Ethylene promotes fruit ripening initiation by downregulating photosynthesis, enhancing abscisic acid and suppressing jasmonic acid in blueberry (Vaccinium ashei).

BACKGROUND: Blueberry fruit exhibit atypical climacteric ripening with a non-auto-catalytic increase in ethylene coincident with initiation of ripening. Further, application of ethephon, an ethylene-releasing plant growth regulator, accelerates ripening by increasing the proportion of ripe (blue) fruit as compared to the control treatment. To investigate the mechanistic role of ethylene in regulating blueberry ripening, we performed transcriptome analysis on fruit treated with ethephon, an ethylene-releasing plant growth regulator. RESULTS: RNA-Sequencing was performed on two sets of rabbiteye blueberry ('Powderblue') fruit: (1) fruit from divergent developmental stages; and (2) fruit treated with ethephon, an ethylene-releasing compound. Differentially expressed genes (DEGs) from divergent developmental stages clustered into nine groups, among which cluster 1 displayed reduction in expression during ripening initiation and was enriched with photosynthesis related genes, while cluster 7 displayed increased expression during ripening and was enriched with aromatic-amino acid family catabolism genes, suggesting stimulation of anthocyanin biosynthesis. More DEGs were apparent at 1 day after ethephon treatment suggesting its early influence during ripening initiation. Overall, a higher number of genes were downregulated in response to ethylene. Many of these overlapped with cluster 1 genes, indicating that ethylene-mediated downregulation of photosynthesis is an important developmental event during the ripening transition. Analyses of DEGs in response to ethylene also indicated interplay among phytohormones. Ethylene positively regulated abscisic acid (ABA), negatively regulated jasmonates (JAs), and influenced auxin (IAA) metabolism and signaling genes. Phytohormone quantification supported these effects of ethylene, indicating coordination of blueberry fruit ripening by ethylene. CONCLUSION: This study provides insights into the role of ethylene in blueberry fruit ripening. Ethylene initiates blueberry ripening by downregulating photosynthesis-related genes. Also, ethylene regulates phytohormone-metabolism and signaling related genes, increases ABA, and decreases JA concentrations. Together, these results indicate that interplay among multiple phytohormones regulates the progression of ripening, and that ethylene is an important coordinator of such interactions during blueberry fruit ripening.

Transcriptomics of different tissues of blueberry and diversity analysis of rhizosphere fungi under cadmium stress.

Blueberry (Vaccinium ssp.) is a perennial shrub belonging to the family Ericaceae, which is highly tolerant of acid soils and heavy metal pollution. In the present study, blueberry was subjected to cadmium (Cd) stress in simulated pot culture. The transcriptomics and rhizosphere fungal diversity of blueberry were analyzed, and the iron (Fe), manganese (Mn), copper (Cu), zinc (Zn) and cadmium (Cd) content of blueberry tissues, soil and DGT was determined. A correlation analysis was also performed. A total of 84 374 annotated genes were identified in the root, stem, leaf and fruit tissue of blueberry, of which 3370 were DEGs, and in stem tissue, of which 2521 were DEGs. The annotation data showed that these DEGs were mainly concentrated in a series of metabolic pathways related to signal transduction, defense and the plant-pathogen response. Blueberry transferred excess Cd from the root to the stem for storage, and the highest levels of Cd were found in stem tissue, consistent with the results of transcriptome analysis, while the lowest Cd concentration occurred in the fruit, Cd also inhibited the absorption of other metal elements by blueberry. A series of genes related to Cd regulation were screened by analyzing the correlation between heavy metal content and transcriptome results. The roots of blueberry rely on mycorrhiza to absorb nutrients from the soil. The presence of Cd has a significant effect on the microbial community composition of the blueberry rhizosphere. The fungal family Coniochaetaceae, which is extremely extremelytolerant, has gradually become the dominant population. The results of this study increase our understanding of the plant regulation mechanism for heavy metals, and suggest potential methods of soil remediation using blueberry.

Impact of Background Fruit Odors on Attraction of Drosophila suzukii (Diptera: Drosophilidae) to Its Symbiotic Yeast.

