Intersectional Hybrids between Darrow's Blueberry (V. darrowii Camp) and Lingonberry (V. vitis-idaea L.).

An initial cross of V. darrowii 'Johnblue' (Darrow's blueberry) × V. vitis-idaea 'Red Sunset' (lingonberry) produced more than 30 true intersectional diploid hybrids as confirmed by molecular markers. The most vigorous of these hybrids was extensively evaluated. This hybrid, US 2535-A, was floriferous and morphologically intermediate to the respective parents. Examination of pollen suggested low male fertility. Numerous crosses using the hybrid as a female reflected similarly low fertility and potential crossing barriers. Stylar examination suggested blockage of pollen tube growth in self-pollinations and significantly retarded growth in backcross pollinations. Nonetheless, two confirmed hybrid offspring were produced using the F1 hybrid as a female in crosses with V. vitis-idaea and V. darrowii, respectively. In a second set of crosses utilizing additional V. darrowii and V. vitis-idaea genotypes, another 23 verified hybrids in seven parental combinations were produced. Hybrids such as the ones presented offer the potential for generating de novo interspecific fruit types in blueberry and/or broadening the adaptation of lingonberry.

Correction: Ehlenfeldt et al. Asymmetric Reciprocal Crossing Behavior of an Andean Blueberry (V. meridionale) × Lingonberry (V. vitis-idaea) Hybrid. Plants 2022, 11, 3152.

In the original publication [...].

Asymmetric Reciprocal Crossing Behavior of an Andean Blueberry (V. meridionale) × Lingonberry (V. vitis-idaea) Hybrid

The fertility and crossing behavior of a tetraploid hybrid of 4x Andean blueberry (V. meridionale) and lingonberry (V. vitis-idaea) was evaluated through a series of crosses. Crosses of the hybrid with highbush blueberry produced divergent results. When used as a female with V. corymbosum males, virtually all offspring were hexaploid, most likely arising from 2n = 4x = 48 female gametes, and 1n = 2x = 24 male gametes. However, when used as a male, tetraploid hybrids were produced, resulting from 1n = 2x = 24 gametes from each parent. To further examine this crossing behavior, the 4x V. meridionale­V. vitis-idaea interspecific hybrid was pollinated with 6x V. virgatum (rabbiteye blueberry). Analogous to the previous crosses, 7x hybrids were produced from the joining of 2n = 4x = 48 female gametes with 1n = 3x = 36 male gametes. Such reciprocal crossing asymmetry is unprecedented. The ability to produce both 6x and 4x offspring from the same V. corymbosum parents allows the potential of bridging a V. meridionale hybrid genotype to both the tetraploid (V. corymbosum) and hexaploid (V. virgatum) commercial crop levels.

There and back again; historical perspective and future directions for Vaccinium breeding and research studies.

The genus Vaccinium L. (Ericaceae) contains a wide diversity of culturally and economically important berry crop species. Consumer demand and scientific research in blueberry (Vaccinium spp.) and cranberry (Vaccinium macrocarpon) have increased worldwide over the crops' relatively short domestication history (~100 years). Other species, including bilberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), and ohelo berry (Vaccinium reticulatum) are largely still harvested from the wild but with crop improvement efforts underway. Here, we present a review article on these Vaccinium berry crops on topics that span taxonomy to genetics and genomics to breeding. We highlight the accomplishments made thus far for each of these crops, along their journey from the wild, and propose research areas and questions that will require investments by the community over the coming decades to guide future crop improvement efforts. New tools and resources are needed to underpin the development of superior cultivars that are not only more resilient to various environmental stresses and higher yielding, but also produce fruit that continue to meet a variety of consumer preferences, including fruit quality and health related traits.

Transcriptome analysis identifies genes related to the waxy coating on blueberry fruit in two northern-adapted rabbiteye breeding populations.

BACKGROUND: Blueberry is of high economic value. Most blueberry varieties selected for the fresh market have an appealing light blue coating or "bloom" on the fruit due to the presence of a visible heavy epicuticular wax layer. This waxy layer also serves as natural defense against fruit desiccation and deterioration. RESULTS: In this study, we attempted to identify gene(s) whose expression is related to the protective waxy coating on blueberry fruit utilizing two unique germplasm populations that segregate for the waxy layer. We bulked RNA from waxy and non-waxy blueberry progenies from the two northern-adapted rabbiteye hybrid breeding populations ('Nocturne' x T 300 and 'Nocturne' x US 1212), and generated 316.85 million RNA-seq reads. We de novo assembled this data set integrated with other publicly available RNA-seq data and trimmed the assembly into a 91,861 blueberry unigene collection. All unigenes were functionally annotated, resulting in 79 genes potentially related to wax accumulation. We compared the expression pattern of waxy and non-waxy progenies using edgeR and identified overall 1125 genes in the T 300 population and 2864 genes in the US 1212 population with at least a two-fold expression difference. After validating differential expression of several genes by RT-qPCR experiments, a candidate gene, FatB, which encodes acyl-[acyl-carrier-protein] hydrolase, emerged whose expression was closely linked to the segregation of the waxy coating in our populations. This gene was expressed at more than a five-fold higher level in waxy than non-waxy plants of both populations. We amplified and sequenced the cDNA for this gene from three waxy plants of each population, but were unable to amplify the cDNA from three non-waxy plants that were tested from each population. We aligned the Vaccinium deduced FATB protein sequence to FATB protein sequences from other plant species. Within the PF01643 domain, which gives FATB its catalytic function, 80.08% of the amino acids were identical or had conservative replacements between the blueberry and the Cucumis melo sequence (XP_008467164). We then amplified and sequenced a large portion of the FatB gene itself from waxy and non-waxy individuals of both populations. Alignment of the cDNA and gDNA sequences revealed that the blueberry FatB gene consists of six exons and five introns. Although we did not sequence through two very large introns, a comparison of the exon sequences found no significant sequence differences between the waxy and non-waxy plants. This suggests that another gene, which regulates or somehow affects FatB expression, must be segregating in the populations. CONCLUSIONS: This study is helping to achieve a greater understanding of epicuticular wax biosynthesis in blueberry. In addition, the blueberry unigene collection should facilitate functional annotation of the coming chromosomal level blueberry genome.

