R. Kasten Dumroese

Molecular Characterization of Fusarium oxysporum and Fusarium commune Isolates from a Conifer Nursery

ABSTRACT Fusarium species can cause severe root disease and damping-off in conifer nurseries. Fusarium inoculum is commonly found in most container and bareroot nurseries on healthy and diseased seedlings, in nursery soils, and on conifer seeds. Isolates of Fusarium spp. can differ in virulence; however, virulence and colony morphology are not correlated. Forty-one isolates of Fusarium spp., morphologically indistinguishable from F. oxysporum, were collected from nursery samples (soils, healthy seedlings, and diseased seedlings). These isolates were characterized by amplified fragment length polymorphism (AFLP) and DNA sequencing of nuclear rDNA (internal transcribed spacer including 5.8S rDNA), mitochon-drial rDNA (small subunit [mtSSU]), and nuclear translation elongation factor 1-alpha. Each isolate had a unique AFLP phenotype. Out of 121 loci, 111 (92%) were polymorphic; 30 alleles were unique to only highly virulent isolates and 33 alleles were unique to only isolates nonpathogenic on conifers. Maximum parsimony and Bayesian analyses of DNA sequences from all three regions and the combined data set showed that all highly virulent isolates clearly separated into a common clade that contained F. commune, which was recently distinguished from its sister taxon, F. oxysporum. Interestingly, all but one of the nonpathogenic isolates grouped into a common clade and were genetically similar to F. oxysporum. The AFLP cladograms had similar topologies when compared with the DNA-based phylograms. Although all tested isolates were morphologically indistinguishable from F. oxysporum based on currently available monographs, some morphological traits can be plastic and unreliable for identification of Fusarium spp. We consider the highly virulent isolates to be F. commune based on strong genetic evidence. To our knowledge, this is the first reported evidence that shows F. commune is a cause of Fusarium disease (root rot and dampingoff) on Douglas-fir seedlings. Furthermore, several AFLP genetic markers and mtSSU sequences offer potential for development of molecular markers that could be used to detect and distinguish isolates of F. oxysporum nonpathogenic to conifers and highly virulent isolates of F. commune in forest nurseries.

Forest Restoration Paradigms

An estimated 2 billion ha of forests are degraded globally and global change suggests even greater need for forest restoration. Four forest restoration paradigms are identified and discussed: revegetation, ecological restoration, functional restoration, and forest landscape restoration. Restoration is examined in terms of a degraded starting point and an ending point of an idealized natural forest. Global change, climate variability, biotechnology, and synthetic biology pose significant challenges to current restoration paradigms, underscoring the importance of clearly defined goals focused on functional ecosystems. Public debate is needed on acceptable goals; one role for science is to inform and help frame the debate and describe feasibility and probable consequences.

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.

Considerations for restoring temperate forests of tomorrow: Forest restoration, assisted migration, and bioengineering

Tomorrow’s forests face extreme pressures from contemporary climate change, invasive pests, and anthropogenic demands for other land uses. These pressures, collectively, demand land managers to reassess current and potential forest management practices. We discuss three considerations, functional restoration, assisted migration, and bioengineering, which are currently being debated in the literature and have the potential to be applied independently or concurrently across a variety of scales. The emphasis of functional restoration is to reestablish or maintain functions provided by the forest ecosystem, such as water quality, wildlife habitat, or carbon sequestration. Maintaining function may call upon actions such as assisted migration—moving tree populations within a species current range to aid adaptation to climate change or moving a species far outside its current range to avoid extinction—and we attempt to synthesize an array of assisted migration terminology. In addition, maintenance of species and the functions they provide may also require new technologies, such as genetic engineering, which, compared with traditional approaches to breeding for pest resistance, may be accomplished more rapidly to meet and overcome the challenges of invasive insect and disease pests. As managers develop holistic adaptive strategies to current and future anthropogenic stresses, functional restoration, assisted migration, and bioengineering, either separately or in combinations, deserve consideration, but must be addressed within the context of the restoration goal.

Fall fertilization of Pinus resinosa seedlings: nutrient uptake, cold hardiness, and morphological development.