Background odors produced by plants in the environment can interfere with the response of insects to a point-releasing attractant, especially when their compositions overlap. In this study, a series of binary choice tests was conducted in a wind tunnel to investigate whether background odors emitted from cherry, blueberry, blackberry, or raspberry fruits would affect the level of Drosophila suzukii (Matsumura) attraction to its symbiotic yeast, Hanseniaspora uvarum (Niehaus) (Saccharomycetales: Saccharomycetaceae). Whether an increase in the intensity of background odors would affect the attractiveness of H. uvarum to D. suzukii was also investigated, either by increasing the number of cherry or raspberry fruit per cup or by increasing the number of fruit cups surrounding the cup baited with the yeast. In wind tunnel assays, background fruit odors interfering with D. suzukii attraction to the yeast varied among fruit types. Raspberry odor inhibited the attractiveness of H. uvarum to the fly the most, followed by blackberry odor, whereas cherry and blueberry odors had no significant impact on the attraction. An increase in the intensity of odors by adding more cherry or raspberry fruit per cup did not increase the impact of fruit odor on the attraction; however, adding more raspberry cups around H. uvarum linearly decreased its attractiveness, suggesting that background host fruit abundance and likely increase in host odor may influence D. suzukii attraction to yeast odor depending on host species.

Differential Morpho-Physiological and Transcriptomic Responses to Heat Stress in Two Blueberry Species.

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, and help in understanding the complex mechanisms involved in heat stress tolerance.

Comparative transcriptome analysis of nonchilled, chilled, and late-pink bud reveals flowering pathway genes involved in chilling-mediated flowering in blueberry.

BACKGROUND: Blueberry cultivars require a fixed quantity of chilling hours during winter endo-dormancy for vernalization. In this study, transcriptome analysis using RNA sequencing data from nonchilled, chilled, and late pink buds of southern highbush blueberry 'Legacy' was performed to reveal genes associated with chilling accumulation and bud break. RESULTS: Fully chilled 'Legacy' plants flowered normally whereas nonchilled plants could not flower. Compared to nonchilled flower buds, chilled flower buds showed differential expression of 89% of flowering pathway genes, 86% of MADS-box genes, and 84% of cold-regulated genes. Blueberry orthologues of FLOWERING LOCUS T (FT) did not show a differential expression in chilled flower buds (compared to nonchilled flower bud) but were up-regulated in late-pink buds (compared to chilled flower bud). Orthologoues of major MADS-box genes were significantly up-regulated in chilled flower buds and down-regulated in late-pink buds. Functional orthologues of FLOWERING LOCUS C (FLC) were not found in blueberry. Orthologues of Protein FD (FD), TERMINAL FLOWER 1 (TFL1), and LEAFY (LFY) were down-regulated in chilled flower buds and in late-pink buds compared to nonchilled flower bud. CONCLUSIONS: The changes from nonchilled to chilled and chilled to late-pink buds are associated with transcriptional changes in a large number of differentially expressed (DE) phytohormone-related genes and DE flowering pathway genes. The profile of DE genes suggests that orthologues of FT, FD, TFL1, LFY, and MADS-box genes are the major genes involved in chilling-mediated blueberry bud-break. The results contribute to the comprehensive investigation of the vernalization-mediated flowering mechanism in woody plants.

Local environment, not local adaptation, drives leaf-out phenology in common gardens along an elevational gradient in Acadia National Park, Maine.

PREMISE OF THE STUDY: Climate-driven changes in phenology are substantially affecting ecological relationships and ecosystem processes. The role of variation among species has received particular attention; for example, variation among species' phenological responses to climate can disrupt trophic interactions and can influence plant performance. Variation within species in phenological responses to climate, however, has received much less attention, despite its potential role in ecological interactions and local adaptation to climate change. METHODS: We constructed three common gardens across an elevation gradient on Cadillac Mountain in Acadia National Park, Maine, to test population-level responses in leaf-out phenology in a reciprocal transplant experiment. The experiment included three native species: low bush blueberry (Vaccinium angustifolium), sheep's laurel (Kalmia angustifolia), and three-toothed cinquefoil (Sibbaldiopsis tridentata). KEY RESULTS: Evidence for local adaptation of phenological response to temperature varied among the species, but was weak for all three. Rather, variation in phenological response to temperature appeared to be driven by local microclimate at each garden site and year-to-year variation in temperature. CONCLUSIONS: Population-level adaptations in leaf-out phenology appear to be relatively unimportant for these species in Acadia National Park, perhaps a reflection of strong genetic mixing across elevations, or weak differences in selection on phenological response to spring temperatures at different elevations. These results concur with other observational data in Acadia and highlight the utility of experimental approaches to understand the importance of annual and local site variation in affecting phenology both among and within plant species.

