Soil-borne Plant Pathogens Research Articles

Physiological and molecular characterization of compost bacteria antagonistic to soil-borne plant pathogens.

Disease suppressive composts have the potential to mitigate the risks associated with chemical pesticides. One of the main characteristics responsible for the suppressive nature of composts is their microbiological populations. To gain insight into the determinants responsible for their suppressive effects, we assayed composts to (i) isolate and identify beneficial antagonistic bacteria, (ii) quantify their antifungal and anti-oomycetal activities, (iii) extract inhibitory compounds produced by the bacteria, and (iv) identify antimicrobial lipopeptides produced by these bacteria. The antagonistic bacteria belonged to the genera Arthrobacter, Pseudomonas, Bacillus, Brevibacillus, Paenibacillus, and Rummeliibacillus and had the ability to antagonise the growth of Fusarium sambucinum, Verticillium dahliae, and (or) Pythium sulcatum. These bacteria produced antimicrobial compounds that affected the mycelial growth and (or) conidial germination of the pathogens. Mass spectrometry analyses showed the presence of various antimicrobial lipopeptides in Bacillus and Bacillus-related spp. extracts, demonstrating that they are responsible, at least in part, for the antagonistic activity of the bacteria. Results from this work provide greater insight into some of the biological, biochemical, and physiological determinants of suppressiveness in composts involved in the control of plant pathogens.

Methods for management of soilborne plant pathogens

Soilborne pathogens cause significant economic losses in agricultural production all over the world. These species can survive for many years in the absence of a host plant by forming persistent structures such as microsclerotia, sclerotia, chlamydospores or oospores. Consequently, soilborne diseases are particularly difficult to predict, detect, diagnose and successfully control. Over the past 30 years, a fumigant, methyl bromide, has been widely used for their control in many crops. In 1992, methyl bromide was listed as an ozone-depleting substance under the Montreal Protocol ? an international treaty to protect the ozone layer. During the phaseout of methyl bromide, problems generated in agricultural production made it clear that dependence on a single method or a single chemical should be avoided. The objective of this review paper was to summarize the current knowledge about different methods of soilborne disease control including: crop rotation, steam soil disinfection, soil amendments, hydroponics and soilless growing systems, soil solarization, grafting, biological control and use of natural compounds, and chemical control. Positive and negative aspects of all available methods were reviewed. Benefits, achieved by simultaneous application of several methods based on different mechanisms of actions, are discussed.

Frequent Applications of Organic Matter to Agricultural Soil Increase Fungistasis

Soil–borne plant pathogens are among the most important limiting factors for the productivity of agro–ecosystems. Fungistasis is the natural capability of soils to inhibit the germination and growth of soil–borne fungi in the presence of optimal abiotic conditions. The objective of this study was to assess the effects of different soil managements, in terms of soil amendment types and frequency of application, on fungistasis. For this purpose, a microcosm experiment was performed by conditioning a soil with frequent applications of organic matter with contrasting biochemical quality (i. e., glucose, alfalfa straw and wheat straw). Thereafter, the fungistasis response was assessed on four fungi (Aspergillus niger, Botrytis cinerea, Pyrenochaeta lycopersici and Trichoderma harzianum). Conditioned soils were characterized by measuring microbial activity (soil respiration) and functional diversity using the BIOLOG EcoPlatesTM method. Results showed that irrespective of the fungal species and amendment types, frequent applications of organic matter reduced fungistasis relief and shortened the time required for fungistasis restoration. The frequent addition of easily decomposable organic compounds enhanced soil respiration and its specific catabolic capabilities. This study demonstrated that frequent applications of organic matter affected soil fungistasis likely as a result of higher microbial activity and functional diversity.

