Biological control of white rot of onion

Summary Interactions of 123 isolates of fungi, 17 of bacteria and 22 of actinomycetes, respectively, withSclerotium cepivorum were studied in agar culture. They were grouped into 4 different types of reactions. Amongst themTrichoderma viride, Fusarium graminearum, Coniothyrium minitans andGliocladium roseum inhibited the growth ofS. cepivorum and later grew over its colony.T. viride showed a characteristic coiling around the hyphae ofS. cepivorum. T. viride andF. graminearum prevented the development of sclerotia.C. minitans was found to parasitize the sclerotia ofS. cepivorum and produced its pycnidia within them.Aleurisma carnis, Cladosporium elatum, Penicillium expansum, P. nigricans, P. notatum, P. piscarium, P. puberulum, P. rolfsii, P. urticae, P. variabile, Tilachlidium humicola andHelminthosporium sp. inhibited the growth ofS. cepivorum at a distance. Eleven isolates of bacteria and 3 ofStreptomyces sp. showed pronounced antagonistic properties againstS. cepivorum.Experiments were carried out to study the effects on white rot development in soil of organisms selected from agar plate tests. None of the antagonistic micro organisms had any deleterious effects on onion growth. Of the organisms testedP. nigricans gave the best results in controlling white rot infection.This work was carried out at the Department of Botany, The University, Birmingham, England. I wish to thank Prof.C. J. Hickman for his invaluable advice and encouragement.     

Inhibition of fungi,actinomycetes and bacteria by Stachybotrys atra

The effect ofStachybotrys atra on fungi, actinomycetes and bacteria was studied in agar culture. Of the 27 genera and 52 species of fungi, 6 species of actinomycetes and 5 genera and 10 species of bacteria tested,S. atra was found to inhibit 95.7 % fungi, all the actinomycetes and 83.3 % of bacteria.Aspergillus ustus, 2 unidentified species ofFusarium, Rhizobium trifolii andPseudomonas spp. were however not inhibited. In experiments in soil,S. atra inhibited the growth ofTrichoderma viride. S atra was ineffective whereT. viride andS. atra were inoculated simultaneously or whereT. viride was added 7 days earlier. Besides,S. atra was not found to controlMacrophomina phaseoli infection on cotton. Soil culture ofS. atra showed phytotoxic effect on cotton seedlings.S. atra produced a dark brown to black thermostable toxic metabolite in the medium. The culture filtrate retarded the growth ofM. phaseoli in agar culture. Despite the importance ofS. atra as a fungistatic agent with wide antimicrobial spectrum, the use ofS. atra as a protectant against plant pathogens seems to be limited because of its phytotoxicity on cotton.     

Comparison ofRhizoctonia solani isolates from web blight and basal canker of cowpea and from soil

Summary Isolates ofRhizoctonia solani from web blight and stem basal canker of cowpea and those obtained from soil had similar linear growth rates on potato dextrose agar (PDA) at various temperatures but differed in other features. The web blight isolates differed from the basal canker and the soil isolates in cultural appearance on PDA and on potato marmite agar (PMA). The web blight isolates readily formed discrete sclerotia on PDA, PMA and soil but the other isolates did not. In greenhouse tests, the former were generally the most virulent in inciting foliage and stem basal necrosis and damping-off of seven crop species. Of the plants tested, the legumes were the most susceptible toR. solani.     

Insecticidal and nematicidal properties of microbial metabolites

Summary Metabolites from 942 microbial isolates were screened for insecticidal and nematicidal properties. The isolates included 302 streptomycetes, 502 novel actinomycetes including representatives of 18 genera, 28 unidentified aerobic actinomycetes, 70 fungi and 40 bacteria other than actinomycetes. When diluted 10-fold, the metabolites from 55 isolates caused nearly 100% mortality in mosquito larvae (Aedes aegypti) within 24 h. These isolates included 27 isolates ofStreptomyces, four ofActinoplanes, three isolates each ofActinomadura andStreptoverticillium, two isolates each ofMicromonospora, Bacillus andPaecilomyces, and one isolate each ofMicropolyspora, Nocardiopsis, Streptosporangium, Oerskovia, Thermomonospora, Chainia, Pseudomonas, Fusarium, Monilia andSyncephalestrum. Two fungal isolates could not be identified to the generie level. Extracts from the culture broth of 18 isolates caused 100% mortality in mosquito larvae within 15 min to 24 h at a concentration of 1000 ppm. The LC₅₀ of partially purified products from two isolates was 1–2.5 ppm and that of the semipurified preparations from four other isolates was 50 ppm. Valinomycin was identified as an active component in the culture broth from one isolate. The culture broth from 15 isolates of aerobic actinomycetes and 4 of fungi were toxic toPanagrellus redivivus (Nematoda); these included 12 isolates with selective nematicidal properties.Michigan Agriculture Experiment Station, Journal Article No. 12321.     