The Cold-Regulated Genes of Blueberry and Their Response to Overexpression of VcDDF1 in Several Tissues.

Expression of blueberry cold-regulated genes (VcCORs) could play a role in the variable cold hardiness of blueberry tissues. In this study, transcriptome comparisons were conducted to reveal expression of VcCORs in non-acclimated leaves, flower buds, and flowers of both non-transgenic and transgenic blueberries containing an overexpressed blueberry DWARF AND DELAYED FLOWERING gene (VcDDF1) as well as in fully chilled flower buds of non-transgenic blueberry. In non-transgenic blueberries, 57.5% of VcCOR genes showed differential expression in at least one of the three pairwise comparisons between non-acclimated leaves, flower buds, and flowers, and six out of nine dehydration-responsive element-binding factors showed differential expression. In addition, expression of VcDDF1 was not cold-inducible in non-transgenic blueberries and had higher expression in flowers than in leaves or non-acclimated flower buds. In transgenic blueberries, overexpression of VcDDF1 resulted in higher VcDDF1 expression in leaves than in flower buds and flowers. VcDDF1 overexpression enhanced expression of blueberry CBF1 and CBF3 in leaves and repressed expression of CBF3 in both flower buds and flowers. Overall, the results revealed tissue-specific expression patterns of VcCORs. The responses of VcCORs to overexpression of VcDDF1 suggest that it is possible to increase plant cold hardiness through overexpression of a non-cold-inducible gene.

Regulation of gene expression in roots of the pH-sensitive Vaccinium corymbosum and the pH-tolerant Vaccinium arboreum in response to near neutral pH stress using RNA-Seq.

BACKGROUND: Blueberries are one of the few horticultural crops adapted to grow in acidic soils. Neutral to basic soil pH is detrimental to all commonly cultivated blueberry species, including Vaccinium corymbosum (VC). In contrast, the wild species V. arboreum (VA) is able to tolerate a wider range of soil pH. To assess the molecular mechanisms involved in near neutral pH stress response, plants from pH-sensitive VC (tetraploid) and pH-tolerant VA (diploid) were grown at near neutral pH 6.5 and at the preferred pH of 4.5. RESULTS: Transcriptome sequencing of root RNA was performed for 4 biological replications per species x pH level interaction, for a total of 16 samples. Reads were mapped to the reference genome from diploid V. corymbosum, transforming ~55% of the reads to gene counts. A quasi-likelihood F test identified differential expression due to pH stress in 337 and 4867 genes in VA and VC, respectively. Both species shared regulation of genes involved in nutrient homeostasis and cell wall metabolism. VA and VC exhibited differential regulation of signaling pathways related to abiotic/biotic stress, cellulose and lignin biosynthesis, and nutrient uptake. CONCLUSIONS: The specific responses in VA likely facilitate tolerance to higher soil pH. In contrast, response in VC, despite affecting a greater number of genes, is not effective overcoming the stress induced by pH. Further inspection of those genes with differential expression that are specific in VA may provide insight on the mechanisms towards tolerance.

Bearing fruit: flower removal reveals the trade-offs associated with high reproductive effort for lowbush blueberry.

