Efficacy of fungicides and essential oils against bacterial diseases of fruit trees

In the framework of the performed studies, the antibacterial activity of the following fungicides was evaluated: Miedzian 50 WG (active substance - a.s. 50% copper oxychloride), Ridomil MZ Gold 68 WG (a.s. 3.8% metalaxyl-M and 64%, mancozeb), Euparen Multi 50 WG (a.s. 50% tolylfluanid), Captan 80 WG [a.s. 80% N-(captan)], Dithane Neotec 75 WG (a.s. 75% mancozeb). The evaluation also concerned the essential oils: lavender, sage, lemon balm, clove, and a preparation based on thyme oil (BioZell). Each preparation and compound was tested against the following bacterial pathogens: Erwinia amylovora, Xanthomonas arboricola pv. corylina, X. arboricola pv. juglandis, Pseudomonas syringae pv. syringae, Agrobacterium tumefaciens (presently Rhizobium radiobacter). Each preparation and compound was tested at a concentration of 1,000 ppm of active substance. Copper oxychloride was also tested at a concentration of 1,500 ppm. Among the tested fungicides, metalaxyl-M with mancozeb, mancozeb alone, and copper oxychloride inhibited all of the tested strains of pathogenic bacteria. Tolylfluanid did not inhibit any of the bacteria used. Out of the investigated essential oils, the strongest inhibitors of bacteria were: sage, cloves, and BioZell. The protective activity of the above mentioned fungicides was also evaluated in vivo. They were assessed against fire blight on apple blossoms and pear fruitlets, against bacterial canker on sweet cherry fruitlets, and against crown gall on sunflower seedlings (the test plant). All fungicides were applied at the same concentrations as those in the in vitro tests. Only copper oxychloride was found to show protective activity against the studied diseases. This result indicates that the antibacterial properties of the other fungicides did not correspond with their activity on the plant organs used in the in vivo experiment

Izolacja, wstępna charakterystyka i wykrywanie bakterii zanieczyszczających kultury roślinne in vitro

In order to limit the contamination problem in plant tissue cultures experiments on selection of media suitable for detection and isolation of bacteria contaminating plant tissue explants, and preliminary characterization of isolates were made. In the first experiment aiming at detection of bacteria in plant explants four strains representing genera most often occurring at our survey of plant tissue cultures, and earlier isolated and identified (Bacillus, Methylobacterium, Pseudomonas and Xanthomonas) were streaked on five bacteriological media (NA, King B, K, R2A and 523) and on the medium used for plant culture initiation – 1 MS with milk albumin (IM). All strains grew on all media but on K and IM at the slowest rate and on 523 medium at the fastest. The IM medium proved to be useful for immediate bacteria detection at the initial stage of culture. In the second experiment, aiming at characterization of isolates on the basis of colony growth and morphology 14 strains (Agrobacterium, Bacillus, Curtobacterium, Flavobacterium, Lactobacillus, Methylobacterium – 2 strains Mycobacterium, Paenibacillus, Plantibacterium, Pseudomonas, Stenotrophomonas, Xanthomonas, and species Serratia marcescens) were streaked on five microbiological media: KB, NBY, YDC, YNA and YPGA. All strains grew on all those media but at different rates. The only exception was the strain of Lactobacillus spp., which did not grow on King B medium. This medium allowed the detection of such characteristic traits as fluorescence (Pseudomonas) and secretion of inclusions (Stenotrophomonas). The third experiment was focussed on assessment of the sensitivity of detection of specific bacteria in pure cultures and in plant tissue cultures using standard PCR and BIO-PCR techniques with genus specific primers and 2 methods of DNA isolation. Results showed that the use of Genomic Mini kit enabled an increase of the sensitivity by 100 times as compared to extraction of DNA by boiling. Moreover, the application of BIO-PCR increased sensitivity of detection from 102 to 105 times over the standard PCR. If looking for unknown cultivable bacteria more effective detection seems to be use of microbiological method enabling detection on bacteriological media single cells in the fragments of explants or in wash liquids, in which fragmented explants were shaken.W pracy określono możliwość wykrywania obecności bakterii najczęściej spotykanych w kulturach roślinnych in vitro, należących do różnych rodzajów, przy pomocy technik mikrobiologicznej i molekularnej. Za najbardziej przydatną do izolacji uznano pożywkę 523 a także pożywkę zawierającą 1 soli mineralnych MS stosowaną do inicjacji kultur roślinnych, z dodatkiem 0,025% albuminy mlecznej. Większą czułość wykrywania bakterii przy pomocy markerowego DNA uzyskano przy zastosowaniu techniki polegającej na preinkubacji badanego materiału na pożywkach bakteriologicznych, a następnie na izolacji DNA ze wzrostu (BIO-PCR). Przy porównaniu sposobów izolacji DNA większą czułość uzyskano stosując zestaw Genomic Mini (A&A Biotechnology) niż po zastosowaniu ekstrakcji w temperaturze wrzenia

Morphological and biochemical characterization of Erwinia amylovora-induced hypersensitive cell death in apple leaves

In attached apple leaves, spot-inoculated with Erwinia amylovora, the phenotypic appearance of the hypersensitive response (HR) and the participation of ethylene, reactive oxygen species (ROS) and of vacuolar processing enzyme (VPE) (a plant caspase-1-like protease) were analysed. The HR in both the resistant and susceptible genotypes expressed a similar pattern of distinguishable micro HR lesions that progressed into confined macro HR lesions. The HR symptoms in apple were compared to those in non-host tobacco. The morphology of dead cells (protoplast shrinkage and retraction from cell wall) in apple leaves resembled necrotic programmed cell death (PCD). Lesion formation in both cv. Free Redstar (resistant) and cv. Idared (highly susceptible) was preceded by ROS accumulation and elevation of ethylene levels. Treatment of infected leaves with an inhibitor of ethylene synthesis led to a decrease of ethylene emission and suppression of lesion development in both cultivars. In the resistant but not in the susceptible apple cultivar an early and late increase in VPE gene expression was detected. This suggests that VPE might be an underlying component of the response to E. amylovora in resistant apple cultivars. The findings show that in the studied pathosystem the cell death during the HR proceeds through a signal transduction cascade in which ROS, ethylene and VPE pathways play a role

Effects of silvicultural techniques on the diversity of microorganisms in forest soil and their possible participation in biological control of Armillaria and Heterobasidion

Effects of different pre-planting soil preparations and post-harvest wood debris applications in a clear-cut Scots pine plantation, on the abundance, diversity, and activity of culturable microorganisms were investigated. The investigation was done 9 years after the re-plantings had been done. This formed part of an investigation of silvicultural practices for conservation and the biological control of Armillaria and Heterobasidion in northern temperate forests (Poland). The treatments being compared, were expected to have altered the soil’s physical and chemical properties, and consequently, its biological properties. Only soft-rot microfungi from the Ascomycota and Zygomycota were detected in the soil. Fungi, including those antagonistic to Armillaria and Heterobasidion, were more abundant after shallow ploughing than after deep ploughing or ridging, and where chipped rather than coarse wood debris was left on the soil surface or incorporated. Scots pine trees had the most biomass and the least mortality after ridging and leaving coarse wood debris on the surface (associated with only a relatively moderate abundance of fungi)