Effects of Pseudomonas aeruginosa on the diversity of culturable microfungi and nematodes associated with tomato: impact on root-knot disease and plant growth

The available information on Pseudomonas biocontrol inoculants on the non-target fungal and nematode community is scant. The current paper addresses this issue and investigates the effects of biocontrol agents Pseudomonas aeruginosa IE-6 and IE-6S⁺ (previously shown to suppress several soil-borne plant pathogens) on soil microfungi and plant-parasitic nematodes as well as on the root-knot development and growth of tomato (Lycopersicon esculentum). Furadan, a granular nematicide was included as a treatment for comparative purposes. Treatments were applied to soil at the start of each 52-day-long tomato growth cycle, and their effects on the composition and diversity of rhizosphere and endophytic microfungi and plant-parasitic nematodes were examined at the end of first and fourth growth cycle. Several diversity indices were employed to assess community diversity. A total of 16 genera comprising 23 microfungal species were isolated from the tomato rhizosphere. The most abundant fungal species belonged to the genera Aspergillus, Fusarium, and Penicillium. With a few exceptions, fungi were neither exclusively inhibited nor specifically promoted by the application of treatments at any of the growth cycles studied. However, Paecilomyces lilacinus, an egg and female parasite of root-knot nematode, though exclusively absent in the controls was isolated from the treatments. Both general diversity and equitability of rhizosphere microfungi were greater at first compared to the fourth growth cycle while species richness remained uninfluenced across the growth cycles and treatments. However, Furadan and IE-6S⁺ treatments considerably abated general diversity and equitability. Of the microfungal species isolated from the rhizosphere seven were also recovered from surface-sterilized root tissue of tomato suggesting that all the endophytes are primarily rhizosphere organisms. Diversity of endophytic fungi was consistently lower compared with that of the rhizosphere. Both general diversity and equitability declined in all three treatments relative to controls in the root tissue but species richness remained unaltered. Diversity and equitability of plant-parasitic nematodes in soil were reduced by all three treatments over the controls at fourth growth cycle whilst species richness did not change at either growth cycle. The biocontrol agents significantly reduced root-knot development and enhanced shoot growth of tomato over the controls. The possible implications of fungal composition and abundance because of biocontrol by Pseudomonas application are discussed.     

Diversity of endophytic actinomycetes in mandarin grown in northern Thailand,their phytohormone production potential and plant growth promoting activity

Abstract

The rapid expansion of mandarin (Citrus reticulata L.) production areas with high agrochemical input in the highland areas of northern Thailand has resulted in negative effects in terms of production, environment, soil quality, and public health. The use of microorganisms as plant growth promoters is an alternative method to reduce agrochemical input. Thus, we studied the diversity of endophytic actinomycetes in mandarin and their potential as plant growth promoters. A total of 252 endophytic actinomycete isolates were recovered from mandarin. Based on spore chain morphology, cell wall type, and 16S rRNA gene sequence, the isolates were classified into six genera: Streptomyces, Nocardia, Nocardiopsis, Spirillospora, Microbispora and Micromonospora. The most frequent isolates recovered were members of Streptomyces (85.3%). Selected isolates (64 isolates) from these genera were evaluated for their indole-3-acetic acid (IAA) production potential in a medium with 2 mg mL^?1 tryptophan, and all the selected isolates showed the potential to produce IAA, with average values of IAA production of 13.34, 3.36, 140.38, 12.55, 1.40, and 6.19 µg IAA mL^?1, respectively. Isolates of genus Nocardiopsis showed a very high ability to produce IAA that was the highest among all the genera, with values ranging from 62.23 to 222.75 µg mL^?1. Twelve isolates selected from these genera were inoculated onto mandarin seedlings, and the results indicated that the shoot height, fresh shoot weight and fresh root weight of the seedlings were promoted by the inoculation of endophytic actinomycetes, with values ranging from 20.2 to 49.1%, 14.9 to 53.6%, and 1.6 to 102% over the control, respectively.     

