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Biocontrol Strain (biocontrol + strain)
Selected AbstractsIdentification and Regulation of Genes from a Biocontrol Strain of Fusarium oxysporumJOURNAL OF PHYTOPATHOLOGY, Issue 9 2007D. R. Fravel Abstract Differential display with three time points revealed that thiram altered expression of numerous genes in the biocontrol fungus Fusarium oxysporum CS-20. Of the 101 bands purified from the differential display gel, 86 were successfully cloned, and 64 sequenced. Based on nucleic acid sequences, homology to known products was found using BLASTn for 26 sequences and homology to hypothetical proteins was found for six sequences, also from Gibberella zeae. One band (BM1 24-1) showed homology to an ABC transporter from three different fungi. Because of its association with detoxification functions, the ABC transporter was selected for further study. Mycelia of CS-20 were exposed to 25 ,g active ingredient (a.i.) thiram in liquid culture for various times from 0 to 8 h. Quantitative real-time PCR was used to evaluate gene expression. At 30 min after treatment with thiram, the ABC transporter was upregulated 20- to 25-fold relative to the control treatment. The ABC transporter was upregulated 15-fold at 1 h after treatment and 10-fold at 2 h. At 8 h after treatment, there was no difference between treated and non-treated for expression of the ABC transporter. Transcription of the gene encoding EST BM1 24-1 is induced in response to thiram treatment and may function in providing resistance in F. oxysporum isolate CS-20 to fungicides and other toxins. Tolerance to toxins may be critical to the successful inclusion of CS-20 in disease control strategies in cropping systems. [source] Use of monoclonal antibodies to quantify the dynamics of ,-galactosidase and endo-1,4-,-glucanase production by Trichoderma hamatum during saprotrophic growth and sporulation in peatENVIRONMENTAL MICROBIOLOGY, Issue 5 2005Christopher R. Thornton Summary Trichoderma species are ubiquitous soil and peat-borne saprotrophs that have received enormous scientific interest as biocontrol agents of plant diseases caused by destructive root pathogens. Mechanisms of biocontrol such as antibiosis and hyperparasitism are well documented and the biochemistry and molecular genetics of these processes defined. An aspect of biocontrol that has received little attention is the ability of Trichoderma species to compete for nutrients in their natural environments. Trichoderma species are efficient producers of polysaccharide-degrading enzymes that enable them to colonize organic matter thereby preventing the saprotrophic spread of plant pathogens. This study details the use of monoclonal antibodies (mAbs) to quantify the production of two enzymes implicated in the saprotrophic growth of Trichoderma species in peat. Using mAbs specific to the hemicellulase enzyme ,-galactosidase (AGL) and the cellulase enzyme endo-1,4-,-glucanase (EG), the relationship between the saprotrophic growth dynamics of a biocontrol strain of Trichoderma hamatum and the concomitant production of these enzymes in peat-based microcosms was studied. Enzyme activity assays and enzyme protein concentrations derived by enzyme-linked immunosorbent assay (ELISA) established the precision and sensitivity of mAb-based assays in quantifying enzyme production during active growth of the fungus. Trends in enzyme activities and protein concentrations were similar for both enzymes, during a 21-day sampling period in which active growth and sporulation of the fungus in peat was quantified using an independent mAb-based assay. There was a sharp increase in active biomass of T. hamatum 3 days after inoculation of microcosms with phialoconidia. After 3 days there was a rapid decline in active biomass which coincided with sporulation of the fungus. A similar trend was witnessed with EG activities and concentrations. This showed that EG production related directly to active growth of the fungus. The trend was not found, however, with AGL. There was a rapid increase in enzyme activities and protein concentrations on day 3, after which they remained static. The reason for the maintenance of elevated AGL probably resulted from secretion of the enzyme from conidia and chlamydospores. ELISA, immunofluoresence and immunogold electron microscopy studies of these cells showed that the enzyme is localized within the cytoplasm and is secreted extracellularly into the surrounding environment. It is postulated that release of oligosaccharides from polymeric hemicellulose by the constitutive spore-bound enzyme leads to AGL induction and could act as an environmental cue for spore germination. [source] Persistence of a biocontrol strain of Phlebiopsis gigantea in conifer stumps and its effects on within-species genetic diversityFOREST PATHOLOGY, Issue 5 2001E. J. Vainio Fungal isolations and