Effect of sewage sludge on suppressiveness to soil-borne plant pathogens

The objective of this study was to evaluate the effect of sewage sludge on soil suppressiveness to the pathogens Fusarium oxysporum f. sp. lycopersici on tomato, Sclerotium rolfsii on bean, Sclerotinia sclerotiorum on tomato, Rhizoctonia solani on radish, Pythium spp. on cucumber, and Ralstonia solanacearum on tomato. Soil samples were collected from an experimental corn field in which sewage sludge had been incorporated once a year, since 1999. Sludge from two sewage treatment stations in Brazil (Franca and Barueri, SP) were applied at the rates of one (1N), two (2N), four (4N) and eight (8N) times the N recommended doses for the corn crop. Soil suppressiveness was evaluated by methods using indicator host plants, baits and mycelial growth. There was no effect of sewage sludge on soil suppressiveness to Fusarium oxysporum f. sp. lycopersici in tomato plants. For S. rolfsii, reduction of the disease in bean was inversely proportional to the dose of Franca sludge. The incidence of dead plants, caused by S. sclerotiorum, was directly proportional to sludge doses applied. For R. solani and R. solanacearum, there was a linear trend with reduction in plant death in soils treated with increasing amounts of sludge from Franca. There was an increase in the pathogen community of Pythium spp., proportional to the amounts of sewage applied. The effects of sewage sludge varied depending on the pathogen, methodology applied and on the time interval between the sewage sludge incorporation and soil sampling.     

Effect of successive cauliflower plantings and Rhizoctonia solani AG 2-1 inoculations on disease suppressiveness of a suppressive and a conducive soil

Disease suppressiveness against Rhizoctonia solani AG 2-1 in cauliflower was studied in two marine clay soils with a sandy loam texture. The soils had a different cropping history. One soil had a long-term (40 years) cauliflower history and was suppressive, the other soil was conducive and came from a pear orchard not having a cauliflower crop for at least 40 years. These two soils were subjected to five successive cropping cycles with cauliflower or remaining fallow in a greenhouse experiment. Soils were inoculated with R. solani AG 2-1 only once or before every crop. Disease decline occurred in all treatments cropped with cauliflower, either because of a decreased pathogen population or increased suppressiveness of the soil. Disease suppressiveness tests indicated that the conducive soil became suppressive after five subsequent cauliflower crops inoculated each cycle with R. solani AG 2-1. Suppressiveness of all treatments was measured in a seed germination test (pre-emergence damping-off) as well as by measuring the spread of R. solani symptoms in young plants (post-emergence damping-off). Results showed that suppressiveness was significantly stimulated by the successive R. solani inoculations; presence of the cauliflower crop had less effect. Suppressiveness was of biological origin, since it disappeared after sterilization of the soil. Moreover, suppressiveness could be translocated by adding 10% suppressive soil into sterilized soil. The suppressive soil contained higher numbers of culturable filamentous actinomycetes than the conducive soil, but treatments enhancing suppressiveness did not show an increased actinomycetes population. The suppressiveness of the soil samples did also not correlate with the number of pseudomonads. Moreover, no correlation was found with the presence of different mycoparasitic fungi, i.e. Volutella spp., Gliocladium roseum, Verticillium biguttatum and Trichoderma spp. The suppressive soil contained a high percentage of bacteria with a strong in vitro inhibition of R. solani. These bacteria were identified as Lysobacter (56%), Streptomyces (23%) and Pseudomonas (21%) spp. A potential role of Lysobacter in soil suppressiveness was confirmed by quantitative PCR detection (TaqMan), since a larger Lysobacter population was present in suppressive cauliflower soil than in conducive pear orchard soil. Our experiments showed that successive cauliflower plantings can cause a decline of the damage caused by R. solani AG 2-1, and that natural disease suppressiveness was most pronounced after subsequent inoculations with the pathogen. The mode of action of the decline is not yet understood, but antagonistic Lysobacter spp. are potential key organisms.     

