Biodiversity amongst microorganisms and its relevance

Concluding remarks from the joint IUBS/IUMS workshop on Biodiversity amongst microorganisms and its relevance held in Amsterdam on 7–8 September 1991. An international microbial ecology programme can be justified in its own right now that appropriate investigative tools have been developed. Microorganisms influence global change, and indicate global health and environmental quality. At the same time, an inventory of the world's microbial species and their properties is required, together with associated culture collections and genomes. Sampling methods need to be standardized, both for species and functions. Extreme environments are a particularly rich source of microbial genomes, and endangered habitats should be sampled as a matter of priority. Cataloguing and conserving the world's microbial biodiversity is justifiable and scientifically important.     

Biodiversity conservation in the Gulf of Guinea islands: taking stock and preparing action

This paper introduces an Action Statement on biodiversity conservation in the four Gulf of Guinea islands arising from a workshop held in Jersey in 1993. The current state of the main habitats on all the islands, pressures acting upon them, existing institutions and actions taken so far are reviewed. The 24-point Action Statement was produced as a framework for future international initiatives in biodiversity research and conservation in this part of Africa. Attention is drawn to the progress made by the governments of the region in legislation and institutional support. The workshop recommended the establishment of a regional programme of research and conservation in conjunction with the Governments of Equatorial Guinea and of São Tomé and Príncipe.     

Effect of temperature and light on the toxicity and growth of the blue-green alga Microcystis aeruginosa (UV-006)

The toxicity and growth of Microcystis aeruginosa (UV-006) from the Hartbeespoort Dam, South Africa were investigated at different temperatures and photon fluence rates under laboratory conditions. Cells harvested in late logarithmic growth phase were most toxic when grown at 20°C (LD₅₀) median lethal dose [IP, mouse]=25.4 mg kg^-1). Toxicity was markedly reduced at growth temperatures above 28° C. Fluence rate had a smaller effect on the toxicity of the cells, but toxicity tended to be less at the very low and high light fluences. Optimal conditions for growth did not coincide with those for toxin production. Well-aerated cultures of this isolate kept at pH 9.5 by CO₂ addition, a temperature of 20–24° C, a fluence rate of 145 mol photons m^-2 s^-1 and harvested in the late logarithmic growth phase yielded the maximum quantity of toxin.Abbreviation LD₅₀ median lethal dose An abstract of this work, presented as a poster at the IUBS symposium on toxins and lectins, held at the CSIR, Pretoria, South Africa during 1982 was published in S. Afr. J. Sci. 78, 375 (1982)     

Elevational and age gradients in hawaiian montane rainforest: foliar and soil nutrients

Summary Soils and plants were sampled along an elevational gradient from 265–1675 m on a 133-and a 3100-year-old lava flow on Mauna Loa, Hawai'i. Soil organic matter and nutrients accumulated more rapidly at low elevation on the young flow, but reached higher levels at higher elevation on the old flow. Foliar nitrogen and phosphorus concentrations were less and specific leaf weight greater for Metrosideros polymorpha leaves collected at high versus low elevations and on the young versus the old flow. Foliar ¹³C was strongly correlated with specific leaf weight across the range of sites sampled.Published as a contribution to the Tropical Mountain Ecosystem Program of the IUBS Decade of the Tropics     

