural and morphological identifications should be confirmed with molecular tools owing to the presence of other Phomopsis species on Vaccinium in Europe. The pathogen can spread via the movement of (asymptomatic) infected or contaminated host plants for planting. No information exists on any methods applied for the control of D. vaccinii in the EU. D. vaccinii does not have the potential to be a quarantine pest as it does not fulfil one of the pest categorisation criteria defined in International Standard for Phytosanitary Measures No 11, that of having a severe impact. Data are not sufficient to conclude on pest categorisation of D. vaccinii based on the criteria of the International Standard for Phytosanitary Measures No 21 because there is no information about the level of potential consequences as a result of the use of infected host plants for planting. © European Food Safety Authority, 2014

Phytochemicals in fruits of Hawaiian wild cranberry relatives

Cranberries (Vaccinium macrocarpon Ait.) contain high levels of phytochemicals such as proanthocyanidins (PACs). These polymeric condensations of flavan-3-ol monomers are associated with health benefits. Our objective was to evaluate phytochemicals in fruit from Hawaiian cranberry relatives, V. reticulatum Sm. and V. calycinum Sm. Normal-phase HPLC coupled with fluorescence and ESI-MS detected PACs; the colorimetric 4-dimethylaminocinnamaldehyde (DMAC) assay was used to determine total PACs. Spectrophotometric tests and reverse-phase HPLC coupled to photodiode array and refractive index detectors evaluated phenolics, sugars, and organic acids. Antioxidant capacity was determined by the ORAC and FRAP assays. Antioxidant capacities of Hawaiian berries were high. The FRAP measurement for V. calycinum was 454.7 ± 90.2 µmol L(-1) Trolox equivalents kg(-1) for pressed fruit. Hawaiian berries had lower peonidin, quinic and citric acids amounts and invert (∼1) glucose/fructose ratio compared with cranberry. Both Hawaiian Vaccinium species were good sources of PACs; they contained phenolics and PAC monomers, A and B-type trimers, tetramers and larger polymers. Vaccinium reticulatum and V. calycinum showed comparable or higher PAC levels than in cranberry. Cranberries had higher percentage of A-type dimers than did V. reticulatum. A and B-type dimers were not differentiated in V. calycinum. The total PACs (as measured by DMAC) for V. calycinum (24.3 ± 0.10 mg catechin equivalents kg(-1) ) were about twice that in cranberry. Berries of V. reticulatum and V. calycinum could serve as a rich dietary source of PACs, comparable to or greater than cranberries. These finding suggest that Hawaiian Vaccinium berries could be a functional food. Additional examination of the phytochemicals in other wild Vaccinium species is warranted.

Antioxidant Activities and Anti-Cancer Cell Proliferation Properties of Natsuhaze (Vaccinium oldhamii Miq.), Shashanbo (V. bracteatum Thunb.) and Blueberry Cultivars.

Antioxidants are abundant in blueberries, and while there are many studies concerning the bioactive compound of fruit, it is only recently that the wild Vaccinium species has attracted attention for their diverse and abundant chemical components. The aim of this study was to investigate the bioactive compounds of blueberry cultivars and wild species found in Japan. Among the five extracts of the Vaccinium species, Natsuhaze (Vaccinium oldhamii Miq.) was found to be the most effective at inhibiting the growth of HL-60 human leukemia cells in vitro. Although all ethanol extracts showed a growth inhibitory effect on HL-60 cells, the degree of the effects differed among the species. The extract of Natsuhaze induced apoptotic bodies and nucleosomal DNA fragmentation in the HL-60 cells. Of the extracts tested, that of Natsuhaze contained the largest amount of total polyphenols and showed the greatest antioxidant activity, but the anthocyanin content of Natsuhaze was similar to that of rabbiteye blueberry (V. virgatum Ait.). The results showed that total polyphenols contributed to the high antioxidant activity and growth inhibitory effect on HL-60 human leukemia cells of Natsuhaze extract.