Soil suppressiveness and functional diversity of the soil microflora in organic farming systems

Arable fields of 10 organic farms from different locations in The Netherlands were sampled in three subsequent years. The soil samples were analysed for disease suppressiveness against Rhizoctonia solani AG2.2IIIB in sugar beet, Streptomyces scabies in radish and Verticillium longisporum in oilseed rape. In addition, a variety of microbial, chemical and physical soil characteristics were assessed. All data were correlated by multiple regression and multivariate analyses with the objective to find correlations between soil suppressiveness and biotic or abiotic soil characteristics. Significant differences in soil suppressiveness were found between the fields for all three diseases. Multiple regression indicated a significant correlation between suppressiveness against Rhizoctonia and the number of antagonistic Lysobacter spp., as well as with % active fungi and bacterial diversity. Grass-clover stimulated Rhizoctonia suppression as well as the presence of antagonistic Lysobacter spp. (mainly L. antibioticus and L. gummosus) in clay soils. Streptomyces suppression correlated with the number of antagonistic Streptomyces spp., % of active fungi and bacterial population size. The presence of antagonistic Streptomyces spp. correlated with a high fungal/bacterial biomass ratio. Verticillium suppression was only measured in 2004 and 2005, due to the inconsistent suppressiveness along the years. Nevertheless, a significant correlation with pH, potential nitrogen mineralization and bacterial biomass was found. Bacterial and fungal PCR-denaturing gel electrophoresis fingerprinting of bacterial and fungal communities, in general, did not significantly correlate with disease suppression. Highly significant explanatory factors of the composition of the dominating bacterial and fungal populations were % lutum, pH, C/N quotient, biomass and growth rate of bacteria. Additionally, the % of organic matter and years of organic farming were explaining significantly the composition of the bacterial population.Thus, significant correlations between several soil characteristics and suppressiveness of different soil-borne pathogens were found. For two of the three pathogens, suppression correlated with biotic soil characteristics combined with the presence of specific bacterial antagonists. Probably the soil suppressiveness measured in the organic fields is a combined effect of general and specific disease suppression.     

Plant monocultures produce more antagonistic soil Streptomyces communities than high-diversity plant communities

Plant–soil feedbacks are important to productivity and plant community dynamics in both natural and managed ecosystems. Among soil bacteria, the Streptomyces possess particularly strong antagonistic activities and inhibit diverse plant pathogens, offering a clear pathway to involvement in plant–soil feedbacks. We hypothesized that feedback effects and the ability of individual host plant species to foster antagonistic Streptomyces populations may be modified by the richness of the surrounding plant community. To test this, we collected soil associated with four different plant species (two C4 grasses: Andropogon gerardii, Schizachyrium scoparium; and two legumes: Lespedeza capitata, Lupinus perennis), grown in communities that spanned a gradient of plant species richness (1, 4, 8, 16, or 32 species). For each of these soils, we characterized the potential of soil Streptomyces to antagonize plant pathogens, using an in vitro plate assay with indicator strains to reveal inhibition. We cultivated each plant species in each conditioned soil to assess feedback effects on subsequent plant growth performance. Surrounding plant richness modified the impacts of particular plant species on Streptomyces antagonistic activity; A. gerardii supported a higher proportion of antagonistic Streptomyces when grown in monoculture than when grown in 32-spp plant communities, and L. capitata supported more strongly antagonistic Streptomyces when grown in 4- or 32-spp plant communities than in 8-spp plant communities. Similarly, the feedback effects of particular plant species sometimes varied with surrounding plant richness; aboveground biomass production varied with plant species richness for A. gerardii in L. perennis-trained soil, for L. capitata in A. gerardii-trained soil, and for L. perennis in L. capitata-trained soil. Streptomyces antagonist density increased with overall Streptomyces density under low but not under high plant richness, suggesting that plant diversity modifies selection for antagonistic phenotypes among soil Streptomyces. This work highlights the complexity of feedback dynamics among plant species, and of plant–microbiome interactions in soil.     