溶磷细菌和丛枝菌根真菌接种对南方红豆杉生长及根际微生物和土壤酶活性的影响

以南方红豆杉实生苗为材料,采用盆栽实验探讨了高效溶磷细菌草木樨中华根瘤菌(Sinorhizobium meliloti)CHW10B与丛枝菌根真菌缩球囊霉(Glomus constrictum)单独和双接种条件下,其植株生长、根际土壤可培养微生物数量、土壤酶活和土壤微生物功能多样性变化,在微生态水平揭示接种对南方红豆杉生长的影响及其机制。结果表明:(1)各接种处理对南方红豆杉幼苗均有促生长作用,接种处理苗高、地径和生物量均较对照显著增加,并以双接种促进效果最好。(2)各接种处理提高了南方红豆杉根际土壤可培养细菌、真菌和放线菌含量,增加了土壤微生物碳源利用率,改变了土壤中物种的丰富度和均一度,增加了土壤中的生物多样性。(3)各接种处理促进了南方红豆杉根际重要土壤酶(酸性磷酸酶、脱氢酶、转化酶)活力的增加,且双接种的促进作用最为明显。可见,溶磷细菌(草木樨中华根瘤菌CHW10B)和丛枝菌根真菌(缩球囊霉)具有协同作用,两者同时接种可显著提高南方红豆杉根际土壤微生物数量及土壤酶活力,提高土壤微生物碳源利用率和土壤肥力,增加土壤中的生物多样性,从而达到间接促进宿主植物南方红豆杉生长的目的。     

The United Kingdom Biodiversity Action Plan moths – selection, status and progress on conservation

As a result of the Earth Summit in 1992, 53 species of moth are covered by the UK Governments Biodiversity Action Plan. The background to the UK Biodiversity Action Plan and the selection of species is discussed. Butterfly Conservations Action for Threatened Moths Project is covered along with its role and approach in overseeing the implementation of the moth Action Plans. A case study on the Straw Belle Aspitates gilvaria (Denis and Schiffermüller) is presented as an example of how an individual Action Plan is being implemented. A subjective consideration of the biological progress with the moth Action Plans is given along with a brief discussion of possible future developments for the UK BAP approach for moths.     

Impact of Soil Applications ofBeauveria bassianaandMariannaeasp. on Nontarget Forest Arthropods

Replicated plots were established in pear thrips-infested sugar maple stands. They were treated withBeauveria bassianaas an emulsifiable concentrate and a nutrient-based granular formulation and withMariannaeasp. as a wettable powder. Postapplication invertebrates moving across plots were collected daily for 10 days in pitfall traps. Collected individuals were identified, held for 10 days, and then evaluated for the presence of fungal infection and other potential pathogenic microorganisms. A total of 3615 invertebrates were collected and identified. Of the invertebrate population in treated plots that fell into pitfall traps, 2.8% became infected with eitherB. bassianaorMariannaeasp. The fungi applied also occur naturally in sugar maple forest soils; therefore, it was impossible to confirm whether the infection we observed in treated plots was a result of contact with the spores we applied. Because we obtained infection among invertebrates collected from control plots, it is likely that these fungi do attack invertebrates under natural forest conditions. Of the 2584 invertebrates examined further for obligate internal pathogens, 2.5% were infected with entomophthoralean fungi, protozoans, and/or nematodes. Results suggest thatB. bassianaandMariannaeasp. could be applied to forest soil without a significant negative impact on the forest-dwelling invertebrate population.     

Quantification of the geocarposphere and rhizosphere effect of peanut (Arachis hypogaea L.)

Roots and pods of field-grown peanut (groundnut) (Arachis hypogaea L.) were sampled at the R3, R5, and R7 developmental stages and examined in comparison to root- and pod-free soil for microbial population densities to assess the geocarposphere and rhizosphere effects. G/ S (no. geocarposphere microorganisms/no. soil microorganisms) and R/S (no. rhizosphere microorganisms/no. soil microorganisms) ratios were calculated for total fungi,Asperigillus flavus, spore-forming bacilli, coryneform bacteria, fluorescent pseudomonads, and total bacteria isolated on low- and high-nutrient media. A clear geocarposphere effect was evidenced by increased population densities of bacteria and fungi associated with developing pods compared to soil. G/S and R/S ratios were generally greater than 1.0 for all groups of microorganisms except bacilli. G/S ratios were greater for total bacteria than for total fungi at two of the three sample times, suggesting that bacteria were stimulated more than fungi in the zone around developing pods. In contrast, R/S ratios, were higher for total fungi than for total bacteria at two of three sample times. The preferential association of fungi and bacteria with early developmental stages of the pod indicates that some microorganisms are particularly well adapted for colonization of the peanut geocarposphere. These microorganisms are logical candidates for evaluation as biological control candiates forA. flavus.     