Survival of Sclerotia of Rhizoctonia solani AG3PT and Effect of Soil‐Borne Inoculum Density on Disease Development on Potato

Sclerotia produced by a single isolate of Rhizoctonia solani AG3PT were buried in small plot experiments to investigate the effects of sclerotial production method, soil type and burial depth on sclerotial viability in field soil. The factor with the greatest effect on sclerotial viability, defined as the percentage of sclerotia germinating on agar following retrieval, in all experiments was the duration of burial. After 18 months, on average across all experiments, 20% of retrieved sclerotia were viable. A comparison between sclerotia produced in vitro on malt yeast extract agar and in vivo using micropropagated tubers in field soil found no significant differences between the two production methods on sclerotial viability. Burial in field soil at 20‐cm depth was found to significantly reduce sclerotial viability to 50% compared to 60% at 5 cm. In two pot experiments, amending the growing medium and soil with increasing inoculum densities of R. solani was found to increase stem number, stem canker and black scurf severity regardless of whether this soil‐borne inoculum was derived from mycelium or sclerotia. Black scurf incidence and severity were assessed 30–32 days posthaulm destruction and found to be similar for a range of sclerotial soil‐borne inoculum densities (1.0 × 10^?1 g/kg d.w. soil to 6 × 10^?3 g/kg d.w. soil). The significance of these findings in relation to pathogen survival, detection in soil and disease development is discussed.     

Enumeration and characterization of the soil microflora from hydrocarbon-contaminated soil sites able to mineralize polycyclic aromatic hydrocarbons (PAH)

The use of a plate screening technique allowed the direct isolation and quantification of polycylic aromatic hydrocarbon (PAH)-degrading bacteria from different soil sites. Bacteria that were able to grow on anthracene, phenanthrene, fluoranthene or pyrene as a sole carbon source were found with numbers between 10³ and 10⁵ colony-forming units (cfu)/g of soil dry weight, but only in samples that originated from PAH-contaminated sites. No isolates were found that could grow on perylene, triphenylene, benzo(a)pyrene or chrysene as sole carbon source. Bacteria that had been selected on the same PAH substrate showed a related degradation pattern for both other PAH and oil compounds and carbohydrate substrates even if they had been collected at distant soil sites. Based on these findings the isolates could be clustered into four different catabolic and taxonomic similarity groups. Taxonomic determination of representative isolates suggested that nocardioform actinomycetes of the genera Mycobacterium, Rhodococcus and Gordona represented a major part of the soil microflora able to mineralize PAH. Three new isolates able to grow on anthracene, pyrene or fluoranthene as the sole carbon source, respectively, have been isolated and identified (Sphingomonas paucimobilis BA2, Gordona sp. BP9, Mycobacterium sp. VF1). The ubiquitous presence of a potent and versatile mineralizing microflora in PAH-contaminated soils indicated that the microflora is not the limiting factor for the degradation of PAH with up to four rings.     

降香黄檀土壤微生物数量的时空动态特征

【目的】研究降香黄檀土壤微生物数量在时间和空间上的动态特征,可为降香黄檀栽培技术的完善提供数据支持。【方法】采用稀释平板涂布法,比较分析降香黄檀根际和非根际土壤微生物三大类群(细菌、放线菌、真菌)数量的季节及垂直分布情况。【结果】(1)降香黄檀根际和非根际土壤均表现为春季细菌、真菌数量较多,冬季细菌数量较少,秋季真菌数量较少;非根际土壤放线菌冬季数量较多,秋季数量较少。(2)细菌的根际效应最为明显,其R/S为0.12~62.96;真菌次之,其R/S为0.22~2.46;放线菌的根际效应较小,其R/S为0.32~1.01。(3)降香黄檀上层土(0~20 cm)的微生物数量普遍高于其他土层;随着土层加深,微生物数量表现为逐渐下降或先下降后升高2种变化趋势。【结论】降香黄檀土壤微生物数量的分布受季节和土层的影响。     