Past studies have shown that taxa from disparate groups often respond similarly to reduced reproductive effort. These common responses imply that high reproductive effort trades off with a consistent set of other life functions for most angiosperms, albeit modulated by their growth form and life history. However, many questions remain about reproductive trade-offs in plants, including just how many other life functions they involve, how diverse these functions may be, and how the severity of these trade-offs may vary through time. To address these questions in a long-lived, iteroparous shrub, we performed flower removal on plots of lowbush blueberry, Vaccinium angustifolium (Ericaceae), over 3 years. We found significant physiological differences between removal and control plots for ten diverse traits. Vegetative phenology was shifted earlier by about 20% in removal plots, and removal plots had about 15% more vegetative biomass by mid-season as well. Removal plots produced about 10% more ripe fruit per reproductive node by harvest than control plots, and reproductive nodes in removal plots produced at least one fruit by harvest about 6% more often. While fruit water content and titratable acidity were increased by removal, other fruit traits, such as sugar content and fresh mass, were not. The strength of the removal effect varied significantly by year for seven traits; for many, such as vegetative mass/stem and ripe fruit production/node, the effect was stronger in years with more stressful abiotic conditions. Our results demonstrate that there are tangible but variable costs to high reproductive effort for flowering plants.

Differential Response of a Local Population of Entomopathogenic Nematodes to Non-Native Herbivore Induced Plant Volatiles (HIPV) in the Laboratory and Field.

Recent work has shown the potential for enhanced efficacy of entomopathogenic nematodes (EPN) through their attraction to herbivore induced plant volatiles. However, there has been little investigation into the utilization of these attractants in systems other than in those in which they were identified. We compared (E)-ß-caryophyllene and pregeijerene in the highbush blueberry (Vaccinium corymbosum) agroecosystem in their ability to enhance the attraction of EPN to and efficacy against the system's herbivore, oriental beetle (Anomala orientalis). The relative attractiveness of (E)-ß-caryophyllene and pregeijerene to a local isolate of the EPN species Steinernema glaseri was tested in a six-arm olfactometer in the laboratory to gather baseline values of attraction to the chemicals alone in sand substrate before field tests. A similar arrangement was used in a V. corymbosum field by placing six cages with assigned treatments and insect larvae with and without compound into the soil around the base of 10 plants. The cages were removed after 72 h, and insect baits were retrieved and assessed for EPN infection. The lab results indicate that in sand alone (E)-ß-caryophyllene is significantly more attractive than pregeijerene to the local S. glaseri isolate Conversely, there was no difference in attractiveness in the field study, but rather, native S. glaseri were more attracted to cages with G. mellonella larvae, no larvae, and cages with the blank control and G. mellonella larvae.

Characterizing Damage of Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) in Blueberries.

Brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is a severe economic pest of growing importance in the United States, Canada, and Europe. While feeding damage from H. halys has been characterized in tree fruit, vegetables, and agronomic crops, less is known about the impacts of stink bugs on small fruits such as blueberries. In this study, we examined H. halys feeding on two representative early and late ripening blueberry cultivars in Oregon and New Jersey. This research examined how different densities of H. halys confined on blueberry clusters for week-long periods affected fruit quality at harvest. After fruit were ripe, we stained and quantified the number of salivary sheaths on berries as an indication of feeding pressure. Feeding by H. halys damaged the fruits by causing increased levels of external discoloration, and internal damage in the form of tissue necrosis. Exposure of berries to H. halys was also associated with decreasing berry weights and lower soluble solids in fruits. However, the different cultivars did not respond consistently to feeding pressure from H. halys. Weekly variability in feeding pressure of two of the cultivars as quantified by the number of stylet sheaths per berry was largely accounted for by environmental variables. We conclude that H. halys does have potential to severely damage blueberries and may become an important economic pest. Characterization of damage is important because correct identification of insect damage is key for successful management.

Assessing the Economic Importance of Dasineura oxycoccana (Diptera: Cecidomyiidae) in Northern Highbush Blueberries.

Infestation by blueberry gall midge, Dasineura oxycoccana Johnson, is common in northern highbush blueberries, but its effects on crop productivity are unknown. We examined whether infestation by blueberry gall midge reduces flower bud production when compared with uninfested shoots, and how infestation at different times affects the crop response. From the fall of 2009 to the spring of 2011, the number of flower buds on infested and uninfested shoots of blueberry bushes was counted and compared. Despite causing branching of vegetative growth, there was no significant effect of infestation on flower bud production. During the summer of 2010, damaged shoots were marked throughout the growing season in June, July, or August. The number of flower buds set per shoot declined with later infestation dates, and shoots damaged in August had significantly fewer buds than those damaged in June and July. We discuss the implications of these findings for management of blueberry gall midge in northern highbush blueberry.

