Electrophoretic techniques for detection of Glypta fumiferanae, an endoparasitoid of western spruce budworm

Parasitoid-free western spruce budworms and western spruce budworm larvae containing the endoparasitoid Glypta fumiferanae (Vier.) (Hymenoptera: Ichneumonidae) were assayed electrophoretically to identify isozyme bands diagnostic for the parasitoid. Bands characteristic of G. fumiferanae were observed with 4 of 15 enzymes tested. These were aconitase, esterase, leucine aminopeptidase and 6-phosphoglucose dehydrogenase. Presence of these isozyme bands is a reliable indication of parasitism by G. fumiferanae. The efficiency of electrophoresis as a tool for routine detection of parasitoids is discussed.

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Zusammenfassung Raupen von Choristoneura occidentalis ohne und mit dem Endoparasitoiden Glypta fumiferanae (Vier.) (Hymenoptera: Ichneumonidae) wurden elektrophoretisch auf Isoenzymbänder untersucht zum Nachweis des Parasitoiden. Für G. fumiferanae typische Bänder wurden in 4 der untersuchten 15 Enzymen beobachtet. Dies waren Aconitase, Esterase, Leucinaminopeptidase und 6-Phosphoglucose-dehydrogenase. Das Vorkommen dieser Isoenzymbänder ist ein zuverlässiges Zeichen der Parasitierung durch G. fumiferanae. Die Brauchbarkeit der Elektrophorese als Mittel zum Routinenachweis von Parasitoiden wird diskutiert.

     

Comparison of some methods for collecting and analyzing data on aboveground net production and diversity of herbaceous vegetation in a northern Utah subalpine context

Our major objective was to compare methods of collecting and analyzing data on herbaceous aboveground net primary production and plant diversity in sequence of sites inferred to represent a meadow aspen fir spruce-sere in the middle Rocky Mountains. These comparisons should help us better interpret the literature and prepare for monitoring vegetation trends due to global climatic changes. Both absolute and relative values for above net primary production were highly dependent on methods used. The magnitude of differences in community diversity also varied greatly depending on the methods used to express plant richness, equitability, heterogeneity and dominance. We conclude that it is impossible to objectively test differences in production and diversity using data from single sampling dates and arbitrary choices of methods. In communities with great inter and intra-seasonal dynamics, an index of dominance that only measures magnitude (e.g. peak standing crop) was shown to have undesirable properties. We have demonstrated how average seasonal phytomass can overcome these problems by employing a measure of residence time as well as magnitude.Abbreviations ANPP = aboveground net primary production - ASP = average seasonal phytomass - NCSC = net aboveground community standing crop - PCSC = peak community aboveground standing crop     

Mismatches between breeding success and habitat preferences in Hen Harriers Circus cyaneus breeding in forested landscapes

During the past century, the upland breeding areas of Hen Harriers in Ireland have been extensively afforested. There is no evidence that this species avoids breeding in heavily forested landscapes and, indeed, young commercial forests in their second rotation are often selected as nest‐sites. However, Hen Harriers have coexisted with these forested areas for only a few decades and it is possible that such landscapes are suboptimal. We examined the relationship between breeding success and habitat using a dataset spanning three years and four study areas in the south and west of Ireland. We assessed whether nest success and fledged brood size were related to habitat type, both at the nest‐site and in the surrounding landscape. Neither measure of breeding productivity was related to total forest cover or to percentage cover of closed canopy forest in the landscape. However, in a subset of areas, high cover of second‐rotation pre‐thicket (young forests planted on land from which a first rotation has already been harvested) in the surrounding landscape was associated with low levels of breeding success. This may be due to factors related to predation, disturbance or prey availability. The fact that second‐rotation pre‐thicket is a preferred habitat for nesting in Ireland suggests that Hen Harriers may be making suboptimal decisions in the landscapes available to them.     