作物土传真菌病害发生的根际微生物机制研究进展

土传病害是制约我国农业生产可持续发展的重要瓶颈。根际是作物养分高效利用的窗口,是植物-土壤-微生物相互作用的关键微域,也是土传病害发生发展的主要场所。在宏基因组学快速发展的今天,了解根际微生物与土传病害互作关系,有利于从微生物种群、功能代谢和抑病物质等研究找出防控土传病害的有效方法。本文综述了根际微生物与土传真菌病害的发生机制关系,探讨了土传病原真菌致害机理,并提出目前研究的不足和未来研究的重点。目前,我国农业生产中土传真菌病害发生严重,很多是由不同病原菌复合侵染的结果,其致害机理较为复杂。定向优化根际微生物群落结构提升植物根际微生态抗性,是防控土传病害途径之一,应加以重视。现有研究多偏向根际促生微生物,而忽视了根际有害细菌与病害发生和作物生长的关系。今后的研究应系统评估土传真菌病害的发生及危害程度、归类土传真菌病原菌类型、深入研究土传真菌病害发生的根际微生态机理以及研发土传真菌病害综合防控技术。     

Biological control: a sustainable and practical approach for plant disease management

ABSTRACT

This review deals with the mechanism of antagonistic action of bacterial and fungal biocontrol agents such as the production of antibiotics, siderophores, enzyme secretion, competition for nutrition, plant growth promotion by rhizosphere microorganism. The utilisation of synthetic pesticides has been the predominant control processor for diseases brought about by phytopathogenic microorganisms. Notwithstanding, their open and improper application in intensive agriculture has realised issues that have prompted ecological contamination, considerable residues in agricultural products and phytopathogen resistance. They are likewise disrupting the quantity of beneficial microorganism which is available in the soil and capable of expanding soil fertility. Along these lines, there is a need to look through the option of synthetic pesticides that are safe, environmental and monetarily feasible to confront this problem. Biocontrol agent’s utilisation is the best alternative method to control the different kinds of diseases, such as nematode infestation, fungal pathogen and bacterial pathogen. Nowadays, biocontrol agents assume a significant role in the field of agriculture. It is a financially savvy, environment-friendly and inhibits the advancement of pathogenic microorganism sustainably. This review emphasises the role of biocontrol agents against different pathogenic microorganisms and their significance potentiality to improve plant growth and enhance defence system of plants.     

Relationship between root-endophytic microbial communities and replant disease in specialized apple growing areas in Europe

Apple replant disorders are one of the causes of the downward trend of land planted with apple orchards in Central Europe. A specific transnational survey was thus conducted on apple root associated microorganisms, aimed at increasing the knowledge regarding crop management to counteract this tendency. Soil health was evaluated using a bioassay test with root cuttings of the clonal M9 rootstock, one of the most commonly used rootstocks in Europe. Plant growth response in replant, fallow and gamma ray-sterilized soil was evaluated using soil samples taken from nine orchards selected (three per country) from specialized apple growing areas of Germany, Austria and Italy. Plant growth significantly differed between treatments but not between countries. Root endophytic fungi were confirmed as one of the main components of growth reduction, while endophytic nematodes (Pratylenchus sp.) were not. The large set of quantitative and qualitative data of root endophytic fungi allowed to clarify that Cylindrocarpon-like fungi (Ilyonectria spp. and Thelonectria sp.) had a major pathogenic role in the three countries, while Pythium spp. prevailed as pathogen only in German orchards. Gamma ray-sterilized soil resulted in a more-than-proportional increase of plant growth (42% and 31% respectively vs. replant and fallow) as compared to that observed between fallow and replant soil (18%). This best performance in gamma ray-sterilized soil, in addition to the lowest root colonization by Cylindrocarpon-like fungi in this treatment, appeared to be due to the different composition of fungal communities as compared to two other treatments (fallow and replant soil), which instead were highly similar to each other. This difference was due to a shift of community composition toward Fusarium and binucleate Rhizoctonia, which prevailed in sterilized soil.     

Status of applied biological control of soil-borne plant pathogens

During the last 15 years a great amount of information has been accumulated on the ecology and physiology of soil-borne plant pathogens around the world. Despite this, very few cases of applied biological or cultural control have been reported. Although soil-borne plant pathogens are widely spread and economically important, only a small fraction of plant pathologists and soil microbiologists has devoted full time in the applied phases of biological control. The number of publications on applied control during the last 10 years in two journals surveyed is exceptionally small. The status of applied biological control of root diseases is considerably less favorable than the situation in entomology where much progress has been made during the last 10 years. The status of biological control (narrowly or broadly defined) of soil-borne pathogens is described here and several examples of applied control are cited. Future prospects for research on the ecology and biological control are also discussed.     

