Abstract
A simple agar diffusion method is developed where pure colony of Chlamydomonas sp. CRP7 was isolated from Chlorella sp. CB4 mixtures by passing through agar migration with a light exposure of 6,000 lux for 7 h. The main concept behind it is that Chlamydomonas has flagella and the rhodopsin pigment is attracted towards light. Thus the above two microalgae species can be separated from the mixtures as eye spot serves as a navigator and flagella serves as a propeller for Chlamydomonas spp. Further the genomic DNA was isolated and purified from the above mentioned two species after the separation from the mixtures. PCR amplification was carried out for ITS1, 5.8S and ITS2 regions. The amplified products were sequenced and the sequence analysis confirmed that they belong to Chlamydomonas sp. and Chlorella sp. This is an important augmentation for isolation and separation of microalgae.
References
Stanier RY, Kunisawa R, Mandel M, Cohen-Bazire G (1971) Purification and properties of unicellular blue-green algae (Order Chroococcales). Bacteriol Rev 35:171–205
Mcdaniel HR, Miiddlebrook JB, Bowman RO (1961) Isolation of pure cultures of algae from contaminated cultures. Appl Microbiol 10:223
Bischoff HW, Bold HC (1963) Phycological studies. IV. Some soil algae from enchanted rock and related algal species. Univ Texas Publ No 6318:1–95
Nguyen TNM, Berzano M, Gualerzi CO, Spurio R (2011) Development of molecular tools for the detection of freshwater diatoms. J Microbiol Methods 84:33–40
Tolvanen, KES, Karp Matti T (2011) Molecular methods for characterizing mixed microbial communities in hydrogen-fermenting systems. Int J Hydrogen Energy 36:5280–5288
Boyer SL, Flechtner VR, Johansen JR (2001) Is the 16S–23S rRNA Internal Transcribed Spacer Region a good tool for use in molecular systematics and population genetics? A case study in Cyanobacteria. Mol Biol Evol 18:1057–1069
Rivas R, Velázquez E, Zurdo-Piñeiro JL, Mateos PF, Molina EM (2004) Identification of microorganisms by PCR amplification and sequencing of a universal amplified ribosomal region present in both prokaryotes and eukaryotes. J Microbiol Methods 56:413–426
Gorman DS and Levine RP (1965) Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardii. Proc Natl Acad Sci USA 54:1665–1669
Alvarez EV, Lago-Leston, A, Pearson GA, Serrao EA, Procaccini G, Duarte CM, Marba N (2006) Genomic DNA isolation from green and brown algae (Caulerpales and Fucales) for microsatellite library construction. J Phycol 42:741–745
Timmins M, Thomas-Hall SR, Darling A, Zhang E, Hankamer B, Marx UC, Schenk PM (2009) Phylogenetic and molecular analysis of hydrogen-producing green algae. J Exp Bot 60:1691–1702
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance and Maximum Parsimony Methods. Mol Biol Evol 28:2731–2739
Proschold T, Harris EH, Coleman AW (2005) Portrait of a species: Chlamydomonas reinhardtii.170:1601–1610
Acknowledgments
This work was done under the Innovative Project Scheme of BIT, Mesra (No. GO/Con Ord/2011-12/4563). Authors acknowledge the Department of Agriculture, Government of Jharkhand for providing infrastructure facilities. CB acknowledges the financial support as JRF from BIT, Mesra
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Banerjee, C., Bandopadhyay, R. & Shukla, P. A Simple Novel Agar Diffusion Method for Isolation of Indigenous Microalgae Chlamydomonas sp. CRP7 and Chlorella sp. CB4 from Operational Swampy Top Soil. Indian J Microbiol 52, 710–712 (2012). https://doi.org/10.1007/s12088-012-0295-6
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DOI: https://doi.org/10.1007/s12088-012-0295-6