5'-Deoxyisopentenyladenosine and other cytokinins in culture filtrates of the bacterium Pantoea agglomerans

A Pantoea agglomerans isolate from barley seeds, selected for inducing growth promotion in tomato test plants, was found to excrete several hormones of the cytokinin class into its culture medium. In addition to isopentenyladenine, isopentenyladenosine and the 2-methylthiol derivates of these, an unknown compound with affinity to anticytokinin antibodies was also isolated. Mass spectroscopy indicated the structure of this to be a deoxyisopentenyladenosine. The structure of 9-(5'-deoxy- β - d -ribofuranosyl)-6-(3-methyl-2-butenylamino)purine was verified after synthesis of standards and analysis with GC–MS. The synthesized 5'-deoxyisopentenyl-adenosine showed activity in the Amaranthus bioassay, specific for cytokinins. To our knowledge this is the first report of a naturally occurring cytokinin containing 5'-deoxyribose.     

Biological substitutes for pesticides

In the 20th century an increasing number of pesticides, based on biocidal molecules, were the means for a substantial increase in food and fibre production and quality. Because of health and environmental concerns continued extensive use of such molecules is intensively debated and substitutes are often urgently required. Beside crop plant resistance, various biological control methods based on natural pest suppressing organisms are regarded as main alternatives. Several approaches and concepts also have been tested and commercial organism-based preparations are steadily increasing. However, further biotechnological efforts are required to give them status of being practical substitutes to pesticides. At present they are not comparable to pesticides in meeting efficacy, market and other expectations, but they still have a promising future, especially where genetically modified organisms can be used.     

Differential reactions of wheat and pea genotypes to root inoculation with growth-affecting rhizosphere bacteria

Spring wheat (Triticum aestivum L.) and pea (Pisum spp.) genotypes were tested for reaction to root inoculation with rhizosphere bacteria affecting plant growth. Plant response was studied in greenhouse experiments after treatment of seedlings with bacteria suspended in nutrient broth. Significant genotype variation was found in both wheat and pea in terms of shoot dry weight and severity of bacteria-induced leaf symptoms. For most bacterial isolates tested, there was good correlation between ratings of leaf symptoms 7 to 14 days after inoculation and growth inhibition measured after four weeks. Interactions between isolates and plant genotypes were significant in both wheat and pea (P=0.0024 and 0.0001, respectively), but genotypes with sensitivity or tolerance to most isolates could be distinguished. In an outdoor pot experiment, two of the bacterial isolates caused delayed plant development and differential decreases in grain yield of wheat genotypes. The hypothesis that the reaction of wheat genotypes to the tested becteria was related to their influence on bacterial establishment in the rhizosphere could not be substantiated.     

Effects of long-term barley monoculture on plant-affecting soil microbiota

Effects of soil microbiota on shoot and root growth of barley were tested in a greenhouse tube-growing system. Tubes were filled with a mixture of pure sand and various percentages of soils sampled from plots in three long-term field experiments measuring effects of various crop rotations on yield. Using 3% soil in the sand-soil mixture, shoot dry weight of barley test plants was reduced by about 35% and root depth by about 40% in soils from monoculture plots as compared to soils from crop-rotation plots. Typical root symptoms on poorly growing barley plants started as distinct dark-brown zones which then rapidly spread over the whole root system until the root tips ceased to grow. As tested in one experiment, the barley monoculture soil also affected wheat and oats, but to a lesser degree than it did barley. Most of the depressing effects of monoculture soil on barley were eliminated when soil samples were treated with metalaxyl or heated to 65°C for 2 hours. A Pythium sp. frequently isolated from barley roots showing typical symptoms affected barley, wheat and oats in the same way as did barley monoculture soil.     

Isolates of Phytophthora cryptogea Pathogenic to Wheat and Some Other Crop Plants

Abstract A fungus of the genus Phytophthora , frequently isolated from diseased spinach roots and also from field-grown wheat plants in an area in the south of Sweden, was identified as P. cryptogea on the basis of morphology, growth characters and cardinal temperatures. Mycelium or zoospores applied as inoculum in a series of pathogenicity tests induced symptoms in spinach, sugarbeet, wheat, cucumber, oil-seed rape, pea and oats. These ranged from death of all (spinach) or some inoculated plants (sugarbeet and wheat), to only slight root symptoms (oats). Successful re-isolations from all plants tested, confirmed infectivity in all cases. This is the first report of the occurrence of P. cryptogea in Sweden and, as far as we know, of pathogenicity of this fungus to wheat, oil-seed rape and oats.     

Colonization Pattern of the Biocontrol Strain Pseudomonas chlororaphis MA 342 on Barley Seeds Visualized by Using Green Fluorescent Protein

Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.     

Agronomic Performance of Partly Clubroot-resistant Spring Oilseed Turnip Rape Lines

In field experiment studies that tested the effects of three partly resistant cultivars of spring oilseed turnip rape ( Brassica campestris ), multiplication of clubroot was moderate. When initial soil inoculum levels were 47–72% of infected test plants in the bioassay, only 2.1–10% of the plants were infected after harvest. The yield of partly resistant lines was 5–10% higher compared with that of the susceptible cultivar SW Kulta, which on average yielded 1.73 t/ha. The average disease severity index (DSI) after harvest in these trials was 12.7 for the nonresistant cultivar SW Kulta and ranged between 2.2 and 6.2 for the partly resistant lines. The study demonstrated that in fields where the soil infestation level gives a DSI of less than 10, or when less than 20% of the bait plants are infected in the bioassay, it is possible to avoid the risk of severe yield losses from clubroot infections by integrating a partly resistant cultivar into the crop rotation.     

AXanthomonas maltophilia isolate tolerating up to 1% sodium azide in Tris/HCl buffer

A bacterium tolerating up to 1% NaN₃ found as a contaminant of Sephadex colums being run with Tris/HCl buffer, was identified asXanthomonas maltophilia. It had low nutrient requirements, was strongly proteolytic and interfered with Sephadex columns run with Tris/HCl buffers.The authors are with the Department of Plant Pathology, Swedish University of Agricultural Sciences, Box 7044, S-750 07 Uppsala, Sweden;     

Colonization pattern of the biocontrol strain Pseudomonas chlororaphis MA 342 on barley seeds visualized by using green fluorescent protein.

Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.