Allelopathic Effects of Tree Species on Some Soil Microbial Populations and Herbaceous Plants

The allelopathic potential of four tree species on soil microbial populations and some herbaceous plants (two understory species and one general biotest species) was investigated. Effects of three nonindigenous tree species, Eucalyptus globulus Labill, Pinus radiata D.Don and Acacia melanoxylon R.Br., on microorganisms participating in the cycle of nitrogen were evaluated, comparing them with those produced by the autochthonous Quercus robur L. Influence of the trees on Lactuca sativa L., Dactylis glomerata L. and Trifolium repens L. was also checked in bioassays. Cell numbers of Nitrosomonas sp. were negatively affected by Acacia and Eucalyptus stands, mainly during spring, when flowers are especially abundant on the ground. Proteolytic microorganisms were also negatively affected by Eucalyptus and Pinus stands, whilst Quercus stand did not show any toxicity. Soil bioassays showed clear inhibitory effects on germination and growth of understory plants, particularly soils from Eucalyptus and Acacia stands. The greatest effects had the soil from Acacia stand, which was phytotoxic during the whole period of germination and growth of understory plants. Allelopathic phenomena could be, at least partially, responsible of the low species diversity in the understory of the nonindigenous tree stands.     

Cytosine deaminase that hydrolyzes creatinine to N-methylhydantoin in various cytosine deaminase-forming microorganisms

Creatinine deimination has been newly detected in the following various cytosine deaminase-forming microorganisms: Escherichia coli, Proteus mirabilis, Pseudomonas aureofaciens, Pseudomonas chlororaphis and Pseudomonas cruciviae. All these microorganisms, except for E. coli, formed cytosine deaminase in a constitutive or repressive way. P. putida 77 and E. coli showed highly increased formation of creatinine deiminase in the presence of creatinine and cytosine. Throughout serial DEAE-Sephacel and Sephacryl S-300 column chromatographies, the cytosine deaminases of these microorganisms, except for that of P. ovalis, were found to hydrolyze both creatinine and cytosine at comparable rates. No concrete evidence was obtained for the presence of any other protein that hydrolyzed creatine and/or cytosine than the cytosine deaminases in the three test microorganisms randomly selected for investigation.Different from P. putida 77, none of the test microorganisms degraded N-methylhydantoin; neither N-methylhydantoin amidohydrolase nor N-carbamoylsarcosine amidohydrolase was formed in the presence of creatinine in these microorganisms. As a result, the wide occurrence of cytosine deaminases in microorganisms was found to be related to the wide distribution of those microorganisms which hydrolyze creatinine to N-methylhydantoin without further degradation.     

Phosphate-solubilizing microorganisms isolated from rhizospheric and bulk soils of colonizer plants at an abandoned rock phosphate mine

The abandoned “Monte-Fresco” rock phosphate mine in Táchira, Venezuela, was sampled to study the biodiversity of phosphate-solubilizing microorganisms (PSM). Rhizosphere and bulk soils were sampled from colonizer plant species growing at a mined site where pH and soluble P were higher than the values found at a near by unmined and shrubby soil. Counting and isolating of PSM choosing strains showing high solubilization halos in a solid minimal medium with hydroxyapatite as phosphate source were evaluated using ammonia or nitrate as nitrogen sources and dextrose, sucrose, and mannitol as carbohydrate sources. A larger number of PSM were found in the rhizospheric than in the bulk soil. Six fungal strains belonging to the genus Penicillium and with high hydroxyapatite dissolution capacities were isolated from bulk soil of colonizer plants. Five of these strains had similar phenotypes to Penicillium rugulosum IR-94MF1 but they solubilized hydroxyapatite at different degrees with both nitrogen sources. From 15 strains of Gram-negative bacteria isolated from the rhizosphere of colonizer plants, 5 were identified as diazotrophic free-living encapsulated Azotobacter species able to use ammonium and/or nitrate to dissolve hydroxyapatite with glucose, sucrose and/or mannitol. Different nitrogen and carbohydrate sources are parameters to be considered to further characterize the diversity of PSM.     

