Societal Impact Statement More than 100 edible native berries grow across Canada's vast territory and are used by over 600 Indigenous Peoples of Canada as a main component of their diet. This research provides critical insights into the ecology, phenology, and cultivation of black huckleberry ( Vaccinium membranaceum ), a species of significant ecological and cultural importance in western North America. This work not only supports biodiversity and ecosystem health but also bolsters Indigenous food sovereignty and cultural heritage. The findings offer practical guidelines for policymakers, conservationists, and Indigenous communities, fostering collaborative efforts to preserve this vital species in the face of climate change and environmental challenges. Summary Conserving native food‐plant species is crucial amid anthropogenic ecosystem disturbances and climate change. In western North America, native Vaccinium spp. (western huckleberries) are vital for sustenance and medicine, historically linking Indigenous communities and European settlers. This review, inspired by Westbank First Nation, synthesizes current knowledge and identifies research gaps concerning the distribution, growth, reproduction, abiotic and biotic stresses, and propagation of V. membranaceum or st̓łqiłəml̓x in Nsyilxcen (black huckleberry), emphasizing its ecological and cultural significance. The distribution and growth of V. membranaceum are shaped by abiotic factors (climate, soil, and fire) and biotic interactions (mutualisms with mycorrhizal fungi, pest and disease pressures). Understanding of how those factors individually and collectively (e.g. through phenology) influence V. membranaceum habitat suitability and berry yield will enhance understanding of the potential for its resilience. Furthermore, the integration of traditional and in vitro propagation techniques offers promising avenues for cultivation and conservation. By highlighting the dynamics shaping V. membranaceum populations, the review emphasizes the need for multidisciplinary research to safeguard this species. Bridging indigenous and contemporary scientific knowledge systems can foster sustainable management practices, ensuring the prosperity of V. membranaceum and the communities and ecosystems dependent on it.

Ensuring genetic fidelity and proper antioxidant regulation is essential for synthetic seed technology and conservation of economically important plant species. In this study, we evaluated the effects of plant growth regulators (PGRs) and storage temperature on the germination and regeneration of encapsulated somatic embryos of Vaccinium membranaceum, while verifying genetic fidelity and physiological competence. The highest somatic embryogenesis was obtained from leaf explants cultured on berry basal medium (BM) containing 4.6 µM thidiazuron (TDZ) and 1.13 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos were encapsulated in 3

Black huckleberry (Vaccinium membranaceum) is a native fruit species of high nutritional, medicinal, ecological, and economic value. The black huckleberries, abundant in bioactive compounds, offer significant antioxidants and anti-inflammatory effects and play a key role in maintaining wildlife and forest ecosystems. Despite its importance, protoplast isolation and gene editing have not been reported in this species. These techniques are essential for functional genomics and crop improvement, but the recalcitrant nature of this species, complex genome, and variable ploidy present significant challenges for cellular and molecular manipulation. This study aimed to establish a reliable protocol for efficient mesophyll protoplast isolation and transient gene expression in V. membranaceum using in vitro-grown leaves. A systematic optimization of enzyme composition, osmotic concentration, antioxidant supplementation, and pH was undertaken to enhance protoplast yield and viability in V. membranaceum. The optimized enzymatic combination of 2

Vaccinium vitis-idaea L. (lingonberry) is a health promoting northern berry crop valued for its antioxidant properties and is commonly propagated through in vitro techniques. However, the success of micropropagation is often hindered by hyperhydricity (HH), a physiological disorder that causes shoots to become water soaked and malformed. This study is the first investigation of HH in the lingonberry cultivar ‘Erntedank’ (Cv1), using a combination of structural, biochemical, and gene expression analyses. Lingonberry shoots grown in both liquid and semi-solid culture media developed typical HH symptoms, including excessive water accumulation and distorted tissues. Treatment with potassium silicate (K₂SiO₃) significantly reduced water content and improved shoot appearance over time. Scanning electron microscopy (SEM) showed distorted stomata, trichomes, and vascular tissues in hyperhydric shoots, which were restored upon silicon treatment. Biochemical profiling, using ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS), showed that HH increased the levels of stress related compounds, such as anthocyanins and phenolic acids, which were reduced after silicon treatment. Gene expression analysis using reverse transcription quantitative polymerase chain reaction (RT-qPCR) revealed an over 30-fold upregulation of antioxidant genes (CAT1, APX, and SOD) and ethylene related genes (ACO1, ACS11, COX2, and ETR1) in HH tissues, which were further downregulated following silicon treatment. Therefore, the findings from this study suggested that hyperhydricity in lingonberry is linked to oxidative and hormonal stress, and that silicon supplementation offers a promising strategy to prevent or recover from this disorder, improving the efficiency and quality of lingonberry micropropagation.

