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Danny Black - Cascade

Vaccinium ovalifolium The fruits of huckleberries and bilberries found in genus Vaccinium section Myrtillus and indigenous to the northwestern United States and western Canada are popular for commercial culinary and botanical products.

Vaccinium ovalifolium The fruits of huckleberries and bilberries found in genus Vaccinium section Myrtillus and indigenous to the northwestern United States and western Canada are popular for commercial culinary and botanical products. Mountain huckleberry synonymous with black huckleberry, thin-leaf huckleberry, or mountain bilberry , and cascade huckleberry synonymous with cascade bilberry or blue huckleberry are tetraploid species Vander Kloet, rich in aroma and flavor chemicals Fellman et al.

These species are also moderately rich in antioxidant compounds, being similar to domestic blueberries Lee et al. Oval-leaf bilberry [synonymous with oval-leaf blueberry or alaska blueberry V.

Although its fruits lack high concentrations of flavor and aroma compounds Fellman et al. Currently, virtually all fruits of these three species are harvested from naturally occurring stands, most of which are on public lands in the northwestern United States.

Oval-leaf bilberry is also harvested from the wild in southeastern Alaska. Demand for these fruits has increased while supplies from wild forest stands have decreased Minore, The strong demand and limited supplies create opportunities for commercial production of these crops Barney, , and efforts are under way at the University of Idaho to produce cascade huckleberry, mountain huckleberry, and oval-leaf bilberry cultivars for field cultivation and managed forest systems.

Vegetative propagation, however, has been problematic. Stark and Baker reported only limited success in propagation of montana huckleberry [V. We have successfully propagated mountain huckleberry from rhizome cuttings D. Barney, unpublished data , but this method was slow and the amounts of stock material limited. A literature search through the U. Department of Agriculture National Agricultural Library failed to reveal research publications related to vegetative propagation of cascade huckleberry or oval-leaf bilberry.

In a new window In vitro culture offers one method for rapidly producing many genetically uniform plants for evaluation and distribution to growers. Little information has been published on in vitro culture of Vaccinium section Myrtillus species, however. Barney reported on the effects of plant growth regulators on in vitro culture and microshoot rooting of mountain huckleberry, although those trials were limited to modified woody plant medium WPM Lloyd and McCown, Reed and Abdelnour-Esquivel reported on in vitro trials involving section Myrtillus diploid species red huckleberry V.

Initiation rates for bilberry were reported to be intermediate on the same medium, but data and additional details were absent from the paper. Hyperhydration and chlorosis were absent on media amended with 2iP, although growth was less than on zeatin- or zeatin riboside-amended media. In combination with ferric ethylenediamine tetraacetate as the sole iron source, however, the latter two growth regulators induced explant hyperhydration and chlorosis.

Adding supplemental Fe in the form of sodium ferric ethylenediamine di- o-hydroxyphenylacetate in combination with zeatin or zeatin riboside dramatically reduced the incidence of hyperhydration and eliminated chlorotic symptoms. A 2iP concentration of Explants collected during the spring established better than those collected in autumn. Extensive work has been published on micropropagation of other Vaccinium species, including highbush blueberry V. Other media evaluated by Wolfe et al. The objectives for our trials were to identify suitable media for micropropagation of cascade huckleberry, mountain huckleberry, and oval-leaf bilberry to facilitate cultivar development and commercialization of these emerging crops, and to establish baselines for additional research on media formulations.

Our approach was to evaluate modifications of MS and WPM media, focusing on mineral nutrient concentrations. Materials and methods Establishment and maintenance of in vitro shoot cultures. Seeds from the three species were collected from geographically distinct sources in Idaho and Washington. This was the only oval-leaf bilberry clone available for the micropropagation trials during both and One vigorous seedling from each seed source was selected for in vitro establishment, and each seedling served as the stock plant for one clone.

The same clones were used for all treatments during both years of the trials. Before the trials described herein, the clones were subcultured at least twice over a minimum of 6 months on modified FSWPM prepared without plant growth regulators to minimize the residual effects of the 2iP and zeatin used during establishment.

Inclusion of zeatin or 2iP in the micropropagation medium induces these species to produce clumps of numerous, short, heavily branched microshoots. All stock cultures exhibited individual, tall, unbranched, or lightly branched microshoots, suggesting no residual effects of these plant growth regulators. The modifications to the MS medium produced Mn and Zn concentrations equivalent to the original formulation.

The modification of the WPM increased the Mn concentration from 7. Composition and concentrations of organic components were the same as used in the original WPM Lloyd and McCown, and were identical for all media tested. The media were adjusted to pH 5. Mineral nutrient concentrations in the media are shown in Table 1. In a new window Table 1. Mineral nutrient concentrations in the Murashige and Skoog MS and woody plant media WPM used for micropropagating cascade huckleberry, mountain huckleberry, and oval-leaf bilberry.

