Genetic analysis of freezing tolerance in blueberry (Vaccinium section Cyanococcus)

An understanding of the genetic control of freezing tolerance (FT) in woody perennials is important for the effective selection and development of plants with a broader climatic adaptation. This study was undertaken to examine the inheritance and gene action of FT in segregating populations of a woody perennial blueberry (Vaccinium, section Cyanococcus). Two backcross populations were derived from interspecific hybrids of the diploid species Vaccinium darrowi and Vaccinium caesariense, which are widely divergent in their FT. The bud FTs of uniformly cold acclimated plants of parental, F₁, and two backcross populations were evaluated with a laboratory controlled freeze-thaw regime, followed by a visual assessment of injury. FT (LT₅₀) was defined as the temperature causing 50% of the flower buds to be injured. Data indicate that the two parents were homozygous for genes for low or high FT. Freezing-tolerance values of the parental and F₁ populations indicate that freeze-sensitivity is a partially dominant trait. Results from reciprocal crosses revealed that there was no significant maternal influence on freezing tolerance. Parental phenotypes were fully recovered in 40–42 plants of each testcross population, suggesting that FT is determined by relatively few genes. The degree of dominance and an analysis of generation means revealed that FT in blueberry is controlled largely by additive gene effects and, to a lesser degree, by dominance gene effects. Testing of various genetic models indicated that FT inheritance can be adequately explained by a simple additive-dominance model; however, two epistatic models involving additive-additive and dominance-dominance interactions also fit the data. Received: 25 May 1999 / Accepted: 16 June 1999     

Blueberry-induced changes in spatial working memory correlate with changes in hippocampal CREB phosphorylation and brain-derived neurotrophic factor (BDNF) levels

Phytochemical-rich foods have been shown to be effective at reversing age-related deficits in memory in both animals and humans. We show that a supplementation with a blueberry diet (2% w/w) for 12 weeks improves the performance of aged animals in spatial working memory tasks. This improvement emerged within 3 weeks and persisted for the remainder of the testing period. Memory performance correlated well with the activation of cAMP-response element-binding protein (CREB) and increases in both pro- and mature levels of brain-derived neurotrophic factor (BDNF) in the hippocampus. Changes in CREB and BDNF in aged and blueberry-supplemented animals were accompanied by increases in the phosphorylation state of extracellular signal-related kinase (ERK1/2), rather than that of calcium calmodulin kinase (CaMKII and CaMKIV) or protein kinase A. Furthermore, age and blueberry supplementation were linked to changes in the activation state of Akt, mTOR, and the levels of Arc/Arg3.1 in the hippocampus, suggesting that pathways involved in de novo protein synthesis may be involved. Although causal relationships cannot be made among supplementation, behavior, and biochemical parameters, the measurement of anthocyanins and flavanols in the brain following blueberry supplementation may indicate that changes in spatial working memory in aged animals are linked to the effects of flavonoids on the ERK-CREB-BDNF pathway.     

In vitro and in vivo evaluation of the prebiotic activity of water-soluble blueberry extracts

The prebiotic effects of water extracts of two blueberry (BBE) cultivars (‘Centurion’ and ‘Maru’) were studied using pure and mixed cultures of human faecal bacteria. The results demonstrated for the first time that addition of BBE from both cultivars to broth media containing pure cultures of Lactobacillus rhamnosus and Bifidobacterium breve resulted in a significant increase (P < 0.05–0.0001) in the population size of these strains. Batch fermentation system was used to monitor the effect of BBE addition on the mixed faecal bacterial populations (obtained from healthy human donors). Addition of BBE from both cultivars to batch cultures inoculated with mixed human faecal cultures resulted in a significant increase in the number of lactobacilli (P < 0.01–0.0001) and bifidobacteria (P < 0.05–0.0001). Furthermore, a significant influence on the population size of lactobacilli and bifidobacteria was observed after administration of extracts from both cultivars to rats daily for 6 days in comparison with the control group. In rats gavaged orally with 4 ml kg⁻¹ day⁻¹ of BBE for 6 days, the population size of lactobacilli (P < 0.05) and bifidobacteria (P < 0.05–0.01) was increased significantly. We hypothesize that BBE could modify the bacterial profile by increasing the numbers of beneficial bacteria and thereby improving gut health.     

Twig pre-harvest temperature significantly influences effective cryopreservation of Vaccinium dormant buds

Cryopreservation of temperate woody-plant material by dormant buds is less expensive than using shoot tips isolated from tissue cultured plants; however currently, dormant buds are used only for preservation of selected temperate tree and shrub species. Using dormant buds could be an efficient strategy for long-term preservation of blueberry (Vaccinium L.) genetic resources. In this study, viability of V. hybrid ‘Northsky’ (PI 554943) dormant buds was evaluated at 30 harvest dates over three consecutive fall/winter seasons to determine the optimal harvest time that promotes high post cryopreservation viability. Twigs with dormant buds were cut into 70 mm segments containing at least two nodes, desiccated, slowly cooled, stored in liquid nitrogen vapor and tested for post-cryopreservation regrowth. The highest regrowth of cryopreserved dormant buds was observed for buds harvested in mid-December and during the first half of January. Pearson's correlation coefficients were computed to evaluate the association between bud characteristics and viability at harvest date and logistic regression models were fit to test the ability of twig characteristics and temperatures to predict post cryopreservation bud viability. Post-cryopreservation viability was negatively correlated (p < 0.05) with average minimum, maximum and daily mean temperature preceding the bud harvest but was not correlated with the dormant bud initial and end moisture content, twig diameter, the number of dormant buds/cm of twig length and the number of days in desiccation. Regression tree analysis suggested post-cryopreservation viability to be between 52 and 80% for dormant buds harvested after a 10 day average maximum air temperature of <11.2 °C. Pre-harvest air temperature was a significant indicator of optimal dormant bud harvest time to produce adequate viability for long term preservation of blueberry genetic resources.     