First Report of Ralstonia pseudosolanacearum Phylotype I Causing Bacterial Wilt on Rodrigues Island, Indian Ocean

The Ralstonia solanacearum species complex (Rssc) encompasses strains that are highly destructive on a wide host range, and some are listed as quarantine organisms in Europe, the United States, and many other countries. This soil-borne bacterial plant pathogen invades the roots and colonizes the xylem vessels, causing a bacterial wilt (BW) disease. Strains are distributed into four phylogenetic groups (phylotypes) linked to their geographical origin of evolution: phylotype I (Asia), phylotype II (America), phylotype III (Africa), and phylotype IV (Australia - Indonesia - Japan). Phylotypes are subdivided into sequevars, encompassing strains that share high homology on the endoglucanase gene partial sequence (750 bp) (Fegan and Prior 2005). Rodrigues is part of the Mascarene Islands in the Southwest Indian Ocean. In February 2016, a survey was conducted to identify the causal agent of BW, encountered for years on this island. Typical BW disease symptoms, including leaf epinasty and browning vessels, were observed on tomato (Solanum lycopersicum L.) in Terre Rouge and on chili pepper (Capsicum frutescens L.) in Nouvelle Decouverte and Montagne Malgache. Seventy-eight stem sections were collected, surface disinfected with 70% ethanol, and crushed into 2 ml of Tris-buffer, then processed for bacterial isolation by plating 50 µl on SM-1 medium (Granada and Sequeira 1983). Among all of the collected stems, typical Ralstonia pseudosolanacearum colonies grew on the semiselective medium from 51 of the 78 samples. Corresponding strains were subcultured and typed as R. pseudosolanacearum phylotype I (Safni et al. 2014), based on the PMX-PCR (Fegan and Prior 2005). A subset of six strains, two from each location, was selected for sequevar identification (Fegan et al. 1998). Strains CFBP8472 and CFBP8465 (Terre Rouge) were identified as sequevar I-31 (GenBank KX242311 and KX242312), but strains CFBP8467 and CFBP8468 (Montagne Malgache) along with CFBP8469 and CFBP8466 (Nouvelle Decouverte) were identified as sequevar I-33 (KX242313 to KX242316). This subset of strains was inoculated on five plants per strain by soil drenching on the susceptible tomato cultivar L390 (Lebeau et al. 2011) with 5 ml of a calibrated suspension (108 CFU/ml in Tris-buffer). Plants were grown in a fully controlled environment at day/night temperatures of 30/25°C ± 1°C under high relative humidity (∼90%). Sterile Tris-buffer inoculated negative controls were grown with the same parameters. Typical BW symptoms were observed within 6 days postinoculation and strains were reisolated and characterized using the same protocol as for the strains that were inoculated, fulfilling Koch's postulates. No symptoms were observed and no Rssc strains could be isolated from negative controls. To our knowledge, this is the first report of R. pseudosolanacearum phylotype I strains causing bacterial wilt on tomato and pepper on Rodrigues. Further studies need to be conducted on the prevalence and diversity of the Rssc strains causing the BW disease, in order to implement adapted control measures, such as the use of local and adapted resistant cultivars. Further surveys on other Southwest Indian Ocean islands such as Mauritius, Reunion, and Seychelles should be achieved to determine the potential origin of the R. pseudosolanacearum phylotype I strains isolated on Rodrigues island. (Texte integral)

Investigating Disease Controlling Ability of Brassica Volatiles and Their Compatibility with Trichoderma harzianum

The present investigation aimed to test the in vitro toxicity of Brassica volatiles against soil-borne plant pathogens (Rhizoctonia solani Kuhn, Sclerotium rolfsii Sacc., Fusarium oxysporum f.sp. ciceris (Padwick) Matuo & K. Sato and Sclerotinia sclerotiorum (Lib.) de Bary) and biocontrol fungus Trichoderma harzianum Rifai. Moreover, the response of T. harzianum to toxic volatiles was also studied in terms of glucanase and chitinase gene expression. A strong fungistatic effect of B. alba treatment was recorded against all the test pathogens. Noticeably, S. sclerotiorum manifested least sensitivity among all the tested pathogens. All T. harzianum isolates were less sensitive as compared to the assayed pathogens. Based on the in vitro study involving 6 T. harzianum isolates, T55 and Th-R showed least, while T39 showed highest sensitivity to the volatiles. Interestingly, the patterns relating to the effect of volatiles on inoculum density of T. harzianum isolates were similar to their effect on fungus growth in vitro. More importantly, expression of chitinase and glucanase genes in different Trichoderma isolates was up-regulated, which could improve the biocontrol activity of T. harzianum. Therefore, here the authors envisage that combining biocontrol and biofumigation has the potential to provide sustainable and cost-effective strategies to manage soil-borne plant pathogens.