Pollen loads and specificity of native pollinators of lowbush blueberry.

The reproduction of lowbush blueberry (Vaccinium angustifolium Aiton) is closely tied to insect pollination, owing to self-incompatibility. Many species are known to have greater pollination efficiency than the introduced Apis mellifera L., commonly used for commercial purposes. In this study, we measured the pollen loads of several antophilous insect species, mostly Apoidea and Syrphidae, present in four lowbush blueberry fields in Lac-St-Jean, Québec. To measure pollen loads and species specificity toward V. angustifolium, we net-collected 627 specimens of pollinators, retrieved their pollen loads, identified pollen taxa, and counted pollen grains. We found that the sizes of pollen loads were highly variable among species, ranging from a few hundred to more than 118,000 pollen grains per individual. Bombus and Andrena species in particular carried large amounts of Vaccinium pollen and thus may have greater pollination efficiency. Also, two species (Andrena bradleyi Viereck and Andrena carolina Viereck) showed nearly monolectic behavior toward lowbush blueberry. Finally, we identified alternative forage plants visited by native pollinators, notably species of Acer, Rubus, Ilex mucronata, Ledum groenlandicum, and Taraxacum. Protecting these flowering plants should be part of management practices to maintain healthy pollinator communities in a lowbush blueberry agroecosystem.

The Vaccinium corymbosum FLOWERING LOCUS T-like gene (VcFT): a flowering activator reverses photoperiodic and chilling requirements in blueberry.

KEY MESSAGE: The blueberry FLOWERING LOCUS T ( FT )-like gene ( VcFT ) cloned from the cDNA of a tetraploid, northern highbush blueberry ( Vaccinium corymbosum L.) is able to reverse the photoperiodic and chilling requirements and drive early and continuous flowering. ABSTRACT: Blueberry is a woody perennial bush with a longer juvenile period than annual crops, requiring vernalization to flower normally. Few studies have been reported on the molecular mechanism of flowering in blueberry or other woody plants. Because FLOWERING LOCUS T (FT) from Arabidopsis thaliana plays a multifaceted role in generating mobile molecular signals to regulate plant flowering time, isolation and functional analysis of the blueberry (Vaccinium corymbosum L.) FT-like gene (VcFT) will facilitate the elucidation of molecular mechanisms of flowering in woody plants. Based on EST sequences, a 525-bpVcFT was identified and cloned from the cDNA of a tetraploid, northern highbush blueberry cultivar, Bluecrop. Ectopic expression of 35S:VcFT in tobacco induced flowering an average of 28 days earlier than wild-type plants. Expression of the 35S:VcFT in the blueberry cultivar Aurora resulted in an extremely early flowering phenotype, which flowered not only during in vitro culture, a growth stage when nontransgenic shoots had not yet flowered, but also in 6-10-week old, soil-grown transgenic plants, in contrast to the fact that at least 1 year and 800 chilling hours are required for the appearance of the first flower of both nontransgenic 'Aurora' and transgenic controls with the gusA. These results demonstrate that the VcFT is a functional floral activator and overexpression of the VcFT is able to reverse the photoperiodic and chilling requirements and drive early and continuous flowering.

Oviposition efficacy of Drosophila suzukii (Diptera: Drosophilidae) on different cultivars of blueberry.

Drosophila suzukii (Matsumura) is an important pest of thin-skinned fruits including blueberry, raspberry, strawberry, and cherry. Blueberry was introduced into Japan in the 1950s, and severe economic losses attributable to D. suzukii were first reported in 2002. The objective of this study was to elucidate whether oviposition behavior varies among blueberry cultivars having different firmness of fruit. Fruit firmness in 12 cultivars of highbush blueberry (Vaccinium corymbosum L.) and rabbiteye blueberry (Vaccinium virgatum Aiton) was determined using a rheometer. More eggs tended to be laid in berries of cultivars possessing softer fruits than in those having firmer fruits. Choice tests, where one female was allowed to oviposit on blueberry fruits with different firmness, showed that softer fruits were more vulnerable to D. suzukii females than firmer fruits.

Assessment of the impact of innovative fertilization methods compared to traditional fertilization in the cultivation of highbush blueberry.

