Effects of the bark beetle nematode, Contortylenchus reversus, on gallery construction,fecundity, and egg viability of the Douglas Fir beetle, Dendroctonus pseudotsugae (Coleoptera: Scolytidae)

Healthy and Contortylenchus reversus infected Douglas Fir beetles were colonized in logs in the laboratory. At 7, 14, and 23 days after introduction of the beetles into the logs, the bark was removed from some of the logs and the length and shape of the primary gallery, the number of eggs laid, and the egg viability were determined for each pair of parent beetles. Parasitism caused a 25, 28, and 27% reduction in the length of the primary egg gallery built by the infected female at 7, 14, and 23 days, respectively, after the start of colonization. In addition, the number of eggs laid by the infected females by 7, 14, and 23 days after the start of colonization were 50, 33, and 45%, respectively, lower than that of healthy female beetles. Despite the differences in gallery length between control and infected females, linear regression analysis showed that the rates of gallery construction were not significantly different. However, infected female beetles showed significantly lower rates of egg-laying than did healthy female beetles. Nematode parasitism did not affect gallery shape and egg viability of female Douglas Fir beetles. Mating of healthy female beetles with infected males affected neither the gallery length nor the fecundity of the uninfected females.     

Procyanidins (Condensed Tannins) in Green Cell Suspension Cultures of Douglas Fir Compared with Those in Strawberry and Avocado Leaves by Means of C(18)-Reversed-phase Chromatography

The procyanidins (the most common type of proanthocyanidin or condensed tannin) from cell suspension cultures derived from cotyledons of Douglas Fir have been compared with those isolated from leaves of strawberry and avocado. Seventy per cent methanol (v/v) extracts from 100 milligrams fresh weight samples were analyzed by a combination of C₁₈-reversed-phase columns with high-performance liquid chromatography, and normal phase paper chromatography. (−)-Epicatechin and its oligomers were generally retarded longer on C₁₈ columns than the corresponding units made of (+)-catechin when eluted with solvents made up of 5% acetic acid alone or mixed with methanol up to 15% (v/v). Douglas fir preparations contained the most complex set of procyanidins and consisted of oligomers of catechin and epicatechin, whereas strawberry and avocado contained mainly (+)-catechin and (−)-epicatechin derivatives, respectively.     

Grazing on bacteria by flagellates and cladocerans in lakes of contrasting food-web structure

We tested the hypothesis that grazing on bacteria would varybetween lakes with differing plankton community structures.Paul and Tuesday lakes (Gogebic County, MI) are respectivelydominated by piscivorous and planktivorous fish. Consequently,zooplankton in Paul are primarily large daphnids, while zooplanktonin Tuesday are primarily small cladocerans and copepods. Wemeasured flagellate grazing on bacteria using a fluorescentminicell method, while cladoceran grazing was estimated fromthe relationship between body length and filtering rate. Wepredicted that cladoceran grazing on bacteria would be higherin Paul, and flagellate grazing would be higher in Tuesday.Cladoceran grazing on bacteria was important in both lakes contraryto our initial expectation. Large populations of the small cladoceran,Bosmina longirostris, in Tuesday exerted a grazing pressure(0.18–35x10⁶ bacteria 1^–1 h^–1) approximatelyequal to that of the large cladoceran, Daphnia pulex, in Paul(0.34–30x10⁶ bacteria 1^–1 h^–1). Flagellategrazing was higher in Tuesday as predicted (range: Paul, 0.1–6x10⁶bacteria 1^–1 h^–1; Tuesday, 0.2–20x10⁶ bacteria1^–1 h^–1). However, there was not a simple relationshipbetween total abundance of flagellates and total grazing rates.High community grazing by flagellates occurred when attachedchoanoflagellates were present. These flagellates had higheringestion rates than free forms. We find no clear evidence thatdifferences in food-web structure between the two lakes influencethe process of grazing on bacteria. Instead, our results emphasizethe significance of cladocerans and attached flagellates asconsumers of bacteria in freshwater ecosystems.     