Current and projected global distribution of Phytophthora cinnamomi, one of the world's worst plant pathogens

Globally, Phytophthora cinnamomi is listed as one of the 100 worst invasive alien species and active management is required to reduce impact and prevent spread in both horticulture and natural ecosystems. Conversely, there are regions thought to be suitable for the pathogen where no disease is observed. We developed a climex model for the global distribution of P.cinnamomi based on the pathogen's response to temperature and moisture and by incorporating extensive empirical evidence on the presence and absence of the pathogen. The climex model captured areas of climatic suitability where P.cinnamomi occurs that is congruent with all available records. The model was validated by the collection of soil samples from asymptomatic vegetation in areas projected to be suitable by the model for which there were few records. DNA was extracted, and the presence or absence of P.cinnamomi was determined by high-throughput sequencing (HTS). While not detected using traditional isolation methods, HTS detected P.cinnamomi at higher elevations in eastern Australia and central Tasmania as projected by the climex model. Further support for the climex model was obtained using the large data set from south-west Australia where the proportion of positive records in an area is related to the Ecoclimatic Index value for the same area. We provide for the first time a comprehensive global map of the current P.cinnamomi distribution, an improved climex model of the distribution, and a projection to 2080 of the distribution with predicted climate change. This information provides the basis for more detailed regional-scale modelling and supports risk assessment for governments to plan management of this important soil-borne plant pathogen.

Screening Arbuscular Mycorrhizal Fungi in Order to Select the Best for Alleviating Wilt Disease Complex of Capsicum

Arbuscular mycorrhizal fungi (AMF) play an important role in protection of host plant from infection by soil-borne plant pathogens. In the present study different AMF were tested for their ability to alleviate wilt disease complex of tomato caused by bacterial pathogen Ralstonia solanacearum, fungal pathogen Phytophthora capsici and root-knot nematode Meloidogyne incognita. Plant growth parameters like plant height, stem girth, biovolume index, root and shoot dry weight, total plant dry weight, disease index and mycorrhizal parameters like per cent root colonization and spore count in the root zone soil were evaluated. In general AMF inoculation increased plant growth parameters. Among 11 AMF screened, Glomus bagyarajii was found to be the best AMF in enhancing plant height, stem girth, total plant dry weight, mycorrhizal root colonization and most importantly exhibited least per cent disease index (50.20 %) as compared to control (82.14 %). The results show the potential of using G. bagyarajii for the biocontrol of wilt disease complex of capsicum.

Synthesis, characterization, anti-bacterial, anti-fungal and nematicidal activities of 2-amino-3-cyanochromenes

Soil-borne plant pathogens such as nematodes, fungi and some bacteria not only affect the plant cultures but also economy and environmental implications. As a result of these pathogens attacks, the yield and quality of crops is affected. Therefore, synthesis of new compounds to control these pathogens is very important and need of time. Chromenes are considered important group of heterocyclic compounds and exhibited significant biological activities. 2-amino-3-cyano chromenes are considered important medicinal scaffolds among the chromenes. We describe three component microwave assisted synthesis of 2-amino-3-cyano chromenes. The versatility of the reaction was examined by varying the aldehydes in the reaction mixture which lead to the synthesis of series of 2-amino-3-cyanochromenes (1–9). The structural elucidations of the compounds were studied by 1H NMR, 13C NMR, HRMS and FTIR spectrometer and results were well correlated. The synthesized compounds were evaluated for their anti-bacterial, anti-fungal and nematicidal activities. Among the synthesized compounds, compound 6 showed good anti-bacterial activity while compound 9 exhibited high anti-fungal activity. In case of nematicidal activity, compound 8 indicated high activity as compare with other compounds followed by compound 9. Results indicate that these synthesized compounds could be used as effective control for soil-borne plant pathogens such as nematodes, fungi and bacteria and can help to improve the plant production.

Tree species effects on pathogen-suppressive capacities of soil bacteria across two tropical dry forests in Costa Rica.