Soil application of insecticides influences microorganisms and plant nutrients

An experiment was conducted under laboratory conditions to investigate the effect of four insecticides, viz. HCH, phorate, carbofuran and fenvalerate at their field application rates (7.5, 1.5, 1.0 and 0.35 kg a.i. ha⁻¹, respectively), on the growth and development of bacteria, actinomycetes and fungi as well as their role in the transformations and availability of some plant nutrients in laterite soil (Typic Orchragualf). All the insecticides in general, and HCH and phorate in particular, significantly increased the population of microorganisms in soil. The most predominant genera of microorganisms, such as Bacillus, Micrococcus and Aspergillus were not affected by most of the insecticides. However, some of the insecticides stimulated the growth and development of Bacillus, Proteus, Corynebacterium, Streptomyces, Fusarium, Trichoderma and Rhizopus. On the other hand, some insecticide exerted deleterious effect on the proportions of Pseudomonas, Staphylococcus, Nocardia, Micromonospora, Aspergillus and Rhizopus. Incorporation of insecticides also significantly stimulated the mineralization and availability of organic C, N and P in soil. Among the tested substances, the stimulations were more pronounced with HCH followed by phorate and fenvalerate.     

连作根系分泌物加剧土传病害的机制和缓解措施研究进展

基于植物-土壤反馈理论,连作体系中的根系分泌物必然在加剧土传病害发生中起重要作用,但相关研究证据尚缺少系统总结。本文梳理了连作导致土传病害加剧的现象以及连作对典型根系分泌物组分的累积。从有利于土传病原菌由土体向根际迁移、增殖和致病（“利病”）、破坏根际有益微生物群落防线（“压益”）和毒害根系免疫系统（“自毒”）等三个方面,揭示连作根系分泌物中某些物质促进土传病原菌入侵的机制。从根系分泌物角度阐述轮作、间作、套作、伴生和嫁接等多样性种植方式缓解连作土传病害的机制。提出鉴定“利病”、“压益”和“自毒”物质以及构建对应的消减技术途径,可为土传病害绿色高效综合防控提供理论和技术支撑。     

利用微生物防除根寄生杂草列当

根寄生杂草列当(Orobanche spp.)已经严重制约全球许多地区的农业发展,寻找有效防除措施迫在眉睫。由于列当具有特殊生活史且与寄主关系密切,常规防除杂草措施难以达到理想防效。目前,尚无既能有效防除列当又不对寄主造成危害且便于大规模推广应用的列当防除措施。在众多列当防除措施中,微生物防除越来越引起关注和重视。本文对微生物防除列当的国内外研究进展及防除机理进行了综述。目前,列当生防微生物的研究主要集中在镰刀菌(Fusarium spp.)等列当病原菌和根瘤菌(Rhizobium spp.)等列当寄主植物共生菌上。微生物防除列当的机制主要包括两方面:一是通过产生代谢产物直接影响列当的萌发和生长,或通过降解列当种子萌发诱导物质间接影响列当的萌发;二是通过提高寄主植物自身对列当的抗性间接影响列当的寄生和生长。此外,本文还重点介绍了植物土传病害的土壤拮抗微生物防除列当杂草的可行性及研究进展。植物土传病害病原菌和列当均首先通过在地下侵染作物的根系进而危害作物正常生长,而作物抗土传病害的机理也与抗列当的机理类似。因此,存在于土壤中具有防治植物土传病害能力的微生物可能也具有防除根寄生杂草列当的功能。本团队前期试验从植物土传病害的土壤拮抗微生物中筛选到在盆栽试验中能够有效防除向日葵列当(O.cumana Wallr.)和瓜列当(O.aegyptiaca Pers.)的放线菌各1株,分别为淡紫褐链霉菌(Streptomyces enissocaesilis Sveshnikova)和密旋链霉菌(Streptomyces pactum Bhuyan B.K)。其中,密旋链霉菌的菌剂在田间试验中既降低了瓜列当的出土数量又增加了番茄的产量。总之,微生物是防除根寄生杂草列当的一条有效途径。     

Synergists and antagonists in the rhizosphere modulate microbial communities and growth of Quercus robur L.

