Effect of Inorganic Nutrients on Relative Contributions of Fungi and Bacteria to Carbon Flow from Submerged Decomposing Leaf Litter

The relative contributions of fungi and bacteria to carbon flow from submerged decaying plant litter at different levels of inorganic nutrients (N and P) were studied. We estimated leaf mass loss, fungal and bacterial biomass and production, and microbial respiration and constructed partial carbon budgets for red maple leaf disks precolonized in a stream and then incubated in laboratory microcosms at two levels of nutrients. Patterns of carbon flow for leaf disks colonized with the full microbial assemblage were compared with those colonized by bacteria but in which fungi were greatly reduced by placing leaf disks in colonization chambers sealed with membrane filters to exclude aquatic hyphomycete conidia but not bacterial cells. On leaves colonized by the full microbial assemblage, elevated nutrient concentrations stimulated fungi and bacteria to a similar degree. Peak fungal and bacterial biomass increased by factors of 3.9 and 4.0; cumulative production was 3.9 and 5.1 times higher in the high nutrient in comparison with the low nutrient treatment, respectively. Fungi dominated the total microbial biomass (98.4 to 99.8%) and cumulative production (97.3 and 96.5%), and the fungal yield coefficient exceeded that of bacteria by a factor of 36 and 27 in low- and high-nutrient treatments, respectively. Consequently, the dominant role of fungi in leaf decomposition did not change as a result of nutrient manipulation. Carbon budgets indicated that 8% of leaf carbon loss in the low-nutrient treatment and 17% in the high-nutrient treatment were channeled to microbial (essentially fungal) production. Nutrient enrichment had a positive effect on rate of leaf decomposition only in microcosms with full microbial assemblages. In treatments where fungal colonization was reduced, cumulative bacterial production did not change significantly at either nutrient level and leaf decomposition rate was negatively affected (high nutrients), suggesting that bacterial participation in carbon flow from decaying leaf litter is low regardless of the presence of fungi and nutrient availability. Moreover, 1.5 and 2.3 times higher yield coefficients of bacteria in the reduced fungal treatments at low and high nutrients, respectively (percentage of leaf carbon loss channeled to bacterial production), suggest that bacteria are subjected to strong competition with fungi for resources available in leaf litter.     

Diversity and Decomposing Ability of Saprophytic Fungi from Temperate Forest Litter

This study was designed to examine saprophytic fungi diversity under different tree species situated in the same ecological context. Further, the link between the diversity and decomposition rate of two broadleaved, two coniferous and two mixed broadleaved-coniferous litter types was targeted. Litter material was decomposed in litter bags for 4 and 24 months to target both early and late stages of the decomposition. Fungal diversity of L and F layers were also investigated as a parallel to the litter bag method. Temperature gradient gel electrophoresis fingerprinting was used to assess fungal diversity in the samples. Mass loss values and organic and nutrient composition of the litter were also measured. The results showed that the species richness was not strongly affected by the change of the tree species. Nevertheless, the community compositions differed within tree species and decomposition stages. The most important shift was found in the mixed litters from the litter bag treatment for both variables. Both mixed litters displayed the highest species richness (13.3 species both) and the most different community composition as compared to pure litters (6.3–10.7 species) after 24 months. The mass loss after 24 months was similar or greater in the mixed litter (70.5% beech–spruce, 76.2% oak–Douglas-fir litter) than in both original pure litter types. This was probably due to higher niche variability and to the synergistic effect of nutrient transfer between litter types. Concerning pure litter, mass loss values were the highest in oak and beech litter (72.8% and 69.8%) compared to spruce and D. fir (59.4% and 66.5%, respectively). That was probably caused by a more favourable microclimate and litter composition in broadleaved than in coniferous plantations. These variables also seemed to be more important to pure litter decomposition rates than were fungal species richness or community structure.     