The aim of the research was to evaluate fertilization technologies for the indicators of the quality and quantity of highbush blueberry yield. In the experiment, a similar level of mineral fertilization was used in all treatments. The experiment was to show the differences between fertilization with biostimulation and without biostimulation. The research was carried out in two seasons (2019-2020) on ´Bluecrop` shrubs growing in the Blueberry Experimental Field in central Poland (51° 55'42.7 "N 20° 59'28.7" E). Shrubs grow at a distance of 1 x 3 m. Plants are rejuvenated every year in spring and irrigated by drip. The experiment was carried out in a random block design (4 fertilizer treatments x 5 replications x 6 bushes). The experiment assessed the effect of fertilization on yield, berry mass, fruit setting, leaf surface and physicochemical parameters of fruit. Based on the conducted research, it was proved that the applied fertilization technologies had a significant impact on the size and quality of the yield of "Bluecrop" highbush blueberry. Particularly noteworthy is the fertilization technology with biostimulation (treatment T4), which has a positive effect on the yield, fruit mass, percentage of setting and firmness of the berries. Analysis of the issue in the light of the results of the conducted research shows that the use of biostimulated products has an important impact on the intensification of production while maintaining good quality of fruits. Through research, the positive effect of fertilization programs with biostimulation (treatment T4) on the most important production parameters of blueberry fruit from the producer's point of view has been proven.

A bZIP transcription factor VabZIP12 from blueberry induced by dark septate endocyte improving the salt tolerance of transgenic Arabidopsis.

Dark septate endophytes (DSEs) have attracted much attention due to their positive roles in plant growth as well as resistance to various abiotic stresses. However, there are no reports on the molecular mechanisms of DSE fungi to improve salt tolerance in plants. In this study, the blueberry seedlings inoculated with T010, a beneficial DSE fungus reported previously, grew more vigorously than the non-inoculated control under salt stress. Physiological indicators showed that T010 inoculation increased antioxidant activities of blueberry roots. To explore its molecular mechanism, we focused on the bZIP TFs VabZIP12, who was highly up-regulated with T010 inoculation under salt stress. Further studies showed that VabZIP12, as a transcription activator, could combine both G-Box 1 and G-Box 2 motifs. Moreover, overexpression of VabZIP12 enhanced salt stress tolerance through increasing the activities of the enzymatic antioxidants in the transgenic Arabidopsis with up-regulation the related genes. These results indicated that the induction of VabZIP12 contribute to improving the tolerance of blueberry to salt stress by T010 inoculation.

Variation in highbush blueberry floral volatile profiles as a function of pollination status, cultivar, time of day and flower part: implications for flower visitation by bees.

BACKGROUND AND AIMS: Studies of the effects of pollination on floral scent and bee visitation remain rare, particularly in agricultural crops. To fill this gap, the hypothesis that bee visitation to flowers decreases after pollination through reduced floral volatile emissions in highbush blueberries, Vaccinium corymbosum, was tested. Other sources of variation in floral emissions and the role of floral volatiles in bee attraction were also examined. METHODS: Pollinator visitation to blueberry flowers was manipulated by bagging all flowers within a bush (pollinator excluded) or leaving them unbagged (open pollinated), and then the effect on floral volatile emissions and future bee visitation were measured. Floral volatiles were also measured from different blueberry cultivars, times of the day and flower parts, and a study was conducted to test the attraction of bees to floral volatiles. KEY RESULTS: Open-pollinated blueberry flowers had 32 % lower volatile emissions than pollinator-excluded flowers. In particular, cinnamyl alcohol, a major component of the floral blend that is emitted exclusively from petals, was emitted in lower quantities from open-pollinated flowers. Although, no differences in cinnamyl alcohol emissions were detected among three blueberry cultivars or at different times of day, some components of the blueberry floral blend were emitted in higher amounts from certain cultivars and at mid-day. Field observations showed that more bees visited bushes with pollinator-excluded flowers. Also, more honey bees were caught in traps baited with a synthetic blueberry floral blend than in unbaited traps. CONCLUSIONS: Greater volatile emissions may help guide bees to unpollinated flowers, and thus increase plant fitness and bee energetic return when foraging in blueberries. Furthermore, the variation in volatile emissions from blueberry flowers depending on pollination status, plant cultivar and time of day suggests an adaptive role of floral signals in increasing pollination of flowers.

