Re-establishment of suppressiveness to soil- and air-borne diseases by re-inoculation of soil microbial communities

The aim of this study was to investigate the potentials and limitations in restoring soil suppressiveness in disturbed soils. Soils from three sites in UK and Switzerland (STC, REC, THE) differing in their level of suppressiveness to soil-borne and air-borne diseases were γ-irradiated and this soil matrix was re-inoculated with 1% (w/w) of either parent native soil or native soil from the other sites (‘soil inoculum’). Suppressiveness to air-borne and soil-borne diseases was quantified by means of the host-pathogen systems Lepidium sativum (cress)-Pythium ultimum, an oomycete causing root rot and seedling damping-off, and Arabidopsis thaliana-Hyaloperonospora parasitica, an oomycete causing downy mildew. Soil microbial biomass, activity and community structure, as determined by phospholipid fatty acid (PLFA) profiles, were measured in native, γ-irradiated, and re-inoculated soils. Both, L. sativum and A. thaliana were highly susceptible to the pathogens if grown on γ-irradiated soils. Re-inoculation completely restored suppressiveness of soils to the foliar pathogen H. parasitica, independently of soil matrix or soil inoculum, whereas suppressiveness to P. ultimum depended on the soil matrix and, to a lesser extent, on the soil inoculum. However, the soil with the highest inherent suppressiveness did not reach the initial level of suppressiveness after re-inoculation. In addition, native microbial populations as defined by microbial biomass, activity and community structure, could not be fully restored in re-inoculated soils. As for suppressiveness to P. ultimum, the soil matrix, rather than the source of soil inoculum was identified as the key factor for re-establishing the microbial community structure. Our data show that soils do not or only slowly fully recover from sterilisation by γ-irradiation, indicating that agricultural soil management practices such as soil fumigation or heat treatments frequently used in vegetable cropping should be avoided.     

Genetic Diversity Among Brazilian Cultivars and Landraces of Tomato Lycopersicon Esculentum Mill. Revealed by RAPD Markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.     

The mycorrhizal fungus Glomus intraradices and rock phosphate amendment influence plant growth and microbial activity in the rhizosphere of Acacia holosericea

Plants inoculated with arbuscular mycorrhizal (AM) fungi utilize more soluble phosphorus from soil mineral phosphate than non-inoculated plants. However, there is no information on the response of soil microflora to mineral phosphate weathering by AM fungi and, in particular, on the catabolic diversity of soil microbial communities.The AM fungus, Glomus intraradices was examined for (i) its effect on the growth of Acacia holosericea, (ii) plant-available phosphate and (iii) soil microbial activity with and without added rock phosphate.After 4-months culture, AM fungal inoculation significantly increased the plant biomasses (by 1.78× and 2.23× for shoot and root biomasses, respectively), while mineral phosphate amendment had no effect in a sterilized soil. After 12-months culture, the biomasses of A. holosericea plants growing in a non-sterilized soil amended with mineral phosphate were significantly higher than those recorded in the control treatment (by 2.5× and 5× for shoot and root biomasses, respectively). The fungal inoculation also significantly stimulated plant growth, which was significantly higher than that measured in the mineral phosphate treatment. When G. intraradices and mineral phosphate were added together to the soil, shoot growth were significantly stimulated over the single treatments (inoculation or amendment) (1.45×). The P leaf mineral content was also higher in the G. intraradices+mineral phosphate treatment than in G. intraradices or rock phosphate amendment. Moreover, the number of fluorescent pseudomonads has been significantly increased when G. intraradices and/or mineral phosphate were added to the soil. By using a specific type of multivariate analysis (co-inertia analysis), it has been shown that plant growth was positively correlated to the metabolization of ketoglutaric acid, and negatively linked to the metabolisation of phenylalanine and other substrates, which shows that microbial activity is also affected.G. intraradices inoculation is highly beneficial to the growth of A. holosericea plants in controlled conditions. This AM symbiosis optimises the P solubilization from the mineral phosphate and affects microbial activity in the hyphosphere of A. holosericea plants.     

Soil inoculation with the biocontrol agent Clonostachys rosea and the mycorrhizal fungus Glomus intraradices results in mutual inhibition, plant growth promotion and alteration of soil microbial communities

Interactions between the biocontrol fungus Clonostachys rosea IK 726 and a tomato/Glomus intraradices BEG87 symbiosis were examined with and without wheat bran, which served as a food base for C. rosea. In soil without wheat bran amendment, inoculation with C. rosea increased plant growth and altered shoot nutrient content resulting in an increase and decrease in P and N content, respectively. Inoculation with G. intraradices had no effect on plant growth, but increased the shoot P content. Dual inoculation with G. intraradices and C. rosea followed the pattern of C. rosea in terms of plant growth and nutrient content. Wheat bran amendment resulted in marked plant growth depressions, which were counteracted by both inoculants and dual inoculation increased plant growth synergistically. Amendment with wheat bran increased the population density of C. rosea and reduced mycorrhizal fungus colonisation of roots. The inoculants were mutually inhibitory, which was shown by a reduction in root colonisation with G. intraradices in treatments with C. rosea and a reduction in colony-forming units (cfu) of C. rosea in treatments with G. intraradices, irrespective of wheat bran amendment. Moreover, both inoculants markedly influenced soil microbial communities examined with biomarker fatty acids. Inoculation with G. intraradices increased most groups of microorganisms irrespective of wheat bran amendment, whereas the influence of C. rosea on other soil microorganisms was affected by wheat bran amendment. Overall, inoculation with C. rosea increased and decreased most groups of microorganisms without and with wheat bran amendment, respectively. In conclusion, despite mutual inhibition between the two inoculants this interaction did not impair their observed plant growth promotion. Both inoculants also markedly influenced other soil microorganisms, which should be further studied in relation to their plant growth-promoting features.     