Saprophytic growth of Rhizoctonia solani Kühn AG2-1 (ZG5) in soil amended with fresh green manures affects the severity of damping-off in canola

Plants of the Brassicaceae contain glucosinolates, the hydrolysis products of which inhibit the growth of many soil-borne fungi that cause plant disease. However, amending soil with green manures of these plants gives inconsistent control of several soil-borne diseases, including those caused by Rhizoctonia solani. To identify factors that contribute to this inconsistency we investigated, in the laboratory and in pot experiments in the glasshouse, the saprophytic behaviour of R. solani AG2-1 (ZG5) in a sandy soil amended with various green manures. Fresh material from either Brassica napus var. Karoo, B. napus B1, B. napus B2, B. nigra, Diplotaxis tenuifolia (a brassicaceous weed) and the non-Brassicaceae species, oat (Avena sativa) or lupin (Lupinus angustifolius) was used at 10 or 100 g of fresh material kg⁻¹ of dry soil in Lancelin sand. At 100 g kg⁻¹ the volatiles of all green manures reduced the hyphal growth of R. solani, except for B. napus B1. D. tenuifolia at 100 g kg⁻¹ inhibited the growth and sclerotial formation of R. solani. Most green manures at 10 g kg⁻¹, and at 40% water holding capacity, stimulated the growth of R. solani for up to 3 months and increased the activity of other microbes. R. solani infected the brassicaceous plants when growing and colonized the residues mixed with soil at 10 g kg⁻¹. This inoculum increased the severity of damping-off in canola, by 27%. Disease was particularly severe when the green manure species, except D. tenuifolia and oat, were grown in situ and residues returned to the pot from which they came, before sowing canola. There is a potential hazard in applying green manures of Brassica species as their residues can, under certain conditions, support the saprophytic activity of R. solani which increases damping-off in canola sown in the amended soils.     

Cause and duration of mustard incorporation effects on soil-borne plant pathogenic fungi

Two fungal plant pathogens, Rhizoctonia solani AG 2-2 and Fusarium oxysporum f.sp. lini, were studied in relation to general responses of soil fungi and bacteria following incorporation of Brassica juncea. Our aim was to understand to what extent the changes in the biological and physicochemical characteristics of the soil could explain the effects on the studied pathogens and diseases, and to determine the temporal nature of the responses. Short-term effects of mustard incorporation (up to 4 months) were investigated in a microcosm experiment, and compared with a treatment where composted plant material was incorporated. In a field experiment, the responses were followed up to 11 months after removal or incorporation of a mustard crop. In general, responses in the variables measured changed more after incorporation of fresh mustard material than after addition of similar amounts of composted plant material (microcosms) or after removal of the mustard crop (field). The soil inoculum potential of R. solani AG 2-2 decreased directly after incorporation of mustard, but increased later to disease levels above those in the untreated soil. Neither of these effects could be explained by changes in the population density of R. solani AG 2-2. Fusarium spp. were less influenced, although an increase in the suppressiveness to Fusarium wilt was observed after mustard incorporation as compared with the treatment where mustard was removed. The microbial responses to mustard incorporation were more pronounced for bacteria than for fungi. After an initial substantial increase, the bacterial density decreased but remained above the levels in the control treatment throughout the experimental periods. The bacterial community structure was modified up to 8 months after mustard incorporation. We conclude that incorporation of fresh mustard influences soil microbial communities, especially the bacteria, and has a potential to control the pathogenic activity of R. solani 2-2 on a short-term perspective. The time dependency in microbial responses is important and should be taken into consideration for the evaluation of the potential of Brassicas to control plant disease on a field scale.     

Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease suppression, located in France (FR), the Netherlands (NL), Sweden (SE) and the United Kingdom (UK). Similarly, two sites from Germany (Berlin, G-BR; and Braunschweig, G-BS) without documentation of disease suppression were sampled. Total bacterial counts were determined by plating serial dilutions from the composite soil samples onto R2A, AGS and King's B media. A total of 1,788 isolates (approximately 100 isolates per medium and site) was screened for antifungal activity, and in vitro antagonists (327 isolates) were found amongst the dominant culturable bacteria isolated from all six soils. The overall proportion of antagonists and the number of isolates with inhibitory activity against F. oxysporum were highest in three of the suppressive soils (FR, NL and SE). Characterization of antagonistic bacteria revealed a high phenotypic and genotypic diversity. Siderophore and protease activity were the most prominent phenotypic traits amongst the antagonists. The composition and diversity of antagonists in each soil was site-specific. Nevertheless, none of the antimicrobial traits of bacteria potentially contributing to soil suppressiveness analyzed in this study could be regarded as specific to a given site.     

