Of 24 Trichoderma isolates, T harzianum Rifai (T24) showed a potential for control of the phytopathogenic basidiomycete Sclerotium rolfsii. When T24 was grown on different carbon sources, growth inhibition of S. rolfsii by the T24 culture filtrate correlated with the activity of extracellular chitinase and beta-1,3-glucanase. The 43-kilodalton (kDa) chitinase and the 74-kDa beta-1,3-glucanase were purified from the T24 culture filtrate in two and three steps, respectively, using ammonium sulphate precipitation followed by hydrophobic interaction chromatography (phenyl-Sepharose) and gel filtration (beta-1,3-glucanase). Km and Kcat were 3.8 g l(-1) and 0.71 s(-1) for the chitinase (chitin) and 1.1 g(-1) and 52 s(-1) for the beta-1,3-glucanase (laminarin). The chitinase showed higher activity on chitin than on less-acetylated substrate analogues (chitosan), while the beta-1,3-glucanase was specific for beta-1,3-linkages in polysaccharides. Both enzymes were stable at 30 degrees C, while at 60 degrees C the chitinase and the beta-1,3-glucanase were rapidly inactivated, showing half-lives of 15 and 20 min, respectively. The enzymes inhibited growth of S. rolfsii in an additive manner showing a promising ED50 (50% effective dose) value of 2.7 microg/ml.
Egypt2 Inoculation of wheat and corn grains with formulations of Azospirillum brasilense significantly increased seedling growth parameters of wheat and corn compared to untreated controls. Inoculation with Azospirillum and supplemental Trichoderma harzianum free or coimmobilized in calcium alginate resulted in significant increase in all plant growth parameters in addition to improving plant nutrient-content [phosphorus (P), potassium (K), and calcium (Ca)]. Grain treatments with T. harzianum alone or in a combination with A. brasilense were protected from invasion by Fusarium in a pot experiment. Nitrogen (N) fixation was investigated by A. brasilense free or double inoculated with T. harzianum in soil amended with different C-sources; also, phosphate solubilization was tested by these two organisms. Single and double inoculation with A. brasilense and/or T. harzianum improved wheat yield growth parameters in addition to seed protein; therefore, immobilized and coimmobilized formulations could be used as biofertilizer and biopesticide, and might be recommended to avoid the extensive use of the agrochemicals.
The antagonistic actions of Trichoderma and Gliocladium species against two phytopathogenic species of Fusarium (a serious disease in cotton production) were studied in vitro. The antagonist induced a substantial lysis of mycelium of the target fungi which became malformed and thickened. Bulging or swollen hyphal strands occurred and sometimes with clamydospores production. Antagonists produced Downloaded by [Temple University Libraries] at 21:28 17 November 2014 chitinase and β-1,3-glucanase in liquid culture contained cell wall of Fusarium spp. as a sole carbon source. Crude enzyme preparations of the antagonists with lytic enzyme activities greatly reduced the germinated conidia of Fusarium spp. Moreover, aberrant morphology of conidia was observed with germ tube lysis. On the other hand, the resulted mycelia showed wilt mycelium, coagulated protoplasm, heavy vacuolization and swelling followed by the complete destruction of the hyphae.
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