Efforts are underway to develop genetic linkage maps for two interspecific blueberry populations (Vaccinium darrowi × V. elliottii and V. caesariense-derived populations). To date, 72 RAPD markers have been mapped, and another 200 markers have been identified as suitable for mapping in the V. darrowi × V. elliottii-derived population. Inheritance of 40 RAPD markers has been followed, and additional 40 RAPD markers have been identified as suitable for mapping in the V. darrowi × V. caesariense population. These two populations are comprised of individual plants that should have a wide range of chilling requirements. At a later date, plants will be classified according to their chilling requirements to identify RAPD markers that cosegregate with chilling requirements. Presently, a bulked-segregant analysis is being performed on a tetraploid breeding population (primarily V. corymbosum) to identify RAPD markers linked to chilling requirement genes.
Eighty-seven highbush blueberry and species-introgressed blueberry cultivars were evaluated for fruit firmness in the 1998-2000 growing seasons with a FirmTech 1 automated firmness tester. Significant differences were observed among cultivars. An average firmness of 136.1 g·mm-1 of deflection (g·mm-1 dfl) was observed across all studied cultivars, and a range of 80.4 g·mm-1 dfl (`Herbert') to 189.0 g·mm-1 dfl (`Pearl River'). Species ancestry was not consistently related to firmness; however, cultivars with higher firmness values often possessed a higher percentage of Vaccinium darrowi Camp and V. ashei Reade ancestry. Conversely, cultivars with softer than average fruit often possessed a higher percentage of lowbush (V. angustifolium Ait.) ancestry. This information may help to identify sources of breeding material for increased firmness in highbush blueberry hybrids.
As a first step in determining the phylogeny of the rabbiteye blueberry (Vaccinium ashei Reade), the potential fertility levels among its purported diploid ancestral species. Vaccinium atrococcum (Gray) Heller, Vaccinium caesariense MacKenzie, Vaccinium darrowi Camp, and Vaccinium tenellum Ait., were determined. Differences existed among species in female fertility as determined by total number of seeds and number of potentially viable seeds. The number of seeds per berry was lower than that reported for highbush and rabbiteye blueberry cultivars. No differences were found among species in potential male fertility as indicated by percent stainable pollen. Pollen diameter was not a good criterion for distinguishing species or ploidy levels. Comparison of fertility of diploid species with that of purported polyploid derivatives is suggested as a possible aid in determining phylogeny in Vaccinium. Unreduced pollen grains were detected in only 2 of the 4 species in this study. The average return from heterploid (4n × 2n) crosses was 3 seedlings per 1000 pollinations.
Progenies and clones of interspecific hybrid blueberries were evaluated for annual fraction of canopy volume (FCYV) and for difference in fraction of canopy volume between control and stressed plants [FCYV(C) - FCYV(S)] in a moderate water-deficit environment. The FCYV(C) - FCYV(S) data were used to determine combining ability effects. In addition, physiological processes of attached leaves of the clones were monitored with a portable photosynthesis apparatus. Specific combining ability (SCA) effects were significant for FCYV(C) - FCYV(S). The clone with the lowest mean for FCYV(C) - FCYV(S) was US75, a hybrid of Vaccinium darrowi Camp × V. corymbosum L. Clone JU64 (V. myrsinites Lamark × V. angustifolium Aiton) also had a low FCYV(C) - FCYV(S) mean, and its two progenies (JU64 × JU11 and G362 × JU64) had low progeny means. Stomatal conductance was lowered when blueberries were exposed to atmospheric and/or soil moisture stress that resulted in lower transpiration and photosynthesis and increased or equal water-use efficiencies (WUE). Blueberry plants adjusted to moisture stress as the season progressed by lowering stomatal conductance and increasing WUE. In particular, stressed plants of US75 and JU64 had equal or higher WUE values than control plants. US226 was the most drought-susceptible clone in the study, and its stomata did not appear to be as responsive to moisture stress as the other clones. Breeding for higher WUE in a dry environment appears possible with the germplasm used in this study.