Bacterial dynamics and distribution during a spring diatom bloom in the Hudson River plume, USA

Bacterial abundance and [³H]thymidine incorporation rates wereused to characterize bacterial distributions and dynamics duringa spring diatom bloom in the coastal plume of the Hudson Riverduring March, 1981. Bacterial abundance did not decline significantlyaway from the plume or across the continental shelf. However,a pronounced gradient was observed for [³H]thymidine incorporation. Following a northeast gale, a bloom of Skeletonema costatumdeveloped in response to coastal upwelling. As the plume becamesaltier, warmer and larger, bacterial abundance averaged 1.3x 10⁹ cells 1^–1. Bacterial incorporation of [³H]thymidineinto cold 5% TCA insoluble materials, bacterial production andspecific growth rates averaged 80–120 pmol 1^–1 d^–1,1.4–2.0 x 10⁹ cells 1^–1 and 1.2–1.5 d^–1.respectively. The mean density of bacteria in the plume didnot change even though growth rates were higher than expectedlosses from sinking, mixing and export. The abundance and production levels of attached bacteria inthe plume were significantly higher than they were in coastalwater outside the plume. In contrast, free bacterial levelswere similar in all regions of the shelf and plume. Bacterialparameters in the plume were not correlated with phytoplanktonpigment concentrations, while significant positive correlationswere found in shelf waters >33 salinity. Thus bacteria werecoupled to phytoplankton in shelf waters but not in the plumewhere allochthonous dissolved matter was probably supportingthe bacteria. ^*Lamont-Doherty Geological Observatory Contribution No. 3459 ²Present address: Department of Microbiology, University ofGeorgia, Athens, GA 30602, USA.     

Trophic links in the lowland River Meuse (Belgium): assessing the role of bacteria and protozoans in planktonic food webs

Trophic interactions within the plankton of the lowland RiverMeuse (Belgium) were measured in spring and summer 2001. Consumptionof bacteria by protozoa was measured by monitoring the disappearanceof ³H-thymidine-labelled bacteria. Metazooplankton bacterivorywas assessed using 0.5-µm fluorescent microparticles (FMPs),and predation of metazooplankton on ciliates was measured usingnatural ciliate assemblages labelled with FMPs as tracer food.Grazing of metazooplankton on flagellates was determined throughin situ incubations with manipulated metazooplankton densities.Protozooplankton bacterivory varied between 6.08 and 53.90 mgC m^–3 day^–1 (i.e. from 0.12 to 0.86 g C^–1bacteria g C^–1 protozoa day^–1). Metazooplankton,essentially rotifers, grazing on bacteria was negligible comparedwith grazing by protozoa (1000 times lower). Predation of rotiferson heterotrophic flagellates (HFs) was generally low (on average1.77 mg C m^–3 day^–1, i.e. 0.084 g C^–1 flagellatesg C^–1 rotifers day^–1), the higher contribution ofHF in the diet of rotifers being observed when Keratella cochleariswas the dominant metazooplankter. Predation of rotifers on ciliateswas low in spring samples (0.56 mg C m^–3 day^–1,i.e. 0.014 g C^–1 ciliates g C^–1 rotifers day^–1)in contrast to measurements performed in July (8.72 mg C m^–3day^–1, i.e. 0.242 g C^–1 ciliates g C^–1 rotifersday^–1). The proportion of protozoa in the diet of rotiferswas low compared with that of phytoplankton (<30% of totalcarbon ingestion) except when phytoplankton biomass decreasedbelow the incipient limiting level (ILL) of the main metazooplantonicspecies. In such conditions, protozoa (mainly ciliates) constituted50% of total rotifer diet. These results give evidence thatmicrobial organisms play a significant role within the planktonicfood web of a eutrophic lowland river, ciliates providing analternative food for metazooplankton when phytoplankton becomesscarce.     

In situ determinations of bacterial selectivity and filtration rates by five cladoceran zooplankters in a hypertrophic subtropical reservoir

Cladoceran in situ feeding rates on natural bacteria labelledwith [methyl-³H] were studied in parallel with feeding ratedeterminations on ¹⁴C-labelled Chlorella in a hypertrophic subtropicalreservoir (Lake Hartbeespoort) through spring and summer (1986/87).Community filtration rates (CFR5) on bacteria and algae weresimilar, but selection for Chlorella (relative to natural bacteria)increased from midsummer in association with declining bacterialdensity and increasing dominance of ‘inedible’ componentsof the natural phytoplankton. Species-specific filtration rates(SSFRs) were determined for Daphnia pulexllongispina, Ceriodaphniareticulata, Diaphanosoma excisum, Bosmina longirostris and Moinamicrura during their respective seasonal occurrence in the studyperiod. SSFRs on algae and bacteria increased with body length(L, mm) in all species apart from Bosmina. Species-specificdifferences in absolute feeding rate (FR, ml animal^–1day^–1), the slope of the FR-L relationship and bacteriaselectivity were evident. The feeding rate of all cladoceranson bacteria is described by the power equation FR 5.231L^1.42FR values on bacteria relative to FR values on algae averaged     