Antibiotic-producing bacteria in the genus Streptomyces can inhibit soil-borne plant pathogens, and have the potential to mediate the impacts of disease on plant communities. Little is known about how antibiotic production varies among soil communities in tropical forests, despite a long history of interest in the role of soil-borne pathogens in these ecosystems. Our objective was to determine how tree species and soils influence variation in antibiotic-mediated pathogen suppression among Streptomyces communities in two tropical dry forest sites (Santa Rosa and Palo Verde). We targeted tree species that co-occur in both sites and used a culture-based functional assay to quantify pathogen-suppressive capacities of Streptomyces communities beneath 50 focal trees. We also measured host-associated litter and soil element concentrations as potential mechanisms by which trees may influence soil microbes. Pathogen-suppressive capacities of Streptomyces communities varied within and among tree species, and inhibitory phenotypes were significantly related to soil and litter element concentrations. Average proportions of inhibitory Streptomyces in soils from the same tree species varied between 1.6 and 3.3-fold between sites. Densities and proportions of pathogen-suppressive bacteria were always higher in Santa Rosa than Palo Verde. Our results suggest that spatial heterogeneity in the potential for antibiotic-mediated disease suppression is shaped by tree species, site, and soil characteristics, which could have significant implications for understanding plant community composition and diversity in tropical dry forests.

Root-mediated signal transmission of systemic acquired resistance against above-ground and below-ground pathogens.

Background and Aims Plants modulate defence signalling networks in response to various biotic stresses via inter-organ communications. The root-mediated transmission of systemic acquired resistance (SAR) against soil-borne and air-borne plant pathogens from SAR-induced plants to neighbouring plants subjected to local chemical and pathogen treatments was evaluated. Methods The first two plants out of ten Nicotiana benthamiana seedlings were pre-treated with the SAR-triggering chemical benzothiadiazole (BTH). All ten seedlings were then challenged with two pathogenic bacteria, i.e. the root (bacterial wilt) pathogen Ralstonia solanacearum and the leaf (wildfire) pathogen Pseudomonas syringae pv. tabaci, at 7 d after SAR induction. Key Results Disease severity was noticeably lower in BTH-pre-treated plants than in the control. Surprisingly, two plants located next to BTH-treated plants exhibited reduced disease symptoms indicating that SAR signal transmission occurred through the root system. Determinant(s) secreted from the root system were search for and it was found that salicylic acid (SA) is a major molecule involved in SAR transmission through the root. Analysis of the expression of the defence-related genes N. benthamiana pathogenesis-related gene 1a (NbPR1a) and NbPR2 confirmed that BTH treatment elicited SAR via root-root transmission between plants. Plants with knock-down of the multiple resistance component SGT1 and SA biosynthesis-related gene ICS1 by Tobacco rattle virus-mediated virus-induced gene silencing exhibited a lack of root-mediated SAR transmission. The biological relevance of this finding was validated by challenge with the SAR-inducing avirulent pathogen P. syringae pv. syringae instead of BTH, which produced similar results. Conclusions Our findings demonstrated that SAR is transmissible through the root system from SAR-triggered plants to neighbouring plants.

ANTIFUNGAL POTENTIAL OF LEAF EXTRACTS OF LEGUMINOUS TREES AGAINST SCLEROTIUM ROLFSII.

Background:Sclerotium rolfsii Sacc. is a destructive soil-borne plant pathogen that infects over 500 plant species and causes significant yield losses in many economically important plant species. Synthetic fungicides used to combat the menace also pollute the environment and cause health hazards. In order to search environmental friendly alternatives from natural resources, methanolic extracts of three leguminous tree species namely Acacia nilotica (L.) Willd. ex Delile subsp. indica (Benth.) Brenan, Prosopis juliflora (Sw.) DC. and Albizia lebbeck (L.) Benth. were evaluated for their antifungal activity against S. rolfsii and A. nilotica subsp. indica exhibited the maximum fungicidal potential.Materials and Methods:Two hundred grams dried leaf material of each of the three test plant species were extracted with methanol for two weeks. After filtration, methanol was evaporated on a rotary evaporator. Malt extract broth was used to make various concentrations of the crude methanolic extracts and their antifungal potential was determined by comparing the fungal biomass in various treatments with control. Chemical composition of methanolic leaf extract of A. nilotica subsp. indica was determined through GC-MS analysis.Results:Methanolic leaf extract of A. nilotica subsp. indica showed the highest fungicidal activity. Fungal biomass was decreased by 17-55% due to various concentrations of this extract over control. Different concentrations of P. juliflora reduced fungal biomass by 3-52%. Fourteen compounds were identified in methanolic extract of A. nilotica subsp. indica. 9,12,15-octadecatrienoic acid, methyl ester, (Z,Z,Z,)- (16.59%) was the most abundant compound followed by 1-pentanol, 2 methyl-, acetate (14.80%); hexanedioic acid, dimethyl ester (13.10%) and cyclotriaconta- 1, 7, 16, 22-tetraone (10.28%).Conclusion:This study concludes that methanolic leaf extract of A. nilotica subsp. indica can be used for management of S. rolfsii.