Establishment of the phylogenetic relationships between the microbial producers of cyclodextrin glucanotransferases using complete amino acid sequences

A phylogenetic tree was constructed on the basis of the amino acid sequences of the known cyclodextrin glucanotransferases (CDGTs), including those deduced from the nucleotide sequences ofBacillus sp. strain 6.6.3 andPaenibacillus macerans IB-7 genes encoding α- and β-CDGTs. The tree clearly demonstrates the existence of distinct phylogenetic groups of CDGT-producing microorganisms and the divergence of the α-, β-, and γ-CDGT produced by microorganisms from the generaBacillus, Paenibacillus, Brevibacillus, andThermoanaerobacter from a common ancestor, whereas the CDGT ofKlebsiella pneumoniae is independent and results from the convergence of different ancestors. The degree of homology of the leader peptide sequences of CDGTs may serve as a criterion of intraspecies relatedness between CDGT-producing microorganisms.     

Occurrence and antagonistic potential of Stenotrophomonas strains isolated from deep-sea invertebrates

Stenotrophomonas maltophilia is known to be of significance as opportunistic pathogen as well as a source of biocontrol and bioremediation activities. S. maltophilia strains have been isolated from rhizospheres, soil, clinical material, aquatic habitats, but little is known about Stenotrophomonas strains recovered from marine environments. During a survey of the biodiversity of Pseudomonas-like bacteria associated with deep-sea invertebrates six Stenotrophomonas strains were isolated from sponge, sea urchin, and ophiura specimens collected from differing Pacific areas, including the Philippine Sea, the Fiji Sea and the Bering Sea. 16S rRNA gene sequence analysis confirmed an assignment of marine isolates to the genus Stenotrophomonas as it placed four strains into the S. maltophilia CIP 60.77^T cluster and two related to the S. rhizophila DSM 14405^T. Together with a number of common characteristics typical of S. maltophilia and S. rhizophila marine isolates exhibited differences in pigmentation, a NaCl tolerance, a range of temperatures, which supported their growth, substrate utilization pattern, and antibiotics resistance. Strains displayed hemolytic and remarkable inhibitory activity against a number of fungal cultures and Gram-positive microorganisms, but very weak or none against Candida albicans. This is the first report on isolation, taxonomic characterization and antimicrobial activity of Stenotrophomonas strains isolated from deep-sea invertebrates.     

Traditional Knowledge and Management of <Emphasis Type="Italic">Feijoa (Acca sellowiana)</Emphasis> in Southern Brazil

Traditional Knowledge and Management of Feijoa (Acca sellowiana) in Southern Brazil. This paper investigates traditional knowledge of the use and management of Acca sellowiana in southern Brazil. Fifty-six informants from three rural communities were assigned to one of four subgroups (“maintainers,” “managers,” “cultivators,” or “users”) based on their responses regarding management and use of A. sellowiana. Traditional knowledge related to use of this species is widespread among rural residents, but traditional knowledge related to management is fragmented depending on whether one uses, manages, or cultivates the species. Knowledge held in rural communities suggests that A. sellowiana could play an expanded role in local economies as well as biodiversity conservation. We suggest that participatory research could stimulate greater local use as well as on-farm conservation of A. sellowiana.     