Microbial dynamics during the summer ice-loss phase in maritime Antarctic lakes

The dynamics of bacterioplankton and protozooplankton in twomaritime Antarctic lakes (Heywood Lake and Sombre Lake, SignyIsland, South Orkneys) were studied during the phase of icebreak-out (December and early January 1994/95). The lakes aresuffering animal-induced (fur seal) eutrophication, though HeywoodLake is most severely affected. Both lakes had morphologicallydiverse bacterial communities which increased during the studyperiod, reaching maxima of 80 x 10⁸ l^–1 in Heywood Lakeand 31.8 x 10⁸ l^–1 in Sombre Lake. Heterotrophic nanoflagellates(HNAN) reached a peak in late December with maxima of 40.6 x10⁸ l^–1 in Sombre Lake and 174 x 10⁵ l^–1 in HeywoodLake. Phototrophic nanoflagellates (PNAN) peaked in late Decemberafter ice loss in Heywood Lake (63 x 10⁵ l^–1), which coincidedwith a peak in a bloom of Chroomonas acuta which reached abundancesof 1.0 x 10⁸ l^–1. In Sombre Lake, ice persisted for alonger period and here PNAN reached their highest density atthe end of the study period (around 70.0 x 10⁵ l^–1). Ciliateabundance reached high levels in Heywood Lake (>60001^–1),while in Sombre Lake maximum abundance was 568l^–1. Protozooplanktondiversity was greater in the less-enriched Sombre Lake. Grazingrates of ciliates averaged 70.6 bacteria indiv.^–1 h^–1in Heywood Lake and 119.3 bacteria indiv.^–1 h^–1in Sombre Lake. The difference was a reflection of the differenttaxonomic make-up of the community in the lakes. HNAN grazingrates varied between 0.51 and 0.83 bacteria indiv.^–1 h^–1in Sombre and Heywood Lakes, respectively. Specific growth rates(r) h^–1 in Sombre Lake were 0.028 for ciliates and 0.013for HNAN, and in Heywood Lake 0.010 for ciliates and HNAN 0.012.These growth rates result in doubling times ranging between38 and 69 h for ciliates and around 55 h for HNAN.HNAN grazingon bacteria was curtailed in Heywood Lake in early January asa result of predation by microcrustacean larvae feeding on theplankton. Thus, for a short phase top-down control was apparentin the dynamics of Heywood Lake, a feature uncommon in Antarcticlake ecosystems. The impact of natural eutrophication on thesesystems is discussed in relation to other unaffected Antarcticlakes.     

Estimation of ammonium regeneration efficiencies associated with bacterivory in pelagic food webs via a 15N tracer method

Phagotrophic protists are major components of pelagic food webs,both as consumers of bacterial and phytoplankton cells, andas regenerators of inorganic nutrients. In this study, we estimatedthe efficiency of ammonium regeneration by protists feedingon bacteria within natural plank-tonic assemblages, using a¹⁵N tracer method, in which the excretion of ¹⁵N-labeled ammoniumdue to grazing on ¹⁵N pre-labeled bacteria was followed overtime. We tested this approach in experiments based on the additionof heat-killed ¹⁵N-labeled bacteria to laboratory cultures andto samples of coastal seawater. During two experiments, variationin abundance of bacterivores and bacterioplankton resulted innon-constant grazing rates. Deterministic computer models thatused abundance of bacteria and protists as variables were developedto estimate best-fit values of grazing mortality (g, h^–1)and of ammonium regeneration efficiency (R_E, fraction of theinitial ¹⁵N label in added bacteria which is released as ammonium).Estimated ammonium R_E were 0.30–0.35 for one trophic linksystems with both a monospecific culture and a mixed speciesassemblage of bacterivorous flagellates. R_E was higher for multi-trophicstep food webs: 0.60 for 5 µm pre-screened coastal seawaterand 0.90 for whole coastal seawater.     

