A positive selection method for isolation of nitrogenase-derepressed mutant strains of a filamentous cyanobacterium, Anabaena variabilis, is described. Mutant strains that are resistant to a glutamate analog, L-methionine-D,L-sulfoximine, were screened for their ability to produce and excrete NH4' into medium.Mutant strains capable of producing nitrogenase in the presence of NH4' were selected from a population of NH4+-excreting mutants. One of the mutant strains (SA-1) studied in detail was found to be a conditional glutamine auxotroph requiring glutamine for growth in media containing N2, N03 , or low concentrations of NH4' (less than 0.5 mM). This glutamine requirement is a consequence of a block in the assimilation of NH4' produced by an enzyme system like nitrogenase. Glutamate and aspartate failed to substitute for glutamine because of a defect in the transport and utilization of these amino acids. Strain SA-1 assimilated NH4' when the concentration in the medium reached about 0.5 mM, and under these conditions the growth rate was similar to that of the parent. Mutant strain SA-1 produced L-methionine-D,L-sulfoximine-resistant glutamine synthetase activity. Kinetic properties of the enzyme from the parent and mutant were similar. Mutant strain SA-1 can potentially serve as a source of fertilizer nitrogen to support growth of crop plants, since the NH4' produced by nitrogenase, utilizing sunlight and water as sources of energy and reductant, respectively, is excreted into the environment.In free-living, nitrogen-fixing organisms, nitrogenase synthesis and activity are regulated by the presence of NH4' in the medium (3,10,31,45). That NH4+-mediated regulation of nitrogenase synthesis is not exerted by NH4+ itself but is actually a consequence of metabolic products of NH4+ assimilation has been revealed by several lines of evidence.(i) Mutant strains incapable of NH4' assimilation were derepressed for nitrogenase synthesis in the presence of NH4+ (28,31,43,44). (ii) Addition of amino acids to the growth medium repressed nitrogenase synthesis even in nitrogenase-derepressed mutant strains (30). (iii) Inhibition of glutamine synthetase, a primary enzyme responsible for NH4+ assimilation, by a substrate (glutamate) analog, Lmethionine-D,L-sulfoximine (MSX) as well as by other inhibitors led to derepression of nitrogenase synthesis in the presence of NH4+ in all organisms tested (8,13,14,21,31,39). The above described derepression of nitrogenase synthesis is achieved by alteration of cellular physiology leading to a decrease in the rate of glutamate production, since addition of amino acids to the medium reversed the effect (22,29). This set of conditions also derepressed other NH4+-as well as glutamate-producing enzyme systems (NO3 assimilation; histidine and proline utilization) in the cell (16,29).Mutant strains that are blocked at the level of NH4+ assimilation were first described in Klebsiella pneumoniae (32), and such mutant strains not only derepress nitrogenase synthesis but also excrete the NH4+ produce...
Leaf rust, caused by Puccinia triticina Eriks., is a common and widespread disease of wheat (Triticum aestivum L.) in Egypt. Host resistance is the most economical, effective, and ecologically sustainable method of controlling the disease. Molecular markers help to determine leaf rust resistance genes (Lr genes). The objective of this study was to identify Lr genes in fifteen wheat cultivars from Egypt. Ten genes, Lr13, Lr19, Lr24, Lr26, Lr34, Lr35 Lr36, Lr37, Lr39, and Lr46, were detected in fifteen wheat cultivars using various molecular markers. The most frequently occurring genes in fifteen Egyptian wheat cultivars were Lr13, Lr24, Lr34, and Lr36 identified in all the cultivars used, followed by Lr26 and Lr35 (93%), Lr39 (66%), Lr37 (53%), and Lr46 (26.6%) of the cultivars, and finally Lr19 was present in 33.3% of cultivars. It is concluded that there was a good variation in Lr genes carried by wheat cultivars commercially grown in Egypt. Therefore, strategies for deploying resistance genes to prolong effective disease resistance are suggested to control wheat leaf rust disease.
The present study evaluated the efficacy of the fungicides Moncut and Rizolex-T and the biocide Bio-Control T34 (Trichoderma asperellum) on suppressing incidence of cotton seedling damping-off caused by Rhizoctonia solani on cotton cultivar Giza 86. The dual treatments of the two fungicides and the biocide also evaluated in vitro and in vivo. In vitro studies showed that Moncut completely inhibited R. solani linear growth when it was applied at a rate of 250ppm, while Rizolex-T completely inhibited R. solani growth when it was applied at a rate of 125ppm. The biocide inhibited the linear growth of R. solani by 59.17%. The biocide growth was not affected by all the applied concentrations of Moncut, while it was partially inhibited by the high concentrations of Rizolex-T. Under greenhouse conditions, the two chemical fungicides suppressed incidence of damping-off, while the biocide was unable to suppress the disease. The use of Moncut before or after the biocide succeeded in reducing the disease incidence. On the other hand, Rizolex-T caused significant reduction in disease incidence only when it was applied before the biocide. The application half or fourth the initial dose of the chemical fungicides before or after the biocide caused significant reduction in the disease incidence. The application of Moncut by the initial dose followed by the biocide caused increases in the activity of polyphenol oxidase (PPO) and phenols and sugars contents of the seedlings. Application half the initial dose of Moncut before or after the biocide caused increases in the previously mentioned variables. The use fourth the initial dose of Moncut caused increases in the same variables when it was followed by the biocide, while the use of this dose after the biocide caused decreases in the same variables.
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