Induced Meiotic Gynogenesis of Paddlefish Polyodon spathula

Viable, diploid gynogenetic (gynogenotes) paddlefish Polyodon spathula were produced by activating eggs with ultraviolet-irradiated shovelnose sturgeon Scaphirhynchus platorynchus spermatozoa and heat shocking. Without irradiation treatment, sturgeon spermatozoa appeared to activate the eggs (up to gastrulation), but did not result in any viable hybrids. Experiment 1 determined that heat-shock treatment of 35 C for a 2-min duration within the interval of 2–22 min post-activation resulted in highest yield of gynogenotes (12–19%) from eggs incubated at 18 C. Experiment 2 applied the heat shock treatment at 35 C from 14.0 to 28.0 min in 2-min intervals after activation at 18 C for a larger scale of gynogenetic production. Both experiments showed that the best yields of gynogenotes were obtained when the heat shock treatment occurred at 16, 18, and 20 min after activation. When these times were expressed in terms of τ₀. units (duration of one mitotic cycle of synchronous cell division related to water temperatures), optimal activations were 0.26, 0.29, and 0.32τ₀ (τ₀@ 18 C = 63.5 min). Experiment 3 tested the utility of τ₀. at two different pre-shock incubation water temperatures of 18 C and 16 C, and determined that there was no significant interaction in percentage of viable gynogenotes between two different incubation temperatures and the mitotic intervals (0.21, 0.26, 0.31, 0.36, 0.41τ₀) tested. Best survival of gynogenotes occurred when eggs held at either pre-shock incubation water temperatures were shocked at 0.26τ₀ All gynogenotes examined were histologically confirmed to have ovarian tissue and were determined to have similar oocyte development to that of normal female (control) paddlefish.     

Spatial scale effects on conservation network design: trade-offs and omissions in regional versus local scale planning

Ecological patterns and processes operate at a variety of spatial scales. Those which are regional in nature may not be effectively captured through the combination of conservation plans derived at the local level, where land use planning frequently takes place. Conversely, regional conservation plans may not identify resources important for conservation of intraregional ecological variation. We compare modeled conservation networks derived at regional and local scales from the same area in order to analyze the impact of scale effects on conservation planning. Using the MARXAN reserve selection algorithm and least cost corridor analysis we identified a potential regional conservation network for the Central Valley ecoregion of California, USA, from which we extracted those portions found within five individual counties. We then conducted the same analysis for each of the five counties. An overlay of the results from the two scales shows a general pattern of large differences in the identified networks. Especially noteworthy are the trade-offs and omissions evident at both scales of analysis and the disparateness of the identified corridors that connect core reserves. The results suggest that planning efforts limited to one scale will neglect biodiversity patterns and ecological processes that are important at other scales. An intersection of results from the two scales can potentially be used to prioritize areas for conservation found to be important at several spatial scales.     

Protection of spruce from colonization by the bark beetle, Ips perturbatus, in Alaska

Field bioassays were conducted in south-central Alaska in a stand of Lutz spruce, Picea × lutzii, to determine whether a semiochemical interruptant (verbenone and trans-conophthorin) and/or a defense-inducing plant hormone (methyl jasmonate, MJ) could be used to protect individual standing trees from bark beetle attack. During two experiments (initiated in May 2004 and 2005, respectively), attacks by Ips perturbatus on standing trees were induced by using a three-component aggregation pheromone (ipsenol, cis-verbenol, and ipsdienol) and prevented by using the interruptant. In 2005, treatments from 2004 were repeated and additional treatments were evaluated by using MJ spray or injection with and without the interruptant. Aggregation began before 3 or 7 June, and attack density was monitored through 3 or 16 August. During both years, tree mortality caused by I. perturbatus was recorded twice (in August, and in May of the following year). In both experiments, attack density was greatest on trees baited with the three-component attractive pheromone, but was significantly reduced by addition of the semiochemical interruptant to trees baited with the attractant. There were no significant differences in attack density between attractant + interruptant-treated trees and unbaited trees. In 2004, mortality was highest among attractant-baited trees, whereas addition of the interruptant significantly reduced the level of initial (10 week post-treatment) and final (54 week post-treatment) mortality. In 2005, no significant reduction in attack density occurred on trees baited with the attractant when MJ was sprayed or injected. The highest initial (10.6 week post-treatment) and final (49.4 week post-treatment) mortality was observed among trees that had been injected with MJ and baited with the attractant. Mortality at the final assessment was significantly lower in all other treatment groups. As in 2004, addition of the interruptant to attractant-baited trees significantly reduced the level of final mortality compared to attractant-baited trees. MJ was not attractive or interruptive to I. perturbatus or associated bark beetles in a flight trapping study. However, MJ-treated trees (sprayed or injected) exuded copious amounts of resin on the bark surface. Anatomical analyses of felled trees from four treatment groups [Tween (solvent)-sprayed, MJ-sprayed, Tween-injected, and MJ-injected + attractant baited] showed that treatment with MJ increased the number and size of resin ducts produced following treatment. These analyses also revealed a reduction in radial growth in MJ-treated trees. Our results show that during both years, treatment with a simple, two-component interruptant system of verbenone and trans-conophthorin significantly reduced I. perturbatus attack density and tree mortality on attractant-baited trees and provided a full year of protection from bark beetle attack.     