Daily variations of highly active bacteria in the Northern Adriatic Sea

Nowadays, it is recognized that only a fraction of aquatic bacteriaare actively growing, but there is little information aboutthe factors constraining their metabolism. Marine bacterioplanktoncan rapidly modify their metabolic activity level in responseto environmental changes. In this study, we focused on the dailychanges in abundance and activity of active bacterial fractionover a 20-day period preceded by intense rainfalls which slightlymodified water column conditions. Cells capable of reducingthe membrane-penetrable dye 5-cyano-2,3-ditolyl tetrazoliumchloride (CTC), estimated by epifluorescence microscopy, areconsidered very active (CTC+ bacteria). Total bacterial abundance(TBA) ranged from 0.8 to 2.4 x 10⁹ cells L^–1, whereasCTC+ bacteria were more variable (1.6–9.2 x 10⁷ cellsL^–1), accounting for 1.2–4.4% of TBA. Bacterialactivity (BA) quantified as the incorporation of [3H]-leucinevaried by more than one order of magnitude over the period (25.0–662.5pmol L^–1 h^–1). BA was strongly related to CTC+ bacteria,suggesting that they were mainly responsible for the bacterialcommunity metabolism. Nevertheless, cell-specific activity,scaled to only CTC+ cells, was very high, suggesting that afraction of cells not detectably CTC+ may be able to assimilate[3H]-leucine. The correlation between salinity and TBA, CTC+bacteria and BA supported the hypothesis of the active roleof freshwater input in enhancing cell activity. Our resultssuggest that freshwater inputs rather than phytoplanktonic bloomsare able to induce shifts in bacterial metabolism over a timescale of days in the area studied.     

Increase in the proportion of metabolically active bacteria along gradients of enrichment in freshwater and marine plankton: implications for estimates of bacterial growth and production rates

It is generally recognized that a fraction of all bacterioplanktoncells enumerated using conventional epifluorescence techniquesis neither growing, dividing nor metabolically active, but thevariation in the proportion of active cells among aquatic systemsis not well understood. Here, we hypothesize that the proportionof metabolically active cells increases systematically alonggradients of enrichment, and to test this hypothesis the numberand proportion of metabolically active planktonic bacteria wereinvestigated during the summer in a set of 24 temperate lakes,which span a considerable range in productivity. The tetrazoliumsalt 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) was usedas an indicator of cells with an active electron transport system.The total number of bacteria ranged from 1.88x10⁶ to 7.70x10⁶ml^–1, whereas the number of active cells was more variableand ranged from 0.37x10⁶ to 2.18x10⁶ ml^–1 in the studylakes. The proportion of metabolically active cells ranged from15 to 33%, and tended to increase with nutrient and chlorophyllconcentrations, but not with dissolved organic carbon (DOC).Data on the number of total and active bacteria culled fromthe literature for marine, estuarine and freshwater systemsshow that the trends we measured in lakes are valid for pelagicsystems in general. Over a broad range of aquatic systems, thetotal number of bacteria varied by three orders of magnitude,whereas the number of active bacteria varied by four ordersof magnitude as system productivity increased. The proportionof active cells increased from ultraoligotrophic openocean areas(<5%) to highly productive estuaries (>50%). Our resultssuggest that in most aquatic systems there is a pool of rapidlygrowmg cells, embedded in a usually larger matrix of inactivebacteria, and that the relative size of the active and inactivepools varies systematically among bacterial populations alonggradients of enrichment.     

Comparative studies on the effects of anions on ATPase and acid phosphatase activities from different sources

The effect of several anions on Mg²⁺-ATPase activity [EC 3.6.1.3 [EC] ]was studied in crude extracts of the following sources: twelvespecies of bacteria, one fungus, one yeast, six species of algae,five higher plant tissues, and two animal tissues. All the materialsexamined exhibited Mg²⁺-ATPase activity at pH 7.5. Except fora few species of bacteria, blue-green alga and etiolated seedlingsof a leguminous plant, Mg²⁺-ATPases of all the materials studiedwere stimulated two- to ten-fold by the addition of Group VIanions such as sulfite, selenite, and chromate. This stimulationseems to be a common characteristic of ATPases of most bacteria,chloroplasts, and mitochondria. On the other hand, the stimulation of acid phosphatase by sulfatewas not observed in any organism other than Thiobacillus thiooxidans. (Received July 27, 1977; )     

The effect of detrital addition on the development of nanoflagellates and bacteria in Lake Kinneret