Diversity of Fungi, Bacteria, and Actinomycetes on Leaves Decomposing in a Stream

Although fungi, bacteria, and specific bacterial taxa, such as the actinomycetes, have been studied extensively in various habitats, few studies have examined them simultaneously, especially on decomposing leaves in streams. In this study, sugar maple and white oak leaves were incubated in a stream in northeastern Ohio for 181 days during which samples were collected at regular intervals. Following DNA extraction, PCR-denaturing gradient gel electrophoresis (DGGE) was performed using fungus-, bacterium-, and actinomycete-specific primers. In addition, fungal and bacterial biomass was estimated. Fungal biomass differed on different days but not between leaves of the two species and was always greater than bacterial biomass. There were significant differences in bacterial biomass through time and between leaf types on some days. Generally, on the basis of DGGE, few differences in community structure were found for different leaf types. However, the ribotype richness of fungi was significantly greater than those of the bacteria and actinomycetes, which were similar to each other. Ribotype richness decreased toward the end of the study for each group except bacteria. Lack of differences between the two leaf types suggests that the microorganisms colonizing the leaf biofilm were primarily generalists that could exploit the resources of the leaves of either species equally well. Thus, we conclude that factors, such as the ecological role of the taxa (generalists versus specialists), stage of decay, and time of exposure, appeared to be more important determinants of microbial community structure than leaf quality.     

Contribution of Fungi and Bacteria to Leaf Litter Decomposition in a Polluted River

The contribution of fungi and bacteria to the decomposition of alder leaves was examined at two reference and two polluted sites in the Ave River (northwestern Portugal). Leaf mass loss, microbial production from incorporation rates of radiolabeled compounds into biomolecules, fungal biomass from ergosterol concentration, sporulation rates, and diversity of aquatic hyphomycetes associated with decomposing leaves were determined. The concentrations of organic nutrients and of inorganic nitrogen and phosphorus in the stream water was elevated and increased at downstream sites. Leaf decomposition rates were high (0.013 day⁻¹ < k < 0.042 day⁻¹), and the highest value was estimated at the most downstream polluted site, where maximum values of microbial production and fungal biomass and sporulation were found. The slowest decomposition occurred at the other polluted site, where, along with the nutrient enrichment, the lowest current velocity and dissolved-oxygen concentration in water were observed. At this site, fungal production, biomass, and sporulation were depressed, suggesting that stimulation of fungal activity by increased nutrient concentrations might be offset by other factors. Although bacterial production was higher at polluted sites, fungi accounted for more than 94% of the total microbial net production. Fungal yield coefficients varied from 10.2 to 13.6%, while those of bacteria were less than 1%. The contribution of fungi to overall leaf carbon loss (29.0 to 38.8%) greatly exceeded that of bacteria (4.2 to 13.9%).     

The Spatial Distribution of Fungi on Decomposing Woody Litter in a Freshwater Stream,Western Ghats,India