genetic fingerprinting were used to determine whether Phlebiopsis gigantea stump treatment against Heterobasidion annosum sl. using a single genotype (Rotstop) would affect the genetic diversity of P. gigantea populations. The survival time of P. gigantea was longer in Norway spruce (Picea abies) stumps compared to Scots pine (Pinus sylvestris) as no isolates were obtained from pine stumps 6 years after treatment, whereas in about half of the spruce stumps the fungus was still present. The usage of Rotstop did not seem to increase the occurrence of the fungus 5 years after the treatment in fresh (1-year-old) untreated stumps within the same forest stands. All the isolates from the 6-year-old treated spruce stumps were identical in genotype with the Rotstop-strain, whereas all isolates from the fresh untreated spruce and pine stumps differed from it. Within the treated pine stand, the biocontrol usage seemed to have caused a slight reduction in genetic markers not related to Rotstop, but there were no statistically significant differences between the marker frequencies and the local natural population. Thus, Rotstop is not likely to cause any immediate threat to the genetic diversity of P. gigantea. Persistance dans les souches de conifères d'un génotype de Phlebiopsis gigantea utilisée en lutte biologique, et effets sur la diversité génétique de l'espèce L'isolement et l'empreinte génétique ont été utilisés pour savoir si le traitement de souches contre Heterobasidion annosum avec un seul génotype de Phlebiopsis gigantea (Rotstop) peut affecter la diversité génétique des populations de P. gigantea. La durée de survie de P. gigantea a été plus longue dans les souches de Picea abies que dans celles de Pinus sylvestris. Chez celles-ci, aucun isolat n'a été obtenu six ans après le traitement alors que chez environ la moitié des souches d'épicéa le champignon était encore présent. Dans les mêmes peuplements, cinq ans après l'application du Rotstop, l'incidence du champignon ne semblait pas avoir augmenté chez les souches fraîches non traitées, âgées de un an. Tous les isolats obtenus à partir des souches traitées six ans auparavant avaient le même génotype que l'isolat du Rotstop, alors que tous ceux issus de souches fraîches non traitées d'épicéa et de pin étaient différents. Chez le peuplement de pins, le traitement biologique semblait avoir causé une légère réduction des marqueurs génétiques non liés au Rotstop, mais leur fréquence n'était pas statistiquement différente de celle de la population naturelle locale. Ainsi, il apparaît peu probable que le Rotstop soit une menace immédiate pour la diversité génétique du P. gigantea. Persistenz eines Phlebiopsis gigantea -Isolates in Koniferenstümpfen und seine Auswirkungen auf die intraspezifische genetische Diversität Mit Isolierungen und genetischem Fingerprinting wurde untersucht, ob eine Stumpfbehandlung mit einem einzelnen Genotyp von Phlebiopsis gigantea (Rotstop) gegen Heterobasidion annosum sl. die genetische Diversität von P. gigantea -Populationen beeinträchtigen könnte. P. giganteaüberlebte auf Stümpfen von Picea abies länger als auf solchen von Pinus sylvestris. Auf Kiefernstümpfen konnte der Pilz sechs Jahre nach der Behandlung nicht mehr nachgewiesen werden, während er auf 50% der Fichtenstümpfe noch vorhanden war. Eine Rotstop-Anwendung 5 Jahre zuvor hatte offenbar keinen Einfluss auf das Vorkommen von P. gigantea in frischen (einjährigen) unbehandelten Stümpfen im gleichen Bestand. Alle Isolate von den sechs Jahre alten behandelten Fichtenstümpfen hatten den gleichen Genotyp wie das Rotstop-Isolat, während alle Isolate von den frischen (unbehandelten) Fichten-und Kiefernstümpfen anderen Genotypen angehörten. In dem behandelten Kiefernbestand war die Frequenz der nicht mit dem Rotstop-Isolat assoziierten genetischen Marker etwas verringert, der Unterschied zur lokalen natürlichen Population war aber statistisch nicht signifikant. Die Anwendung von Rotstop dürfte somit keine kurzfristige Bedrohung der genetischen Diversität von P. gigantea darstellen. [source] Enhanced biocontrol activity of Trichoderma virens transformants constitutively coexpressing ,-1,3- and ,-1,6-glucanase genesMOLECULAR PLANT PATHOLOGY, Issue 4 2007SLAVICA DJONOVI SUMMARY Evidence for the role of chitinases, proteases and ,-1,3- and ,-1,6-glucanases in mycoparasitism by Trichoderma species has been well documented. Moreover, constitutive over-expression of genes encoding individual cell-wall-degrading enzymes (CWDEs) has been shown to improve the potential of biological agents. In this study, we generated transformants of T. virens in which ,-1,3- and ,-1,6-glucanase genes, TvBgn2 and TvBgn3, respectively, were constitutively coexpressed in the same genetic T. virens Gv29.8 wild-type background. The double over-expression transformants (dOEs) grow and sporulate slower than the wild-type (WT). However, the