Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).     

Compost Tea as a Container Medium Drench for Suppressing Seedling Damping-Off Caused by Pythium ultimum

ABSTRACT Compost tea is being used increasingly in agricultural production to control plant diseases. However, there has been limited investigation relating disease control efficacy to various compost tea production methods, particularly compost tea produced with active aeration and additives to increase microbial population densities in compost tea. Aerated compost tea (ACT) and nonaerated compost tea (NCT), produced with or without additives, was investigated for the suppression of damping-off of cucumber caused by Pythium ultimum. Compost tea was used to drench soilless container medium inoculated with P. ultimum; effect on damping-off ranged from not suppressive to consistently suppressive depending on the method used to produce the tea. The most consistent formulation for damping-off suppression was ACT produced with kelp and humic acid additives. Producing ACT with a molasses-based additive inconsistently suppressed damping-off; evidence suggests that residual nutrients can interfere with disease suppression. Heating or diluting compost tea negated suppression. Across all compost tea samples, there was no significant relationship of bacterial populations, measured as active cells, total cells, or CFU, to disease suppression. However, for all ACT produced without the molasses-based additive, there was a threshold of bacterial population density (6 log(10) active cells per ml, 7.48 log(10) total cells per ml, or 7 log(10) CFU per ml) above which compost teas were suppressive.     

Induction of Soil Suppressiveness Against Rhizoctonia solani by Incorporation of Dried Plant Residues into Soil

ABSTRACT Suppressive effects of soil amendment with residues of 12 cultivars of Brassica rapa on damping-off of sugar beet were evaluated in soils infested with Rhizoctonia solani. Residues of clover and peanut were tested as noncruciferous controls. The incidence of damping-off was significantly and consistently suppressed in the soils amended with residues of clover, peanut, and B. rapa subsp. rapifera 'Saori', but only the volatile substance produced from water-imbibed residue of cv. Saori exhibited a distinct inhibitory effect on mycelial growth of R. solani. Nonetheless, disease suppression in such residue-amended soils was diminished or nullified when antibacterial antibiotics were applied to the soils, suggesting that proliferation of antagonistic bacteria resident to the soils were responsible for disease suppression. When the seed (pericarps) colonized by R. solani in the infested soil without residues were replanted into the soils amended with such residues, damping-off was suppressed in all cases. In contrast, when seed that had been colonized by microorganisms in the soils containing the residues were replanted into the infested soil, damping-off was not suppressed. The evidence indicates that the laimosphere, but not the spermosphere, is the site for the antagonistic microbial interaction, which is the chief principle of soil suppressiveness against Rhizoctonia damping-off.     

Suppression of Seedling Damping-Off Caused by Pythium ultimum, P. irregulare, and Rhizoctonia solani in Container Media Amended with a Diverse Range of Pacific Northwest Compost Sources

ABSTRACT Suppression of seedling damping-off disease caused by Pythium spp. and Rhizoctonia solani is a potential benefit of formulating soilless container media with compost. Thirty-six compost samples from Pacific Northwest commercial composting facilities were analyzed for a number of physical, chemical, and biological properties, including suppression of damping-off caused by Pythium ultimum, P. irregulare, and R. solani. The samples were produced from diverse feedstocks and composting technol ogies; this was reflected in a large degree of variability in the measured properties. When mixed with sphagnum peat moss and inorganic aggregates, 67% of the compost samples significantly suppressed P. irregulare damping-off of cucumber, 64% suppressed P. ultimum damping-off of cucumber, and 17% suppressed damping-off of cabbage caused by R. solani. Suppression of Pythium damping-off was related to the potential of compost to support microbial activity and a qualitative index of ammonia volatilization. Suppression of Rhizoctonia damping-off was not related to any one compost factor. Currently available compost products potentially could provide commercially acceptable control of damping-off caused by Pythium spp., but it is necessary to fortify composts with microbial antagonists for the control of R. solani.     