We mapped filamentous fungal association with mechanically “hard” and “soft” woody litter naturally deposited in a stream of the Western Ghats of India. Using a durometer (rubber hardness tester), the toughness of surface of wood collected from stream was determined by considering durometer reading from 60–72 to 30–37 as hardwood and softwood, respectively. From each wood (1.5 cm diameter), two segments each of 3 cm length were excised and vertically cut into nine sections comprising eight marginal and one central section. From three stream locations, hardwood and softwood sections were assessed for the occurrence of lignicolous and Ingoldian fungi. A first set of wood sections was incubated in damp chambers up to 4 months with periodical screening (every 2 weeks) for lignicolous fungi. Another set was incubated in bubble chambers up to 72 h to ascertain colonization of Ingoldian fungi. In hardwood sections, 17 lignicolous fungi (ascomycetes, four; mitosporic fungi, 13; mean, 6.8; range, 6–8/section) and ten Ingoldian fungi (mean, 2; range, 0–4/section) comprising nine lignicolous (11.1–40.7%) and three Ingoldian (11.1–14.8%) fungi as core-group taxa were recovered. In softwood, ten lignicolous fungi (ascomycetes, 0; mitosporic fungi, ten; mean, 3.8; range, 2–5/section) and 26 Ingoldian fungi (mean, 8.1; range, 5–10/section) comprising six lignicolous (11.1–85.2%) and 12 Ingoldian (11.1–88.9%) fungi as core-group taxa were recovered. The ratio of lignicolous fungi/Ingoldian fungi was higher in hardwood than softwood (1.7 vs. 0.4). The spore output of Ingoldian fungi was higher in softwood (mean, 901 g⁻¹; range, 80–2546 g⁻¹) than hardwood (mean, 21 g⁻¹; range, 0–140 g⁻¹). The Shannon diversity of lignicolous fungi was higher in hardwood than softwood (3.604 vs. 2.665), whereas it was opposite for Ingoldian fungi (3.116 vs. 3.918). The overall fungal diversity was higher in softwood than hardwood (4.413 vs. 4.219). The range of Jaccard’s index of similarity among wood sections was higher in lignicolous fungi (8–71% and 13–75%) than Ingoldian fungi (0–50% and 8–55%) in hardwood and softwood. The rarefaction indices of expected number of taxa against hardwood sections revealed higher and persistent lignicolous fungi than the Ingoldian fungi, while the Ingoldian fungi were persistent in softwood sections, although they were lower than lignicolous fungi. Our study demonstrated the dominance of lignicolous fungi and Ingoldian fungi in hardwood and softwood, respectively.     

Antibiosis between Ruminal Bacteria and Ruminal Fungi

Cellulose digestion, bacterial numbers, and fungal numbers were monitored over time in vitro by using a purified cellulose medium with and without antibiotics (penicillin and streptomycin). All fermentations were inoculated with a 1:10 dilution of whole rumen contents (WRC). Without antibiotics, cellulose digestion was higher (P < 0.01) at 24, 30, 48, and 72 h; fungi had almost disappeared by 24 h, while bacterial concentrations increased over 100-fold in 24 h and then decreased gradually up to 72 h. In those fermentations with added antibiotics, fungal concentrations increased 4-fold by 30 h and up to 42-fold at 72 h; bacterial concentrations were markedly reduced by 24 h and remained low through 72 h. Similar results were obtained with ground alfalfa as a substrate. In further studies, the in vitro fermentation of purified cellulose without antibiotics was stopped after 18 to 20 h, and the microbial population was killed by autoclaving. Antibiotics were added to half of the tubes, and all tubes were reinoculated with WRC. After 72 h, extensive cellulose digestion had occurred in those tubes without antibiotics, as compared to very low cellulose digestion with added antibiotics. The extent of this inhibition was found to increase in proportion to the length of the initial fermentation period, suggesting the production of a heat-stable inhibitory factor or factors. The inhibitory activity was present in rumen fluid, could be extracted from lyophilized rumen fluid (LRF) with water, and was stable in response to proteolytic enzymes. In addition, the water-extracted residue of LRF was found to contain growth factor activity for rumen fungi in vitro.     

Influence of Environmental Factors on Antagonism of Fungi by Bacteria in Soil: Nutrient Levels

The addition of 0.25, 0.5, or 1.0% glucose to a soil (K) amended with either 6% kaolinite (K6K) or montmorillonite (K6M) or the adjustment of the C/N ratio of the soils from 23/1 to 10/1 with NH₄NO₃ eliminated the inhibition of Aspergillus niger by Serratia marcescens, regardless of whether the fungus and bacterium were inoculated into the same or separate sites in the soils. The adjustment of the C/N ratio to 15/1 or of the C/P ratio from 1,000/1 to 100/1 with KH₂PO₄ did not eliminate the antagonism. However, with the higher glucose and NH₄NO₃ amendments, S. marcescens died out in the K and K6K (but not in the K6M) soils, apparently due to reductions in pH that resulted from the increased metabolism induced by added nutrients. In soils amended with CaCO₃, S. marcescens did not die out, but the inhibition of A. niger by S. marcescens or Agrobacterium radiobacter was eliminated or reduced by the addition of glucose, but not of NH₄NO₃, and was influenced by the clay mineralogy and pH of the soils. When NH₄NO₃ was added to the soils adjusted with CaCO₃ to pH values above 6.0, growth of A. niger was inhibited, regardless of whether bacteria were present or not, as a result of the volatilization of NH₃. Bacillus cereus and another species of Bacillus did not inhibit A. niger under any of the environmental conditions. There was a direct correlation between the degree of inhibition and the rate of glucose utilization by the various bacteria, indicating that the antagonism of A. niger by some bacteria in soil was the result primarily of a competition for carbon and that this competition was influenced by other environmental factors, such as pH and clay mineralogy.     

