Thirty-one bacterial isolates out of 133 isolates, were obtained from rhizosphere of Egyptian clover plants, and had variant capability for starch degradation on starch agar medium. The isolate E109 was the most potent being 72.5 U ml À1 and 2.5 for amylase activity and starch hydrolysis ratio (SHR), respectively, at 50°C. The potent isolate E109 was identified based on phenotypic characteristics, phylogenetic positions based on 16S rRNA gene analysis and base sequences (submitted to NCBI Gen Bank). 16S rRNA gene analysis confirmed that this isolate belonged to the genus Bacillus and it was most closely related to B. amyloliquefaciens (95% similarity). For the production of amylases, nine agro-industrial residues were added as carbon sources to the basal medium. The medium supplemented with potato starchy waste as the sole carbon source enhanced the enzyme activity more than soluble starch as control for a, b and c amylases activity, as it increased by B. amyloliquefaciens about 1.26 & 4 and 8-fold, respectively after 48 h at 50°C using rotary shaker at 150 rpm. B. amyloliquefaciens gave the maximum values of a, b and c amylases activity on medium supplemented with 2% potato starchy waste after 30, 30 & 36 h of fermentation periods at 50°C using shake flasks technique as a batch culture. These values were 155.2 U ml À1 (R 2 = 0.93), 1.0 U ml À1 (R 2 = 0.94) and 2.4 U ml À1 (R 2 = 0.95), respectively. It could be stated that productive medium supplemented with 2% potato starchy waste as a low price substrate could be more favorable than basal medium containing 1% starch for amylases production in submerged fermentation, as it increased a, b and c amylase activity by 1.98, 7.69 and 12-fold than that produced in basal medium (control), respectively. ª 2015 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams University.
The efficacy of eight plant extracts (garlic, clove, garden quinine, Brazilian pepper, anthi mandhaari, black cumin, white cedar and neem) in controlling leaf rust disease of wheat was investigated in vitro and in vivo. In vitro, all treatments inhibited spore germination by more than 93%. Neem extract recorded 98.99% inhibition of spore germination with no significant difference from the fungicide Sumi-8 (100%). Under greenhouse conditions, seed soaking application in neem extract (at concentration of 2 ml/L) resulted in 36.82% reduction in the number of pustules/leaf compared with the untreated control. Foliar spraying of plant extracts on wheat seedlings decreased the number of pustules/leaf. Foliar spraying of plant extracts four days after inoculation led to the highest resistance response of wheat plants against leaf rust pathogen. Spray application of wheat seedlings with neem, clove and garden quinine extracts, four days after inoculation with leaf rust pathogen completely prevented rust development (100% disease control) and was comparable with the fungicide Sumi-8. Foliar spray application of wheat plants at mature stage with all plant extracts has significantly reduced the leaf rust infection (average coefficient of infection, ACI) compared with the untreated control and neem was the most effective treatment. This was reflected on grain yield components, whereas the 1000-kernel weight and the test weight were improved whether under one-or two-spray applications, with two-spray application being more effective in this regard. Thus, it could be concluded that plant extracts may be useful to control leaf rust disease in Egypt as a safe alternative option to chemical fungicides.
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