Suppression of Rhizoctonia solani on Impatiens by Enhanced Microbial Activity in Composted Swine Waste-Amended Potting Mixes

ABSTRACT Peat moss-based potting mix was amended with either of two composted swine wastes, CSW1 and CSW2, at rates from 4 to 20% (vol/vol) to evaluate suppression of pre-emergence damping-off of impatiens (Impatiens balsamina) caused by Rhizoctonia solani (anastomosis group-4). A cucumber bioassay was used prior to each impatiens experiment to monitor maturity of compost as the compost aged in a curing pile by evaluating disease suppression toward both Pythium ultimum and R. solani. At 16, 24, 32, and 37 weeks after composting, plug trays filled with compost-amended potting mix were seeded with impatiens and infested with R. solani to determine suppression of damping-off. Pre-emergence damping-off was lower for impatiens grown in potting mix amended with 20% CSW1 than in CSW2-amended and nonamended mixes. To identify relationships between disease suppression and microbial parameters, samples of mixes were collected to determine microbial activity, biomass carbon and nitrogen, functional diversity, and population density. Higher rates of microbial activity were observed with increasing rates of CSW1 amendment than with CSW2 amendments. Microbial biomass carbon and nitrogen also were higher in CSW1-amended mixes than in CSW2-amended potting mixes 1 day prior to seeding and 5 weeks after seeding. Principal component analysis of Biolog-GN2 profiles showed different functional diversities between CSW1- and CSW2-amended mixes. Furthermore, mixes amended with CSW1 had higher colony forming units of fungi, endospore-forming bacteria, and oligotrophic bacteria. Our results suggest that enhanced microbial activity, functional and population diversity of stable compost-amended mix were associated with suppressiveness to Rhizoctonia damping-off in impatiens.     

Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).     

Field management effects on damping-off and early season vigor of crops in a transitional organic cropping system

Transitioning farmland to certified organic vegetable production can take many paths, each varying in their costs and benefits. Here, the effects of four organic transition strategies (i.e., tilled fallowing, mixed-species hay, low-intensity vegetables, and intensive vegetable production under high tunnels), each with and without annual compost applications for 3 years prior to assessment, were characterized. Although transition cropping strategies differed in soil chemistry (P < 0.05), the magnitude of the changes typically were marginal and pairwise comparisons were rarely significant. In contrast, the compost amendment had a much greater impact on soil chemistry regardless of cropping strategy. For example, percent C and total P increased by 2- to 5-fold and K increased from 6- to 12-fold. Under controlled conditions, damping-off of both edamame soybean (cv. Sayamusume) and tomato (cv. Tiny Tim) was reduced from 2 to 30% in soils from the mixed-hay transition. In the field, damping-off of both crops was also significantly lower in plots previously cropped to hay (P < 0.05). Although not always significant (P < 0.05), this pattern of suppression was observed in all four of the soybean experiments and three of the four tomato experiments independent of compost application. The compost amendments alone did not consistently suppress damping-off. However, plant height, fresh weight, and leaf area index of the surviving seedlings of both crops were greater in the compost-amended soils regardless of the transitional cropping treatment used (P < 0.05 for most comparisons). These data indicate that mixed-hay cropping during the transition periods can enhance soil suppressiveness to damping-off. In addition, although compost amendments applied during transition can improve crop vigor by significantly enhancing soil fertility, their effects on soilborne diseases are not yet predictable when transitioning to certified organic production.     

Comparison of an in vitro and a damping-off assay to test soils for suppressiveness to Pythium aphanidermatum

Testing of soil samples in greenhouse assays for suppressiveness to soilborne plant pathogens requires a considerable investment in time and effort as well as large numbers of soil samples. To make it possible to process large numbers of samples efficiently, we compared an in vitro growth assay with a damping-off assay using Pythium aphanidermatum as the test organism on tomato seedlings. The in vitro test compares the radial growth or relative growth of the fungus in soil to that in autoclaved soil and reflects suppressiveness of soils to the pathogen. We used soils from a field experiment that had been farmed either organically or conventionally and into which a cover crop (oats and vetch in mixture) had been incorporated 0, 10, 21, and 35 days previously. We obtained a significant, positive correlation between damping-off severities of tomato seedlings in damping-off assays and both relative and radial growth in vitro. In addition, radial and relative growth of P. aphanidermatum in the in vitro assay were positively correlated with several carbon and nitrogen variables measured for soil and incorporated debris. We did not find differences between the two farming systems for either growth measures of P. aphanidermatum or disease severities on tomato at different stages of cover crop decomposition. The in vitro assay shows potential for use with any fungus that exhibits rapid saprophytic growth, and is most suitable for routine application in suppressiveness testing.     