Somatic embryogenesis triggers epigenetic changes that influence catechin accumulation in lingonberry, helping to improve phytochemical quality in micropropagated plants. Vaccinium vitis-idaea L. (lingonberry), a boreal superfruit valued for its high antioxidant content, exhibits high potential for regeneration via somatic embryogenesis (SE). However, in-vitro culture often induces epigenetic variations that can influence phytochemical stability. To find out whether DNA methylation plays a role during SE, we compared DNA methylation levels in the regenerants of two lingonberry genotypes, Erntedank (Cv1) and a hybrid designated as “H1” (identity undisclosed). We demonstrate that SE in lingonberries induces epigenetic modifications, particularly epigenetic memory and biological imprinting that contribute to enhanced catechin accumulation, as revealed by highly sensitive DNA methylation profiling using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) and catechin analysis via proton nuclear magnetic resonance (1H NMR) across four experimental groups: in-vitro-derived shoots, in-vitro and ex-vitro SE-regenerated plants, and controls. The optimized UHPLC protocol enabled robust quantification of 5-methylcytosine (5mC), revealing significantly elevated DNA methylation in in-vitro-derived shoots (Cv1-TC: 84.02

Blueberry (Vaccinium spp.) is one of the most important small fruit crops due to its taste and its antioxidant capacity. Methods and Results: To understand the genetic basis of these antioxidant properties, we evaluated the antioxidant activity of 156 diverse blueberry genotypes, including cultivars, hybrids, and wild clones. Phenotypic characterization revealed diverse antioxidant activities ranging from 0.088 to 1.338 mg gallic acid equivalent per gram of fresh leaf weight. On average, 94.51

Understanding the physiological characteristics of hyperhydric plantlets is ultimately necessary since hyperhydricity results in financial loss for in vitro plants from a commercial perspective. Although many studies report the possible causes and symptoms of hyperhydricity, knowledge of it remains limited. This review aims to provide an integrated overview of this phenomenon and outline the perspectives for its prevention. First, we summarize the factors of in vitro hyperhydricity, including gelling agents, growth regulators, vessel ventilation and gas exchange, light, and osmotic conditions. Second, we describe physiological and internal changes commonly observed in hyperhydric plants, such as ROS/ethylene imbalance, altered antioxidant capacity, defects in the cell wall, and lignification. Third, we outline ultrastructural characteristics and accumulate HPLC findings to recognize the metabolite profiles of hyperhydric plantlets. Fourth, we introduce emerging AI-assisted MLM (machine learning model) approaches to detect and optimize the culture parameters to prevent hyperhydricity. Finally, we evaluate the strategies for the protection of the culture from hyperhydric conditions. This structured overview intends to reduce hyperhydricity in commercial and research settings.

Vaccinium vitis-idaea L. (lingonberry), globally recognized as a superfruit for its medicinal properties, has long been cultivated and consumed by Canadian Indigenous communities. This study introduces an AI-powered surveillance system that leverages an optimized You Only Look Once (YOLOv12) architecture to revolutionize yield estimation, phenomic profiling, and genomic/epigenomic analysis in micropropagated lingonberry. A custom multi-class annotated dataset was developed to evaluate model performance under real-world conditions. The YOLOv12 model, built on a RELAN backbone with flash-attention mechanisms, excelled in global context modeling, enabling accurate detection of berries and regenerated shoots in both ex vitro and in vitro environments. In contrast, YOLOv8 and YOLOv9, which rely on CNN-based feature extraction, demonstrated computational efficiency but suffered from overfitting and reduced operational robustness. In multi-class detection scenarios, YOLOv12 achieved the highest mean Average Precision with 67.3 % mAP@50 in yield detection, 1.0–99.5 % mAP@50 in micropropagated plant trait detection (shoots, berries, flowers), and 32.2–74 % accuracy in gel electrophoresis band detection. These results reflect a 22 % increase in throughput and a 38 % reduction in error rates compared to conventionally human-monitored methods, significantly reducing labor cost for plant breeders and agricultural biotechnologist. The integrated system enables simultaneous monitoring of phenotypic traits across growth stages and precise molecular band analysis, establishing a new paradigm for precision agriculture and lingonberry improvement.This work establishes YOLOv12 as the first unified framework for micropropagated lingonberry phenotyping across biological scales, demonstrating labor reduction in breeding programs while maintaining operational reliability. The technology's mobile compatibility and cloud-integration potential offer immediate applications for the global $2.3B lingonberry market, particularly in precision nurseries and nutraceutical production.