Survival of Macrophomina phaseoli (Maubl.) Ashby on cucurbit roots

Summary Experiments on the survival ofM. phaseoli on cucurbit root pieces were carried out. The sclerotia ofM. phaseoli were almost unaffected by 10 months dry storage in the laboratory. Under dry to moderately moist soil conditions it survived for 2 months with little loss of viability. In wet soils however, the buried root pieces were colonized by other soil microorganisms which prevented the growth ofM. phaseoli.     

Application of percoll density gradients in studies of the adhesion ofStreptococcus pyogenes to human epithelial cells

A new assay was used to study the adhesion ofStreptococcus pyogenes strains to epithelial cells. [³H]thymidine-labeled bacteria were incubated with standardized preparations of epithelial cells collected from oral-pharyngeal surfaces of human volunteers. The mixtures were then centrifuged in 50% Percoll to form a density gradient. Epithelial cells with attached bacteria formed a band near the top of the tube, whereas unattached bacteria were located near the bottom. The epithelial cells were collected on membrane filters, and the number of adherent bacteria was then determined by scintillation counting.The abilities of M-protein-positive (M⁺) and M-protein-negative (M^–) strains ofS. pyogenes to attach to human pharyngeal, buccal, and tongue epithelial cells were compared. The results obtained confirmed the significant difference previously shown to exist between the attachment of M⁺ and M^– strains to human epithelial cells. M⁺ strains ofS. pyogenes exhibited a much greater ability to bind to pharyngeal epithelial cells than did M^– variants. Also, M⁺ strains were bound in higher numbers to pharyngeal epithelial cells than to buccal or tongue epithelial cells. The adhesion ofS. pyogenes strains to epithelial cells was time dependent, and a significant increase in the adhesion of M⁺ strains occurred after 3–4 h of exposure of the bacteria to epithelial cells.The adsorption ofS. pyogenes strains to epithelial cells was described by a Langmuir isotherm. With this model, the number of binding sites and the affinities of the streptococci for epithelial cells were estimated. Significantly higher numbers of binding sites were calculated to be present on pharyngeal epithelial cells for M⁺ strains ofS. pyogenes than on buccal cells. However, the affinity of the organisms was similar for both types of cells.Adsorption of M⁺ strains to human pharyngeal epithelial cells was inhibited by certain galactosides and fucose, but not by glucose or xylose. This suggests that saccharide moities play a role in the binding of M⁺ strains ofS. pyogenes to human pharyngeal epithelial cells.     

华西雨屏区天然常绿阔叶林土壤可培养微生物数量对模拟氮沉降的响应

为了解氮沉降对华西雨屏区天然常绿阔叶林土壤微生物数量的影响,从2013年11月至2014年12月,通过野外模拟N(NH_4NO_3)沉降,氮沉降水平分别为对照(CK 0 kg N hm~(-2)a~(-1))、低氮沉降(L 50 kg N hm~(-2)a~(-1))、中氮沉降(M 150 kg N hm~(-2)a~(-1))和高氮沉降(H 300 kg N hm~(-2)a~(-1)),研究了氮沉降对天然常绿阔叶林0—10cm和10—20cm土层土壤可培养微生物数量的影响。结果表明:华西雨屏区天然常绿阔叶林0—10cm土层的细菌、真菌和放线菌数量均显著大于10—20cm土层,氮沉降未改变原有垂直分布格局。L处理对0—10cm和10—20 cm土层土壤微生物总量无显著影响,M和H处理则显著降低了土壤微生物总量。氮沉降降低了0—10cm和10—20cm土层的细菌数量,且抑制作用随氮沉降量的增加而增强。氮沉降降低了0—10cm土层的真菌数量,但下降幅度与氮沉降量之间无明显规律;在10—20cm土层,M和H处理在夏季显著增加了真菌数量,表明适量氮沉降能有效缓解夏季土壤真菌的氮限制状态。氮沉降对0—10cm土层放线菌数量的影响表现为先促进再抑制,L和M处理增加了放线菌数量,H处理降低了放线菌数量;氮沉降增加了10—20cm土层的放线菌数量,其中M处理的促进作用最大。氮沉降对土壤微生物数量的影响随土壤深度的增加而减弱。     