玉米几种重要病害蛋白质组学研究进展

 本文介绍了玉米镰孢菌和黄曲霉穗腐病、腐霉菌苗期病害抗性相关蛋白, 以及在我国曾严重发生的玉米弯孢霉叶斑病菌致病性相关蛋白质组和寄主抗性相关蛋白质组的研究进展, 探讨了利用蛋白质组学技术深入研究玉米-病原菌互作的优势和重要方向。     

枯草芽孢杆菌G3str菌株在有机硫熏蒸土壤中的繁殖及其对病原真菌的抑制作用

生防菌枯草芽孢杆菌G3^str。菌株与营养添加剂混配使用，在经有机硫土壤熏蒸剂“大扫灭”熏蒸过的土中有一定的增殖，增殖量大于不熏蒸的自然土。同时，在加入菌剂的前6d，灭菌土中脂肽和伊枯草菌素检测量高于自然土。抑制水稻纹枯病菌菌核形成试验表明，G3^str菌增殖过的灭菌土比不增殖的对照有更强的抑菌作用。盆栽试验结果表明，土壤中添加G3^str。菌显著减轻由腐霉引起的茄子猝倒病，土壤熏蒸加强防病效果。     

Control of damping-off in cress and sugar-beet by commercial seed-coating with Pythium oligandrum

Seeds of cress and sugar-beet were coated with oospores of Pythium oligandrum using commercial seed-pelleting or film-coating procedures. Following either procedure approximately 10⁴ oospores were recovered from both seed types, achieving 75.94% of the targeted dose. Oospore germination (9.19%) was unaffected by the coating treatments. Both types of treatment reduced damping-off of cress caused by P. ultimum in artificially infested sand and potting compost and by Rhizoctonia solani in artificially infested sand. In some cases, the level of control was equivalent to fungicide drenches. In general, pelleting of P. oligandrum on cress gave better control than film-coating treatments. P. oligandrum also reduced damping-off of sugar-beet in soil naturally infested with Aphanomyces cochlioides and Pythium spp. Control was equivalent to that achieved with hymexazol fungicide seed-coating treatments and was related to the inoculum potential of A. cochlioides in the soil; neither standard hymexazol coatings nor P. oligandrum treatments gave control at high inoculum potentials. P. oligandrum was not rhizosphere competent on cress or sugar-beet.     

Identification and Quantification of Pathogenic Pythium spp. from Soils in Eastern Washington Using Real-Time Polymerase Chain Reaction

ABSTRACT Traditional methods of quantifying Pythium spp. rely on the use of selective media and dilution plating. However, high variability is inherent in this type of enumeration and counts may not be representative of the pathogenic population of Pythium spp. Variable regions of the internal transcribed spacer of the rDNA were used to design species-specific primers for detection and quantification of nine Pythium spp. from soils in eastern Washington. Primer pairs were designed for Pythium abappressorium, P. attrantheridium, P. heterothallicum, P. irregulare group I, P. irregulare group IV, P. paroecandrum, P. rostratifingens, P. sylvaticum, and P. ultimum and used with real-time polymerase chain reaction. Standard curves were generated for each of the species using SYBR Green I fluorescent dye for detection of amplification. Seventy-seven isolates of Pythium were screened to confirm specificity of each primer set. DNA was extracted from soil and standard curves were generated for P. irregulare group I, P. irregulare group IV, and P. ultimum to correlate populations of each species in the soil with quantities of DNA amplified from the same soil. Examination of raw field soils revealed results similar to those observed in previous studies. This new technique for the quantification of Pythium spp. is rapid and accurate, and will be a useful tool in the future study of these pathogenic Pythium spp.     

Short-term Cover Crop Decomposition in Organic and Conventional Soils: Soil Microbial and Nutrient Cycling Indicator Variables Associated with Different Levels of Soil Suppressiveness to Pythium aphanidermatum

Stages of oat–vetch cover crop decomposition were characterized over time in terms of carbon and nitrogen cycling, microbial activity and community dynamics in organically and conventionally managed soils in a field experiment and a laboratory incubation experiment. We subsequently determined which variables describing soil microbial community dynamics, C and N cycling could be used as predictors of Pythium aphanidermatum damping-off incidence and relative growth. Disease incidence and relative growth of P. aphanidermatum were measured in growth chamber assays and in vitro growth tests. No significant differences were detected between the conventional and organic farming system with respect to either relative growth or disease incidence. Stepwise discriminant analysis on three classes of disease incidence or relative growth led to selection of qualitatively similar variables. Only one soil microbial variable, total biomass of actinomycetes, was selected. Total C and N content of debris extracted from soil as well as NH₄-N content of soil were selected most consistently and show promise for assessment of potential damping-off incidence by P. aphanidermatum for young seedlings.     