The “superfruit” lingonberry (Vaccinium vitis-idaea L.) is well known for its enormous health benefits and rich phytochemical contents. However, its regeneration and propagation face significant challenges due to the woody nature of the plant and poor proliferation rates using conventional methods. By using juvenile leaf explants of two V. vitis-idaea genotypes, this study introduces a novel and efficient somatic embryogenesis (SE) protocol for lingonberry micropropagation. The berry basal medium supplemented with 5.5 µM thidiazuron was optimized for the SE protocol where 92

Hyperhydricity is a significant challenge in the tissue culture of blueberry plantlets, affecting their propagation, survival and quality, which results in economic losses for industrial blueberry micropropagation. The in vitro liquid propagation of two half-highbush blueberry hybrids, HB1 and HB2, showed that a Growtek stationary bioreactor culture system containing a liquid medium exhibited a higher hyperhydricity percentage than a Sigma glass culture system with a semi-solid medium. The percentage of hyperhydricity (75.21 ± 1.89%) and water content (72%) of HB2 was more than that of HB1. A scanning electron microscopy study revealed that hyperhydric plantlets from both genotypes developed slowly, had closed stomata, and displayed enlarged intercellular spaces between the palisade and spongy parenchyma layers. Disrupted vascular bundles, underdeveloped sieve elements and a weak connection between phloem and xylem tissue were also observed in hyperhydric plantlets. An analysis of mesophyll and stem tissues highlighted a compressed adaxial epidermis, which led to compact palisade parenchyma, with irregularly shaped mesophyll cells. Hyperhydric plants showed strong nuclear magnetic resonance (NMR) signals in the aliphatic, aromatic, and sugar regions, specifically at peaks of 2.0, 2.5, 4.0, 4.5, 6.0, and 6.7 ppm. These signals were attributed to the presence of catechin (C15H14O6), a flavonoid compound, suggesting its significant role or accumulation in these plants under hyperhydric conditions. Despite the negative effects of hyperhydricity on commercial propagation, hyperhydric plants were found to contain higher levels of valuable untargeted metabolites, such as β-P-arbutin, chlorogenic acid, quercetin-3-O-glucoside, epicatechin, 2-O-caffeoyl arbutin, various fatty acids, β-glucose, linolenic acid, and acetyl than both in vitro and ex vitro conditions. The enrichment of bioactive compounds in blueberry enhances its antioxidant properties, nutritional profile, and potential health benefits, making them significant for plant defense mechanisms and stress adaptation.

Plant tissue culture has been recognized as an essential technology in plant science research. This process is widely used to regenerate and conserve phenotypically and genetically identical plant resources. The advancements in tissue culture methods have become a feasible option for the micropropagation of plants at the commercial level. The success of commercial micropropagation necessitates genetic stability among regenerated plants. Sometimes, in vitro-grown plants show genetic and epigenetic alterations due to stressful artificial culture conditions, media compositions, and explant types. As a result, it is essential to ensure genetic stability among tissue culture-derived plantlets at a very early stage. Somaclonal variations can be detected by phenotypic assessment, cytogenetic, DNA-based molecular markers, bisulfite sequencing, and RNA sequencing. This review aims to describe the causes behind somaclonal variation, the selection of somaclonal variants, and their uses in crop and plant improvement at the commercial level. This study discusses the optimization processes of undesirable genetic and epigenetic variation among micropropagated plants and their application in global horticulture, agriculture, and forestry.

