David L. Wenny

Efficacy of Trichoderma harzianum as a biological control of Fusarium oxysporum in container-grown Douglas-fir seedlings

Inoculating a soilless medium with encapsulated Trichoderma harzianum did not affect any aspect of Douglas-fir (Pseudotsuga menziesii var. glauca [Beissn.] Franco) seed germination or subsequent growth. Results of inoculating medium with a known pathogenic isolate of Fusarium oxysporum alone, or concurrently with T. harzianum, were the same: high levels of damping-off, low amounts of hypocotyl and root disease in midsummer, and significant reductions in height growth. When seedling roots grew through T. harzianum-inoculated medium before growing into a mixture of T. harzianum-F. oxysporum-inoculated medium, mortality was reduced about 50%. Although contamination by resident Fusarium occurred, subsequent root colonization was significantly reduced in T. harzianum-amended growing medium.

First year field growth of chemically root pruned containerized seedlings

Containers deform seedling root systems and have a potential to inhibit tree growth after outplanting. Ponderosa pine (Pinus ponderosa Dougl. ex Laws. var. ponderosa), western white pine (Pinus monticola Dougl. ex D. Don), and Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) were grown in containers coated with latex paint containing different concentrations of cupric carbonate and outplanted in a forest. Treatment effects on seedling growth and root morphology were determined after one growing season in the field. Seedling height was unrelated to treatment and stem diameter was not significantly affected (p≤0.05). New root growth in the upper portion of the root plug was significantly increased (p≤0.05). The optimal concentration of cupric carbonate for seedling root growth in the upper portion of the root plug was estimated by multiple regression.

Gliocladium virens in an alginate prill ineffective as a biological control of Fusarium root disease in container-grown Douglas-fir

An alginate prill formulation of Gliocladium virens (GL-21) was added as a top-dressings (54 g per m2) or incorporated into medium (1.2 kg per m3) used to grow Douglas-fir seedlings in styrofoam containers. Seedlings in the top-dress treatment were similar to control seedlings; infection and colonization by naturally-occurring Fusarium was unaffected by treatment. Incorporated G. virens reduced seedling growth and increased occurrence and colonization intensity of Fusarium. In a laboratory experiment, inoculating Douglas-fir seedlings with G. virens (10% w/w) prior to inoculation with Fusarium increased survival time when compared to concurrent inoculations of fungi.

Minimizing growth regulators in shoot culture of an endangered plant,Hackelia venusta (Boraginaceae)

SummaryHackelia venusta (Boraginaceae) is an endangered perennial herb endemic to the interior northwestern United States. Because of seed scarcity, micropropagation (anex situ conservation strategy) could produce true-to-type plantlets suitable for reintroduction. We hypothesized that clones of predetermined size could be rapidly produced by supplementing multiplication and rooting media with minimal levels of cytokinin and auxin. Microshoots derived from shoot expants were cultured on Murashige and Skoog (1962) media supplemented with 1% (wt/vol) agar and 0.0001 to 10 μM benzyladenine. Inverse regression estimates on 3 genotypes predicted that a target of 2.5 axillary microshoots per explant would require a minimal level of 0.04±0.02 μM benzyladenine. Culture of 25 genotypes with 0.04 μM benzyladenine resulted in an average of 2.3±0.1 axillary microshoots per explant. Elongated microshoots were transferred to media supplemented with 0.1 to 25 μM indoleacetic acid. Clones rooted from 36% to 100% success after 4 wk in 2.0 μM indoleacetic acid. Plantlets transplantedex vitro with three or more roots survived at 84% versus 46% of plantlets with fewer roots. Up to 84% of the plantlets survived in a planting trial. The data suggest that shoot culture ofHackelia venusta, with minimal growth regulators, can produce axillary microshoots for reintroduction.

Fusarium root infection of container-grown Douglas-fir: effect on survival and growth of outplanted seedlings and persistence of the pathogen

Greenhouse-grown Douglas-fir (Pseudotsuga menziesii var.glauca [Beissn.] Franco) seedlings with roots infected with eitherFusarium oxysporum Schlect. orFusarium proliferatum (Matsushima) Nirenberg were outplanted on a forest site in northern Idaho, U.S.A. No residentFusarium populations were detected in the forest soil.Fusarium persisted the first four years on roots initiated during the greenhouse phase, but occurred sparingly or was absent on roots that grew after outplanting. Height growth was unaffected, and mortality was not often associated withFusarium. A seasonal pattern ofFusarium activity was observed. Low levels (10–40%) of initial root infection apparently have little adverse effect on outplanting performance of Douglas-fir seedlings.

Interactions among Streptomyces griseoviridis, Fusarium root disease, and Douglas-fir seedlings

In a laboratory experiment, Douglas-fir (Pseudotsuga menziesii var. glauca [Beissn.] Franco) seedlings had similar disease ratings when treated with known Fusarium isolates or concurrently with Fusarium and Streptomyces griseoviridis. When tested under greenhouse conditions and against known Fusarium isolates, more seeds germinated and survived as seedlings in control medium than survived in S. griseoviridis-inoculated medium or when S. griseoviridis and Fusarium were added together. A series of applications of S. griseoviridis as a soil drench to a crop of Douglas-fir seedlings did not affect seedling morphology. However, against resident levels of Fusarium, S. griseoviridis reduced Fusarium infection by 16%, but increased infection by F. oxysporum and F. proliferatum, two potentially pathogenic fungi, by 40%.

Effects of nursery environment on needle morphology of Pinus monticola Dougl. and implications for tree improvement programs.

Statistically significant differences were found in 14 needle traits of western white pine (Pinus monticola Dougl.) seedlings grown from the same seed orchard source in the three nurseries in northern Idaho. Traits with significant variation included needle length and width, number of stomatal rows, number of stomata per row, total stomata per needle, adaxial surface area, stomatal density, major axes of stomata, stomatal shape, stomatal area, stomatal occlusion, epistomatal wax degradation, weight of wax per dry weight of needle, and the contact angles of water droplets placed on adaxial needle surfaces. Wax crystallites on needle surfaces were hollow and tubular and the amount of surface wax appeared to be associated with surface wettability. Our results may have important implications for tree improvement programs that require successful inoculation of nursery-grown seedlings with spores of Cronartium ribicola J. C. Fisch. ex Rabenh. to reliably screen white pines for resistance to blister rust.

Inducing long-shoot growth for vegetative propagation of western larch

This study evaluated methods to release from suppression both short shoots on lower stems and developing buds on leaders of western larch (Larix occidentalis Nutt.) seedlings, in order to increase the number of long shoots suitable for stem-tip cuttings. Treatments included application of the cytokinin N-6-benzyladenine (BA), horizontal stem bending, and decapitation on container-grown 1.5-year-old seedling ortets. The rate of release of suppressed growth of short shoots on the basal stem segments increased from 15% to 83% after spraying with 200 ppm BA and doubled on unsprayed horizontally-bent stems. Syllepsis on the upper stems increased by 19% following decapitation and by 32% after a combination of decapitation and BA treatments. Although these treatments resulted in large increases in the numbers of long shoots available for stem cuttings, the rootability of BA-treated shoots needs assessing before use in operational programs.