Proteomic analysis of membrane preparations from developing Pinus radiata compression wood

For coniferous gymnosperms, few data exist as to the contribution of the membrane-associated proteome to cell wall and wood formation. In this study, we begin to address this knowledge deficiency by examining the proteomic profile of Golgi-enriched membrane preparations derived from developing Pinus radiata compression wood. These membrane preparations were generated by a combination of discontinuous sucrose gradient centrifugation and Triton X-114-based phase separation. Fractionation by phase separation removed contaminating proteins associated with the cytoskeleton and enabled the discrimination between soluble and membrane-bound/integral proteins. The proteomic analysis of the resulting aqueous and detergent phases using high-performance liquid chromatography-tandem mass spectrometry resulted in the identification of 175 proteins. The majority of the identified proteins were membrane bound/integral and originated from cellular components such as the nucleus, plastids, endoplasmic reticulum, plasma membrane and Golgi vesicles. On the basis of bioinformatic analysis, many of the identified proteins were predicted to be involved either in the regulation of wood formation or in cell wall biosynthesis, which indicated that the proteomic analysis of non-cytosolic proteins in developing xylem is a useful strategy to investigate the molecular aspects of wood formation in pine.     

Predicting survival of planted Douglas-fir and ponderosa pine seedlings on dry,low-elevation sites in southwestern Oregon

Four equations were developed for predicting the probability of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] and ponderosa pine (Pinus ponderosa Dougl. ex Laws.) survival for the first (0–1) and first to third (1–3) growing seasons after applying mulching, scalping, or artificial shading (shade cards) treatments in plantations in southwestern Oregon, U.S.A. Variables describing conifer size, levels of competing vegetation, presence of silvicultural treatments, site factors, and climate factors were collected from 13 sites ranging from 0 to 6 years after planting and examined as potential predictors of survival. Age, stem diameter, a competition index for shrubs, severity of growing season at time of treatment, average annual precipitation, aspect, and slope angle were predictors of Douglas-fir survival during 0–1 and 1–3 growing seasons after treatment; the presence of silvicultural treatments was also a predictor only during the first growing season after treatment. Age, aspect, and slope angle were predictors of ponderosa pine survival over both 0–1 and 1–3 growing seasons after treatment; height-diameter ratio, competition indices for herbs, shrubs, and hardwoods, silvicultural treatment, severity of growing season at time of treatment, and average annual precipitation were also predictors only during the first growing season after treatment; crown width was a predictor of survival only during 1–3 growing seasons after treatment. When significant in the models, predicted probability of survival increases with treatments, less severe weather conditions, diameter, crown width, age, and precipitation; probability decreases with increasing height-diameter ratio and competition indices for herbs, shrubs, and hardwoods.     

Physiological variation among western redcedar (Thuja plicata Donn ex D. Don) populations in response to short-term drought

? Variation in the ability of western redcedar (Thuja plicata Donn ex D. Don) populations to withstand water stress may exist because this species is found in coastal and interior biogeoclimatic subzones representing the full range of precipitation regimes in British Columbia, Canada.

? Seven western redcedar populations from locations in British Columbia, representing a wide range of habitat types, were assessed for their gas exchange and water relations response to controlled drought.

? Before drought, population variation occurred in stomatal conductance, net CO₂ assimilation rate and intrinsic water use efficiency and the relative water content at turgor loss point. During drought, populations had different responses of net CO₂ assimilation to decreasing predawn shoot water potential. After drought, populations differed in stomatal conductance and intrinsic water use efficiency, plus osmotic potential at turgor loss point, osmotic potential at saturation and apparent cuticular transpiration. Western redcedar populations from drier-inland habitats had a lower osmotic potential at turgor loss point, lower relative water content at turgor loss point and lower apparent cuticular transpiration in response to drought than populations from coastal origin with temperate maritime habitat.

? Reduction of cuticular water loss and adjustments of cellular water relations in response to drought was found to occur among seven western redcedar populations originating along a precipitation gradient while; there were minimal population differences in the gas exchange response to drought.