Distribution of pathogenic vibrios and other bacteria in imported frozen shrimps and their decontamination by gamma-irradiation

The distribution of pathogenic vibrios and other bacteria in eight samples of imported frozen shrimps and the effect of irradiation on these bacteria were investigated. Total aerobic bacteria were at 2×10⁴ to 4×10⁶/g. Coliforms consisted mainly ofEnterobacter. No salmonella were detected. A total of 66 isolates, includingVibrio parahaemolyticus, V. mimicus, V. alginolyticus, V. vulnificus, V. fluvialis and a few ofListeria monocytogenes, were obtained. The gamma-radiation dose needed to reduce by 10^–4 the number of vibrio isolates andAeromonas hydrophila was about 3 kGy in frozen shrimps, whereas about 3.5 kGy was required forL. monocytogenes.     

Adherence of clinical isolates ofPseudomonas aeruginosa to hamster trachael epithelium in vitro

The adherence to hamster tracheal epithelium, of mucoid and nonmucoid clinical isolates ofPseudomonas aeruginosa from cystic fibrosis patients, was studied using tracheal organ cultures. Tracheal cultures were infected with 10⁷ colony-forming units per ml of either mucoid or nonmucoid clinical isolates ofP aeruginosa. The tracheal explants were rinsed at various time intervals to remove nonadherent bacteria, fixed, and prepared for transmission-and scanning-electron microscopy. Mucoid isolates were seen adhering to the ciliated epithelium as early as 4 h after initiation of infection, whereas nonmucoid isolates were only observed adhering at 6 to 8 h after infection. Mucoid organisms were found as clusters of bacteria embedded in an extensive extracellular matrix. The nonmucoid isolates were generally found as single organisms with no evidence of an extracellular matrix. These results suggest that the prevalence of mucoid isolates ofP. aeruginosa in cystic fibrosis may be due to adherent properties of the mucoid organism.     

Nitrate production in the rhizosphere of Plantago species

Summary The production of nitrate in an old established dune grassland soil and its uptake by plants was studied by comparing amounts of mineral nitrogen and numbers of nitrifying bacteria in the rhizosphere on the one hand, and on the other accumulated nitrate and levels of nitrate reductase (NaR) of individual plants of three Plantago species,i. e., P. major, P. lanceolata andP. coronopus. For these three Plantago species andP. media basal levels of NaR in the absence of nitrate were determined in plants grown in culture solutions. The basal NaR levels ofP. major andP. media (species occurring on nutrient-rich soils) were significantly higher than those ofP. lanceolata andP. coronopus (species found on nutrient-poor soils). NaR activity increased in the presence of nitrate and was suppressed by ammonium.From the numbers of nitrifying bacteria in the rhizosphere and NaR activity in the leaves it was concluded that nitrate was produced in the root environments of the three Plantago species and that the compound was taken up by the plants. NaR activities and numbers of nitrifying bacteria were higher for individuals ofP. major than for those ofP. lanceolata andP. coronopus. No correlation was found between the ammonium levels and the numbers of nitrifying bacteria in the soil, and no indications of inhibition of nitrifying bacteria in the rhizosphere were obtained. For individuals ofP. lanceolata a correlation was found between the numbers of nitrifying bacteria in the soil and NaR activity in the leaves. The results are discussed in relation to the ecological habitats of the three species.Grassland Species Research Group Publication No.38.     

Changes in soil microflora associated with control of Sclerotium Rolfsii by Furfuraldehyde

The efficacy of 2‐furfuraldehyde for control of Sclerotium rolfsii was studied in laboratory and greenhouse experiments. Mycelial growth of the fungus was reduced proportionally with concentrations of 0.1–0.5 ml furfuraldehyde l^‐1 agar medium, and viability of sclerotia diminished on exposure to 2‐furfuraldehyde vapours. Detectable populations of bacteria and fungi, including Trichoderma spp., were reduced significantly (9=0.05) when furfuraldehyde was added to the agar used for soil dilution plates of untreated soil. Repeated treatments of natural soil with the fumigant significantly increased populations of Trichoderma spp. and bacteria, but diminished numbers of actinomycetes. Increasing dosages applied to soil artificially infested with S. rolfsii caused a reduction of disease on lentil, Lens culinaris. Results indicate that the compound, when applied to field soil, changes the composition of soil microflora and has potential for integrated control of S. rolfsii.     

