Acid-catalyzed steam pretreatment of lodgepole pine and subsequent enzymatic hydrolysis and fermentation to ethanol

Utilization of ethanol produced from biomass has the potential to offset the use of gasoline and reduce CO(2) emissions. This could reduce the effects of global warming, one of which is the current outbreak of epidemic proportions of the mountain pine beetle (MPB) in British Columbia (BC), Canada. The result of this is increasing volumes of dead lodgepole pine with increasingly limited commercial uses. Bioconversion of lodgepole pine to ethanol using SO(2)-catalyzed steam explosion was investigated. The optimum pretreatment condition for this feedstock was determined to be 200 degrees C, 5 min, and 4% SO(2) (w/w). Simultaneous saccharification and fermentation (SSF) of this material provided an overall ethanol yield of 77% of the theoretical yield from raw material based on starting glucan, mannan, and galactan, which corresponds to 244 g ethanol/kg raw material within 30 h. Three conditions representing low (L), medium (M), and high (H) severity were also applied to healthy lodgepole pine. Although the M severity conditions of 200 degrees C, 5 min, and 4% SO(2) were sufficiently robust to pretreat healthy wood, the substrate produced from beetle-killed (BK) wood provided consistently higher ethanol yields after SSF than the other substrates tested. BK lodgepole pine appears to be an excellent candidate for efficient and productive bioconversion to ethanol.     

Anatomical defences against bark beetles relate to degree of historical exposure between species and are allocated independently of chemical defences within trees

Conifers possess chemical and anatomical defences against tree‐killing bark beetles that feed in their phloem. Resins accumulating at attack sites can delay and entomb beetles while toxins reach lethal levels. Trees with high concentrations of metabolites active against bark beetle‐microbial complexes, and more extensive resin ducts, achieve greater survival. It is unknown if and how conifers integrate chemical and anatomical components of defence or how these capabilities vary with historical exposure. We compared linkages between phloem chemistry and tree ring anatomy of two mountain pine beetle hosts. Lodgepole pine, a mid‐elevation species, has had extensive, continual contact with this herbivore, whereas high‐elevation whitebark pines have historically had intermittent exposure that is increasing with warming climate. Lodgepole pine had more and larger resin ducts. In both species, anatomical defences were positively related to tree growth and nutrients. Within‐tree constitutive and induced concentrations of compounds bioactive against bark beetles and symbionts were largely unrelated to resin duct abundance and size. Fewer anatomical defences in the semi‐naïve compared with the continually exposed host concurs with directional differences in chemical defences. Partially uncoupling chemical and morphological antiherbivore traits may enable trees to confront beetles with more diverse defence permutations that interact to resist attack.     

Soil-surface carbon dioxide efflux and microbial biomass in relation to tree density 13 years after a stand replacing fire in a lodgepole pine ecosystem

The effects of fire on soil‐surface carbon dioxide (CO₂) efflux, F_S, and microbial biomass carbon, C_mic, were studied in a wildland setting by examining 13‐year‐old postfire stands of lodgepole pine differing in tree density (< 500 to > 500 000 trees ha^?1) in Yellowstone National Park (YNP). In addition, young stands were compared to mature lodgepole pine stands (～110‐year‐old) in order to estimate ecosystem recovery 13 years after a stand replacing fire. Growing season F_S increased with tree density in young stands (1.0 µmol CO₂ m^?2 s^?1 in low‐density stands, 1.8 µmol CO₂ m^?2 s^?1 in moderate‐density stands and 2.1 µmol CO₂ m^?2 s^?1 in high‐density stands) and with stand age (2.7 µmol CO₂ m^?2 s^?1 in mature stands). Microbial biomass carbon in young stands did not differ with tree density and ranged from 0.2 to 0.5 mg C g^?1 dry soil over the growing season; C_mic was significantly greater in mature stands (0.5–0.8 mg C g^?1 dry soil). Soil‐surface CO₂ efflux in young stands was correlated with biotic variables (above‐ground, below‐ground and microbial biomass), but not with abiotic variables (litter and mineral soil C and N content, bulk density and soil texture). Microbial biomass carbon was correlated with below‐ground plant biomass and not with soil carbon and nitrogen, indicating that plant activity controls not only root respiration, but C_mic pools and overall F_S rates as well. These findings support recent studies that have demonstrated the prevailing importance of plants in controlling rates of F_S and suggest that decomposition of older, recalcitrant soil C pools in this ecosystem is relatively unimportant 13 years after a stand replacing fire. Our results also indicate that realistic predictions and modeling of terrestrial C cycling must account for the variability in tree density and stand age that exists across the landscape as a result of natural disturbances.     

