Forty two strains of Trichoderma sp. were isolated from cultivated lands around Bangalore and analyzed for their antagonistic potential against Sclerotium rolfsii and Fusarium ciceri. The potential of biocontrol agents ultimately lies in their capacity to control pathogens in vivo. Bioefficacy studies were hence conducted using chickpea (Cicer argentums c.v. Annigeri) as an experimental plant by the roll paper towel method. Overall the isolates T40, T35, T30 and T25 showed better antagonistic potential in addition to enhancing plant growth. The production of chitinases to break down the mycelial cell walls of fungal plant pathogens has been implicated as a major cause of biocontrol activity (Inbar and Chet, 1995). In order to study the mechanism of biocontrol, ten better performing strains were plated on media, amended with colloidal chitin and Sclerotium rolfsii cell wall extract. All the isolates showed chitinolytic activity on day three as well as day five. Production of endochitinase and exochitinase were assayed in liquid media using colloidal chitin amended broth. Strains T35 and T6 displayed maximum endochitinase and exochitinase activity. Although all strains exhibited cellulase activity, the quantum of enzyme produced was higher in T35 and T6. The results also indicate a positive correlation between enzyme production and bioefficacy.
An attempt was made to develop the best method to extract DNA from four selected varieties of the ornamental and medicinal plant, Hibiscus rosa-sinensis which can be distinguished one from the other morphologically. DNA extraction was carried out in several sets and each time there were more modifications that have been documented to bring out an efficient method to extract DNA from this plant of the Mavlacea family, which have a high content of gummy, mucilaginous substances. That apart, the DNA extracted was qualified and quantified based on the Beer-Lambert's law using UV-Visible Spectrophotometer. The results were not up to the mark of purity but further modifications can be made in this respect. Then a comprehensive protocol to isolate the DNA from Hibiscus sp was developed. The plant genomic DNA was extracted and also restriction digested by several combinations of restriction enzymes and conditions of incubation. The best one was found to completely digest the genomic DNA and this was the combination of three restriction enzymes Eco RI, Bam HI, Hind III at 37ºC for 15-18 hours. The dual success in developing a standardized condition for restriction digestion of genomic DNA from this species as well as the obtaining of completely digested plant genomic DNA can be further used in DNA Analysis of the related cultivars, and their RAPD analysis can be carried out which helps to identify and characterise the plants within this species as part of germplasm conservation.
This study sought to utilize indigenous soil micro-organisms to suppress wilt-causing fungal pathogens of the banana. Fungal pathogens were isolated from wilt-affected rhizospheric soil, and potential antagonistic bacterial strains were isolated from healthy rhizospheric soil in the same area from which fungal pathogens were isolated. The antifungal activity of isolated micro-organisms against fungal pathogens was studied both in vitro and in vivo against fungal pathogens. It was found that Fusarium oxysporum and Alternaria sp. were pathogenic, while Penicillium sp. Bacillus velezensis and Bacillus subtilis were antagonistic. Moreover, it was seen that B. velezensis, B. subtilis and Penicillium sp. Inhibited the growth of the two fungal pathogens in both in vitro and in vivo experiments. An antagonistic consortium isolated in this study has demonstrated remarkable potential for controlling fungal diseases caused by Fusarium sp. and Alternaria sp. Therefore, the use of indigenous microflora to improve disease suppression of banana plants against soil-borne pathogens is a preferable approach.
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