Baby food jars mL capacity and 96 mm tall; Sigma Chemical Co. Louis containing 50 mL medium served as culture vessels. Translucent polyethylene lids were sealed to the jars with self-cling plastic film. Tetraploid species in Vaccinium section Myrtillus often grow slowly in vitro, and the extended cultivation time was required to develop sufficient new microshoot tissues for evaluation.

The cultures were destructively harvested and percentage survival, percentage of explants forming roots, shoot dry weight less original explants , and numbers and lengths of microshoots were recorded. For microshoot measurements, we counted and measured initial shoots arising from the original explants as well as secondary lateral shoots without distinguishing between the two.

The species studied in these trials often develop adventitious roots on original explants and new microshoots, even in the absence of root-promoting plant growth regulators in the media. Roots at or below the medium surface are typically plump and white.

Aerial roots arising at leaf axils above the medium are thin, brown, and wiry. For these trials we considered a culture to have rooted if either root form could be visually distinguished, without magnification, on the original explant or new microshoots. Quantities of roots formed were usually insufficient for accurate quantitative measurement. Separate studies are underway to evaluate the effects of root-promoting growth regulators on in vitro and ex vitro rooting of these species.

Experimental design and data analysis. The experiment was set up as a randomized complete block design. An experimental unit consisted of one baby food jar containing five explants of a single clone. Each clone and treatment combination was replicated with three jars per block. Within each block, the three jars representing a clone—treatment combination were kept together in a randomly assigned location within that block.

The experiment was repeated in and Survival and rooting percentages, dry weights, and shoot lengths were normally distributed. Data for the number of microshoots were square root transformed and analyzed by the same method. Previous Section Next Section Results and discussion Results for all species and clones were generally similar for and trials. Although parameter values differed significantly between years for all species and most parameters, the patterns of plant responses to the treatments were similar for both years and the data were combined for analyses Table 2.

The differences in growth between the 2 years may have been the result of aging of the stock cultures and variations in laboratory environmental conditions. In a new window Table 2. Explant and microshoot survival, growth, and rooting of cascade huckleberry, mountain huckleberry, and oval-leaf bilberry as affected by media formulations and concentrations.

Survival was consistently lower on FSMS during both years for all species and clones. As for survival, the dry weights of shoots formed after placing explants onto the test media were significantly lower on FSMS than on other media for all species, clones, and years Table 2.

Culturing cascade huckleberry and mountain huckleberry explants on FSMS reduced the number of new shoots compared with the other media for all clones and years Table 2. The latter three media produced generally similar numbers of new shoots from mountain huckleberry explants. Average shoot lengths were significantly reduced on FSMS compared with all other media for cascade huckleberry and oval-leaf bilberry Table 2. Although root-promoting plant growth regulators were excluded from the media, we determined the percentages of explants and newly formed shoots that formed roots Table 2.

Adventitious root formation percentages were significantly reduced on FSMS compared with other media for all species, clones, and years except for mountain huckleberry in when the two MS formulations produced similar rooting percentages data not shown. For cascade huckleberry and mountain huckleberry, we observed genotypic differences in all evaluation criteria except survival percentages.

The differences were not always consistent or significant for both years. Only a single oval-leaf bilberry clone was available for the full course of the 2-year study. Despite the observed genotypic differences in growth parameter values, the overall responses of all clones to the test media were generally similar. Variability in responses of Vaccinium genotypes to in vitro formulations has also been observed in rabbiteye V.

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The results we observed were generally similar to those obtained by Wolfe et al. The salt concentrations in FSMS may be too high for optimal in vitro survival and growth of cascade huckleberry, mountain huckleberry, and oval-leaf bilberry. Electrical conductivity values for the media were 1.

Eck noted that blueberries are less tolerant of high soluble salt levels than most other crops, with high chloride concentrations causing toxicity symptoms. Ballinger suggested that a high chloride-to-S ratio favored Ca uptake in blueberries, noted as being calcifuges, whereas high S-to-chloride ratios favored blueberry growth. The MS media in our trials had a S-to-chloride ratio of 0. The higher agar concentration in FSMS was unlikely to have affected survival and growth. Several marked differences in mineral nutrient concentrations between the media may have contributed to the different growth responses observed Table 1.

The ratio of NH4 to NO3 was similar for all media: For related Vaccinium species, results have been mixed in trials comparing physiological and growth responses with NH4 and NO3 forms of N. Perhaps pertinent to our results was the observation by Herath and Eaton that high NO3 concentrations caused marginal necrosis and leaf abscission in container-grown highbush blueberry.

Nitrate concentrations were particularly high in the FSMS treatment, in which we observed explant necrosis and death. Eck concluded that highbush blueberry can use either form of N at soil pH values less than 5. The relationship between medium pH and N utilization in micropropagated Vaccinium has not been determined. The media pH in our trials was adjusted to 5. Although the K concentration in FSMS was comparatively high, Eck noted that blueberries tolerate high K concentrations without toxicity.

Although the WPM formulations generally produced higher rooting percentages and longer microshoots than the HSMS, all three formulations produced suitable results for in vitro propagation of cascade huckleberry. Both WPM formulations provided acceptable results.

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