Effects of Zn,Fe and Mn on Leaf Litter Breakdown by Aquatic Fungi: a Microcosm Study

We investigated the effects of heavy metals on leaf litter decomposition in streams. Leaves were immersed (10 days) at a reference (R) and a metal‐impacted (I) site and exposed in microcosms with increased Zn, Mn or Fe content, and to stream water from site R or I. Fungal biomass was higher in microcosms with leaves colonized at I and water from R. Fungal sporulation was higher in microcosms with leaves and water from R. Concentrations of 4.9, 9.6 and 5 ppm of Zn, Mn and Fe decrease fungal sporulation. The number of fungal species (spore counts and DGGE fingerprints) was lower in leaves colonized at site I. Cluster analyses of DGGE showed that Fe was the metal that most altered the structure of fungal community. Our results suggest that metal pollution affect leaf‐associated fungi depending on metal identity and concentration, and effects appear to be less pronounced in metal‐adapted communities. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Plant Reactions to Inoculation of Roots with Fungi and Bacteria

The potential of 120 isolates of fungi and bacteria from plant rhizospheres to interfere with plant development and growth was studied in greenhouse experiments. The pure cultured isolates were obtained from plant roots in the field and applied as suspensions to the roots of eight test plant species. 10–20% of the isolates caused distinct symptoms on shoots, growth retardations without other symptoms or growth promotions. Responses of treated plants ranged from death of plants soon after treatment to up to about 40% higher shoot fresh weight than in control plants. Two bacterial isolates induced strong reactions in most of the plant species tested while other isolates showed a more or less pronounced specificity by giving reactions in only some of the plant species tested.     

Influence of Environmental Factors on Antagonism of Fungi by Bacteria in Soil: Clay Minerals and pH

The soil replica plating technique was used to evaluate the influence of clay minerals and pH on antagonistic interactions between fungi and bacteria in soil. In general, the antagonistic activity of bacteria towards filamentous fungi was greater in soil than on agar. The spread of Aspergillus niger through soil was inhibited by Serratia marcescens when the organisms were inoculated into separate sites in soil, and this antagonistic effect was maintained when the soil was amended with 3, 6, 9, or 12% (vol/vol) montmorillonite, whereas the addition of kaolinite at a concentration of 3% reduced the antagonism and at 6, 9, or 12% totally eliminated it. Similar results were obtained with the inhibition of A. niger by Agrobacterium radiobacter and of Penicillium vermiculatum by either S. marcescens or Nocardia paraffinae. When A. niger and S. marcescens were inoculated into the same soil site, A. niger was inhibited in all soils, regardless of clay content, although the extent of inhibition was greater as the concentration of montmorillonite, but not of kaolinite, increased. A. niger was inhibited more when inoculated as spores than as mycelial fragments and when inoculated 96 h after S. marcescens, but a 1% glucose solution reduced the amount of inhibition when the fungus was inoculated 96 h after the bacterium. When the pH of the soil-clay mixtures was altered, the amount of antagonism usually increased as the pH increased. Antagonism appeared to be related to the cation-exchange capacity and the pH of the soil-clay mixtures. Bacillus cereus and another species of Bacillus showed no activity in soil towards A. niger under any of the environmental conditions tested, even though the Bacillus sp. significantly inhibited A. niger and seven other fungi on agar.     