Genetic Determinants for Pyomelanin Production and Its Protective Effect against Oxidative Stress in Ralstonia solanacearum.

Ralstonia solanacearum is a soil-borne plant pathogen that infects more than 200 plant species. Its broad host range and long-term survival under different environmental stress conditions suggest that it uses a variety of mechanisms to protect itself against various types of biotic and abiotic stress. R. solanacearum produces a melanin-like brown pigment in the stationary phase when grown in minimal medium containing tyrosine. To gain deeper insight into the genetic determinants involved in melanin production, transposon-inserted mutants of R. solanacearum strain SL341 were screened for strains with defective melanin-producing capability. In addition to one mutant already known to be involved in pyomelanin production (viz., strain SL341D, with disruption of the hydroxphenylpyruvate dioxygenase gene), we identified three other mutants with disruption in the regulatory genes rpoS, hrpG, and oxyR, respectively. Wild-type SL341 produced pyomelanin in minimal medium containing tyrosine whereas the mutant strains did not. Likewise, homogentisate, a major precursor of pyomelanin, was detected in the culture filtrate of the wild-type strain but not in those of the mutant strains. A gene encoding hydroxyphenylpyruvate dioxygenase exhibited a significant high expression in wild type SL341 compared to other mutant strains, suggesting that pyomelanin production is regulated by three different regulatory proteins. However, analysis of the gene encoding homogentisate dioxygenase revealed no significant difference in its relative expression over time in the wild-type SL341 and mutant strains, except for SL341D, at 72 h incubation. The pigmented SL341 strain also exhibited a high tolerance to hydrogen peroxide stress compared with the non-pigmented SL341D strain. Our study suggests that pyomelanin production is controlled by several regulatory factors in R. solanacearum to confer protection under oxidative stress.

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation.

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10(8) spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10(8) spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.

Anaerobic Soil Disinfestation and Brassica Seed Meal Amendment Alter Soil Microbiology and System Resistance

ABSTRACTBrassica seed meal amendments and anaerobic soil disinfestation (ASD) control a spectrum of soil-borne plant pathogens via a diversity of mechanisms. Transformations in microbial community structure and function in certain instances were determinants of disease control and enhanced plant performance. For instance, in strawberry field soils, increased strawberry yields in response to ASD were attained in a carbon-source input-dependent manner. ASD conducted with rice bran as the carbon input, but not molasses, resulted in significantly greater yields. Effective ASD treatments, but not ineffective treatments, resulted in increased abundance of bacteria in phylum Bacteroidetes in treated soils, specific genera of which are known to provide biological disease control. Brassica seed meal amendment resulted in development of a soil system suppressive towards disease incited by the root pathogen Macrophomina phaseolina. Brassica seed meal treated orchard soil systems exhibit resistance to re-infestation by soil-borne pathogens, including the plant parasitic nematode Pratylenchus penetrans. System resistance is associated with significant and prolonged changes in rhizosphere microbiology and specifically amplification of specific organisms with potential to parasitize P. penetrans.