Brassica napus seed meal soil amendment modifies microbial community structure, nitric oxide production and incidence of Rhizoctonia root rot

A low glucosinolate content (21.8 μmol g⁻¹) Brassica napus seed meal (RSM) applied to orchard soils altered communities of both pathogenic and saprophytic soil micro-organisms. RSM amendment reduced infection by native and introduced isolates of Rhizoctonia spp. and recovery of Pratylenchus spp. from apple roots. Root infection by Rhizoctonia solani AG-5 was also suppressed in split-root assays where a portion of the root system was cultivated in RSM-amended soils and the remainder grown in the presence of the pathogen but lacking RSM. R. solani hyphal growth was not inhibited by RSM amendment. Suppression of Pratylenchus was attained to an equivalent extent by amending soils with either RSM or soybean meal (SM) when applied to provide a similar N content. Thus, glucosinolate hydrolysis products did not appear to have a significant role in the suppression of Rhizoctonia spp. or Pratylenchus spp. obtained via RSM amendment. RSM amendment elevated populations of Pythium spp. and of ammonia-oxidizing bacteria that release nitric oxide but suppressed fluorescent pseudomonad numbers. Streptomyces spp. soil populations increased significantly in response to RSM but not SM amendment. The vast majority of Streptomyces spp. recovered from the apple rhizosphere produced nitric oxide and possessed a nitric oxide synthase homolog. We propose that transformations in the bacterial community structure are associated with the observed control of Rhizoctonia root rot, with NO production by soil bacteria potentially having a role in the induction of plant systemic resistance.     

Microbial populations involved in the suppression of Rhizoctonia solani AG1-1B by lignin incorporation in soil

Rhizoctonia solani causes worldwide losses in numerous crops. Sclerotia of R. solani remain viable for several years in soil and are an important source of primary infection. In this study the effect of soil incorporation of Kraft pine lignin, a side product of the paper industry, on viability of R. solani AG1-1B sclerotia was investigated. The efficacy of lignin was assessed in a sandy loam (Oppuurs) and a silt loam soil (Leest) collected from commercial fields in Belgium. Evaluating sclerotial viability after 4 weeks incubation in the two soils amended with 1% (w/w) Kraft pine lignin demonstrated a soil-dependent effect. In Leest soil the addition of lignin resulted in a significantly reduced sclerotial viability, together with an increased mycoparasitism by Trichoderma spp.; in Oppuurs soil, on the other hand, only a slight and insignificant reduction in sclerotial viability was observed. Based on phospholipid fatty acid analysis, different changes in microbial community structure upon lignin amendment were detected in the two soils. Both amended soils showed a significant increase in Gram negative bacteria. In Leest soil this increase was accompanied with a significantly higher increase in fungi and actinomycetes compared with Oppuurs soil. In addition, Kraft pine lignin resulted in both soils in a small but significant increase in manganese peroxidase activity and this increase tended to be higher in Leest soil. Manganese peroxidase produced by lignin-degrading basidiomycetes has previously been shown to degrade melanin, which protects the sclerotia against biotic and abiotic stress. We hypothesize that lignin-degrading fungi increased the susceptibility of the sclerotia to sclerotial antagonists such as Trichoderma, Gram negative bacteria and actinomycetes. Clearly, the effect observed here did not rely on the stimulation of one microbial group, but is the result of an interaction of different groups.     

Suppressiveness of 18 composts against 7 pathosystems: Variability in pathogen response

Compost is often reported as a substrate that is able to suppress soilborne plant pathogens, but suppression varies according to the type of compost and pathosystem. Reports often deal with a single pathogen while in reality crops are attacked by multiple plant pathogens. The goal of the present study was to evaluate the disease suppression ability of a wide range of composts for a range of plant pathogens. This study was conducted by a consortium of researchers from several European countries. Composts originated from different countries and source materials including green and yard waste, straw, bark, biowaste and municipal sewage. Suppressiveness of compost-amended (20% vol./vol.) peat-based potting soil was determined against Verticillium dahliae on eggplant, Rhizoctonia solani on cauliflower, Phytophthora nicotianae on tomato, Phytophthora cinnamomi on lupin and Cylindrocladium spathiphylli on Spathiphyllum sp., and of compost-amended loamy soil (20% vol./vol.) against R. solani on Pinus sylvestris and Fusarium oxysporum f. sp. lini on flax. From the 120 bioassays involving 18 composts and 7 pathosystems, significant disease suppression was found in 54% of the cases while only 3% of the cases showed significant disease enhancement. Pathogens were affected differently by the composts. In general, prediction of disease suppression was better when parameters derived from the compost mixes were used rather than those derived from the pure composts. Regression analyses of disease suppression of the individual pathogens with parameters of compost-amended peat-based mixes revealed the following groupings: (1) competition-sensitive: F. oxysporum and R. solani/cauliflower; (2) rhizosphere-affected: V. dahliae; (3) pH-related: P. nicotianae; and (4) specific/unknown: R. solani/pine, P. cinnamomi and C. spathiphylli. It was concluded that application of compost has in general a positive or no effect on disease suppression, and only rarely a disease stimulating effect.     