Abstract• Fall fertilization may increase plant nutrient reserves, yet associated impacts on seedling cold hardiness are relatively unexplored.• Bareroot red pine (Pinus resinosa Ait.) seedlings in north-central Minnesota, USA were fall fertilized at the end of the first growing season with ammonium nitrate (NH4NO3) at 0, 11, 22, 44, or 89 kg N ha−1. Seedling morphology and cold hardiness [assessed by freeze induced electrolyte leakage (FIEL)] were evaluated six weeks after fertilization and following the second growing season.• Seedling height and number of needle primordia increased with fertilizer rate for both sampling years. Seedlings fertilized with 44 and 89 kg N ha−1 attained target height (15 cm) after the second growing season. Shoot and root N concentration increased after the first growing season in fall fertilized seedlings compared to controls. Fall fertilized seedlings had lower FIEL (i.e., increased cold hardiness) compared to controls when tested at −40 °C after the first growing season, but no significant differences in FIEL of control and fertilized seedlings were observed after the second growing season.• Results suggest that fall fertilization of red pine seedlings can help render desired target height in the nursery, while maintaining or increasing cold hardiness levels.Résumé• La fertilisation automnale peut augmenter les réserves en éléments nutritifs des plants, mais les répercussions sur la rusticité au froid des semis sont encore relativement inexplorées.• Des plants à racines nues de Pinus resinosa Ait.dans le centre-nord du Minnesota (USA) ont été fertilisés à l’automne à la fin de la première saison de croissance avec du nitrate d’ammonium (NH4NO3) à 0, 11, 22, 44, ou 89 kg N ha−1. La morphologie des plants et la rusticité au froid [estimée par la fuite d’électrolyte (FIEL) induite par le gel] ont été évaluées six semaines après la fertilisation et à la suite dans la deuxième saison de croissance.• La hauteur des plants et le nombre de primordiums d’aiguilles ont augmenté avec le taux de fertilisation pour les deux années d’échantillonnage. Les plants fertilisés avec 44 et 89 kg N ha−1 ont atteint l’objectif de hauteur (15 cm) après la deuxième saison de croissance. La concentration en N des tiges et des racines a augmenté après la première saison de croissance chez les plants fertilisés à l’automne par rapport aux témoins. Les plants fertilisés à l’automne ont eu un plus faible FIEL (c’est-à-dire, une augmentation de rusticité), comparativement aux témoins lors du test à −40 °C après la première saison de croissance, mais aucune différence significative de FIEL entre plants fertilisés et témoins n’a été observée après la deuxième saison de croissance.• Les résultats suggèrent que la fertilisation d’automne des plants de Pinus resinosa Ait. peut aider à obtenir l’objectif de hauteur souhaité dans la pépinière, tout en maintenant ou en augmentant les niveaux de rusticité au froid.

Root diseases in bareroot and container nurseries of the Pacific Northwest: epidemiology, management, and effects on outplanting performance

Abstract.In forest and conservation nurseries in the Pacific Northwest USA, seedling production can be limited by root diseases caused by fungi in the genera Fusarium Link:Fr., Cylindrocarpon Wollenw., Phytophthora de Barry, and Pythium Pringsh. Fusarium, Cylindrocarpon, and Pythium are the most ubiquitous, whereas incidence of Phytophthora is mostly associated with coastal bareroot nurseries. All of these root pathogens are encouraged by water saturated soils or media. Seedlings infected with Fusarium, Phytophthora, or Pythium often appear chlorotic or necrotic with extensive root decay. Cylindrocarpon often causes serious root decay without shoot symptoms. The best approach to mitigate losses from these diseases is to use a holistic integrated pest management program. This program should combine chemical controls with cultural practices, particularly those that increase soil permeability and drainage and reduce potential sources of inoculum, especially by disinfesting seeds and containers reused for crops. In general, we found that seedlings meeting nursery specifications for outplanting on forest soil (proper height, root collar diameter, healthy shoot color, lack of disease symptoms) but having these disease organisms on their root systems perform as well as non-infected seedlings.