     

Early growth and ecophysiological responses of Koa (Acacia koa A. Gray) seedlings to reduced water and phosphorus

Sites in need of restoration typically have one or more environmental factors that limit seedling establishment. Identifying ecophysiological responses to environmental stressors can provide important insights into mitigating measures that would allow seedlings to overcome such constraints to survival. Koa (Acacia koa A. Gray) is a nitrogen-fixing tree species endemic to Hawai?i that is highly valued in restoring degraded forest ecosystems, which are often limited in available water and phosphorus. This study examined how koa seedlings respond to conditions of reduced water (65 W) and no phosphorus (0P). After 17 weeks, seedlings subjected to 65 W or 0P accumulated less biomass, smaller root-collar diameters, and lower nitrogen and phosphorus contents. Combined reductions in water and P resulted in seedlings with increased root to shoot dry biomass and shorter shoots. Seedlings subjected to 65 W also had lower instantaneous rates of CO₂ assimilation, but higher instantaneous water-use efficiencies following irrigation, suggesting that koa responds to water deficits by decreasing water loss via reduced stomatal conductance. Seedlings subjected to 0P had similar rates of CO₂ assimilation relative to those grown with adequate P, suggesting that koa is able to employ strategies to avoid physiological impairment from conditions of inadequate P. Future research should assess whether subjecting koa seedlings to reduced water before planting on water-limited sites cues increased drought resistance and whether uptake and storage of P by seedlings in the nursery better supports growth following outplanting, particularly on sites with anticipated low plant-available water.

     

Resistance to amitraz in the parasitic honey bee mite Varroa destructor is associated with mutations in the β-adrenergic-like octopamine receptor

Varroa destructor is considered a major reason for high loss rate of Western honey bee (Apis mellifera) colonies. To prevent colony losses caused by V. destructor, it is necessary to actively manage the mite population. Beekeepers, particularly commercial beekeepers, have few alternative treatments other than synthetic acaricides to control the parasite, resulting in intensive treatment regimens that led to the evolution of resistance in mite populations. To investigate the mechanism of the resistance to amitraz detected in V. destructor mites from French and U.S. apiaries, we identified and characterized octopamine and tyramine receptors (the known targets of amitraz) in this species. The comparison of sequences obtained from mites collected from different apiaries with different treatment regimens, showed that the amino acid substitutions N87S or Y215H in the OctβR were associated with treatment failures reported in French or U.S. apiaries, respectively. Based on our findings, we have developed and tested two high throughput diagnostic assays based on TaqMan technology able to accurately detect mites carrying the mutations in this receptor. This valuable information may be of help for beekeepers when selecting the most suitable acaricide to manage V. destructor.

     

Computational Comparison of Stand-Centered Versus Cover-Constraint Formulations

The area restriction model for harvest scheduling problems can be formulated using mixed integer programs. The three different formulation types are cluster packing, cell aggregation, and the assignment formulation. Within the cell aggregation subgroup there is an exact formulation, the cover constraint (CC) approach; and an inexact formulation, the stand-centered approach. The cover constraint formulation has significantly more constraints than the stand-centered formulation. A computational comparison between these two methods is completed using cutting planes and constraint combinations using the CPLEX solver package. The CC approach as cutting planes was superior based on solution time. If formulation time were included with solution time, then the CC approach remains superior. The maximum harvest area is a significant factor of solution time. In addition, the percentage of total cutting planes used to solve the problem was examined.     

Coast live oak, Quercus agrifolia, susceptibility and response to goldspotted oak borer, Agrilus auroguttatus, injury in southern California

Oak mortality is often associated with a complex of decline factors. We describe the morphological and physiological responses of coast live oak, Quercus agrifolia Née, in California to an invasive insect, the goldspotted oak borer (GSOB), Agrilus auroguttatus Schaeffer (Coleoptera: Buprestidae), and evaluate drought as a potential inciting factor. Morphological traits of 356 trees were assessed and physiological traits of 70 of these were monitored intensively over one growing season. Morphological characteristics of tree health included crown thinning and dieback; bole staining resulting from larval feeding; density of GSOB adult exit holes; and holes caused by woodpecker feeding. These characteristics were used to rank GSOB infestation/injury into four classes, and taken together, they explained 87% of the variation in a principal component analysis. Drought stress on various size/age and infestation classes of Q. agrifolia was measured by assessing branchlet pre-dawn and solar noon xylem water potential, leaf cell turgor potential, and water use efficiency over one growing season. Both morphological and physiological traits were highly variable in mature and old growth trees. Early summer plant water status (branchlet xylem water potential and water use efficiency) was similar between uninfested and newly colonized trees, suggesting that GSOB are not pre-selecting drought-stressed Q. agrifolia for oviposition. By late summer, leaf water and cell turgor potentials were lower in infested than in uninfested mature trees, suggesting that GSOB infestation causes drought stress in these trees. Among the tree size/age classes, infested old growth trees exhibited the greatest change in water use efficiency over the growing season, and showed greater morphological injury symptoms of decline than infested mature trees. Morphological attributes of decline in Q. agrifolia associated with GSOB were correlated weakly with increasing physiological drought stress among infestation classes of trees. We propose that the collection of morphological responses of Q. agrifolia to GSOB described here can be used to monitor the future expansion of the GSOB distribution as well as the GSOB-induced decline of Q. agrifolia in California.