增铵营养对番茄幼苗生长和有机酸含量的影响

A hydroponic experiment was conducted to study the effect of partial replacement of NO(3^-)-N by NH -N on theseedling growth and organic acid content of tomato (Lycopersicon esculentum Mill.). A completely randomized designwas established with three replications and five treatments, i.e., NO(3^-)-N/NH(4^ )-N of 100/0, 75/25, 50/50, 25/75 and0/100. Results showed that 25% replacement of NO(3^-)-N by NH(4^ )-N significantly (P = 0.05) improved fresh and dryweight, revealing that a proper percentage of NH -N was important for tomato nitrogen nutrition. This could increasethe plant growth even though tomato was a crop that preferred nitrate nutrition. Also an increase in the proportion ofNH -N in the nutrient solution led to a significant decrease (P=0.05) in malate, citrate and fumarate. However, the25% NH(4^ )-N plus 75% NO(3^-)-N treatment had no significant effect (P = 0.05) on the 2-ketoglutarate, succinate or oxalicacid content, showing that only some organic acids in tomato plants were affected. Only pyruvate increased significantly(P=0.05), and it only increased for 25% and 50% replacement of NO(3^-)-N by NH(4^ )-N. Metabolism of these organicacids, especially malate, citrate and fumarate, should be further studied at the molecular level in vegetables applied withdifferent nitrogen forms.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Long-term effects of organic and synthetic soil fertility amendments on soil microbial communities and the development of southern blight

The effects of tillage and soil fertility amendments on the relationship between the suppressiveness of soils to southern blight and soil physical, chemical and biological factors were examined in experimental station plots in North Carolina. Main plots were either tilled frequently or surface-mulched after one initial tillage. Organic soil amendments including composted cotton gin trash, composted poultry manure, an incorporated rye-vetch green manure, or synthetic fertilizer were applied to subplots in a split-plot design experiment. Incidence of southern blight was lower in surfaced-mulched than tilled soils. Incidence of southern blight was also lower in soils amended with cotton gin trash than those amended with poultry manure, rye-vetch green manure or synthetic fertilizer. Soil water content was negatively correlated with the incidence of disease in both years. Disease incidence was negatively correlated with the level of potassium, calcium, cation exchange capacity (CEC), base saturation (BS) and humic matter in 2002, and net mineralizable nitrogen in 2001. Although, populations of thermophilic organisms were significantly higher in soils amended with cotton gin trash than the other three fertility amendments in each year, there was no significant correlation between the populations of thermophiles and incidence of the disease. Bacterial community diversity indices based on community-level physiological profiling (CLPP) and denaturing gradient gel electrophoresis (DGGE) were significantly higher in soils amended with cotton gin trash than those amended with poultry manure, green manure or synthetic fertilizer. There was a significant negative correlation between the incidence of southern blight, and CLPP and DGGE diversity indices. Greater differences in the richness of bacterial functional groups than genotypes were observed. These results demonstrate that organic soil fertility amendments and cotton gin trash in particular, reduced the development of the disease and affected soil physical, chemical and biological parameters.     

The effect of acidity on the production of signal molecules by Medicago roots and their recognition by Sinorhizobium

The Medicago sativa-Sinorhizobium symbiosis is challenged by acidity, resulting in generally poor nodulation and production. Medicago murex, however, can nodulate and grow at low pH. The effect of low pH on signal exchange in the Sinorhizobium-Medicago symbiosis was studied to gain a greater understanding of the basis for poor nodulation of M. sativa compared to M. murex. Root exudates from M. sativa and M. murex grown in buffered nutrient solution at pH 4.5, 5.8 and 7.0, were collected to measure the expression of nodB induction in Sinorhizobium. A nodB-gusA fusion was constructed and inserted into Sinorhizobium medicae strains WSM419 (acid tolerant) and CC169 (acid sensitive). We identified greater induction by root exudates from both Medicago spp. collected at pH 4.5 than at pH 5.8 and 7.0, less induction by M. murex than M. sativa and less induction of WSM419 than CC169. The same major inducing compounds, 4′,7-dihydroxyflavanone (liquiritigenin), 4′,7-dihydroxyflavone, and 2′,4′,4-trihydroxychalcone (isoliquiritigenin), were identified in exudates of M. murex and M. sativa at all pH values, although in increasing amounts at lower pH. Poor nodulation of M. sativa relative to M. murex under acid conditions is not the consequence of decreased induction of Sinorhizobium nodB by chemical inducers present in the root exudates of both species at low pH.     