The synergistic and antagonistic interactions among biotic components in the rhizosphere play a crucial role in plant defence against soil-borne pathogens. We investigated if the rhizosphere helper bacterium Streptomyces sp. AcH 505 (HB) indirectly protects the plant from the parasitic nematode Pratylenchus penetrans by modifying the rhizosphere microbial community structure and whether these interactions are dependent on the growth stage of oaks. Changes in the abundance of Streptomyces sp. AcH 505 and the phospholipid fatty acid (PLFA) composition of the rhizosphere soil as well as oak shoot and root biomass were assessed. Investigated were the bud resting stage A and the bud swelling stage B with maximal root elongation of oak microcuttings at two successive harvest times. The deleterious effect of P. penetrans on oak biomass was dependent on plant development, being limited to oak microcuttings growing at the stage B. In comparison to control and HB inoculated soils, shoot biomass decreased by about 33% and 41%, and root biomass by about 33 and 48%, respectively. The antagonistic effect of Streptomyces against the nematode was linked to shifts in the rhizosphere microbial community. The Streptomyces AcH505 strain promoted growth of oak microcuttings at bud swelling stage B during maximal root elongation and enhanced the abundance of saprophytic and ectomycorrhizal fungi in the rhizosphere by 158% with respect to controls. Our results highlight the importance of Streptomyces for counteracting the damage of nematodes and promoting plant growth in natural ecosystems such as forests.     

Selective influence on populations of rhizosphere or rhizoplane bacteria and actinomycetes by mycorrhizas formed by Glomus fasciculatum

The influence of infection by the vesicular-arbuscular (VA) mycorrhizal fungus Glomus fasciculatum on populations of general taxonomic and functional groups of naturally-occurring rhizosphere and rhizoplane bacteria and actinomycetes associated with roots of sweet corn (Zea mays var. rugosa) and subterranean clover (Trifolium subterraneum L.) was assayed on selective media. Total numbers of bacteria, but not actinomycetes, on the rhizoplane increased on plants with VA mycorrhizas (VAM) compared to plants without VAM. Bacteria and actinomycete populations were not affected quantitatively in the rhizosphere soil of VAM plants. However, VAM affected specific groups of bacteria and actinomycetes in both the rhizosphere soil and rhizosplane. Rhizosphere soil of mycorrhizal plants contained more facultative anaerobic bacteria, had fewer fluorescent pseudomonads, but had the same number of Gram-negative bacteria as non-mycorrhizal plants. Of the actinomycetes assayed, populations of both Streptomyces spp and chitinase-producing actinomycetes decreased in the rhizosphere, but not in the rhizoplane of mycorrhizal plants.Leachates of VAM and non-VAM rhizosphere soil were also compared for the presence or activity of bacteria that could influence sporulation by the root pathogen Phytophthora cinnamomi Rands. Fewer sporangia and zoospores were produced by P. cinnamomi in leachates of rhizosphere soil from VAM plants than from non-VAM plants, suggesting that sporangium-inducing microorganisms had declined or sporangium-inhibitors had increased.Since assays for specific functional groups of microorganisms revealed changes even when total numbers seemed the same, we conclude that the microbial equilibrium had been altered by formation of VA mycorrhizas.     

Biochemical quality of organic amendments affects soil fungistasis

Soilborne plant pathogens are among the most important limiting factors for the productivity of agro-ecosystems. Identifying reliable and effective control methods is crucial for efficient biological control. Soil fungistasis is the capability of soils to inhibit the germination and growth of soil-borne fungi in presence of optimal abiotic conditions. The aim of this study is to clarify the relationships between soil amendments with plant residues spanning a wide variety of biochemical quality and soil fungistasis. Microcosms experiments were performed with 42 different plant residues and the effect on soil fungistasis was assessed by using four different fungi (Aspergillus niger, Botrytis cinerea, Pyrenochaeta lycoperici and Trichoderma harzianum). We measured soil respiration and FDA enzymatic activity and compared classic litter proximate chemical analysis with ¹³C-CPMAS NMR spectroscopy. Results showed that quality of organic amendments is a major controlling factor of soil fungistasis. The dramatic relief of soil fungistasis when soil was amended with lignin poor, but labile C rich, substrates gives strong support to the competition-based hypothesis. The positive correlation between soil respiration and fungal growth further supports the competition hypothesis. Finally, ¹³C NMR results showed a relationship between soil fungistasis and the biochemical quality of plant residues, and provided a quantitative assessment of the time required for fungistasis restoration after organic materials application.     