Immunoaffinity columns for isolation of abscisic acid in conifer seedlings

Spleen cells of mice immunized with cis, trans (±) abscisic acid (ABA) conjugated to bovine serum albumin (BSA) were fused with myeloma cells to produce hybridomas. Hybridomas secreting antibodies to cis, trans (+) ABA were detected by enzyme-linked immunoassay and injected into the peritoneal cavity of mice to produce ascites fluid. Antibodies purified by size-exclusion chromatography were linked to cyanogen bromide-activated Sepharose beads and used to prepare immunoaffinity columns (IAC). Semi-purified conifer seedling (Douglas-fir [ Pseudotsuga menziesii (Mirb) Franco] lodgepole pine ( Pinus contorta Dougl), cedar ( Thuja plicata D. Don), and spruce [ Picea glauca (Moench) Voss)]) extracts were passed through IAC, washed thoroughly with water and then eluted with methanol. ABA in the methanol eluate was analysed by high performance liquid chromatography with UV detection. UV absorption of extracts before and after purification through IAC indicated an excellent purification of extract by IAC. The identity of the ABA peak was confirmed by gas chromatography-mass spectrometry.     

A comparative kinetic analysis of nitrate and ammonium influx in two early-successional tree species of temperate and boreal forest ecosystems

Root NO₃ ^? and NH₄ ⁺ influx systems of two early‐successional species of temperate (trembling aspen: Populus tremuloides Michx.) and boreal (lodgepole pine: Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) forest ecosystems were characterized. NO₃ ^? and NH₄ ⁺ influxes were biphasic, consisting of saturable high‐affinity (HATS) and constitutive non‐saturable low‐affinity transport systems (LATS) that were evident at low and relatively high N concentrations, respectively. NO₃ ^? influx via HATS was inducible (IHATS); nitrate pre‐treatment resulted in 8–10‐fold increases in the V_max for influx in both species. By contrast, HATS for NH₄ ⁺ were entirely constitutive. In both species, V_max values for NH₄ ⁺ influx were higher than those for NO₃ ^? uptake; the differences were larger in pine (6‐fold) than aspen (1·8‐fold). In aspen, the K_m for NH₄ ⁺ influx by HATS was approximately 3‐fold higher than for IHATS NO₃ ^? influx, while in pine the K_m for IHATS NO₃ ^? influx was approximately 3‐fold higher than for NH₄ ⁺ influx. The aspen IHATS for NO₃ ^? influx appeared to be more efficient than that of pine (V_max values for aspen being approximately 10‐fold higher and K_m values being approximately 13‐fold lower than for pine). By contrast, only small differences in values for the NH₄ ⁺ HATS were evident between the two species. The kinetic parameters observed here probably result from adaptations to the N availabilities in their respective natural habitats; these may contribute to the distribution and niche separation of these species.     

Differential selection of baculovirus genotypes mediated by different species of host food plant

Recent studies have demonstrated high levels of genotypic and phenotypic variation in populations of parasites, even within individual hosts. Several genetic, immunological and epidemiological mechanisms have been postulated as promoters of such variation, but little empirical work has addressed the role of host ecology. A nucleopolyhedrovirus that attacks larvae of the pine beauty moth, Panolis flammea , exists as a complex mixture of genotypes within individual host larvae. We demonstrate that the food plant species eaten by the host (Scots pine vs. lodgepole pine) differentially affects the pathogenicity and productivity of two virus genotypes originally purified from a single host individual. We hypothesize that such food plant-mediated differential selection will promote genotypic variation between baculovirus populations, and that subsequent remixing of virus genotypes could maintain genotypic variation within individual hosts. Our results provide a tritrophic explanation for the genotypic and phenotypic complexity of host–parasite interactions with complex ecologies.     

Promotion of flowering in the Pinaceae by gibberellins

Significant female flowering of 6- to 11-year-old seedlings and grafted ramets of sexually mature scions of lodgepole pine (Pinus contorta Dougl.) was promoted by both topical and spray applications of a gibberellin (GA) A_4/7 mixture (1.6 to c. 5 mg per plant in total) during that period (June to September) when sexual differentiation of lateral primordia would be expected to take place. Girdling was used in most experiments to enhance the GA_4/7 effect, as was the auxin, naphthaleneacetic acid (NAA). Average frequency of flowering branches on treated plants over all experiments ranged from 27 to 59% (control ranged from 0 to 36%) and average number of female strobili was increased from 2- to 6-fold by growth regulator treatment, relative to controls. Within an experiment, clonal or family frequency of flowering for treated plants ranged from 11 to 67% (controls were 0 to 28%), and number of female strobili was increased from 2- to 14-fold by growth regulator treatment, relative to controls. Movement of the flowering stimulus from the point of application was apparent in several experiments, the response in adjacent branches being correlated positively with increasing dosage of GA_4/7. Significant male flowering occurred only in one experiment, girdling and GA_4/7 treatment being promotive factors. The use of spray applications of GA_4/7+ NAA is warranted to induce early and enhanced flowering in lodgepole pine seedlings and vegetative propagules for genetic improvement programs.     

