Seventy bacterial isolates from the rhizosphere of tomato were screened for antagonistic activity against the tomato foot and root rot-causing fungal pathogen Fusarium oxysporum f. sp. radicis-lycopersici. One isolate, strain PCL1391, appeared to be an efficient colonizer of tomato roots and an excellent biocontrol strain in an F. oxysporum/tomato test system. Strain PCL1391 was identified as Pseudomonas chlororaphis and further characterization showed that it produces a broad spectrum of antifungal factors (AFFs), including a hydrophobic compound, hydrogen cyanide, chitinase(s), and protease(s). Through mass spectrometry and nuclear magnetic resonance, the hydrophobic compound was identified as phenazine-1-carboxamide (PCN). We have studied the production and action of this AFF both in vitro and in vivo. Using a PCL1391 transposon mutant, with a lux reporter gene inserted in the phenazine biosynthetic operon (phz), we showed that this phenazine biosynthetic mutant was substantially decreased in both in vitro antifungal activity and biocontrol activity. Moreover, with the same mutant it was shown that the phz biosynthetic operon is expressed in the tomato rhizosphere. Comparison of the biocontrol activity of the PCN-producing strain PCL1391 with those of phenazine-1-carboxylic acid (PCA)-producing strains P. fluorescens 2-79 and P. aureofaciens 30-84 showed that the PCN-producing strain is able to suppress disease in the tomato/F. oxysporum system, whereas the PCA-producing strains are not. Comparison of in vitro antifungal activity of PCN and PCA showed that the antifungal activity of PCN was at least 10 times higher at neutral pH, suggesting that this may contribute to the superior biocontrol performance of strain PCL1391 in the tomato/F. oxysporum system.
Host responses of elms susceptible and resistant to Dutch elm disease were histologically examined. In a time course study the susceptible elm clone Ulmus × hollandica 'Belgica' and U. × hollandica '390', a clone which shows a high degree of resistance to non-aggressive isolates and a moderate degree of resistance to aggressive isolates of Ophiostoma ulmi, were inoculated in twig or trunk with either an aggressive or a non-aggressive isolate of O. ulmi. For purposes of comparison, the susceptible elm U. americana and the more resistant clones U. × hollandica 'Groeneveld', U. 'Lobel' and U. 'Sapporo Autumn Gold' were included. Depending on clone-isolate compatibility, infected twigs reacted by a walling off process, by barrier zone formation, or failed to resist the infection and died. Trees inoculated into the trunk reacted comparably but in the case of a compatible combination they always formed a barrier zone and the cambium never died in the year of inoculation.
SUMMARYAntagonism of a selected group of bacteria against Ophiostoma (=Ceratocystis) ulmi, the Dutch elm disease pathogen, was determined on agar media. Four promising bacterial isolates, all fluorescent Pseudomonads, were used for field experiments and for further tests on in vitro antagonism against several fungi. It was shown that only slight differences existed in antagonism against different O. ulmi isolates. Also Ceratocystis fagacearum and C. fimbriata were inhibited similarly to O. ulmi.In field experiments bacteria were applied to elm trees by low pressure injection or by injection with a specially developed ‘gouge‐pistol’. Elms treated only with bacteria remained healthy throughout two growing seasons. Elms inoculated with O. ulmi developed severe Dutch elm disease symptoms. Trees, inoculated first with O. ulmi and treated subsequently with bacteria also developed severe Dutch elm disease symptoms. Trees treated with bacteria first and inoculated subsequently with O. ulmi showed significantly fewer symptoms, especially those where treatments were carried out with the gouge‐pistol.
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