Dataset of de novo assembly and functional annotation of the transcriptome of blueberry (Vaccinium spp.).

Blueberry is an economically important berry crop. Both production and consumption of blueberries have increased sharply worldwide in recent years at least partly due to their known health benefits. The development of improved genomic resources for blueberry, such as a well-assembled genome and transcriptome, could accelerate breeding through genomic-assisted approaches. To enrich available transcriptome data and identify genes potentially involved in fruit quality, RNA sequencing was performed on fruit tissue from two northern-adapted hybrid blueberry breeding populations. RNA-seq was carried out using the Illumina HiSeqTM 2500 platform. Because of the absence of a reference-grade genome for blueberry, a transcriptome was de novo assembled from this RNA-seq data and other publicly available transcriptome data from blueberry downloaded from the National Center for Biotechnology Information (NCBI) Short Read Archive (SRA) using Trinity. After removing redundancy, this resulted in a dataset of 91,861 blueberry unigenes. This unigene dataset was functionally annotated using the NCBI-Nr protein database. All raw reads from the breeding populations were deposited in the NCBI SRA with accession numbers SRR6281886, SRR6281887, SRR6281888, and SRR6281889. The de novo transcriptome assembly was deposited at NCBI Transcriptome Shotgun Assembly (TSA) database with accession number GGAB00000000. These data will provide real expression evidence for the blueberry genome gene prediction and gene functional annotation and a reference transcriptome for future gene expression studies involving blueberry fruit.

Simultaneous LC-MS quantification of anthocyanins and non-anthocyanin phenolics from blueberries with widely divergent profiles and biological activities.

A sensitive and straightforward LC-IT-TOF-MS method was validated for the profiling and simultaneous quantification of anthocyanins, flavan-3-ols, flavonols, phenolic acids, and resveratrol in blueberry genotypes with fruit color ranging from deep purple (Vaccinium angustifolium) to various shades of pink (crosses of V. corymbosum, V. darrowii, and V. ashei). Standard calibration curves were linear for all analytes with correlation coefficients >0.99. The relative standard deviation for intra- and inter-day precision was lower than 10%. The method allowed an easy and selective identification and quantification of phenolics in blueberries with divergent profiles. The in vitro antioxidant assay results were strongly correlated with total phenolics and total anthocyanin content. Lowbush blueberry extracts (50 µg/mL) reduced ROS and NO production, and inhibited the transcription of the proinflammatory cytokines IL-6ß, COX2, iNOS, and IL-6 in the in vitro assays at much lower concentrations than pink fruited berries (250 µg/mL).

Molecular Detection and Discrimination of Blueberry red ringspot virus Strains Causing Disease in Cultivated Blueberry and Cranberry.

Blueberry red ringspot virus (BRRV) causes red ringspots on the stems, leaves, and ripening fruit of infected highbush blueberry (Vaccinium corymbosum) plants. The disease was originally observed in New Jersey and has now been reported in other blueberry growing regions in the United States, as well as several locations in Europe. A disease with similar symptoms occurs in American cranberry (V. macrocarpon), but BRRV has never been confirmed as the causal agent. Serological detection of BRRV in infected plants has been unsatisfactory. Using a primer set designed for routine detection (RRSV3/RRSV4), we successfully amplified a fragment of the virus from all tissues of infected highbush blueberry plants. Using the same primer set, we confirmed natural infection of BRRV in rabbiteye (V. virgatum) blueberry cultivars and the rabbiteye × V. constablaei hybrid cultivar Little Giant. These species have not been previously reported as hosts for this virus. Viral fragments were cloned from representative blueberry and cranberry plants exhibiting ringspot symptoms. Phylogenetic analysis of sequence data showed that cranberry strains of BRRV are precursors to the more derived blueberry strains. The techniques reported in this paper are being used to evaluate strain variation in Vaccinium species and to identify the as yet unknown vector(s) of this virus.

Anthracnose Fruit Rot Resistance in Blueberry Cultivars.

Anthracnose fruit rot (causal agent, Colletotrichum acutatum) is an important disease in most blueberry growing regions of North America. Losses caused by the disease are usually seen as a postharvest rot with orange spore masses appearing on the surface of affected fruit. One hundred cultivars/selections of blueberry were screened for resistance to fruit rot between 1993 and 2003 by inoculating container-grown plants bearing green fruit. Visible rot symptoms on ripe fruits were evaluated after a 1-week incubation at room temperature. Our analyses revealed that infection levels were affected by mean May temperatures in New Jersey, generally increasing as temperatures increased; however, this effect was not consistent among all cultivars. A generalized linear mixed model was developed to predict resistance at the historic mean May temperature, conservatively explaining 59% of the variance in resistance. Percent infection ranged from 9 to 91% with a mean of 51% across all cultivars. Results for common cultivars corresponded well with field reports of their relative susceptibilities. An estimate of narrow-sense heritability of 0.32 suggested additive inheritance of resistance. Since very high inoculum loads were used in this study, cultivars exhibiting a low percentage of fruit rot are predicted to show superior field resistance to the disease and will be incorporated into an ongoing breeding program.