Vaccinium membranaceum (black huckleberry) is known for its high content of bioactive compounds. This study introduces a novel approach for bioreactor micropropagation using stationary (Growtek) and temporary immersion (RITA) bioreactor systems using a liquid nutrient medium to enhance the in vitro propagation of black huckleberry. Adventitious shoot regeneration from leaf explants reached 80

Lingonberry ( Vaccinium vitis-idaea L.) produces tiny red berries that are tart and nutty in flavour. It grows widely in the circumpolar region, including Scandinavia, northern parts of Eurasia, Alaska, and Canada. Although cultivation is currently limited, the plant has a long history of cultural use among indigenous communities. Given its potential as a food source, genomic resources for lingonberry are significantly lacking. To advance genomic knowledge, the genomes for two subspecies of lingonberry ( V. vitis-idaea ssp. minus and ssp. vitis-idaea var. ‘Red Candy’) were sequenced and de novo assembled into contig-level assemblies. The assemblies were scaffolded using the bilberry genome ( V. myrtillus ) to generate chromosome-anchored reference genome consisting of 12 chromosomes each with total length 548.07 Mbp (contig N50 = 1.17 Mbp, BUSCO (C%) = 96.5%) for ssp. vitis-idaea , and 518.70 Mbp (contig N50 = 1.40 Mbp, BUSCO (C%) = 96.9%) for ssp. minus . RNA sequencing based gene annotation identified 27,243 genes on the ssp. vitis-idaea assembly, and transposable element detection methods found that 45.82% of the genome was repeats. Phylogenetic analysis confirmed that lingonberry is most closely related to bilberry and is more closely related to blueberries than cranberries. Estimates of past effective population size suggested a continuous decline over the past 1–3 MYA, possibly due to the impacts of repeated glacial cycles during Pleistocene leading to frequent population fragmentation. The genomic resource created in this study can be used to identify industry relevant genes (e.g., flavonoid genes), infer phylogeny, and call sequence-level variants (e.g., SNPs) in future research.

The lingonberry (Vaccinium vitis-idaea L.), recognized for its nutritional value and adaptability to cold climates, faces cultivation challenges, particularly in soil pH and fertility optimization. In a greenhouse study, lingonberry transplants were grown in media with pH levels of 6.5 (3:1:1 PRO-MIX BX/peat moss/perlite) and 5.2 (2:1 peat moss/perlite). Seven months post-exposure to different media pH, various fertility treatments (NPK) were tested, including a control (0–0–0), a balanced 5–5–5 kg ha−1 rate, a standard 36–24–48 kg ha−1 rate, and both higher (up to 54–36–72 kg ha−1) and lower (down to 9–6–12 kg ha−1) rates, applied every three weeks for fifteen weeks across six replications with a standard micronutrient rate. Results showed that media pH significantly affected plant height and volume, with plants at pH 6.5 growing 27% taller and larger than plants at pH 5.2. Fertility levels influenced plant volume, peaking at a moderate fertility rate (18–12–24 kg ha−1) before declining at higher rates. Interactions between pH and fertility significantly impacted shoot biomass, where higher fertility rates (above 36–24–48 kg ha−1) had a more pronounced negative effect on shoot biomass at pH 6.5 compared to pH 5.2. Root dry biomass was consistently 1.2–2.3 times greater than shoot dry biomass and less influenced by the treatments. Shoot death rates increased sharply at fertility rates above 18–12–24 kg ha−1, peaking at 21–35%. Nitrogen concentration in shoots and roots increased with higher fertilizer rates, peaking at 1.74% in the 45–30–60 kg ha−1 treatment. Fertility treatments raised growing media’s electrical conductivity (EC, 1:20 ratio), with a maximum of 1.41 dS m−1 in the 54–36–72 kg ha−1 treatment, though pH remained unchanged. Growing media nitrate levels increased with higher N rates, while ammonium levels were unaffected. Shoot death rates rose significantly with higher nitrate concentrations, particularly above 17.5 mg L−1, but showed no link to ammonium levels. Lingonberries can survive and thrive across a wide range of pH levels. These results indicate that lingonberries are resilient and low maintenance, requiring modest nutrient levels, and excessive fertilization hampers their growth.

Lingonberry (Vaccinium vitis-idaea L.) is a dwarf shrub that is native to Northern climates. The berries are harvested from wild plants and the disease incidence in the crop is generally low. However, disease pressure may increase since the surface area dedicated to the crop is expected to expand in the future. In 2021 and 2022, brown necrotic leaves and stem dieback symptoms were observed on lingonberry plants grown at the St. John's Research and Development Center, Newfoundland and Labrador, Canada. Pathogens were isolated from the diseased plant tissues and identified as Neopestalotiopsis rosae (n = 2) and Neopestalotiopsis zimbabwana (n = 2) using colony and conidia morphology as well as sequencing of the Internal Transcribed Spacer (ITS), beta-tubulin gene (beta-tub), and the translation elongation factor 1-alpha gene (tef1). Pathogenicity assays in detached lingonberry and strawberry leaves as well as lingonberry plants indicated that all four isolates were able to cause disease. This is the first report of N. rosae and N. zimbabwana causing leaf spot and stem dieback of lingonberry. Further monitoring of this disease is important to understand its prevalence in berry crops, as Neopestalotiopsis spp. in Canada have been reported to occur since 2018 in strawberry and other small fruits. Control measures and strategies are required to mitigate the potential risk from the disease, as there are no registered fungicides available to the growers.