Subirrigation Reduces Water Use, Nitrogen Loss, and Moss Growth in a Container Nursery

With about half the amount of water, subirrigated Metrosideros polymorpha Gaud. (Myrtaceae) grown 9 mo in a greenhouse were similar to those irrigated with an existing fixed overhead irrigation system; moss growth was about 3X greater in the fixed overhead system after 3 mo. Moss growth was affected by the rate of preplant controlled release fertilizer added (more fertilizer, less moss) and moss maturity, quantified as presence or absence of sporangia, was slowed with subirrigation. About 5 g nitrogen (N) leached per m2 (0.02 oz/ft2 ) of greenhouse bench under the fixed irrigation system, whereas none was lost from subirrigation. Besides Metrosideros macropus, the USDA Forest Service and Purdue University are evaluating subirrigation for nursery production of other species. To date, the results indicate subirrigation may be a useful technique for growing native plants with large canopies where conventional irrigation systems are less effective, or where water use or other environmental concerns are paramount.

NURSERY RESPONSE OF ACACIA KOA SEEDLINGS TO CONTAINER SIZE, IRRIGATION METHOD, AND FERTILIZATION RATE

Planting koa (Acacia koa A. Gray) in Hawai‘i, USA aids in restoration of disturbed sites essential to conservation of endemic species. Survival and growth of planted seedlings under vegetative competition typically increases with initial plant size. Increasing container size and fertilizer rate may produce larger seedlings, but high fertilization can lead to surface and groundwater pollution and relatively low fertilizer use efficiency. Subirrigation systems may help mitigate this problem. Our study objective was to evaluate koa seedling growth with overhead or subirrigation over a range of container volumes (50 to 656 mL) and fertilizer rates (0 to 9.6 kg·m−3). Increasing container volume from 50 to 656 mL yielded koa seedlings with 200% more height and stem diameter growth. Subirrigation resulted in less nutrient leaching losses and yielded seedlings of similar vigor as overhead irrigated seedlings. Subirrigation helps optimize fertilizer delivery, which may improve fertilizer use efficiency and reduce environmental contamination.

Short-day treatment alters Douglas-fir seedling dehardening and transplant root proliferation at varying rhizosphere temperatures

We tested effects of shortened day length during nursery culture on Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco) seedling development at dormancy release. Seedlings from a 428N source were grown either under ambient photoperiods (long-day (LD)) or with a 28 day period of 9 h light: 15 h dark photoperiods (short-day (SD)). Seedlings were periodically removed from freezer storage from January to May. Sensitivity of plant tissues to cold temperatures was investigated via electrolyte leakage at nine test temperatures ranging from 2 to -40 8C. New root growth was assessed with rhizosphere temperatures of 10, 15, 20, and 25 8C. From 2 to -13 8C, there was no difference between treatments in cold hardiness. However, at or below -18 8C, LD seedlings exhibited higher indices of damage than SD seedlings. The LT50 (temperature at which 50% cell electrolyte leakage occurred) was consistently lower for SD than LD seedlings. Rhizosphere temperature differentially influenced new root proliferation: LD seedlings had greater new root pro- duction than SD seedlings at 20 8C, whereas the opposite response was detected at 10 8C. Our results confirm photoperiod sensitivity of Douglas-fir sources from relatively low (i.e., <458N) latitudes. Increased spring cold hardiness and greater rooting at lower rhizosphere temperatures may improve field performance potential of SD-treated seedlings.

Ostryopsis davidiana seedlings inoculated with ectomycorrhizal fungi facilitate formation of mycorrhizae on Pinus tabulaeformis seedlings

Reforestation in China is important for reversing anthropogenic activities that degrade the environment. Pinus tabulaeformis is desired for these activities, but survival and growth of seedlings can be hampered by lack of ectomycorrhizae. When outplanted in association with Ostryopsis davidiana plants on reforestation sites, P. tabulaeformis seedlings become mycorrhizal and survival and growth are enhanced; without O. davidiana, pines often remain without mycorrhizae and performance is poorer. To better understand this relationship, we initiated an experiment using rhizoboxes that restricted root and tested the hypothesis that O. davidiana seedlings facilitated ectomycorrhizae formation on P. tabulaeformis seedlings through hyphal contact. We found that without O. davidiana seedlings, inocula of five indigenous ectomycorrhizal fungi were unable to grow and associate with P. tabulaeformis seedlings. Inocula placed alongside O. davidiana seedlings, however, resulted in enhanced growth and nutritional status of O. davidiana and P. tabulaeformis seedlings, and also altered rhizosphere pH and phosphatase activity. We speculate that these species form a common mycorrhizal network and this association enhances outplanting performance of P. tabulaeformis seedlings used for forest restoration.