Uptake and Assimilation of Nitrate and Iron in Cultivated and Wild Vaccinium Species

Growth of cultivated Vaccinium species on soils with a pH above 6.0 is inhibited and production can be extended to these types of soils only after extensive soil modifications. V. arboreum is a wild species that grows on soils with higher pH values. The aim of the present study was to characterize differences in nitrate and iron assimilation in cultivated vs. wild Vaccinium. Genotypes from V. corymbosum and V. arboreum were grown hydroponically with different nitrate and iron concentrations. Nitrate and iron uptake were significantly greater in V. arboreum compared with V. corymbosum. This was correlated with increased nitrate reductase and ferric chelate reductase activities and increased tissue concentrations of reduced nitrogen and iron in V. arboreum. The wild species, V. arboreum, clearly exhibits an increased ability to assimilate nitrate and iron compared with V. corymbosum. This greater capacity for assimilation may play a significant role in this species' broader soil adaptation.

Resistance to Blighting by Monilinia vaccinii-corymbosi in Diploid and Polyploid Vaccinium Species

Resistance to blighting by Monilinia vaccinii-corymbosi (Reade) Honey was evaluated under greenhouse conditions in multiple populations of the diploid species Vaccinium boreale Hall &amp; Aalders, V. corymbosum L., V. darrowi Camp, V. elliottii Chapm., V. myrtilloides Michx., V. myrtillus L., V. pallidum Ait., and V. tenellum Ait., as well as in accessions of the polyploid species 4x V. hirsutum Buckley and 6x V. corymbosum f. amoenum Aiton. Significant species differences were found in mean blighting levels averaged over 2 years, with values ranging from 3.5% for V. boreale to 49.2% for 2x V. corymbosum, compared with 27.5% for the resistant 4x V. corymbosum check, `Bluejay', and 64.3% for the susceptible 4x V. corymbosum check, `Blueray'. Wild Vaccinium species may serve as new sources of resistance to blighting, if resistance can be transferred easily and horticultural type recovered.

The Investigation and Cultivation ofVacciniumSpecies in the Soviet Union

This paper catalogues work in the Soviet Union on selection and breeding of various Vaccinium species and on the attempts underway to develop commercial industries for the highbush blueberry, bog whortlebeny, European and American cranberries, lingonberry, and other wild Vaccinium species.

Improved drought tolerance and root development as components of a scheme to breed blueberries for mineral soil adaptability

Blueberry breeding in North America started at the beginning of the 20th century and present cultivars are adapted to irrigated, high organic, acidic, sandy soils. However, sites meeting these requirements are limited, which has forced breeders to develop germplasm that is adapted to unamended mineral soils. Wild Vaccinium species can be found growing on a number of different soil types under a variety of climatic conditions, which indicates that genetic variability for mineral soil adaptability is present in Vaccinium germplasm. However, for a number of reasons progress in the development of mineral soil-adapted blueberry cultivars has been slow. In this paper a breeding scheme is presented that would allow the user to more efficiently access the variability that exists for mineral soil adaptability. The scheme is composed of two different seedling screening techniques (drought and seedling rooting screen) that are designed to eliminate the less vigorous individuals in a population before field testing. The field environment would be used primarily to evaluate individuals for fruit quality and yield characteristics. In blueberries, both general and specific combining ability can be fully exploited because they are vegetatively propagated which immediately fixes any unique combination of genes.

Virus and Mycoplasmalike Diseases of Blueberry

Abstract Although the fruit of wild Vaccinium species has long been harvested by man, a commercial fruit industry was not established until the early 1920’s with the introduction of highbush blueberry cultivars of V. australe Small and V. corymbosum L. Plants were at first apparently free of serious disease problems, but the new industry was soon threatened by stunt, which was considered a virus disease until the discovery of mycoplasmalike organisms (MLO’s) in plants. Since 1954, several virus diseases have been recognized. Researchers have had the unique opportunity to follow the development of virus and MLO diseases while a native wild plant was modified by plant breeders and brought under cultivation. Information acquired before 1970 and illustrations of symptoms may be found in earlier reviews (10, 11, 12).