The effect of adding dissolved substrates derived from algalcells on the patterns of nutrient cycling and growth of bacteria,heterotrophic nanoflagellates (HNAN) and photoautotrophs wasdetermined in samples of near-surface waters from Lake Kinneret.Supplementation of substrates always resulted in an increasedpeak of HNAN numbers and had little effect on bacterial numbers.HNAN-mediated nutrient remineralization of nitrogen and phosphoruswas also stimulated. In light-incubated samples the remineralizednutrients were taken up by photoautotrophic cells. Maximum growthrates observed for HNAN ranged from 0.03 to 0.11 h^–1,clearance rates for bacteria 1.1–7.3 nl HNAN^–1 h^–1and remineralization rates 6.4–8.4 µg N mg dry wt^–1h^–1 and 0.37–0.99 µg P mg dry wt^–1 h^–1.     

Time Course of Cell Biological Events Evoked in Legume Root Hairs by Rhizobium Nod Factors: State of the Art

In many common legumes, when host-specific nodule bacteria meettheir legume root they attach to it and enter through root hairs.The bacteria can intrude these cells because they instigatein the hairs the formation of an inward growing tube, the infectionthread, which consists of wall material. Prior to infectionthread formation, the bacteria exploit the cell machinery forwall deposition by inducing the hairs to form a curl, in whichthe dividing bacteria become entrapped. In most species, Nodfactor alone (a lipochito-oligosaccharide excreted by bacteria)induces root hair deformation, though without curling, thusmost aspects of the initial effects of Nod factor can be elucidatedby studying root hair deformation. In this review we discussthe cellular events that host-specific Nod factors induce intheir host legume root hairs. The first event, detectable onlya few seconds after Nod factor application, is a Ca²⁺influxat the root hair tip, followed by a transient depolarizationof the plasma membrane potential, causing an increase in cytosolic[Ca²⁺] at the root hair tip. Also within minutes, Nod factorschange the cell organization by acting on the actin cytoskeleton,enhancing tip cell wall deposition so that root hairs becomelonger than normal for their species. Since the remodellingof the actin cytoskeleton precedes the second calcium event,Ca²⁺spiking, which is observed in the perinuclear area, we proposethat the initial cytoskeleton events taking place at the hairtip are related to Ca²⁺influx in the hair tip and that Ca²⁺spikingserves later events involving gene expression. Copyright 2001Annals of Botany Company Review, Nod factor, tip growth, root hair, Rhizobium, legume, cytoskeleton, calcium, symbiosis     

Ca(2+)-dependent activation of c-jun NH(2)-terminal kinase in primary rabbit proximal tubule epithelial cells

Previous work from this laboratorydemonstrated that arachidonic acid activates c-junNH₂-terminal kinase (JNK) through oxidative intermediatesin a Ca²⁺-independent manner (Cui X and Douglas JG.Arachidonic acid activates c-jun N-terminal kinase throughNADPH oxidase in rabbit proximal tubular epithelial cells. ProcNatl Acad Sci USA 94: 3771-3776, 1997.). We now report thatJNK can also be activated via a Ca²⁺-dependent mechanism byagents that increase the cytosolic Ca²⁺ concentration(Ca²⁺ ionophore A₂₃₁₈₇, Ca²⁺-ATPaseinhibitor thapsigargin) or deplete intracellular Ca²⁺stores [intracellular Ca²⁺ chelator1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid(BAPTA)-AM]. The activation of JNK by BAPTA-AM occurs despite adecrease in cytosolic Ca²⁺ concentration as detected by theindicator dye fura 2, but appears to be related to Ca²⁺metabolism, because modification of BAPTA with two methyl groups increases not only the chelation affinity for Ca²⁺, butalso the potency for JNK activation. BAPTA-AM stimulates Ca²⁺ influx across the plasma membrane, and the resultinglocal Ca²⁺ increases are probably involved in activation ofJNK because Ca²⁺ influx inhibitors (SKF-96365, nifedipine)and lowering of the free extracellular Ca²⁺ concentrationwith EGTA reduce the BAPTA-induced JNK activation.

