Direct and Specific Assessment of Colonisation of Wheat Rhizoplane by Pseudomonas fluorescens Pf29A

The efficacy of fluorescent pseudomonads as suppressors of soil-borne diseases is linked to their ability to colonise plant roots. Monitoring the dynamics of biocontrol agents in the rhizosphere should improve the irreliability. We designed a pair of Sequenced Characterised Amplified Region (SCAR) primers specific to Pseudomonas fluorescens Pf29A, based on a specific 700 bp RAPD product selected in a previous work. Primer specificity was tested with DNA samples extracted from rhizospheric soil and rhizoplane of wheat plants grown in two different non-sterile soils. We assessed the total population of Pf29A by PCR and the culturable population by counting a tetracycline-resistant Pf29A transformant producing Green Fluorescent Protein (GFP), on selective medium 5 days after inoculation of non-sterile soil. SCAR primers were specific for Pf29A in both soils. We evaluated the limit of detection to 14.2 fg of target DNA, equivalent to 242 Pf29A cells per cm of wheat root. Culturable populations of Pf29A transformant accounted for 13% and 4% of the total populations 5 days after treatment with 10³ and 10⁷ CFU of transformed Pf29A per gram of soil. The SCAR derived sequence is a good candidate to develop a strain specific and sensitive PCR-quantification of Pf29A available for population dynamic studies in fields. We confirm that only a small proportion of the total Pf29A rhizosphere population is culturable.     

Influence of soil type and pH on the colonisation of sugar beet seedlings by antagonistic Pseudomonas and Bacillus strains,and on their control of Pythium damping-off

In five different soils originating from Scotland (Craibstone and Cruden Bay), Germany (Magdeburg and Uelzen) and Greece (Tymbaki), Pseudomonas fluorescens B5 reached higher population sizes (4.7–5.7logCFU/plant) on 12-day-old sugar beet seedlings than Bacillus subtilis MBI 600 (4.1–4.8logCFU/plant). Total population size per plant was not affected by soil type. In all five soils, the antagonists reached highest population densities in the hypocotyl and the upper 2cm root section (P. fluorescensB5: 5.2–6.8log₁₀CFU/g plant fresh weight, Bacillus subtilisMBI 600: 5.2–6.1log₁₀CFU/g plant fresh weight) and declined to 0–3log₁₀CFU below 4cm root depth. Colonisation by P. fluorescens B5 down the root was slightly increased in the soils from Craibstone, Magdeburg, and Uelzen compared to the sandy clay loam from Tymbaki. In lux-marked P. fluorescens B5, population density was positively correlated with light emission in all soils; the light emission indicated physiological activity of the strains. However, P. fluorescens B5 reduced Pythium damping-off (measurement after 14 days plant growth) only in three of the five soils (Craibstone, Cruden Bay and Magdeburg). Co-inoculation of B. subtilis MBI 600 increased downward colonisation of the root by P. fluorescens B5, but not the total population ofP. fluorescens B5 per plant. Bacillus subtilisMBI 600 did not reduce Pythium damping-off in any of the soils nor did it influence the efficiency of co-inoculated P. fluorescens B5; its population consisted mainly of physiologically inactive spores. In Craibstone soil, pH did not affect population density, distribution along the root or biocontrol activity against P. ultimum of P. fluorescens B5 or B. subtilis MBI 600.     

Implication of Systemic Induced Resistance in the Suppression of Fusarium Wilt of Tomato by Pseudomonas fluorescens WCS417r and by Nonpathogenic Fusarium oxysporum Fo47

Fluorescent pseudomonads and nonpathogenic Fusarium oxysporum have been shown to suppress fusarium wilts. This suppression has been related to both microbial antagonism and induced resistance.The aim of the present study was to assess the relative importance of systemic induced resistance in the suppression of fusarium wilt of tomato in commercial-like conditions by a reference strain of each type of microorganism (P. fluorescens WCS417r and nonpathogenic F. oxysporum Fo47). The spatial separation of the pathogen and the biocontrol strains excluded any possible microbial antagonism and implicated the involvement of the systemic induced resistance; whereas the absence of any separation between these microorganisms allowed the expression of both mechanisms. Since systemic induced resistance has often been associated with the synthesis of PR-proteins, their accumulation in tomato plants inoculated with WCS417r or with Fo47 was determined.The analysis of the results indicates that the suppression of fusarium wilt by P. fluorescens WCS417r was ascribed to systemic induced resistance without any detection of the PR-proteins tested (PR-1 and chitinases). In contrast, the suppression achieved by nonpathogenic F. oxysporum Fo47 appeared to be mainly ascribed to microbial antagonism but also to a lesser extent to systemic induced resistance. This induced resistance could be related to the accumulation of PR-1 and chitinases.The possible relationship between the ability of Fo47 to suppress fusarium wilt more efficiently than WCS417r and its ability to show both mechanisms is discussed.     

