Cyanobacterial Biodiversity T.Bhuvaneshwari, Research Scholar, NFMC.

Cyanobacteria – An Introduction Cyanobacteria - morphologically distinct group of Oxygenic Photosynthetic organisms Gram negative prokaryote - low state of cell organization The cell lacks well defined nucleus and the DNA floats in the protoplast Cyanobacteria exist in all known photic habitats and can thrive in extreme levels of humidity, light, salinity, temperature, availability of oxygen or cabon di oxide and solar radiation

Cyanobacterial Taxonomy Botanical Approach Morphological and cytological (Bornet and Flahaut, Gomont, Geitler and desikachary) Bacteriological Approach Polyphasic Approach- physiological,Cytological, and biochemical (Stanier, waterberry and Ripka) Molecular Approach Polyphasic Approach - phenotypic and Genotypic characters (Anagnostidis and Komarek ) Identification at the Genetic level

Project Objectives Upgradation and maintanence of marine cyanobacteria in the Repository Axenization of the Marine cyanobacteria Identification of the strains by adapting morphological and Molecular Methods Revalidation of the strains for their identify, lipid and fatty acid content Supply of cultures to the needy researchers Bioprospecting of marine cyanobacteria for Biofuel production  Screening of cultures for lipid and fatty acid profile  Optimization of physico chemical parameters for high lipid production.

Aim of MY Study To maintain the Repository of Marine cyanobacteria To upgradate the strength of the Repository by isolating new strains To identify the strains Morphologically To Characterise the isolated cyanobacteria using Molecular approach such as  16SRNA sequencing  ITS (internal transcribed spacer region)  cpc region

Repository Maintanence Marine cyanobacterial strains of the Repository were maintained in both Solid and Liquid culture. Liquid cultures were subcultured regularly at the interval of 15 days and incubated at 25±2ºC under continuous low light illumination. For long Preservation, the cultures were also maintained in Agar Slants

Isolation of marine Cyanobacterium Samples are taken from different environments covering Rocky shores, Sandy shores, Saltpans, Backwater and Estuarine areas of Rameshwaram, Krusadai, Cuddalore, Vizhingam, Jalgoan, Tuticorn and Andaman They are processed immediately and pure cyanobacteria were isolated using Standard Microbiological Protocols

Isolate NoOrganismSample AreaSample No 1Chroococcus speciesVizhingam03 2Gleocapsa crepidinumGujarat12 3Myxosarcina burmensisVizhingam01 4Aphanothece saxicolaGujarat08 5Aphanothece saxicolaGujarat09 6Oscillatoria probosideaeTuticorn153 7Oscillatoria speciesTuticorn34 8Oscillatoria speciesCuddalore16 9Phormidium speciesRameshwaram05 10Phormidium speciesKrusadai01 11Phormidium tenueKrusadai11 12Nostoc calcicolaGujarat06 Further 8 pure isolated strains yet to be identified isolated strains so far

Chroococcus species

Gleocapsa crepidinum

Myxosarcina burmensis

Aphanothece saxicola

Oscillatoria probosideae

Oscillatoria species

Phormidium tenue

Phormidium species

Nostoc calcicola

Molecular Approaches

Molecular work carried out so far DNA were isolated for ten organism by Xanthogenate method Among them, Pcr was carried out for four samples using the Primer 16s-1F and 16s 740 R for amplifying the regions of 16s rDNA It was then Send for sequencing the results obtained were not satisfactory as the chromatogram was not good and also the sequence matches with certain bacterial 16s rDNA sequence when performed blast analysis with the available nucleotide sequence of NCBI.

Other markers to be studied Mcy (Microcystin synthesizing gene) Gyr B (Gyrase B) rpo C (RNA Polymerase B) rpo D (RNA Polymerase D)

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