Molecular cloning and characterization of Polygalacturonase-Inhibiting Protein and Cinnamoyl-Coa Reductase genes and their association with fruit storage conditions in blueberry (Vaccinium corymbosum)

Blueberry is a widely grown and easily perishable fruit crop. An efficient post-harvest handling is critical, and for that purpose gene technology methods have been part of ongoing programmes to improve crops with high food values such as blueberry. Here we report the isolation, cloning, characterization and differential expression levels of two cDNAs encoding Polygalacturonase-Inhibitor Protein (PGIP) and Cinnamoyl-Coa Reductase (CCR) from blueberry fruits in relation to various storage conditions. The open reading frame of PGIP and CCR encodes a polypeptide of 329 and 347 amino acids, respectively. To assess changes in the expression of blueberry PGIP and CCR after harvest, a storage trial was initiated. The northern blots hybridization showed a clear differential expression level of PGIP and CCR between freshly harvested and stored fruits as well as between fruits stored under various storage conditions. Although the prospects of exploiting such a strategy for crop improvement are limited, the results provide further insight into the control of the quality over the storage period at the molecular level.     

Occurrence of unreduced pollen in diploid blueberry species,Vaccinium sect. Cyanococcus

Summary A total of 1475 individuals belonging to 43 natural populations of seven diploid (2x) blueberry species (Vaccinium section Cyanococcus) and two natural interspecific 2x hybrid populations were evaluated for unreduced pollen production. Significant differences were found in the frequency of unreduced pollen producers between species and within and between populations of the same species. Individuals with 1% or more unreduced pollen were considered unreduced pollen producers. The average frequency of unreduced pollen producers in these diploid species was 13.5%, ranging from 7.4% (V. corymbosum) to 18.4% (V. darrowii). The frequency of unreduced pollen grains in individual clones varied from <-1% to 28.6%. The production of unreduced pollen was not associated with male fertility. The widespread occurrence of unreduced pollen in the diploid species should allow the introgression of this germ plasm to the tetraploid level via unilateral sexual polyploidization.New Jersey Agricultural Experiment Station, Publication No. D-12163-18-91, supported by State and U.S. Federal funds, USDA-ARS Specific Cooperative Agreement No. 58-3615-9-068     

Identification of Fungi in the Botryosphaeriaceae Family Associated with Stem Blight of Vaccinium spp. in the Southeastern United States

Stem blight is a major disease of blueberry caused by Botryosphaeriaceae fungi. Chemical and cultural management options are limited, putting emphasis on breeding efforts to identify sources of resistance. The efficacy and durability of host resistance could be impacted by the species composition of the pathogen population in a region and by the isolates employed in the screenings used to identify the resistance. Samples (365) were collected from southern highbush (SHB) and rabbiteye blueberry (REB) cultivars from 28 sites in the southeastern US (AL, FL, GA, NC, and SC). Colony morphology identified 86% of the isolates as Botryosphaeriaceae. Conidia morphology and Maximum Likelihood analysis of the Internal Transcribed Spacer rDNA regions (ITS), translation elongation factor one alpha (tef1-α), and β-tubulin were used to identify isolates at genera or species level. A PCR-restriction fragment length polymorphism (PCR-RFLP) test was used to identify isolates to genus. Neofusicoccum and Lasiodiplodia were the predominant genera. N. kwambonambiense, N. ribis, L. theobromae and L. pseudotheobromae were the most common species isolated. Phylogenies conducted with a limited number of isolates indicated non-clonal and potentially diverse populations occur on blueberry that warrant additional study. Botryosphaeria corticis, B. dothidea, and Diplodia seriata were isolated infrequently.     

Cinnamyl alcohol: An attractant of the flower thrips Frankliniella intonsa

Flower-inhabiting thrips find hosts using olfactory and visual cues. In this study, we report the identification of a plant-produced attractant of the flower thrips Frankliniella intonsa (Trybom), an important agricultural pest worldwide. GC–MS analysis of solid-phase microextraction samples from blueberry flowers, Vaccinium corymbosum L., that mediate the attraction of adult F. intonsa revealed that the major component was cinnamyl alcohol, followed by cinnamyl acetate, cinnamaldehyde, germacrene D, β-bourbonene, β-caryophyllene, and benzyl benzoate. The biological activity of the floral compounds was investigated using commercial cinnamaldehyde, cinnamyl alcohol, β-caryophyllene, cinnamyl acetate, and benzyl benzoate in hot pepper (Capsicum annuum L.) fields. Significantly more F. intonsa males and females were caught in red delta traps with cinnamyl alcohol than in all other traps. Cinnamaldehyde and cinnamyl acetate attracted adult F. intonsa but were not as attractive as cinnamyl alcohol. β-Caryophyllene and benzyl benzoate were not attractive. Furthermore, the addition of four minor components to cinnamyl alcohol did not result in increased trap catches relative to cinnamyl alcohol alone, indicating that cinnamyl alcohol is responsible for attracting adult F. intonsa toward blueberry flowers. Therefore, this phenylpropanoid could be used as an effective lure for monitoring and controlling F. intonsa.