Effects of rhizosphere remediation and bioaugmentation on carbofuran removal from soil

Rhizosphere soil contains important sources of nutrients for microorganisms resulting in high number of microorganisms capable of degrading various types of chemicals in the soil. Thus, this study investigated a carbofuran dissipation in rhizosphere soils of 6 weeds namely, umbrella sedge (Cyperus iria L.), fuzzy flatsedge (C. pilosus V.), small flower umbrella plant (C. difformis L.), tall-fringe-rush hoorah grass (Fimbristylis miliacea V.), cover fern (Marsilea crenata P.), and water primrose (Jussiaea linifolia V.). Rhizosphere soil of fuzzy flatsedge showed the shortest half-life (t_1/2) of carbofuran (15 days) among other soils. So, it was selected to be used in the bioaugmentation experiment using carbofuran degrader namely Burkholderia cepacia, PCL3, as inoculum in order to examine whether they would improve carbofuran degradation in soil. The results showed that the addition of PCL3 into rhizosphere soil did not improve carbofuran degradation suggesting that microorganisms in rhizosphere soil might be capable enough to remove carbofuran from soil. The number of carbofuran degraders in the rhizosphere soils was greater than in bulk soil 10–100 times which might be responsible to a rapid degradation of carbofuran in rhizosphere soils without the addition of PCL3. The ability of PCL3 to degrade carbofuran was evident in bulk soil (t_1/2 of 12 days) and autoclaved soils (t_1/2 13–14 days) when compared to soils without an inoculation (t_1/2 of 58 days) indicated that the addition of a degrader was useful in improving carbofuran degradation in soil.     

Microorganisms asssociated with Withania somnifera leaves

Microorganisms including bacteria, actinomycetes and fungi were recovered from the leaves of Withania somnifera, which were collected from two altitudinal ranges (0–300 m and 1700–2000 m) in the Asir region, Saudi Arabia. Types and numbers of microorganisms varied according to the altitude and the month of collection. The number of microorganisms was higher on old leaves than that on young ones in most cases. Low altitude exhibited more microorganisms than high altitude. The relationship between meteorological factors and type and number of the recovered microorganisms is discussed. Inoculation of detached healthy leaves of Withania by all recovered fungal species revealed only Alternaria solani as a pathogen of this plant. To confirm pathogenicity, scanning and transmission electron microscopic examination revealed the colonization of this pathogen inside the leaf tissue. Penetration of Withania leaves by the fungus occurred only through stomata, and the invading hyphae were located in the intercellular spaces of leaf tissues. Ultrastructural changes noted in infected cells included changes in chloroplasts and the invagination of the host plasma membrane.     

Effect of time of inoculation on in vitro ectomycorrhizal colonization and nodule initiation in Acacia holosericea seedlings

The complex interactions that occur in systems with more than one type of symbiosis were studied using one isolate of Bradyrhizobium sp. and the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch inoculated on to the roots of Acacia holosericea A. Cunn. ex G. Don in vitro. After a single inoculation with Bradyrhizobium sp., bacteria typically entered the roots by forming infection threads in the root hair cells via the curling point of the root hair and/ or after intercellular penetration. Sheath formation and intercellular penetration were observed on Acacia roots after a single inoculation with Pisolithus tinctorius but no radial elongation of epidermal cells. Simultaneous inoculation with both microorganisms resulted in nodules and ectomycorrhiza on the root system, occasionally on the same lateral root. On lateral roots bearing nodules and ectomycorrhiza, the nodulation site was characterized by the presence of a nodule meristem and the absence of an infection thread; sheath formation and Hartig net development occurred regularly in the region of the roots adjacent to nodules. Prior inoculation with Bradyrhizobium sp. did not inhibit ectomycorrhizal colonization in root segments adjacent to nodules in which nodule meristems and infection threads were clearly present. Conversely, in ectomycorrhizae inoculated by bacteria, the nodule meristem and the infection thread were typically absent. These results show that simultaneous inoculation with both microorganisms inhibits infection thread development, thus conferring an advantage on fungal hyphae in the competition for infection sites. This suggests that fungal hyphae can modify directly and/or indirectly the recognition factors leading to nodule meristem initiation and infection thread development.     