Relation between oak tree phenology and the secretion of organic matter degrading enzymes by Lactarius quietus ectomycorrhizas before and during bud break

The ectomycorrhizas (ECM) formed by Lactarius quietus, an ECM fungus specifically associated with Quercus spp., are abundant all the year long. Root and stem growth, beginning before bud break in oak, are strong carbon sinks partially fulfilled with carbohydrate reserves. We hypothesized that L. quietus contributes to providing trees with carbon at bud break through enzymatic activities before photosynthesis begins. Activities of eight secreted enzymes (xylosidase, glucuronidase, cellobiohydrolase, β-glucosidase, N-acetyl-glucosamine, leucine aminopeptidase, acid phosphatase and laccase) relevant to carbon cycling and the release of phophorus and nitrogen from soil organic matter were measured on L. quietus ECMs before, during and after the bud break. Phenological, climatic and pedoclimatic parameters were also measured. Laccase, glucuronidase, cellobiohydrolase and β-glucosidase activities proved to be significantly related to tree reactivation and climate. All these activities can help the formation of new tissues by supplying carbon. L. quietus can behave saprotrophically, using soil organic matter as substrate. This is consistent with the hypothesis that it provides the oak trees with carbon when demand is high and photoassimilates are not yet available.     

Nurse shrubs increased the early growth of Cupressus seedlings by enhancing belowground mutualism and soil microbial activity

The influence of shrubs used as nurse plants was tested on the growth of Cupressus atlantica, on microbial activity and on arbuscular mycorrhizal (AM) soil potential in a Mediterranean environment. An experimental plantation was conducted combining uninoculated, arbuscular mycorrhized Cypress seedlings and an association between Lavandula stoechas planted close to newly planted C. atlantica seedlings. After three years plantation, this association between C. atlantica and L. stoechas lead to a higher growth of C. atlantica and better soil microbial characteristics compared to the control treatment. AM mycelium network, total microbial activity, dehydrogenase activity, phosphate-solubilizing fluorescent pseudomonads and N, P nutrient uptake by C. atlantica, were significantly higher in the presence of L. stoechas than those recorded in the other treatments. This pioneer shrubs facilitates the early establishment of Cypress seedlings by improving soil microbial characteristics and AM fungus community development. Given that the facilitative effect of one plant species to another increases with abiotic stress, the benefits of this technique would be useful in reforestation programs undertaken to rehabilitate degraded areas in Mediterranean region.     

Hybridization mechanisms between the endangered marble trout (Salmo marmoratus) and the brown trout (Salmo trutta) as revealed by in-stream experiments

Populations of the marble trout (Salmo marmoratus) are critically declining due to introgression by brown trout (S. trutta) strains. Hybrids between the two forms are fertile and presently predominant in most rivers of the species’ range.The involved hybridization mechanisms have been studied through two large scale in-stream experiments (Driselpoh and Stopnikarca) as 50% of each species have been stocked at the age of one year in fishless streams, each fish being individually marked. Diagnostic molecular markers were applied to test a partial reproductive isolation between the two species. Emphasis was put on survival and growth patterns of stocked fish (parental generation) as well as on fish hatched within the stream (F₁ generation).No evidence of a partial reproductive barrier between the two species was observed. Survival of the parental generation depended on the year in both streams, as well as on the species in Stopnikarca, but was identical for both species in Driselpoh. In both streams survival of instream hatched individuals measured from 0+ to 2+ was lower for brown trout. In Driselpoh, F₁ 0+, 1+ and 2+ hybrids were larger than pure individuals. Larger hybrids were only observed in Stopnikarca when analyses focused on individuals in inter-specific competition suggesting that heterosis and stress effects may explain the observed size differences.Our results point out important ecological differences between marble and brown trout and have shown that hybridization can easily take place. The findings indicate that high F₁ hybrid presence, survival and heterosis effects may impede marble trout rehabilitation in the area.     

The adverse effects of soil salinization on the growth of Trifolium alexandrinum L. and associated microbial and biochemical properties in a soil from Iran

The aim of this study was to determine the effects of increasing concentrations of salt solutions (including 0.12, 2, 6, and 10 dS m⁻¹) on the growth of berseem clover (Trifolium alexandrinum L.) and related soil microbial activity, biomass and enzyme activities. Results showed that the dry weights of root and shoot decreased with an increase in the concentrations of salt solutions. Soil salinization depressed the microbiological activities including soil respiration and enzyme activities. Substrate-induced respiration was consistently lower in salinized soils, whereas microbial biomass C did not vary among salinity levels. Higher metabolic quotients (qCO₂) and unaffected microbial biomass C at high EC values may indicate that salinity is a stressful factor, inducing either a shift in the microbial community with less catabolic activity or reduced efficiency of substrate utilization. Acid phosphatase and alkaline phosphatase activities decreased with increasing soil salinity. We found significant, positive correlations between the activities of phosphatase enzymes and plant's root mass, suggesting that any decrease in the activities of the two enzymes could be attributed to the reduced root biomass under saline conditions.