Potential Physiological Activities of Fungi and Bacteria in Relation to Plant Litter Decomposition along a Gap Size Gradient in a Natural Subtropical Forest

Abstract The dynamics of fungal and bacterial potential physiological activities during leaf, branch, and bark litter decomposition along a gap size gradient in a subtropical forest was determined using substrate-induced respiration (SIR) with antibiotics selective for fungi and bacteria, respectively. A gap size gradient (1) was under closed canopy; (2) had small gaps with a diameter (≤5m); (3) had small to intermediate gaps (5–15 m diameter); (4) had intermediate to large gaps (15–30 m diameter); and (5) had large gaps (≥30 m diameter). Litter decomposition was studied using a litter bag technique. Fungi had higher SIR than bacteria for each type of litter in any size class of gaps. Gaps 1, 2, and 3 had higher fungal and bacterial SIRs than gaps 4 and 5. Moreover, decomposing leaf litter exhibited higher fungal and bacterial SIRs than branch, and branch higher than bark. Simple correlation analysis indicated that fungal SIR was a reliable index of decomposition rates. Fungal SIR was significantly and positively correlated with soil moisture, whereas bacterial SIR was not significantly correlated with soil moisture. The relationships among microclimatic factors, fungal and bacterial physiological activities, and rates of plant litter decomposition suggest that, in subtropical ecosystems, fungal community activities were strongly and directly regulated by the environmental heterogeneity within gaps, and an important regulator of rates of plant litter decomposition rates. Received: 13 January 1997; Accepted 28 March 1997     

小麦内生细菌的分离及其对小麦纹枯菌的拮抗作用

利用涂布平板法从小麦根系中分离出8株内生细菌,从中筛选出1株对小麦纹枯菌(Rhizoctonia cerealis)具有拮抗作用的内生菌。室内测定该菌株培养液对小麦纹枯病菌的抑制作用,结果发现,小麦纹枯病菌在培养液中生长缓慢,培养6d后菌丝量与对照相比下降了89％,同时发现病菌菌丝生长畸形,出现断裂和细胞壁瓦解。双抗标记法测定该拮抗菌在小麦根系中的定殖情况,发现该菌能够在根系中长期定殖。初步的鉴定结果表明该菌为蜡样芽孢杆菌。     

深海细菌及其活性物质防控植物病原真菌的研究进展

近年来,从天然产物中开发生物农药成为了研究者关注的热点,海洋微生物因其独特的生存环境,产生了许多从陆地微生物中未曾发现的生物活性物质,为新型生物农药开发带来了佳音。从微生物多样性、抑菌物质的种类与结构和抑菌机理3个方面阐述深海细菌防控植物病原真菌的研究进展,并对其发展前景进行了展望,以期为抗植物病原真菌的海洋微生物的开发利用提供参考。     

High Diversity of Fungi may Mitigate the Impact of Pollution on Plant Litter Decomposition in Streams

We investigated how a community of microbial decomposers adapted to a reference site responds to a sudden decrease in the water quality. For that, we assessed the activity and diversity of fungi and bacteria on decomposing leaves that were transplanted from a reference (E1) to a polluted site (E2), and results were compared to those from decomposing leaves either at E1 or E2. The two sites had contrasting concentrations of organic and inorganic nutrients and heavy metals in the stream water. At E2, leaf decomposition rates, fungal biomass, and sporulation were reduced, while bacterial biomass was stimulated. Fungal diversity was four times lower at the polluted site. The structure of fungal community on leaves decomposing at E2 significantly differed from that decomposing at E1, as indicated by the principal response curves analysis. Articulospora tetracladia, Anguillospora filiformis, and Lunulospora curvula were dominant species on leaves decomposing at E1 and were the most negatively affected by the transfer to the polluted site. The transfer of leaves colonized at the reference site to the polluted site reduced fungal diversity and sporulation but not fungal biomass and leaf decomposition. Overall, results suggest that the high diversity on leaves from the upstream site might have mitigated the impact of anthropogenic stress on microbial decomposition of leaves transplanted to the polluted site.     

