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 ApproachPolyphasic Approach- physiological,Cytological, and biochemical (Stanier, waterberry and Ripka)Molecular ApproachPolyphasic 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 CyanobacteriumSamples are taken from different environments covering Rocky shores , Sandy shores, Saltpans, Backwater and Estuarine areas of Rameshwaram, Krusadai, Cuddalore, Vizhingam, Jalgoan, Tuticorn and AndamanThey are processed immediately and pure cyanobacteria were isolated using Standard Microbiological Protocols

Isolate No Organism Sample Area Sample No

1 Chroococcus species Vizhingam 03

2 Gleocapsa crepidinum Gujarat 12

3 Myxosarcina burmensis Vizhingam 01

4 Aphanothece saxicola Gujarat 08

5 Aphanothece saxicola Gujarat 09

6 Oscillatoria probosideae Tuticorn 153

7 Oscillatoria species Tuticorn 34

8 Oscillatoria species Cuddalore 16

9 Phormidium species Rameshwaram 05

10 Phormidium species Krusadai 01

11 Phormidium tenue Krusadai 11

12 Nostoc calcicola Gujarat 06

Further 8 pure isolated strains yet to be identified

isolated strains so far

Chroococcus species

Gleocapsa crepidinum

Myxosarcina burmensis

Aphanothece saxicola

Aphanothece saxicola

Oscillatoria probosideae

Oscillatoria species

Oscillatoria species

Oscillatoria species

Phormidium tenue

Phormidium species

Phormidium species

Nostoc calcicola

Molecular Approaches

Molecular work carried out so farDNA were isolated for ten organism by Xanthogenate methodAmong them, Pcr was carried out for four samples using the Primer 16s-1F and 16s 740 R for amplifying the regions of 16s rDNAIt 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)

• Thank

ThaNK you