Use of bacteria, proteins or peptides derived from clones of Bacillus subtilis, deposited at american type culture collection, as stimulators of entire natural plant, against pathogen of fungal, bacterial and/or viral type cultures

摘要

Use of bacteria, proteins or peptides derived from: clones of Bacillus subtilis, deposited at ATCC (american type culture collection) under number of 55614, and 6633; clones of Bacillus cereus, deposited at ATCC under number of 21928, 55609, 21929, 53522 and 55609; and clones of Bacillus pumilus, deposited at ATCC under numbers of 31132 and 31340, and at northern regional research laboratory collection under number of B-30087, as stimulators of entire natural plant, against pathogen of fungal, bacterial and/or viral type cultures, is claimed. An independent claim is included for a process for obtaining a composition containing bacteria, proteins or peptides, comprising: culturing the bacteria using a culture medium, which is relatively simple (MS), where the composition comprises potassium dihydrogen phosphate (2.5 g/l), dipotassium dihydrogen phosphate (2.5 g/l), diammonium hydrogen phosphate ((NH 4) 2HPO 4) (1 g/l), magnesium sulfate heptahydrate (0.2 g/l), manganese sulfate heptahydrate (7 mg/l), ferrous sulfate heptahydrate (0.01 g/l), and calcium chloride (10 mg/l), adding standard saline solution of 1% (p/v) D-glucose and adjusting the pH 7, sterilizing the medium by autoclaving at 120[deg] C for 20 minutes, inoculating a preculture of bacteria in MS medium for 15 hours at 30[deg] C under agitation, and placing a flask containing the bacteria in culture at 37[deg] C under gentle rotative agitation (150 rpm) for 5-7 days; and (b) separating bacteria and proteins, performed at 4[deg] C comprising centrifuging the contents of each flask at 4500 rpm for 30 minutes, washing the pellet in 1M sodium chloride solution, twice with distilled water containing 1 mM phenylmethylsulfonyl fluoride and 10 mM EDTA and then resuspended in a volume of demineralized water and purifying the protein part of a discontinuous sucrose gradient by method of Thomas and Ellar (1983) [ultracentrifugation at 25000 rpm and 48[deg] C for 16 hours], concentrating into protein, which is determined by the Bradford method after alkaline solubilization of the extract and electrophoresis (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) denaturing conditions for detecting proteins using Thomas and Ellar (1983) method. ACTIVITY : Fungicide; Virucide; Antibacterial. MECHANISM OF ACTION : None given.