策勒县流动性沙地沙拐枣属的引选和造林研究

策勒县南倚昆仑山,北入塔克拉玛干沙漠腹地,属极端干旱的荒漠地区。沙拐枣属在这里选优获得成功,筛选出的主要种:东疆沙拐枣,头状沙拐枣、乔木状抄拐枣。本文研究了沙拐枣属抗干旱、抗风蚀、沙埋等的内在机制,以及抗逆性造林技术。     

A review of biocontrol agents in controlling late blight of potatoes and tomatoes caused by Phytophthora infestans and the underlying mechanisms

Phytophthora infestans causes late blight on potatoes and tomatoes, which has a significant economic impact on agriculture. The management of late blight has been largely dependent on the application of synthetic fungicides, which is not an ultimate solution for sustainable agriculture and environmental safety. Biocontrol strategies are expected to be alternative methods to the conventional chemicals in controlling plant diseases in the integrated pest management (IPM) programs. Well-studied biocontrol agents against Phytophthora infestans include fungi, oomycetes, bacteria, and compounds produced by these antagonists, in addition to certain bioactive metabolites produced by plants. Laboratory and glasshouse experiments suggest a potential for using biocontrol in practical late blight disease management. However, the transition of biocontrol to field applications is problematic for the moment, due to low and variable efficacies. In this review, we provide a comprehensive summary on these biocontrol strategies and the underlying corresponding mechanisms. To give a more intuitive understanding of the promising biocontrol agents against Phytophthora infestans in agricultural systems, we discuss the utilizations, modes of action and future potentials of these antagonists based on their taxonomic classifications. To achieve a goal of best possible results produced by biocontrol agents, it is suggested to work on field trials, strain modifications, formulations, regulations, and optimizations of application. Combined biocontrol agents having different modes of action or biological adaptation traits may be used to strengthen the biocontrol efficacy. More importantly, biological control agents should be applied in the coordination of other existing and forthcoming methods in the IPM programs. © 2023 Society of Chemical Industry.     

Siderophore-mediated competition for iron and induced resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp

The mechanisms of suppression of fusarium wilt of carnation by two fluorescentPseudomonas strains were studied.Treatments of carnation roots withPseudomonas sp. WCS417r significantly reduced fusarium wilt caused byFusarium oxysporum f. sp.dianthi (Fod). Mutants of WCS417r defective in siderophore biosynthesis (sid^–) were less effective in disease suppression compared with their wild-type. Treatments of carnation roots withPseudomonas putida WCS358r tended to reduce fusarium wilt, whereas a sid^– mutant of WCS358 did not.Inhibition of conidial germination of Fod in vitro by purified siderophores (pseudobactins) of bothPseudomonas strains was based on competition for iron. The ferrated pseudobactins inhibited germination significantly less than the unferrated pseudobactins. Inhibition of mycelial growth of Fod by bothPseudomonas strains on agar plates was also based on competition for iron: with increasing iron content of the medium, inhibition of Fod by thePseudomonas strains decreased. The sid^– mutant of WCS358 did not inhibit Fod on agar plates, whereas the sid^– mutants of WCS417r still did. This suggests that inhibition of Fod by WCS358r in vitro was only based on siderophore-mediated competition for iron, whereas also a non-siderophore antifungal factor was involved in the inhibition of Fod by strain WCS417r.The ability of thePseudomonas strains to induce resistance against Fod in carnation grown in soil was studied by spatially separating the bacteria (on the roots) and the pathogen (in the stem). Both WCS417r and its sid^– mutant reduced disease incidence significantly in the moderately resistant carnation cultivar Pallas, WCS358r did not.It is concluded that the effective and consistent suppression of fusarium wilt of carnation by strain WCS417r involves multiple mechanisms: induced resistance, siderophore-mediated competition for iron and possibly antibiosis. The less effective suppression of fusarium wilt by WCS358r only depends on siderophore-mediated competition for iron.