Relationship between respiratory depletion of sugars and loss of cold hardiness in coniferous seedlings over-wintering at raised temperatures: indications of different sensitivities of spruce and pine

Long-term effects of elevated winter temperatures on cold hardiness were investigated for Norway spruce (Picea abies L. Karst.), lodgepole pine (Pinus contorta Dougl.) and Scots pine (Pinus sylvestris L.). Two-year-old seedlings with the same pre-history of growth and cold hardening in the field were maintained from early December to late March at two field sites in northern Sweden and in a cold room. The temperatures at these locations averaged –13·5, –8·9 and 5·5°C, respectively. Following treatments, carbohydrate contents and cold tolerances were assessed. Needle respiration was also analysed during the 5·5°C treatment. Cold tolerance of lodgepole pine and Scots pine was much reduced following the 5·5°C treatment. Cold tolerance was somewhat reduced in lodgepole pine following the –8·9 °C treatment, but was essentially maintained in spruce throughout all treatments. The cold tolerance of needles was strongly correlated with their soluble sugar contents. Spruce maintained cold hardiness by having larger reserves of sugars and lower rates of respiration which decreased more rapidly as sugars were depleted. Tolerance of lodgepole pine to frost desiccation was also much reduced following the 5·5°C treatment.     

Life table for Leptoglossus occidentalis Heidemann (Heteroptera: Coreidae) and prediction of damage in lodgepole pine seed orchards

Abstract 1 Fecundity, egg hatch and nymphal survivorship of Leptoglossus occidentalis Heidemann were determined and used to develop a damage prediction formula for seed bugs in lodgepole pine seed orchards. 2 A single overwintered L. occidentalis female may lay up to 80 eggs during early and mid‐season cone development. 3 Up to 83.4% of eggs that were attached to branches with tape hatched successfully, compared with 53.5% of eggs attached to needles with wire. Egg parasitism was only observed in egg masses attached with wire. 4 Egg masses attached with wire suffered approximately 30% mortality from parasitism, primarily by Gryon pennsylvanicum (Ashmead). 5 Approximately 15.1 and 2.5% of first instars survived to adulthood in 2000 and 2001, respectively. Survival of caged nymphs that were protected from predators was 14.5 and 17.8%, respectively. 6 A composite survival table indicated that 9.3% of eggs laid by L. occidentalis are expected to survive to adulthood. 7 Based on previously determined damage estimates for each life stage of feeding during three periods of cone development, a hypothetical density of one seed bug per tree early in the season will result in an expected seed loss of approximately 310 seeds.     

Temptations of weevil: feeding and ovipositional behaviour of Hylobius warreni Wood on host and nonhost bark in laboratory bioassays

• 1 Warren root collar weevil Hylobius warreni Wood (Coleoptera: Curculionidae) is a long‐lived, flightless insect native to coniferous forests across northern North America. Girdling by larval feeding causes significant mortality on young trees. The insect poses considerable challenges to reforestation.
• 2 Adult weevils feed on all life stages of a variety of coniferous hosts prior to oviposition. Their relative feeding preferences, however, have not been quantified. Moreover, it is not known whether host bark influences oviposition behaviour.
• 3 Feeding preferences of adult weevils were tested in both choice and no‐choice laboratory bioassays using small branches from three conifers (lodgepole pine Pinus contorta var. latifolia, interior hybrid spruce Picea glauca×engelmannii, and Douglas‐fir Pseudotsuga menziesii) and one deciduous tree (trembling aspen Populus tremuloides). Measurements included the surface area of bark consumed, rate of consumption, the number of days of feeding, and, in the no‐choice assay, the number of eggs oviposited.
• 4 Bark consumption was greatest on pine and Douglas‐fir, followed by spruce. Little to no feeding occurred on aspen. Consumption did not vary between male versus female insects for any of the feeding metrics quantified.
• 5 The presence of aspen branches did not inhibit feeding on any of the other species in the choice bioassays.
• 6 The number of eggs laid by female insects did not differ significantly among tree species in the no‐choice assay. Eggs were laid indiscriminately in the presence of all four host types.
• 7 Results and opportunities for future research are discussed in the context of formulating new integrated pest management strategies for this insect, which is increasingly important in the period of reforestation subsequent to the mountain pine beetle epidemic in western Canada.