The study describes the development and employment of plant tests based on artificial inoculation of seeds or the potting substrate for evaluating the potential of microorganisms to control seedling blight of maize caused by seed- and soil-borne fusaria. Nine strains of Fusarium were isolated from maize kernels and identified morphologically and by molecular methods as belonging to the species Fusarium verticillioides, F. subglutinans, F. cerealis, F. poae and F. proliferatum. In order to determine pathogenicity, maize kernels were inoculated by immersion in suspensions of conidia of these isolates and sown in a pasteurized substrate in seed trays. Based on plant dry weight, the isolates of F. verticillioides and F. subglutinans were more pathogenic than the other isolates. Using an isolate of F. subglutinans, the efficacy of a set of 25 potential fungal and bacterial antagonists was assessed using inoculation of maize kernels by placement in mixtures of the pathogen and the antagonists. The results obtained with this methodology indicate the potential of a number of different microorganisms applied as seed treatments, including some reported previously as biocontrol agents, to control seed-borne seedling blight of maize. In order to develop a method for the testing of biocontrol agents against soil-borne attack, isolates of F. subglutinans, F. cerealis and F poae from maize kernels together with isolates of F. avenaceum, F. culmorum and F. graminearum originating from maize silage and wheat were used to artificially inoculate the potting substrate. The results showed large differences in pathogenicity, with the most aggressive isolates belonging to F. culmorum and F. graminearum.
Plant-based screening experiments were conducted with the aim of identifying biocontrol bacteria and fungi for seed treatment of maize. Candidate microorganisms were evaluated for their protective effects against soilborne infections by species of Fusarium, Globisporangium (syn. Pythium) and Rhizoctonia. The microorganisms tested were bacteria and fungi from maize roots or other sources, including some active microbial components of commercial biocontrol products. Due to the method of isolation chosen, the majority of bacteria from maize roots were spore formers, most of them species of the genera Bacillus,Brevibacillus and Paenibacillus. In pot tests with the potting substrate inoculated with F. culmorum, the level of control provided by seed treatment with the most efficacious bacterial and fungal isolates was comparable or close to the chemical reference seed treatment thiram. The most effective bacteria were species of Pseudomonas, Burkholderia and Streptomyces. Among a subset of approx. 100 bacteria studied, the in vivo and in vitro activities against F.culmorum were only weakly correlated, although some strains deviated from this pattern. The most effective fungi were two strains of Clonostachys rosea and isolates of Trichoderma. The latter and a strain of Gliocladium virens provided also protection against R.solani. Activity against Globisporangium ultimum was recorded for one isolate of Trichoderma and the two strains of C. rosea. A reduction in the impact of seedborne F. culmorum was also observed after seed treatment with two strains of F. oxysporum f. sp. strigae. The results are discussed in relation to previous reports on rhizosphere bacteria of maize and their use in biocontrol of plant pathogens or for plant growth promotion.
Pythium isolates from diseased and dead bait plants of maize and cress grown in compost or various soils (maize fields, parkland under deciduous trees, grassland) were characterised and tested for pathogenicity to maize (Zea mays L.). In pot tests performed under controlled conditions, pathogenicity of the isolates to maize was apparent by reduction of root and shoot growth, whereas damping-off of maize seedlings was less frequent. Contrarily, pea seedlings were killed by pathogenic Pythium isolates. Pythium isolates from diseased maize seedlings and pathogenic strains from other gramineous plants (P. phragmitis, P. aff.phragmitis, P. catenulatum) were not necessarily more virulent to maize compared to isolates originating from dicotyledonous plants (cress). The most virulent isolates originated from compost and caused a reduction of maize shoot growth of up to 60%. Phylogenetic analysis revealed that they were very closely related to P. ultimum var. ultimum and P. arrhenomanes, respectively. Isolates originating from maize fields, grassland and parkland under deciduous trees, a reference culture of P. arrhenomanes and strains of P. phragmitis, P. aff. phragmitis and P. catenulatum with known pathogenicity on reed were non-pathogenic on maize. Isolates from compost, and from maize fields generally had a higher temperature optimum for mycelial growth (30 °C) and a faster growth rate (1.5–2.0 mm h−1) compared to the isolates from parkland under deciduous trees and grassland soil (20–25 °C, ~1.0 mm h−1), respectively. This study indicates a potential impact of pathogenic Pythium on maize plants even in the absence of visible symptoms.
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