     

Factors affecting the bacteria-heterotrophic nanoflagellate relationship in oligo-mesotrophic lakes

The coupling between bacteria and heterotrophic nanoflagellates(HNF) was examined in nine lakes of low productivity for evidenceof the effects of various metazooplankton (i.e. rotifers, cladoceransand copepods) on this relationship. We considered the size ofcladocerans and, in contrast to most previous across-systemstudies, the three strata of the water column (i.e. epilimnion,metalimnion and hypolimnion). Rotifers were numerically dominantin all lakes and accounted for 45–84% of total metazooplanktonabundance, while the abundance of large cladocerans was relativelylow, ranging from 0.066 to 15.2 ind. L^–1. The across-lakerelationship between bacteria and HNF was significant in thedeeper strata (meta- and hypolimnion) but not in the epilimnionand in the two groups of lakes separated on the basis of theiraverage number of large cladocerans (<5 and >5 ind. L^–1,respectively). The results confirmed the negative impacts oflarge cladocerans on HNF, but also showed that rotifers, probablythrough grazing on HNF, may be an important factor causing variationin the bacteria–HNF relationship in unproductive waters.Quadratic models best described the relationships between metazooplanktonand the ratio of bacteria to HNF. This ratio seemed to be aresult of complex interactions between several factors, includingthe zooplankton composition and abundance and the depth of thelake. Indeed, this ratio significantly decreased across lakes,with increase in depth. In addition, shallower lakes (having<5 large cladocerans L^–1 and fewer Polyarthra vulgaris)tended to have more bacteria and HNF and a higher ratio of bacteriato HNF than deeper lakes (which had >5 large cladoceransL^–1 and substantial proportions of P. vulgaris). We suggestthat the epilimnion, metalimnion and hypolimnion of lakes betaken into account when analysing the bacteria–HNF relationshipas well as the cascading effects of zooplankton on microbialcommunities.     

Some Effects of Douglas Fir Terpenes on Certain Microorganisms

The Douglas fir terpene α-pinene was shown to inhibit the growth of a variety of bacteria and a yeast. Other terpenes of the Douglas fir, including limonene, camphene, and isobornyl acetate, were also inhibitory to Bacillus thuringiensis. All terpenes were inhibitory at concentrations normally present in the fir needle diet of Douglas fir tussock moth larvae. The presence of such terpenes in the diet of these insects was found to strongly influence the infectivity of B. thuringiensis spores for the Douglas fir tussock moth larvae. The terpene α-pinene destroyed the cellular integrity and modified mitochondrial activity in certain microorganisms.     

甘草根茎乙醇提取物抗菌活性研究

本实验采用琼脂扩散法和微量肉汤稀释法,研究了甘草根茎乙醇提取物对5种细菌(表皮葡萄球菌、金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌和绿脓杆菌)和2种真菌(白色念珠菌和黑曲霉)的抗菌活性。结果表明,甘草根茎乙醇提取物对革兰氏阳性菌非常敏感,而对革兰氏阴性菌和真菌不敏感,80%乙醇提取物对革兰氏阳性菌的MIC范围为0.156～0.312 mg·mL^-1,而10%乙醇提取物对革兰氏阳性菌的MIC范围为0.625～1.250 mg·mL^-1,表明甘草根茎抗菌活性成分在高浓度乙醇中溶解度较大,为临床上应用甘草根茎醇提物作为抗菌制剂提供了科学依据。     

Nitrogen Fixation by Free-living Bacteria in the Soil and in the Canopy of Douglas Fir

Nitrogen fixing bacteria have been isolated from the soil andthe leaves of Douglas fir. Measurements, taken at monthly intervalsusing ¹⁵N, have shown that small quantities of nitrogen arefixed on the leaves and in the various soil layers and thatthe highest rates of fixation occur in the spring. Further studieswith ¹⁵N have shown that the products of nitrogen fixation inDouglas fir soils are available for growth of seedlings andfor denitrification.     

Nitrogen Fixation in the Canopy of Temperate Forest Trees: A Re-examination

¹⁵N₂ studies and acetylene reduction assays of leaves and shootsof Douglas fir and other forest trees do not confirm previousreports that extensive nitrogen fixation occurs on leaf surfacesand it is concluded that the importance of nitrogen fixationin the canopy of forest trees has been exaggerated. The presenceof nitrogen-fixing bacteria on the leaves of trees is confirmed,however, and they have been identified as Enterobacter agglomerans,Clostridium butyricum and Bacillus sp. Their distribution onleaves is fortuitous since dead oak leaves and artificial leavesbecome colonized to the same extent as living oak leaves. nitrogen fixation, acetylene reduction, Enterobacter agglomerans, Clostridium butyricum, Bacillus sp, Douglas fir, Pseudotsuga menziensii, larch, Larix x oak, Quercus petraea.