Biological control of damping-off caused by Rhizoctonia solani using chitinase-producing Paenibacillus illinoisensis KJA-424

A bacterium having strong chitinolytic activity was isolated from a coastal soil in Korea and identified as Paenibacillus illinoisensis KJA-424 on the basis of the nucleotide sequence of a 16S rRNA gene. By activity staining after SDS-PAGE, three major chitinase bands with chitinolytic activity, approximate molecular weight of 63, 54 and 38 kDa were detected. On co-culture Rhizoctonia solani with KJA-424, abnormal swelling and deformation of R. solani hyphae were observed, where the release of N-acetyl-d-glucosamine was detected. The bacterium suppressed the symptom of damping-off cucumber seedlings caused by R. solani, in greenhouse trial.     

Preferential spread of the pathogenic fungus Rhizoctonia solani through structured soil

Most studies on soil fungi have been carried out with little explicit characterisation of soil structure within which fungi spread and biotic interactions occur. In this paper we use a combination of epidemiological (colonisation efficiency) and soil bio-physical (thin sectioning) techniques to investigate the role of macropores in soil on the spread of a fungal colony. Macropores, in the form of gaps orientated in various directions, were artificially introduced in replicated samples of sand and a sandy loam. The pathogenic fungus Rhizoctonia solani AG4 was introduced on the surface (encountering a gap whilst the colony expands over the surface) or within soil (encountering a gap whilst the colony spreads through the bulk soil). Depending on the orientation, location and width, gaps were demonstrated to act as preferential pathways (increasing the colonisation efficiency of R. solani), or as a partial barrier (reducing the colonisation efficiency). Within bulk soil, R. solani preferentially followed larger pores, enabling the fungus to by-pass more densely areas. Study of soil thin sections revealed that hyphal densities were greater in gaps than in the surrounding bulk soil. We use the results to discuss how macropore structure in soil can either enhance or reduce the parasitic spread and saprotrophic invasion of soil by fungi.     

Plant growth responses to biochar addition: an Australian soils perspective

The use of biochar as an agricultural amendment has attracted much attention owing to its potential to improve soil condition and plant growth; however, production outcomes are often uncertain. Although soil type is a major driver of plant productivity, there are relatively few biochar studies that directly compare plant growth responses across a range of soil types. We tested the wheat growth response to biochar derived from poultry litter and from wheat straw applied at 1, 5 and 10 t ha^?1 (approximately 0.13, 0.67 and 1.33 % w/w) in four soils representing major agricultural regions in Australia: an acidic arenosol (Western Australian cereal belt), an acidic rhodic ferralsol (Northern New South Wales), a neutral vertisol (Queensland cropping) and an alkaline haplic calcisol (Eyre Peninsula in South Australia). In the neutral vertisol, where plant growth was vigorous in the control treatments, biochar had little impact, whereas in the alkaline calcisol, there was a small significant increase in shoot biomass at high (10 t ha^?1) application rates. Plant growth responses in the acidic soils were most evident but demonstrated a strong contrast to one another. In the acidic arenosol, negative growth impact correlated with increasing electrical conductivity, while in the acidic ferralsol a small rate-dependent increase in pH correlated with relatively large gains in biomass, possibly due to improved phosphorus nutrition and alleviated Al toxicity. Moving towards effective integration of biochar as a management tool will not only require stratification based on soil types, but wider consideration of the main plant production constraints, such as pH, pertinent to a particular system.     