On the mechanism of apoplastic H<Subscript>2</Subscript>O<Subscript>2</Subscript> production during lignin formation and elicitation in cultured spruce cells—peroxidases after elicitation

A cell culture of Picea abies (L.) Karst. was used for studies of H₂O₂ generation during constitutive extracellular lignin formation and after elicitation by cell wall fragments of a pathogenic fungus, Heterobasidium parviporum. Stable, micromolar levels of H₂O₂ were present in the culture medium during lignin formation. Elicitation induced a burst of H₂O₂, peaking at ca. 90 min after elicitation. Of exogenous reducing substrates that may be responsible for the synthesis of H₂O₂ from O₂, NADH stimulated H₂O₂ production irrespective of elicitation. Cysteine (Cys) and glutathione (GSH) partially scavenged the constitutive H₂O₂, but usually increased or prolonged elicitor-induced H₂O₂ formation. Culture medium peroxidases were not able to generate H₂O₂ in vitro with Cys or GSH as reductants. These thiols, however, generated H₂O₂ non-enzymically at pH 4.5. [³⁵S]Sulphate feeding to spruce cells showed that endogenous sulphur-containing compounds (including GSH, GSSG and cysteic acid) existed in the culture medium. The apoplastic levels of these were, however, undetectable by the monobromobimane method suggesting that their contribution to apoplastic H₂O₂ formation is probably minor. Azide, an inhibitor of haem-containing enzymes, slightly inhibited constitutive H₂O₂ generation but strongly delayed the elicitor-induced H₂O₂ accumulation. Diphenylene iodonium, an inhibitor of flavin-containing enzymes, efficiently inhibited H₂O₂ production irrespective of elicitation. Elicitation led to downregulation of the expression of several peroxidase genes, and peroxidase activity in the culture medium was slightly reduced. Expression of three other peroxidase genes and a respiratory burst oxidase homologue (rboh) gene were upregulated. These data suggest that both peroxidases and rboh may contribute to H₂O₂ generation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Growth characteristics of Sakhalin spruce (Picea glehnii) planted on the northern Japanese hillsides exposed to strong winds

Sakhalin spruce (Picea glehnii), a native species typically found in northern Japan, has been used in reforestation on hillsides exposed to strong winds. In the reforestation areas, there are south-facing (S-slope) and northwest-facing slopes (NW-slope). Climatic conditions are contrasting between the two slopes, with shallower snow cover on the S-slopes. We compared growth responses of the spruce to micro-environment between the S- and NW-slopes through soil nutrients, needle longevity, water status, photosynthesis, and nutrients in the needles. These parameters were measured in needles exposed above the snow in winter and in lower needles protected by snow cover. High-position needles suffered from drought stress, especially in winter, and needles were shed early in the year on both slopes. Low-position needles did not suffer from drought stress, and maintained a high photosynthetic rate on both slopes. However, needle longevity was reduced on the S-slope, and concentrations of nitrogen, phosphorus, and potassium in the needles decreased with needle age. Soil nutrient concentrations were low on the S-slope, which suggests that the needles on the S-slope may remobilize nutrients from aged needles to younger needles prior to shedding. This characteristic is probably an adaptation in Sakhalin spruce to poor soil conditions.     

The relation among soil microorganism, enzyme activity and soil nutrients under subalpine coniferous forest in Western Sichuan

Soil microorganism and enzymes are important parts of forest ecosystem and sensitive to environmental changes. They have many critical functions in energy conversion and material cycle of forest soil. However, there are few studies about soil biological properties under subalpine coniferous forest, in particular, a serial of spruce plantation chronosequences following clear-cutting of natural coniferous forest in western Sichuan. We measured the quantity of soil microorganism (including bacteria, fungi and actinomyces), enzyme activity and soil nutrients under spruce plantation chronosequences in western Sichuan to investigate soil biological properties and their relationship with soil nutrients. The results showed that soil microorganism, enzyme activity and soil nutrients of the mature spruce plantation were significantly lower than those of the young spruce plantation and secondary broad-leaved forest. Soil fertility degraded greatly with the increasing of spruce plantation age and was mainly affected by forest micro-environment. There were significant correlation between the amounts of soil microorganisms, soil enzyme activities and nutrients (e.g. soil organic matter, total N, total P, alkali-hydrolyzable N and available K). Therefore soil biological indices can be used to evaluate soil fertility. In order to accelerate the course of restoration and rehabilitation of degraded pure plantation, the strategy and measures were put forward, including application of thinning rationally for existing dense plantations and establishment of mixture forest of coniferous and broad-leaved trees for new afforestations, which would create good forest micro-environment for plant growth.     