Physiological studies on Phymatotrichum omnivorum VI. Lipid composition of mycelium and sclerotia

The lipid composition of the mycelium and sclerotia ofPhymatotrichum omnivorum was compared. The lipids of the mycelium contained 47.9 % polar lipids as compared to 21.4 % in the sclerotia. Sterols represented 10 % of the lipids in sclerotia as contrasted to 3.6 % of the mycelium. More monoglycerides (17.5 %) were detected in the sclerotia as compared to the mycelium (1.6 %). Fatty acid analysis indicated that linoleic acid was the predominant fatty acid in the total fatty acids fraction in both the mycelium and the sclerotia. Palmitic acid was the major free fatty acid in the mycelium, whereas myristic acid was the predominant free fatty acid in the sclerotia. In the fatty acids of the diglycerides of sclerotia, palmitic acid represented 71 % of that fraction, as compared to 6.6 % of the fatty acids of the diglycerides in the mycelium. The major fatty acid in the diglycerides of the mycelium was oleic acid.     

Diversity and Antimicrobial Activity of Actinomycetes Isolated from Lut Desert: The Extremely Arid Climatic Zones of Iran

Lut desert is situated in one of the extremely arid climatic zones of Iran and is one of the hottest deserts in our plant with the extreme fluctuation of temperature over a day. The main objective of this study is to characterize the diversity of the culturable actinomycetes and preliminary evaluation of their extracts as antimicrobial components on drug resistant pathogens. Twenty-four soil samples were collected, successively diluted and inoculated into the different culture media to support the growth of most culturable bacteria including actinomycetes. Phenotypic and molecular methods were used for accurate identification of recovered isolates particularly actinomycetes at the genus and species levels. The isolates were also evaluated for their inhibitory activities against drug resistant Acinetobacter baumannii, Enterococcus faecium, Klebsiella pneumoniae and Staphylococcus aureus. A total of 56 isolates recovered from the samples. Based on phenotypic tests, 41 isolates were identified as actinomycetes, amongst them 8 isolates were active against drug resistant pathogens. Our study revealed Lut desert, as one of the hottest deserts in the world, is the habitat to diverse taxa of bacteria particularly actinomycetes which have potential novel antimicrobial components.

     

Evaluation of bio-aerosols at an animal feed manufacturing industry: A case study

Both total and biological particles (totalculturable bacteria, Gram negative bacteria,mold and actinomycetes) were measured at ananimal feed manufacturing industry. Suspendedparticle concentration ranged from 1.72 to2.3 mg m^–3 with a mean value of1.97 mg m^–3. Airborne microorganisms weredetected in lower concentrations than thoseassociated with suspended dust andfeed-materials. Bacterial concentrations weretwo to three times higher than concentrationsof mold and actinomycetes. Bacterialconcentrations averaged4.86 × 10³ cfu m^–3; 2.6 × 10⁴cfu m^–3 and 3.96 × 10⁷ cfu g^–1 inair, associated with suspended dust andfeed-materials, respectively, whereas moldconcentrations averaged 7.33 × 10²cfu m^–3; 1.97 × 10³ cfu m^–3 and7 × 10⁵ cfu g^–1 of the correspondingenvironments, respectively. Enterobacterspp and Klebsiella spp were the mostabundant Gram negative bacteria, whereas Bacillus species. were the most dominant Grampositive bacteria. Aspergllius niger,other Aspergillus species and Penicillium were the dominant mold isolates.Acremonium was only detected in feedmaterials, whereas Aspergillus fumigatuswas only detected in air. The animal feedindustry environment has a significantbio-contamination and many of microorganismsimplicated in respiratory problems weredetected in this environment.     

