|SELECTIVE ELIMINATION OF CHLOROPLASTIDIAL DNA FOR METAGENOMICS OF BACTERIA ASSOCIATED WITH THE GREEN ALGA CAULERPA TAXIFOLIA (BRYOPSIDOPHYCEAE)(1).
|Aires, T, Marba, N, Serrão, EA, Duarte, CM, Arnaud-Haond, S
|Year of Publication
Molecular analyses of bacteria associated with photosynthetic organisms are often confounded by coamplification of the chloroplastidial 16S rDNA with the targeted bacterial 16S rDNA. This major problem has hampered progress in the characterization of bacterial communities associated to photosynthetic organisms and has limited the full realization of the potential offered by the last generation of metagenomics approaches. A simple and inexpensive method is presented, based on ethanol and bleach treatments prior to extraction, to efficiently discard a great part of chloroplastidial DNA without affecting the characterization of bacterial communities through pyrosequencing. Its effectiveness for the description of bacterial lineages associated to the green alga Caulerpa taxifolia (M. Vahl) C. Agardh was much higher than that of the preexisting enrichment protocols proposed for plants. Furthermore, this new technique requires a very small amount of biological material compared to the other current protocols, making it more realistic for systematic use in ecological and phylogenetic studies and opening promising prospects for metagenomics of green algae, as shown by our data.