Selection of phosphorus solubilizing bacteria with biocontrol potential for growth in phosphorus rich animal bone charcoal

Bacteria with the ability to solubilize phosphorus (P) and to improve plant health were selected and tested for growth and survival in P-rich animal bone charcoal (ABC). ABC is suggested to be suitable as a carrier for biocontrol agents, offering them a protected niche as well as delivering phosphate to plants, meanwhile re-using P from waste of the food chain. Ninety-seven bacterial isolates from different soils were tested for their potential to dissolve P from ABC. Of these isolates, 60% showed positive scores; they belonged to the genera Arthrobacter, Bacillus, Burkholderia, Collimonas, Paenibacillus, Pseudomonas, Serratia, and Streptomyces. Twelve isolates from different taxonomic groups were selected for further research on growth ability and survival in ABC, and on their potential to control plant pathogens. The highest concentrations of P were dissolved by Pseudomonas chlororaphis and Bacillus pumilus, followed by Paenibacillus polymyxa, Burkholderia pyrrocinia and three Streptomyces isolates. P. chlororaphis and P. polymyxa showed strongest growth inhibition of plant pathogenic Pythium and Fusarium sp., followed by the Streptomyces spp. isolates.     

Rice (Oryza sativa L.) plant protection using dual biological control and plant growth-promoting agents: Current scenarios and future prospects

Various microorganisms live in association with different parts of plants and can be harmful, neutral, or beneficial to plant health. Some microbial inhabitants of plants can control plant diseases by contesting with, predating on, or antagonizing plant pathogens and by inducing systems for plant defense. A range of methods, including plant growth-promoting microorganisms(PGPMs) as biological control agents(BCAs)(BCA-PGPMs) are used for the biological management and control of plant pathogens. S...     

Trichoderma–plant–pathogen interactions

Biological control involves the use of beneficial organisms, their genes, and/or products, such as metabolites, that reduce the negative effects of plant pathogens and promote positive responses by the plant. Disease suppression, as mediated by biocontrol agents, is the consequence of the interactions between the plant, pathogens, and the microbial community. Antagonists belonging to the genus Trichoderma are among the most commonly isolated soil fungi. Due to their ability to protect plants and contain pathogen populations under different soil conditions, these fungi have been widely studied and commercially marketed as biopesticides, biofertilizers and soil amendments. Trichoderma spp. also produce numerous biologically active compounds, including cell wall degrading enzymes, and secondary metabolites. Studies of the three-way relationship established with Trichoderma, the plant and the pathogen are aimed at unravelling the mechanisms involved in partner recognition and the cross-talk used to maintain the beneficial association between the fungal antagonist and the plant. Several strategies have been used to identify the molecular factors involved in this complex tripartite interaction including genomics, proteomics and, more recently, metabolomics, in order to enhance our understanding. This review presents recent advances and findings regarding the biocontrol-resulting events that take place during the Trichoderma–plant–pathogen interaction. We focus our attention on the biological aspects of this topic, highlighting the novel findings concerning the role of Trichoderma in disease suppression. A better understanding of these factors is expected to enhance not only the rapid identification of effective strains and their applications but also indicate the potentials for improvement of natural strains of Trichoderma.     