Management of soil-borne diseases of organic vegetables

AbstractWith the rising awareness of the adverse effects of chemical pesticides, people are looking for organically grown vegetables. Consumers are increasingly choosing organic foods due to the perception that they are healthier than those conventionally grown. Vegetable crops are vulnerable to a range of pathogenic organisms that reduce yield by killing the plant or damaging the product, thus making it unmarketable. Soil-borne diseases are among the major factors contributing to low yields of organic produce. Apart from chemical pesticides there are several methods that can be used to protect crops from soil-borne pathogens. These include the introduction of biocontrol agents against soil-borne plant pathogens, plants with therapeutic effects and organic soil amendments that stimulate antagonistic activities of microorganisms to soil-borne diseases. The decomposition of organic matter in soil also results in the accumulation of specific compounds that may be antifungal or nematicidal. With the growing interest in organic vegetables, it is necessary to find non chemical means of plant disease control. This review describes the impact of soil-borne diseases on organic vegetables and methods used for their control.

Role and its utilization of beneficial micro-organisms for sustainable crop production

Soil micro-organisms are important component of integrated nutrient management and soil biodiversity system. They play a pivotal role in the functioning of plants by influencing their physiology and development. It is very important role in biogeo-chemical cycles and has been used for crop production for decades. Plant-bacterial interactions in the rhizosphere are the determinants of plant health and soil fertility. Soil bacteria which are beneficial to plant growth, referred to plant growth promoting rhizobacteria (PGPR), which are capable of promoting plant growth by colonizing the plant root. Symbiotic nitrogen-fixing bacteria include the Cyanobacteria of the genera Rhizobium, Brady rhizobium, Azorhizobium, Allorhizobium, Sinorhizobium and Mesorhizobium . Plant growth promotion and development can be facilitated both directly and indirectly. Indirect plant growth promotionincludes the prevention of the deleterious effects of phytopathogenic organisms. This can be achieved by the production of siderophores, i.e. small metal-binding molecules. Biological control of soil-borne plant pathogens and the synthesis of antibiotics have also been reported in several bacterial species. Another mechanism by which PGPR can inhibit phytopathogens is the production of hydrogencyanide (HCN) and/or fungal cell wall degrading enzymes, e.g., chitinase and β-1,3-glucanase. Direct plant growth promotion includes symbiotic and non-symbiotic PGPR which function through production of plant hormones suchas auxins, cytokinins, gibberellins, ethylene and abscisic acid. Production of indole-3-ethanol or indole-3-acetic acid(IAA), PGPR also help in solubilisation of mineral phosphates and other nutrients, enhance resistance to stress, stabilize soil aggregates, and improve soil structure and organic matter content. PGPR retain more soil organic N, and other nutrients in the plant-soil system, thus, reducing the need for fertilizer.

The Effects of Fungicide, Soil Fumigant, Bio-Organic Fertilizer and Their Combined Application on Chrysanthemum Fusarium Wilt Controlling, Soil Enzyme Activities and Microbial Properties.

Sustained monoculture often leads to a decline in soil quality, in particular to the build-up of pathogen populations, a problem that is conventionally addressed by the use of either fungicide and/or soil fumigation. This practice is no longer considered to be either environmentally sustainable or safe. While the application of organic fertilizer is seen as a means of combating declining soil fertility, it has also been suggested as providing some control over certain soil-borne plant pathogens. Here, a greenhouse comparison was made of the Fusarium wilt control efficacy of various treatments given to a soil in which chrysanthemum had been produced continuously for many years. The treatments comprised the fungicide carbendazim (MBC), the soil fumigant dazomet (DAZ), the incorporation of a Paenibacillus polymyxa SQR21 (P. polymyxa SQR21, fungal antagonist) enhanced bio-organic fertilizer (BOF), and applications of BOF combined with either MBC or DAZ. Data suggest that all the treatments evaluated show good control over Fusarium wilt. The MBC and DAZ treatments were effective in suppressing the disease, but led to significant decrease in urease activity and no enhancement of catalase activity in the rhizosphere soils. BOF including treatments showed significant enhancement in soil enzyme activities and microbial communities compared to the MBC and DAZ, evidenced by differences in bacterial/fungi (B/F) ratios, Shannon–Wiener indexes and urease, catalase and sucrase activities in the rhizosphere soil of chrysanthemum. Of all the treatments evaluated, DAZ/BOF application not only greatly suppressed Fusarium wilt and enhanced soil enzyme activities and microbial communities but also promoted the quality of chrysanthemum obviously. Our findings suggest that combined BOF with DAZ could more effectively control Fusarium wilt disease of chrysanthemum.