Three blueberry (Vaccinium section Cyanococcus) genotypes, that have different chilling requirements and levels of cold hardiness, were studied. Depth of dormancy was evaluated and water status was determined, using nuclear magnetic resonance (NMR), throughout the accumulation of chilling that leads to release from dormancy. Among the two highbush cultivars studied, `Bluecrop' (Vaccinium corymbosum L.) was the most dormant and `Gulfcoast' (Vaccinium corymbosum L. x Vaccinium darrowi Camp) was the least dormant. The rabbiteye cultivar `Tifblue' (Vaccinium ashei Reade) had an intermediate dormancy. It appeared that the cultivar with the deepest dormancy had also the highest chilling requirement (CR). The NMR results showed that `Bluecrop' buds had the lowest relaxation times (T2), indicating that water was relatively more bound in `Bluecrop' buds than in the buds of the two other cultivars. Whatever the cultivar, no significant variation of T2s and water content of the buds was noted throughout the accumulation of chilling, even after CRs were satisfied. Within 1 day of forcing (24 °C, long day), there was a shift towards freer water but no change in the water content. Forcing was ineffective in freeing water until the plants received enough chilling to satisfy their CRs.
To determine if the net CO2 assimilation and water use efficiency (WUE) of highbush blueberry under high temperature can be improved genetically, gas exchange determinations were made for a selection of Vaccinium darrowi Camp (Florida 4B), a highbush cultivar (Bluecrop) (V. corymbosum L.), their F1 hybrid (US75), and two crosses of the F1 hybrid to another improved genotype (US239 and US245). All genotypes responded parabolically to increasing temperature at vapor pressure deficits <1 kPa. Maximum CO2 assimilation of US75 (15 µmol·s–1·m–2) was 30% to 40% higher than either parent. Carbon dioxide assimilation of US75 and Florida 4B was optimum at 30°C and that of ‘Bluecrop’ at 20°. The optimum for US239 was similar to ‘Bluecrop’, and that of US245 to Florida 4B. Florida 4B had higher WUEs than ‘Bluecrop’ at both 20° (5.64 µmol CO2/mmol H2O to 4.01) and 30° (3.73 to 2.53). US239 and US245 had significantly (P < 0.05) higher WUEs at 30° than did ‘Bluecrop’. Residual conductance to CO2 (gr) decreased in ‘Bluecrop’ when temperature was raised from 20° to 30°, but increased in all other genotypes. Due to the favorable gas exchange properties of US75 and US245 at 30°, we suggest that the high temperature tolerance of V. darrowi may be heritable and that US245 may be used to improve the heat tolerance of highbush blueberry.
Open-pollinated species, interspecific hybrid seedling populations, and selected clones propagated by softwood cuttings and tissue culture were used to further evaluate the use of dry soil to screen blueberry seedlings for drought resistance. These different propagation types of Vaccinium (subgenus Cyanococcus) were screened for drought resistance in a Galestown fine sandy loam soil. The soil was permitted to dry to progressively higher soil tension levels to a maximum of 0.8 MPa. The plants were evaluated by scoring them on a 1 to 9 shoot damage rating scale and by determining the fraction of total biomass partitioned as roots. Drought resistance in blueberries is heritable and there is a high degree of genetic diversity within and among Vaccinium spp. for resistance to water deficits. Southern species (Vaccinium darrowi Camp, V. elliottii Chapman, and V. ashei Reade) were more drought-resistant than northern species (V. corymbosum L., V. vacillans Torrey, and V. myrtilloides Michaux), which demonstrated the reliability of this soil screening method. Clones with half their germplasm from southern species were usually drought-resistant. This screening method can be used to select for shoot and root vigor and drought resistance in 6- to 7-month-old blueberry seedlings.
A portable, handheld, mechanical shaking device was developed and its effects on fruit detachment in rabbiteye bluebbery (Vaccinium ashei Reade) and southern highbush blueberry (hybrids of mostly Vaccinium corymbosum L. and Vaccinium darrowi Camp.) were evaluated. The instrument facilitated effective detachment of fruit within a branch, especially in rabbiteye blueberry (greater than 75%). Approximately 3 to 4 seconds of shaking was sufficient to detach the majority of the fruit. Differences in the extent of fruit detachment were observed across the genotypes, especially among southern highbush blueberry genotypes. The majority of fruit detachment in rabbiteye blueberry and most of the southern highbush blueberry genotypes occurred at the point of attachment of the pedicel to the berry, although a significant portion detached with the stem intact, resulting in stemmy fruit. Although only a small proportion of the detached fruit was immature in the rabbiteye blueberry genotypes, up to 23% of the detached fruit was immature in the southern highbush blueberry genotypes. Application of the abscission agents methyl jasmonate (MeJa; 20 mm) and ethephon (1000 mg·L−1) reduced the time required for fruit detachment on mechanical shaking by up to 5-fold. Together, these data indicate that the mechanical shaking device developed here is an effective tool for studying fruit detachment in blueberry. This instrument has potential applications in blueberry research programs evaluating fruit production. It can be used in breeding programs to aid in the selection of genotypes with fruit detachment characteristics that are potentially better suited for mechanical harvesting, and also in programs involving the screening and evaluation of abscission agents in blueberry.
