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Subject: Agricultural Biotechnology × Author: Amruta Gokul Alone × End date: 2017 × Start date: 2010 ×
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    ThesisItemOpen Access
    Functional and Molecular Characterization of Marine and Freshwater Microorganisms
    (UAS, Dharwad, 2012) Amruta Gokul Alone; Narayan Moger
    Marine environment which is an enormous pool of biodiversity resources covering 70% surface of earth harbors microorganisms that have unique properties which has attracted many researchers to harvest their potential. Twenty four marine and freshwater samples were collected from different regions of Karnataka and west coast of India, to isolate and characterize the microbial diversity. Out of 876 total isolates, 233 purified colonies including 101 bacteria, 60 actinomycetes and 72 fungi were randomly screened for functional diversity such as mineral phosphate solubilization, chitinolytic activity, salinity tolerance activity and antagonistic activity against five plant pathogens. Among forty positive phosphate solubilizers, four bacteria, five fungi and four actinomycetes isolates were found efficient to solubilize tricalcium phosphate. The screening of chitinolytic activity of microbial isolates showed complete hydrolysis of chitin by three bacteria, three fungi and one actinomycete than reference strains. The results of salinity tolerance by marine and freshwater microbes revealed highest salinity tolerance (20% NaCl) was only by marine bacteria and actinomycetes. The analysis of antagonistic activity of test bacteria, fungi and actinomycetes revealed poor antibacterial activity against Ralstonia solanacearum and Xanthomonas auxanopodis pv. punicae but showed strong inhibition against fungal pathogens such as Sclerotium rolfsii, Colletotricum capsici and Fusarium oxysporum. The strains possessing efficient functional activity were further selected for identification through cloning and sequencing of 16s rDNA and ITS region. The molecular diversity study among potent microbial isolates through rep-PCR fingerprinting technique was found to be more discriminatory at strain level than 16s rDNA and ITS region characterization. The cloning and sequencing analysis of ~850 bp of ectoine gene which is involved in cell homeostasis, PCR fragment showed no homology with available sequence of NCBI database. The BlastX results of cloned fragment showed 96% similarity with ATP dependent Clp protease ATP binding subunit of Halomonas boliviensis.