Competitive Interaction between Two Aquatic Hyphomycete Species and Increase in Leaf Litter Breakdown

Aquatic hyphomycete species produce large numbers of conidia which rapidly colonize the leaf litter that falls into rivers during autumn. Our objective was to understand how a species which produces many fewer conidia than another in laboratory conditions can nevertheless be codominant in a natural setting. In microcosm studies with two pioneer dominant species, Flagellospora curvula and Tetrachaetum elegans, inoculated on alder leaves, we first verified that the ratio of the conidium production of both species (6 to 7:1) was inverse to that of individual conidial masses (1:7) as previously described. Calculating the percentage of leaf mass loss that corresponds to 1Â mg of conidial mass produced, the combination of the two species produced 2.9-fold more loss than the mean of each species. By contrast, the reproductive biomasses of F. curvula and T. elegans were 5.2- and 2.6-fold lower, respectively. As a result, the conidium production of F. curvula in the combination was only 3.2-fold that of T. elegans instead of 6- to 7-fold in pure culture. In a mixed culture of the two species, T. elegans conidia had a high germination potential (>90%) whereas the proportion of germinated F. curvula conidia was only 50%. Moreover, T. elegans reduced the area on which F. curvula could grow on poor and rich solid media. These results indicate that the dominance of F. curvula conidia in the river may be partly controlled by T. elegans and suggest that a negative interaction between microfungi may have a positive effect on the ecosystem functioning.     

核桃凋落叶分解对萝卜生长和生理的影响

为考察核桃(Juglans regia L.)与冬性作物复合种植模式中凋落叶分解对作物生长的影响,该试验采用盆栽法,以冬性作物萝卜(Raphanus sativus L.)为受体,种入混有0、60、120和180g/pot核桃凋落叶的土壤中,并在萝卜生长过程中测定其形态、生理及生殖相关指标。结果表明:(1)萝卜地上部分的生长在凋落叶分解初期受到显著抑制,而在后期得到明显恢复。(2)在核桃凋落叶分解的影响下萝卜出现了明显的胁迫响应,特别是在分解初期和较高凋落叶量(120、180g/pot)处理下,光合色素含量受到明显抑制,叶片内活性氧含量增加,并在一定程度上受到渗透胁迫。(3)在测定时期内,萝卜叶片内MDA含量并未显著增加,即萝卜可通过自身调节抵御胁迫造成的危害。(4)萝卜在较高凋落叶剂量处理下,产出的种子颜色较深,且在180g/pot处理下产出的种子萌发率较低,种子质量受到明显影响。研究认为,土壤中核桃凋落叶分解初期能对萝卜造成化感胁迫,但长久来看不会降低萝卜产量;将两者复合种植时,可保留地面凋落叶,且最好在叶片分解一段时间后种植作物。     

Aquatic Hyphomycete Communities Associated with Decomposing Alder Leaf Litter in Reference Headwater Streams of the Basque Country (northern Spain)

The community of aquatic hyphomycetes associated with decomposing alder leaf litter was studied during autumn-winter in nine headwater reference streams of the Basque Country (northern Spain). In order to study the spatial variability in composition and community structure, three streams from each of three different river basins were compared. The colonization dynamics and community changes throughout the decomposition process were also followed in three of the rivers (one per basin). The taxonomic richness and community structure of these fungi varied among rivers, including similar streams of a given watershed. However, neither species diversity nor total abundance was statistically related to environmental variables. Only the conidial production of two of the species, Flagellospora curvula and Lunulospora curvula appeared to be enhanced by nitrate availability in the water. The taxonomic richness and the reproductive activity (sporulation rate) were positively related to the leaf litter decomposition rate. The changes in conidial production along the process were similar for all the streams and helped explain leaf litter quality dynamics.