Assessment of the impact of the fumigant dimethyl disulfide on the dynamics of major fungal plant pathogens in greenhouse soils

Soil fumigants constitute a major tool for the control of soil borne fungal plant pathogens in protected crops. Dimethyl disulfide (DMDS) is a novel soil fumigant used either alone or in combination with other fumigants for the control of soil borne pests and diseases. In a commercial greenhouse for tomato production, we evaluated the impact of DMDS, comparatively to the alternative fumigant metam sodium, on the population of the dominant fungal plant pathogens in the study soil via q-PCR. Prior to soil fumigation, estimation of the fungal diversity in the studied soil via clone libraries identified Fusarium oxysporum and Rhizoctonia solani as the most abundant soil borne plant pathogens, while Cladosporium spp., known as opportunistic airborne tomato pathogens, were the most dominant fungi and based on this their dynamics upon fumigation was also studied. DMDS, at two dose rates, induced a drastic reduction in the population of F. oxysporum and R. solani, which lasted for the whole cultivation season. On the contrary, metam sodium exhibited an inhibitory effect on F. oxysporum that was alleviated at 120 d post fumigation. Both DMDS and metam sodium induced only a temporal reduction in the soil population of Cladosporium sp. which recovered by 60 days post fumigation. Our data suggest that DMDS even at the low dose rate (56.4 g m−2) could drastically reduce the population of the major soil borne tomato pathogens F. oxysporum and R. solani. Establishment of population thresholds as determined by q-PCR could represent a valuable tool for the estimation of the risk for disease severity and crop yield losses.

Expression of salicylic acid-related genes in Brassica oleracea var. capitata during Plasmodiophora brassicae infection.

Brassica oleracea var. capitata (cabbage) is an important vegetable crop in Asian countries such as Korea, China, and Japan. Cabbage production is severely affected by clubroot disease caused by the soil-borne plant pathogen Plasmodiophora brassicae. During clubroot development, methyl salicylate (MeSA) is biosynthesized from salicylic acid (SA) by methyltransferase. In addition, methyl salicylate esterase (MES) plays a major role in the conversion of MeSA back into free SA. The interrelationship between MES and methytransferases during clubroot development has not been fully explored. To begin to examine these relationships, we investigated the expression of MES genes in disease-susceptible and disease-resistant plants during clubroot development. We identified three MES-encoding genes potentially involved in the defense against pathogen attack. We found that SS1 was upregulated in both the leaves and roots of B. oleracea during P. brassicae infection. These results support the conclusion that SA biosynthesis is suppressed during pathogen infection in resistant plants. We also characterized the expression of a B. oleracea BSMT gene, which appears to be involved in glycosylation rather than MeSA biosynthesis. Our results provide insight into the functions and interactions of genes for MES and methyltransferase during infection. Taken together, our findings indicate that MES genes are important candidates for use to control clubroot diseases.

Seed Treatment with Ethanol Extract of Serratia marcescens is Compatible with Trichoderma Isolates for Control of Damping-off of Cucumber Caused by Pythium ultimum.

Environmentally friendly control measures for soilborne plant pathogens are needed that are effective in different soils when applied alone or as components of an integrated disease control strategy. An ethanol extract of Serratia marcescens N4-5, when applied as a cucumber seed treatment, effectively suppressed damping-off caused by Pythium ultimum in potting mix and in a sandy loam soil. Plant stand associated with this treatment was similar to that of seed treated with the chemical pesticide Thiram in the sandy loam soil. The N4-5 ethanol extract did not consistently provide significant disease control in a loam soil. The N4-5 ethanol extract was compatible with two Trichoderma isolates, not affecting in vitro or in situ colonization of cucumber by these biological control fungi. Control of damping-off of cucumber was never diminished when this ethanol extract was applied as a seed treatment in combination with in-furrow application of the Trichoderma isolates, and disease control was improved in certain instances with these combinations in the loam soil. Data presented here indicate that the N4-5 ethanol extract is compatible with certain beneficial fungi, suggesting that this extract can be used as a component of integrated disease control strategies featuring biological control fungi.