Mungbean, a socioeconomically important legume, is a key player in the pursuit of food security and sustainable agriculture demanding attention for developing a highly efficient transformation system to support functional genomics and translational research. This study was focused to fill in the gap using both Agrobacterium-mediated and biolistic methods by experimenting with variables of the procedure to attain higher regeneration and transformation efficiencies. The study found comparable regeneration and transformation efficiencies between direct and indirect methods of gene transfer under selection pressure. Biolistic transformation of embryonic axis explants yielded an average transformation efficiency of 14.22

Dietary intake of Vaccinium berries has demonstrated significant potential in preventing many risk factors associated with metabolic syndromes in the human population. In recent years, a multitude of research has shown the role of antioxidants derived from Vaccinium berries on chronic diseases such as cardiovascular disorders, diabetes, obesity, and cancer. Several studies have also investigated the effect of Vaccinium berry consumption on their ability to modulate the risk factors associated with oxidative stress, vascular function, inflammation, and lipid metabolism. Regarding cancer, studies showed that the consumption of berries reduces inflammation, inhibits angiogenesis, protects against DNA damage within the cell, and controls apoptosis and proliferation rates in malignant tumours. However, which components are responsible for the health benefits is still unclear. Reports show that whole berry consumption usually confers positive effects on human health, and the health-promoting potentials are likely due to the presence of polyphenols with antioxidant activities. Among these polyphenols, various Vaccinium berry species have been reported to contain anthocyanins and flavonoids. These two polyphenolic compounds are known to have higher antioxidant activity and are beneficial for human health. There are now several studies and human clinical trials documenting the beneficial effects of Vaccinium berries, and these findings suggest that they may be promising for preventing and treating neurodegenerative diseases. This review focuses primarily on dietary Vaccinium berries consumption effects on human health and their potential role as therapeutic agents.

Abstract A proficient plant propagation technique using somatic embryogenesis was successfully established in lingonberry (Vaccinium vitis-idaea L.), a small evergreen shrub. The formation of callus and somatic embryos was observed in two lingonberry genotypes during the process. The most favorable conditions for embryogenic callus induction were observed on a nutrient medium supplemented with 5.5 µM thidiazuron. To confirm the development of somatic embryos, scanning electron microscopy analysis was conducted. The embryos were further cultured on a nutrient medium with 4.0 μM zeatin to facilitate their transformation into plantlets. The growth chamber-maintained ex vitro plants were compared with in vitro-grown somatic embryonic plants to determine whether the regenerants exhibited clonal stability. This was done by using 3 EST-SSR, 3 EST-PCR, and 3 genomic SSR primers. All in vitro- and ex vitro-grown plants displayed similar monomorphic amplification profiles in both genotypes, indicating that they maintained genetic fidelity. The total flavonoid and anthocyanin contents, antioxidant activity, and the catalase and superoxide dismutase enzyme activities were higher in SE plants than ex vitro-grown plants. On the other hand, total phenolic and proanthocyanin contents and glutathione reductase enzyme activity was more in the ex vitro plants compared to SE plants.

Berry crops of the genera Fragaria (strawberry; Rosaceae), Rubus (brambles: raspberry and blackberry; Rosaceae) and Vaccinium (blueberry, cranberry and lingonberry; Ericaceae) are of commercial importance worldwide. They play important roles in anti-tumor, anti-ulcer, anti-oxidant and anti-inflammatory activities. In vitro culture has become an important tool for the rapid propagation of berry crops, although clonal fidelity is a major concern in commercial micropropagation. In vitro-derived variations (somaclonal variations) are both heritable (genetic) and nonheritable (epigenetic), and depend on a number of factors, including genotype, chimeric tissue availability, media type, explant origin and type, plant growth regulator concentration, culture environment and duration. Tissue culture berry crops may show increased growth, berry production, and antioxidant activity, which may be of practical benefit to growers. The present review provides critical information for a better understanding of micropropagation and its epigenetic effects on morphology and antioxidant properties, along with DNA methylation in berry crops, and fills the prevailing gap in the literature.