Grape marc compost: microbial studies and suppression of soil-borne mycosis in vegetable seedlings

Compost suppression of soil-borne diseases in horticultural crops has been attributed to the activities of antagonistic microorganisms. A great diversity of microorganisms, capable of suppressing pathogens naturally colonize compost. A large number of microbes appeared in microbiological analyses of grape marc compost. Most microorganisms were bacteria. Average percentages were 31% mesophilic and 28% thermophylic bacteria, 16% mesophilic actinomycetes and 20% thermophylic actinomycetes. Only a few mould and yeast morphologies were obtained, 4% and 1% respectively. Antagonist in vitro assays were performed with 432 microbial morphologies isolated from grape marc compost. The microbes isolated were extremely effective antagonists in in vitro assays against all the fungal pathogens tested. Seven microorganisms were selected for further bioassay with Rhizoctonia solani in radish, Fusarium oxysporum f. sp. radicis-cucumerinum in melon, and Phytophthora parasitica in tomato and two microorganisms with Pythium aphanidermatum in cucumber. Those experiments indicate that grape marc compost reduces the severity of Pythium damping-off in cucumber, but does not reduce the severity of Phytophthora root rot in tomato, Fusarium oxysporum f. sp. radicis-cucumerinum in melon and Rhizoctonia solani in radish. Better suppressive effects were not demonstrated by either compost or vermiculite amended with microbes selected from grape marc compost.     

Effects of the infection of toxigenic fungi and an antagonistic Streptomyces strain on wheat spikes

The objective of the present study was to determine the effect on infection of wheat spikes by toxigenic fungi (Aspergillus parasiticus NRRL 2999, Fusarium tricinctum NRRL 3299, Fusarium graminearum CEREMIC 136/92) and a strain of Streptomyces sp. that is antagonistic to the above-mentioned fungi. Wheat grains (variety GRANERO INTA) were sown in 8 pots containing natural soil and kept in a greenhouse chamber. In the period of the early anthesis the wheat spikes were inoculated with conidial suspensions of each of the fungi in the presence or absence of Streptomyces. Each pot was assigned a different treatment. After an incubation of 100 days and when the wheat plants had attained maturity, the spikes were separated and the following items were determined: (a) number of grains obtained with each treatment, (b) weight of the grains, (c) average weight of the grains/treatment, (d) average number and weight of the grains/spike, and (e) invasion of the caryopses by the microorganisms determined by the analysis of the caryopses in seriate histological sections.There was a significant decrease (p<0.01) in the average weight of the caryopses and in the weight and number of grains/spike in the presence F. graminearum. The wheat grains were invaded by of F. graminearum and A. parasiticus, an effect which was partially attenuated by the presence of antagonist Streptomyces sp. Nevertheless, the effect was not strong enough to prevent the degenerative consequences on the size and weight of the grains produced by F. graminearum.     

The perfect invader: a parthenogenic crayfish poses a new threat to Madagascar’s freshwater biodiversity

In 2007 an unusual crayfish found in food markets in the capital of Madagascar was preliminarily identified as Procambarus ‘Marmorkrebs’: a new world taxa and the only decapod known to reproduce by parthenogenesis. We present information on the identity, distribution and ecology of this recent invader and attempt to evaluate the threat it poses to Madagascar’s biodiversity and to livelihoods. The species appears to be currently limited to the area close to Antananarivo, but is being sold alive on major transport routes. We present molecular evidence of its taxonomic relationships and confirm that the Procambarus present in Madagascar is indeed the parthenogenic taxa. We investigate its reproductive ecology and find Procambarus ‘Marmorkrebs’ to have an extremely high fecundity; more than six times that of the native crayfish Astacoides. The limited evidence we have suggests that this species poses a serious threat to freshwater biodiversity and that it is likely to damage human livelihoods (through its impact on fishing and possibly rice agriculture). More research is urgently needed but in the meantime action is needed to reduce the rate of spread before it is too late.