Isotope ratio mass spectrometry identifies soil microbial biocontrol agents having trophic relations with the plant pathogen Armillaria mellea

An understanding of the types of interactions that take place between plant pathogens and other microorganisms in the natural environment is crucial in order to identify new potential biocontrol agents. The use of microorganisms labelled with stable isotopes is a potentially useful method for studying direct parasitisation of a given pathogen or assimilation of the pathogen's metabolites by microorganisms. A microorganism labelled with a stable isotope can be monitored in the environment and isotope ratio mass spectrometry can detect whether it is directly parasitised or its metabolites are used by other microorganisms. In this study, we isolated 158 different species of fungi and bacteria from soil and assayed their biocontrol potential against a plant pathogen (Armillaria mellea) by coupling a dual-culture test with mass spectrometry analysis of the ¹³C isotope in the microorganisms in presence of ¹³C-labelled A. mellea. The microorganisms affected the pathogen by means of antibiosis phenomena (total or partial inhibition of pathogen growth, alteration of its morphology) and by antagonism, probably resulting from competition for space and nutrients or from mycoparasitism. Isotope ratio mass spectrometry was used to identify direct trophic interactions between microorganisms and the pathogen as in dual cultures as in soil microcosms. Six fungi and one bacterium were found to display the best active trophic behaviour against the pathogenin dual cultures; three microorganisms were discarded due to their plant pathogen potential. Trichoderma harzianum, Pseudomonas fluorescens and Rhodosporidium babjevae were selected to carry out the experiments. T. harzianum inhibited pathogen development (rate of inhibition 80 ± 0.19%) and its δ ¹³C values increased (244.03 ± 36.70‰) in contact with ¹³C-labelled A. mellea. Lower levels of antagonism and correspondingly lower assimilation of ¹³C were detected in P. fluorescens and R. babjevae. Only T. harzianum maintained mycoparasitic activity in the soil microcosm, showing a δ ¹³C value of 1.97 ± 2.24‰ after one month in co-presence with the labelled pathogen. This study provides support for the use of isotope ratio mass spectrometry as an additional tool in screening for potential biocontrol agents.     

Studies on the ecology of actinomycetes in soil—VIII: Distribution and characteristics of acidophilic actinomycetes

Acidophilic actinomycetes, growing between pH 3.5 and 6.5, were isolated from 17 natural soils and mine wastes. Most isolates were Streptomyces spp. and selected ones were compared with some established neutrophilic species by assessment of overall similarity. Acidophilic and neutrophilic strains clustered separately, differing in a number of cultural and physiological characters, in addition to pH requirements. The possible importance of acidophilic actinomycetes in soil is discussed.     

生防链霉菌配施棉秆炭对连作棉田土壤微生物区系的影响

棉花是重要的经济作物,长期连作能引起棉花土壤微生态的失衡、土传病害加重、进而导致产量和品质的下降,影响棉花产业的健康发展。本文以连作棉田土壤为研究对象,进行室内培养试验,在施用生防放线菌黄三素链霉菌(Streptomyces flavotricini)的基础上添加不同量的棉秆炭[0 g·kg~(-1)(CK)、25.0 g·kg~(-1)、50.0 g·kg~(-1)、100.0 g·kg~(-1)],采用微生物计数和16S rDNA基因序列分析的方法,研究两者配施对连作棉田土壤中生防菌数量、微生物数量和种类的影响,为棉花黄萎病的生物防治提供新的思路。研究结果表明:(1)生防放线菌配施棉秆炭对连作棉田土壤中微生物区系有显著的影响。与单施生防放线菌菌剂的处理相比,两者配施显著增加了土壤中细菌、放线菌和真菌数量,其中配施25.0 g·kg-1棉秆炭处理使土壤中细菌/真菌数量比(B/F)、放线菌/真菌数量比(A/F)分别增加了5 271.2%和30.8%(P0.05)。(2)土壤中生防放线菌数量随着棉秆炭施用量增加而显著增加,配施100.0 g·kg~(-1)棉秆炭处理显著增加了2 672.8%(P0.05)。棉秆炭具有作为生防放线菌良好载体的潜力。(3)生防放线菌配施棉秆炭也改变了土壤中优势微生物的数量和比例,尤其提高了细菌中芽孢杆菌的数量和所占的比例;100.0 g·kg~(-1)棉秆炭与菌剂配施使土壤中链霉菌的数量及比例显著高于对照,但降低了小单孢菌数量;增加了真菌中米曲梅、黑曲霉和木霉的数量,但使其所占比例降低。由此可以看出,生防放线菌配施棉秆炭能提高连作棉田土壤中生防放线菌的数量,增强生防菌制剂的防病促生作用,改善连作棉田土壤微生物群落结构,在防控棉花连作障碍上具有较大的应用潜力。