Survey of human pathogenic actinomycetes and fungi in soil from Rome and other Italian areas

As part of a study sponsored by the Ministry of Health of Italy, a research program on pathogenic actinomycetes, keratinophilic and pathogenic fungi in soil was carried out. Two hundred soil samples, collected from different areas of the city of Rome, Calabria, Emilia Romagna, Latium, Apulia, Sardinia, Sicily, Tuscany and Umbria, were examined by several techniques to detect the widest possible variety of pathogenic actinomycetes and fungi. Seven isolates ofNocardia asteroides, four ofActinomadura madurae and one ofNocardiopsis dassonvillei were isolated for the first time from soil in Italy. In addition, numerous isolates ofPetriellidium boydii, Aspergillus fumigatus, A. flavus, A. niger and keratinophilic fungi of the generaMicrosporum, Trichophyton andChrysosporium were also recovered.Presented at the VII Congress of the International Society for Human and Animal Mycology, Jerusalem, Israel, 11–16 March, 1979.     

Increasing phosphorus supply in subsurface soil in northern Australia: Rationale for deep placement and the effects with various crops

Three field experiments involving wheat, lucerne or cotton were established at different sites in the semiarid cropping regions of northern Australia, to test whether the deep placement of P fertiliser improved P availability, compared to the conventional practice of placing the fertiliser beside or adjacent to the seed. At Mulga View, near St George in southern Queensland on a red Kandosol soil with a Colwell soil test value of 19 mg P kg soil⁻¹ in the top 10 cm, there was no response to 10 kg P ha⁻¹ applied in the 5–7 cm layer. However, increasing the depth of placement of 10 kg P ha⁻¹ from 5–7 to 10–15 cm resulted in increased shoot growth and grain yield of spring wheat (Triticum aestivum) by 43 and 30%, respectively. A further grain yield increase of 43% to 3.2 t ha⁻¹ resulted when the deep P rate was increased from 10 to 40 kg P ha⁻¹. At Roma, in southern Queensland, on a grey/brown Vertosol with a Colwell soil test value of 15 mg P kg soil⁻¹, there was no difference in the winter growth of lucerne (Medicago sativa) when P fertiliser had been applied at 5–7 cm depth at rates of 10 and 40 kg P ha⁻¹. Shoot dry matter yields were around 2 t ha⁻¹. However dry matter yields increased significantly to 2.6 and 3.7 t ha⁻¹ when 10 and 40 kg P ha⁻¹, respectively were applied at the 10–15 cm depth. The third experiment was carried out on a grey Vertosol at Kununurra in Western Australia. Significant increases in the yield of seed cotton (Gossypium hirsutum) occurred when 50 kg P ha⁻¹ was applied at depth (10–15 and 25–30 cm), compared with the conventional placement at 7–10 cm, with maximum yield response to deep placement occurring with DAP, and the minimal response with MAP. The cotton was grown on raised beds and the crop was irrigated according to district practice. The response to deep P at all sites was attributed to the rapid drying of the soil surface layers, reducing the availability of soil or fertiliser P in these layers. The deep fertiliser P remained available during the growing season and alleviated the P deficiency that appears to be a feature of these soils when the surface layers become dry.     

Survival of Sclerotium cepivorum Sclerotia and Fusarium oxysporum Chlamydospores in Soil Amended with Cruciferous Residues

The use of cruciferous plant residues to reduce the amount of sclerotia of Sclerotium cepivorum and chlamydospores of Fusarium oxysporum f. sp. lycopersici in soil was investigated. Air‐dried and crushed mustard (Brassica juncea) added to the soil effectively reduced the viability of fungal propagules. Consequently, the reduction of white rot of onion, caused by S. cepivorum and wild of tomato caused by F. oxysporum was observed. The addition of rapeseed (Brassica. napus cv. Bolko and B. napus cv. Gorczanski) residues to soil also resulted in a significant decrease of number of S. cepivorum sclerotia but the effect on F. oxysporum chlamydospores was variable. Introduction of the plant material increased the total number of bacteria, spore‐forming bacteria, fluorescent pseudomonads, actinomycetes, and fungi in soil. One year after the soil amendment, the amount of sporeforming bacteria in treatments with cruciferous residues was higher as compared to the control soil without plant residues. The possible contribution of the decomposition of plant residues and soil micro‐organisms to the reduction of the pathogens population is discussed.