Four chimeric bialaphos resistance (bar) genes driven by different promoters were evaluated for production of herbicide-resistant ‘Legacy’ blueberry plants (73.4% Vaccinium corymbosum L. and 25% Vaccinium darrowi Camp) through Agrobacterium tumefaciens (Smith & Towns.) Conn.-mediated transformation. When the bars were used as selectable marker genes, different promoters yielded different transformation frequencies. Three chimeric bar genes with the promoter nopaline synthase (nos), cauliflower mosaic virus (CaMV) 35S, or CaMV 34S yielded transgenic plants, whereas a synthetic (Aocs)3AmasPmas promoter did not lead to successful regeneration of transgenic plants. In addition, herbicide resistance in bar-expressing plants was influenced by the promoter strength. Under controlled environmental conditions, 3-month-old plants from six single-copy transgenic events with 35S∷bar or nos∷bar, as well as those nontransgenic plants, were sprayed with herbicide glufosinate ammonium (GS) at five levels (0, 750, 1500, 3000, and 6000 mg·L−1). Evaluations on leaf damage 2 weeks after spraying indicated that all transgenic plants exhibited much higher herbicide resistance than nontransgenic plants. Additionally, the transgenic plants with the 35S∷bar showed a higher herbicide resistance than those with the nos∷bar. After application of 6000 mg·L−1 GS, over 90% of the leaves from plants with the 35S∷bar and 19.5% to 51.5% of the leaves from plants with the nos∷bar showed no symptom of herbicide damage, whereas only 5% of leaves from the nontransgenic had no damage. One-year-old, field-grown plants from four transgenic events with the nos∷bar were evaluated for herbicide resistance after spraying with 750 mg·L−1 GS. Transgenic plants survived with variations in the level of foliar damage; in contrast, all nontransgenic plants died. This study is the first investigation of different promoters for engineering transgenic blueberry plants.
Two abscission agents, ethephon and methyl jasmonate, were investigated in five studies to determine their potential for increasing fruit detachment during harvest in rabbiteye (Vaccinium ashei Reade) and southern highbush (hybrids based largely on Vaccinium corymbosum L. and Vaccinium darrowi Camp.) blueberry. In the first study with a rabbiteye blueberry genotype, T-451, ethephon applications up to 1000 mg·L−1 did not affect fruit drop but reduced fruit detachment force (FDF) by up to 21%. In the second study with two southern highbush blueberry genotypes, ethephon (up to 1500 mg·L−1) and methyl jasmonate (MeJa; up to 10 mm) applications resulted in significant fruit drop in ‘Star’ but neither of the growth regulators affected the fruit detachment characteristics of ‘Farthing’. In a third study with rabbiteye blueberry genotypes, MeJa applications of 10, 20, and 30 mm displayed an increasing linear trend in fruit drop in ‘Climax’ and linear and quadratic trends in fruit drop in ‘Powderblue’. In a fourth study with ‘Powderblue’, MeJa (20 mm) and ethephon (1000 mg·L−1) applications resulted in rapid and significant fruit drop. The fruit drop induced by MeJa in this study was attenuated by the coapplication of aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor, suggesting that MeJa induced fruit detachment partly through its effects on ethylene biosynthesis. In another study with the southern highbush blueberry genotype, O’Neal, MeJa applications (20 mm) induced significant fruit drop but ethephon (1000 mg·L−1) applications did not affect fruit detachment. Overall, MeJa applications (20 mm or greater) generally induced rapid and extensive fruit abscission, often within 1 day after treatment. Applications of MeJa resulted in leaf yellowing and necrosis of leaf tips and margins, especially at high rates of application (20 mm or greater). Ethephon applications resulted in the abscission of mature and immature berries. Both ethephon and MeJa applications resulted in the detachment of the pedicel along with the fruit. Together, these data suggest that although ethephon and MeJa have the potential to be used as harvest aids in blueberry, the rates of application require further optimization to minimize potential phytotoxicity. Additionally, effective de-stemming of the berries may be essential if these compounds are to be used as harvest aids.