reduction in growth did not seem to affect their mycoparasitic and biocontrol capabilities, as dOEs displayed much higher levels of total ,-1,3- and ,-1,6-glucanase activity than the WT. This higher enzymatic activity of dOEs positively correlated with observed in vitro inhibition of Pythium ultimum and Rhizoctonia solani mycelia, and with enhanced bioprotection of cotton seedlings against P. ultimum, R. solani and Rhizopus oryzae. Besides effective biocontrol of all pathogens at an original inoculum level, the performance of dOEs was highly enhanced (up to 312% of WT performance) when pathogen pressure was greater (i.e. concentration of inoculum was higher or pathogens applied in combination). These results demonstrate that the strategy of introducing multiple lytic enzyme-encoding genes through transformation of a given biocontrol strain can be successfully used to achieve better biocontrol. [source] Biocontrol and Plant Pathogenic Fusarium oxysporum -Induced Changes in Phenolic Compounds in Tomato Leaves and RootsJOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2007Y. Panina Abstract The biocontrol fungus Fusarium oxysporum strain CS-20 was previously shown to reduce the incidence of Fusarium wilt of tomato through an uncharacterized host-mediated response. As phenolic compounds are involved in the defence response of tomato to pathogens and other stressors, this work was undertaken to determine whether biocontrol strains induced changes in phenolic compounds in leaves and roots of tomato seedlings in the presence and absence of pathogenic F. oxysporum f. sp. lycopersici. Roots of intact tomato seedlings were placed in water or aqueous fungal spore suspensions. Two biocontrol F. oxysporum strains [CS-20 (host-mediated mechanism) and 85SK-1 (control mechanism unknown)] and two plant pathogenic strains of F. oxysporum f. sp. lycopersici Race 1 were used. After 24 or 72 h exposure, phenolic compounds were extracted from leaves and roots before identification by HPLC. There were significant qualitative and quantitative differences between the two sampling times. Compared with the control treatment, strain CS-20 significantly altered (usually increasing) the ferulic, caffeic and vanillic acid contents, and concentrations once unidentified phenolic compounds recovered from leaves and roots. In another experiment, tomato seedlings growing in sterile sand were drenched with spores of strain CS-20 the day before treating them with varying concentrations of spores of the pathogen for 24 or 72 h. The amount of pathogen present did not significantly affect the plant phenolic response to the presence of strain CS-20. This work demonstrates that tomato responds within 24 h to the presence of the biocontrol strain CS-20 by alterations in secondary metabolism that are typical of resistance responses in tomato. [source] Recombination and lineage-specific gene loss in the aflatoxin gene cluster of Aspergillus flavusMOLECULAR ECOLOGY, Issue 23 2009GEROMY G. MOORE Abstract Aflatoxins produced by Aspergillus flavus are potent carcinogens that contaminate agricultural crops. Recent efforts to reduce aflatoxin concentrations in crops have focused on biological control using nonaflatoxigenic A. flavus strains AF36 (=NRRL 18543) and NRRL 21882 (the active component of afla-guard®). However, the evolutionary potential of these strains to remain nonaflatoxigenic in nature is unknown. To elucidate the underlying population processes that influence aflatoxigenicity, we examined patterns of linkage disequilibrium (LD) spanning 21 regions in the aflatoxin gene cluster of A. flavus. We show that recombination events are unevenly distributed across the cluster in A. flavus. Six distinct LD blocks separate late pathway genes aflE, aflM, aflN, aflG, aflL, aflI and aflO, and there is no discernable evidence of recombination among early pathway genes aflA, aflB, aflC, aflD, aflR and aflS. The discordance in phylogenies inferred for the aflW/aflX intergenic region and two noncluster regions, tryptophan synthase and acetamidase, is indicative of trans-species evolution in the cluster. Additionally, polymorphisms in aflW/aflX divide A. flavus strains into two distinct clades, each harbouring only one of the two approved biocontrol strains. The clade with AF36 includes both aflatoxigenic and nonaflatoxigenic strains, whereas the clade with NRRL 21882 comprises only nonaflatoxigenic strains and includes all strains of A. flavus missing the entire gene cluster or with partial gene clusters. Our detection of LD blocks in partial clusters indicates that recombination may have played an important role in cluster disassembly, and multilocus coalescent analyses of cluster and noncluster regions indicate lineage-specific gene loss in A. flavus. These results have important implications in assessing the stability of biocontrol strains in nature. [source] | ||||||||||