Calcium concentrations of soil affect suppressiveness against Aphanomyces root rot of pea

The potential for field soils to cause Aphanomyces root rot of pea (Pisum sativum) was estimated for a large number of samples from commercial pea fields over a period of 5 years, using a greenhouse bioassay. The aim of the research project was to gain a mechanistic understanding of soil suppressiveness to the disease. Regression analysis showed that of the measured soil variables (Ca, Mg, K, P, pH), soil Ca concentrations had the strongest (negative) correlation with disease prevalence, and also a significant negative correlation with disease severity in samples with confirmed presence of the disease. Greenhouse bioassays using a set of non-infested soils inoculated with artificially produced oospore inoculum of the casual organism Aphanomyces euteiches, showed a similar negative correlation between soil Ca content and disease severity. Disease severity was not consistently affected by soil sterilisation, but was lowered by the addition of two different Ca salts. In contrast, addition of sodium bicarbonate to two soils lowered the content of water-soluble Ca in the soils and increased disease severity. Studies of cultures of A. euteiches exposed to varying Ca concentrations in vitro showed that zoospore production was inhibited at submillimolar concentrations, while mycelial growth was stimulated or unaffected. We conclude that free Ca is a major variable controlling the degree of soil suppressiveness against A. euteiches, and that inhibition of zoospore production from oospores is a possible mechanism.     

A nonpathogenic species of binucleate Rhizoctonia inhibits the formation of infection structures caused by Rhizoctonia solani on cucumber

Binucleate Rhizoctonia (BNR) W7 was evaluated for the efficacy to control damping-off disease in cucumber seedlings. Results revealed that Lobate appresoria and dome-shaped infection cushion were produced by virulent Rhizoctonia solani C9 AG4 on the surface of the cucumber seedling hypocotyls, causing damping-off, 12 h after inoculation. A significant positive correlation was observed between disease severity rating and number of infection cushions (r = 0.94) and appresoria (r = 0.97) on the hypocotyls of seedlings inoculated with virulent R. solani C9 alone. Inoculation of a nonpathogenic species of BNR prior to virulent R. solani significantly reduced the number of appresoria on hypocotyls surface and inhibited the formation of infection cushions that accompanied by highly significant (P ≤ 0.01) reduction in disease severity in comparison with seedling inoculated with R. solani C9 alone. Formation of mucilaginous material upon recognition of the nonpathogenic isolate of BNR followed by hyphal lysis of BNR W7 6 h after inoculation was responsible for reduction in appresorium formation and complete inhibition of infection cushion formation. However, the number of infection structures produced on cucumber was not significantly different for hypocotyls of seedlings inoculated with R. solani C9 with and without the nonpathogenic BNR W7.     

New insights into the origins and evolution of rhizobia that nodulate common bean (Phaseolus vulgaris) in Brazil

It is generally accepted that there are two major centers of genetic diversification of common beans (Phaseolus vulgaris L.): the Mesoamerican (Mexico, Colombia, Ecuador and north of Peru, probably the primary center), and the Andean (southern Peru to north of Argentina) centers. Wild common bean is not found in Brazil, but it has been grown in the country throughout recorded history. Common bean establishes symbiotic associations with a wide range of rhizobial strains and Rhizobium etli is the dominant microsymbiont at both centers of genetic diversification. In contrast, R. tropici, originally recovered from common bean in Colombia, has been found to be the dominant species nodulating field-grown common-bean plants in Brazil. However, a recent study using soil dilutions as inocula has shown surprisingly high counts of R. etli in two Brazilian ecosystems. In the present study, RFLP-PCR analyses of nodABC and nifH genes of 43 of those Brazilian R. etli strains revealed unexpected homogeneity in their banding patterns. The Brazilian R. etli strains were closely similar in 16S rRNA sequences and in nodABC and nifH RFLP-PCR profiles to the Mexican strain CFN 42^T, and were quite distinct from R. etli and R. leguminosarum strains of European origin, supporting the hypothesis that Brazilian common bean and their rhizobia are of Mesoamerican origin, and could have arrived in Brazil in pre-colonial times. R. tropici may have been introduced to Brazilian soils later, or it may be a symbiont of other indigenous legume species and, due to its tolerance to acidic soils and high temperature conditions became the predominant microsymbiont of common bean.     

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.     