Ecological impacts of the N-viro biosolids land-application for wild blueberry (Vaccinium angustifolium. Ait) production in Nova Scotia.

Land application of biosolids from processed sewage sludge may deteriorate soil, water, and plants. We investigated the impact of the N-Viro biosolids land-application on the quality of the soil water that moved through Orthic Humo-Ferric Podzols soil of Nova Scotia (NS) at the Wild Blueberry Research Institute, Debert, NS Canada. In addition, the response of major soilproperties and crop yield was also studied. Wild blueberry (Vaccinium angustifolium. Ait) was grown under irrigated and rainfed conditions in 2008 and 2009. Four experimental treatments including (i) NI: N-Viro irrigated, (ii) NR: N-Viro rainfed, (iii) FI: inorganic fertilizer irrigated, and (iv) FR: inorganic fertilizer rainfed (control) were replicated 4 times under randomized complete block design. Soil samples were collected at the end of each year and analyzed for changes in cation exchange capacity (CEC), soil organic matter (SOM), and pH.Soil water samples were collected four times during the study period from the suction cup lysimeters installed within and below crop root zone at 20 and 40 cm depths, respectively. The samples were analyzed for a range of water quality parameters including conductance, hardness, pH, macro- and micronutrients, and the infectious pathogens Escherichia coli (E. coli) and salmonella. Berries were harvested for fruit yield estimates. Irrigation significantly increased CEC during 2008 and the soil pH decreased from 4.93 (2008) to 4.79 (2009). There were significant influences of irrigation, fertilizer, and their interaction, in some cases, on most of the soil water quality parameters except on the infectious bacteria. No presence of E. coli or salmonella were observed in soil and water samples, reflecting the absence of these bacteria in biosolids used in this experiment. Nutrient concentration in the soil water samples collected from the four treatments were higher in the sequence NI > NR > FI > FR. The irrigation treatment had significant effect on the unripe fruit yield. We conclude that the comparable performance of N-Viro biosolids and the increasing prices of inorganic fertilizers would compel farmers to use economically available N-Viro biosolids that, coupled with the supplemental irrigation, did not deteriorate the studied soil properties, soil water quality, and the wild blueberry yield during this experiment.

Effects of water and fertilizer coupling on the physiological characteristics and growth of rabbiteye blueberry.

Understanding the impact of irrigation and fertilizer on rabbiteye blueberry (Vaccinium virgatum) physiology is necessary for its precision planting. Here, we applied varied irrigation and fertilizer under completely randomized experimental design to see its impact on the physiological characteristics and bush growth of rabbiteye blueberries. A comprehensive evaluation of the membership function was used to establish the best water-fertilizer coupling regimes. Rabbiteye blueberry enhanced the net photosynthetic rate, stomatal conductance and transpiration rate of leaf and improved its photosynthetic capacity at maximum level of irrigation water and fertilizer application (F3W4). The high fertilizer-medium water treatment (F3W3) increased leaf-soluble protein contents. The medium fertilizer-medium water treatment (F2W3, F2W2) increased leaf- soluble sugar, superoxide dismutase, and chlorophyll contents; decreased the malondialdehyde content; and enhanced leaf resistance and metabolism. It also promoted the growth of flower buds and new shoots. Combined membership function and cluster analyses revealed that the optimal water and fertilizer conditions for promoting rabbiteye blueberry plant growth were the medium fertilizer-medium water [(NH4)2SO4:Ca(H2PO4)2:K2SO4 at 59:10:20 g plant-1; 2.5 L water plant-1], medium fertilizer-medium-high water [(NH4)2SO4:Ca(H2PO4)2:K2SO4 at 59:10:20 g plant-1; 3.75 L water plant-1], and high fertilizer-medium-high water [(NH4)2SO4:Ca(H2PO4)2:K2SO4 at 118:20:40 g plant-1; 3.75 L water plant-1] treatments. The findings of this study could be used in improving the precision and efficacy of rabbiteye blueberry planting in Guizhou, China. Such an approach can increase the productivity and profitability for local fruit farmers.