The global regulator GacA of Pseudomonas fluorescens CHA0 is required for suppression of root diseases in dicotyledons but not in Gramineae

Pseudomonas fluorescens strain CHA0 suppresses various plant diseases caused by soil-borne fungi. The pseudomonad produces the antimicrobial metabolites 2,4-diacetylphloroglucinol (Phl), pyoluteorin (Plt) and hydrogen cyanide, which are important for disease suppression, as well as the siderophores pyoverdine (Pvd), salicylic acid (Sal) and pyochelin (Pch). In the current work, a derivative of CHA0 with a mutation in the global regulator gene gacA (GacA⁻), which is unable to produce Phl, Plt and HCN, failed to protect the dicotyledonous plants cress and cucumber against damping-off caused by Pythium ultimum . In contrast, the GacA⁻ mutant could still protect the Gramineae wheat and maize against damping-off mediated by the same strain of P. ultimum , and wheat against take-all caused by Gaeumannomyces graminis . However, the GacA⁻ mutant overproduced Pch and Pvd. To gain more insight into disease protection afforded by the GacA⁻ mutant, a GacA⁻ Pvd⁻ double mutant (strain CHA496) was constructed by gene replacement. Strain CHA496 overproduced Pch and Sal compared with CHA0 and protected wheat against P. ultimum and G. graminis , whereas cress and cucumber were not protected. Addition of FeCl₃ repressed Pch and Sal production by strain CHA496 in vitro and impaired the protection of wheat in soil microcosms. In conclusion, a functional gacA gene was necessary for the protection of dicotyledons against root diseases, but not for that of Gramineae. Results indicated also that Pch and/or Sal were involved in the ability of the GacA⁻ Pvd⁻ mutant of CHA0 to suppress root diseases in Gramineae.     

Lessons from 10 years of stakeholder adoption of a soil bioassay for assessing the risk of spinach Fusarium wilt

Although the maritime Pacific Northwest (PNW) is the only region of the United States suitable climatically for spinach seed production, the acidic soils are highly conducive to spinach Fusarium wilt caused by Fusarium oxysporum f. sp. spinaciae. A soil bioassay developed to quantify the risk of spinach Fusarium wilt in fields has been offered to seed growers annually since 2010. Soil sampled from growers' fields each winter was planted with highly susceptible, moderately susceptible, and partially resistant spinach inbred lines, and the plants rated weekly to calculate a Fusarium wilt severity index (FWSI) and the area under the disease progress curve (AUDPC). Results for 147 soils tested from 2010 to 2013 have been published. This study examined results for an additional 248 soils tested from 2014 to 2019 with the bioassay modified to include an option of agricultural limestone amendment to the soils tested. FWSI and AUDPC were affected significantly (p < .001) by the main effects of soil and spinach inbred line, and the interaction of these factors. Correlation analyses showed a range in degree of association of FWSI and AUDPC with spinach seed crop rotation duration and soil properties, depending on the spinach inbred line (r = −.255 to –.267, n = 172 soils with characteristics suitable for correlation analyses). Stepwise regression models for 172 soils with relevant parameters for regression analyses identified spinach seed crop rotation interval, rate of agricultural limestone amendment, soil pH, and soil Fe, Mn, and Zn concentrations as most strongly associated with FWSI and AUDPC. However, the models accounted for ≤33.4% (R²) of the variability in Fusarium wilt risk. The soil bioassay remains a primary tool for spinach seed growers to select fields with low risk of Fusarium wilt.     