Rhizosphere colonization by Serratia marcescens for the control of Sclerotium rolfsii

Two-hundred and three different strains of bacteria were isolated from the rhizosphere of bean, peanut and chickpea plants grown in Sclerotium rolfsii infested soil. A bacterium, identified as Serratia marcescens, was found to be the best biocontrol agent of the pathogen, under greenhouse conditions (up to 75% disease reduction). Populations of 10⁵ or 10⁶ CFU g⁻¹ soil were the most effective in disease control. The drench and drip application of S. marcescens suspension were more effective in controlling S. rolfsii than spraying, mixing in soil or seed coating. This bacterium significantly reduced damping-off incidence of bean, caused by Rhizoctonia solani, by 50%, but was not effective against Pythium aphanidematum in cucumber. A natural mutant of S. marcescens, resistant to the antibiotic rifampicin, was isolated. The mutant, effective as the wild type, was used to study rhizosphere colonization. The highest population density of the bacteria was found on the proximal portion of the root, decreasing significantly until the tips, where they increased again.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

Soil texture and irrigation influence the transport and the development of Pasteuria penetrans, a bacterial parasite of root-knot nematodes

The transport of the spores of Pasteuria penetrans was studied in three contrasted textured soils (a sandy, a sandy-clay and a clay soils), cultivated with tomato, inoculated with juveniles of Meloidogyne javanica and watered with 25 or 150 mm day⁻¹. One month after inoculation of the nematodes, 53% of the spores inoculated were leached by water flow in the sandy soil but only 14% in the sandy-clay soil and 0.1% in the clay soil. No nematodes survived in the clay soil, while the population was multiplied both in the sandy and in the sandy-clay soils. But juveniles of M. javanica were more infected by P. penetrans in the sandy-clay soil than in the sandy soil. Comparing different combinations of bare soils containing 1.1-57% of clay showed that the best spore percolation and retention balance occurred in soils amended with 10-30% clay. However, the spore recoveries decreased when the soil was enriched with more than 30% clay. The role of clay particles on the extractability of spores and on their availability to attach to the nematode cuticle in the soil is discussed.     

Rhizosphere microbial communities associated with Rhizoctonia damage at the field and disease patch scale

Rhizoctonia solani AG-8 is a major root pathogen in wheat (Triticum aestivum L.) systems worldwide and while natural disease suppression can develop under continuous cropping, this is not always the case. The main aim of our work was to elucidate the rhizosphere microbial community underlying a Rhizoctonia suppressive soil (Avon, South Australia) and to investigate how this community may develop in agricultural soils conducive to disease and of different soil type (Galong and Harden, New South Wales). The Avon suppressive soil community included Asaia spp. and Paenibacillus borealis, which were absent from a paired non-suppressive site. At Galong, soil taken from inside and outside disease patches showed no evidence of suppression, and disease suppression could not be transferred from the suppressive soil to the conducive soil from a different soil type and climatic area. 16S rRNA microarray analysis revealed Pseudomonas spp. were significantly more abundant inside than outside three disease patches at Galong. However, a survey of 32 patches across a range of stubble and tillage treatments at a nearby site showed no correlation between Pseudomonas and disease incidence. R. solani levels were significantly lower when stubble was retained rather than burnt or when nutrients (N, P and S) were incorporated with stubble during the non-crop period. Our results suggest soil type is an important factor for suppressive capability and that where specific disease suppression is absent, agronomic practice to increase soil carbon can encourage a non-specific microbial response that limits disease severity.     

Abiotic soil properties and the occurrence of Rhizoctonia crown and root rot in sugar beet

Since 1993, Rhizoctonia crown and root rot (Rhizoctonia solani AG 2–2 IIIB) has represented an increasing problem for sugar beet production in Germany. Up to now, the outbreak of the infection and the spread of the disease within a field cannot be predicted and effective countermeasures are not available. Although little is known about the living conditions of R. solani in soils, abiotic soil properties are likely to influence the disease occurrence. Investigations were carried out based on 60 pairwise comparisons, each consisting of a disease‐affected and an adjacent nonaffected patch on farmers' fields in 2002 and 2003. Soil samples from the top soil layer (0–30 cm) were collected before harvest, and eight of the most frequently mentioned soil properties potentially influencing Rhizoctonia crown and root rot infection were examined: bulk density, texture, carbonate carbon, potassium, phosphorus, organic carbon, total nitrogen, and pH. The occurrence of the disease was significantly related to the soil C : N ratio, indicating the influence of soil organic matter on the disease occurrence. Examinations of soil thin sections showed that organic‐matter particles in the soil serve as a substrate for R. solani. All other soil physical and chemical properties examined did not differ between the disease‐affected and nonaffected patches and seem to be of minor importance.