NMR structural determination of dissolved O-acetylated galactoglucomannan isolated from spruce thermomechanical pulp

Water-soluble O-acetylated galactoglucomannan (GGM) isolated from spruce thermomechanical pulp (TMP) by hot-water extraction was characterized by 1D and 2D (homo- and heteronuclear) NMR analysis. The backbone was found to consist of (1-->4)-linked mannopyranosyl and glucopyranosyl units in a ratio of 10:1.9-2.6. The mannopyranosyl units were acetylated at C-2 and C-3 with a degree of acetylation around 0.28-0.37 as determined by NMR. A slightly larger amount of 2-O-acetylated mannopyranosyl was detected when compared to the 3-O-acetylated component. Approximately every 10th mannopyranosyl unit was substituted at C-6 by a single alpha-galactopyranosyl unit. Fine structure determination based on sequence-specific chemical shift variations showed that the distribution of glycosyl residues is random. Small amounts of other minor polysaccharide species including xylans and galactans could also be identified by NMR.     

Changes in the understory of Canadian southern boreal forest after fire

Abstract. We investigated changes in the composition and abundance of understory species after fire in the southern boreal forest around Lake Duparquet, Québec. Ten plots of 100 m² were sampled in each of eight sites varying in post-fire age from 26 to 230 yr, with 20 1-m² quadrats in each of these 80 plots. Variation in the understory was described by DCA ordination and interpreted as a regeneration succession series. Thickness of the organic layers, stand age and canopy composition were all correlated with vegetational change. This change was not constant throughout succession; some old sites showed an increase in the diversity and abundance of certain pioneer species. This was partly related to openings in the canopy resulting from a major outbreak of spruce budworm, which affected sites dominated by Abies balsamea. The ordinations were performed on both the 100-m² plots and the 1-m² quadrats. Heterogeneity within sites was larger at the 1-m² scale and there was a great deal of overlap in the position of the quadrats in ordination space. At the smaller scale of analysis, stand age and thickness of the organic layers were not correlated with the changes observed in the understory.     

Changes in the forest floor of Canadian southern boreal forest after disturbance

Abstract. The concentrations and contents of organic matter and nutrients in organic deposits on the forest floor were estimated along a 231-yr chronosequence following fire at the southern limit of the boreal forest in eastern Canada. The sampling design was stratified to take into account the variability related to the presence of the principal tree species as well as to the presence of large gaps created by a recent spruce budworm (Choristoneura fumiferana) outbreak. The forest floor showed a steady accumulation of organic matter and total nutrients with time-since-fire and a 50 % decrease in the concentrations of available P and K, but not N (as determined by aerobic incubation). The increase in forest-floor weight was accompanied by an increased storage of available N, Ca and Mg. The availability of N and Ca was more strongly affected by tree species and gaps than by time-since-fire. A high N-availability was observed under Betula papyrifera and in gaps, while high a Ca-availability was found near Populus tremuloides and Thuja occidentalis. In old sites, the forest floor of gaps, created by a recent spruce budworm outbreak, had a necromass similar to that of a young forest, but the low concentrations of available P and K of an old forest.     

Species-specific nuclear and chloroplast single nucleotide polymorphisms to distinguish Picea glauca, P. mariana and P. rubens

 Picea rubens (red spruce) and P. mariana (black spruce) are closely related species which are difficult to differentiate morphologically. They are sympatric with P. glauca (white spruce) in the northern portion of their ranges. In order to identify potential interspecific polymorphisms, the chloroplast trnK intron and rpl33-psaJ-trnP region were sequenced, and the nuclear-encoded ITS region of the rDNA repeat was partially sequenced. Thirteen chloroplast and 12 nuclear candidate interspecific single nucleotide polymorphisms (SNPs) were identified. The species-specificity of several SNPs was determined by surveying DNAs amplified from trees representing range-wide provenance tests; these included 46 red spruce from 11 provenances, 84 black spruce from 30 provenances and 90 white spruce from 22 provenances. Two SNPs (1 chloroplast and 1 nuclear), which distinguish black spruce from red and white spruce, were consistent among 96–100% of the trees surveyed. Five SNPs (4 chloroplast and 1 nuclear), which distinguish white spruce from red and black spruce, were consistent among 100% of surveyed trees. These species-specific SNPs were used to identify anonymous spruce samples in a blind test, and their utility for small amounts of tissue, as little as single needles, was demonstrated. Scoring these SNPs is much less labor intensive than previous molecular methods for taxa differentiation (restriction fragment length polymorphisms or random amplified polymorphic DNAs), therefore they can be applied to large population studies. Received: 16 December 1998 / Accepted: 5 January 1999