An improved enrichment broth for the sensitive detection of Ralstonia solanacearum (biovars 1 and 2A) in soil using DAS–ELISA

A reliable, sensitive, low-cost and easy-to-use technique is described for the detection of Ralstonia solanacearum (the causal organism of bacterial wilt, BW) in soil. A total of 273 potato isolates belonging to five different biovars (Bv), originating from 33 countries worldwide, were tested and successfully detected by antibodies produced at the International Potato Center (CIP). Isolates of R. solanacearum belonging to Bv1 and Bv2A were successfully detected by double antibody sandwich–enzyme-linked immunosorbent assay (DAS–ELISA) at low population levels after incubation of soil suspensions for 48 h at 30°C in a new semiselective broth containing a potato tuber infusion. Detection thresholds of 20 and 200 CFU g⁻¹ inoculated soil were obtained for Bv1 and Bv2A, respectively. Sensitivity of detection of Bv2A was similar or even higher in five different inoculated soil types. No cross-reactions were obtained in DAS–ELISA after enrichment of soil suspensions (i) prepared from 23 different soils sampled in BW-free areas in six departments of Peru; and (ii) inoculated with 10 identified bacteria and 136 unknown isolates of soil microbiota isolated from eight different locations. Only the blood disease bacterium gave a low-level reaction after enrichment. In naturally infested soils, average sensitivities of 97·6 (SE 14·8) and 100·9 (SE 22·6) CFU g⁻¹ were obtained for biovars 1 and 2A, respectively. By making serial dilutions of the soil suspension before enrichment, densities of R. solanacearum could be determined in a semiquantitative way. Results also showed that composite samples of five soils could be analysed to assess field soil populations without reducing detection sensitivity.     

Short-term Cover Crop Decomposition in Organic and Conventional Soils: Characterization of Soil C, N, Microbial and Plant Pathogen Dynamics

Stages of oat–vetch cover crop decomposition were characterized over time in terms of carbon and nitrogen cycling, microbial activity and damping-off pathogen dynamics in organically and conventionally managed soils in a field and a controlled incubation experiment. A measurement of relative growth consisting of radial growth of a fungal colony over non-sterilized soil divided by that over sterilized soil was used as an assay of suppressiveness. No differences in relative growth of Pythium aphanidermatum and Rhizoctonia solani were detected between organic and conventionally managed soils amended with cover crop residue. Significant effects of cover crop decomposition stage on the relative growth of both pathogens were obtained. Relative growth of P. aphanidermatum was highest just after incorporation and decreased 3 weeks after incorporation. Relative growth of R. solani was highest about 20 days after incorporation, and decreased 2 weeks later in the organic system, but continued to increase in the conventional system. In both experiments, the N or C content, C:N ratio or dry weight of retrieved debris were significantly correlated with relative growth of P. aphanidermatum. Relative growth of R. solani was significantly correlated with the C:N ratio of soil or the C or N content of debris. Microbial activity was not consistently associated with relative growth of either pathogen.     

人参病原菌拮抗细菌的分离筛选与鉴定

为明确土壤拮抗细菌对人参常见病原菌的生防价值,从吉林省抚松县人参根际土壤中分离、纯化获得113株细菌,采用平板对峙法测定菌株的拮抗能力,通过温室盆栽试验研究其在土壤及人参根部的定殖规律,并通过形态、培养特征及16S r DNA序列分析进行分类鉴定。对峙试验结果表明,2株细菌对供试人参病原菌具有良好的拮抗广谱性,且多次验证其抑菌效果稳定。定殖研究结果表明,活性菌株SZ-56与SZ-60的定殖能力均达到显著水平,在土壤中的最大定殖菌量分别为每克土7.51×106CFU和7.28×106CFU,30 d后定殖菌量仍保持在105CFU;21 d后在人参根部的定殖菌量仍保持在每克根105CFU水平。根据形态、培养特征、生理生化指标和16S r DNA序列分析,确定SZ-56为内生芽胞杆菌Bacillus endophyticus,SZ-60为解淀粉芽胞杆菌B.amyloliquefaciens。     

进境玉米中玉米内州萎蔫病菌的PCR检测

 为了快速准确检测进境玉米样品中的玉米内州萎蔫病菌Clavibacter michiganensis subsp. nebraskensis(Cmn), 根据GenBank中Cmn的16S-23S序列设计引物CM1/CM4和引物PSM1/CM3。引物PSM1/CM3仅能从供试的4株Cmn菌株中扩增获得208 bp的预期产物, 而其他36株对照菌株均不能扩增出预期条带。灵敏度测试结果表明引物CM1/CM4和PSM1/CM3组合的巢式PCR方法的检测灵敏度高于常规PCR, 检测灵敏度可达40 fg DNA或6.8 CFU目标细菌。常规PCR和巢式PCR方法对进境美国玉米样品的阳性检出率分别为8%和24%, 试验结果表明所建立的PCR方法可用于玉米样品中Cmn的快速检测。