CAPÍTULO 2: Low-coverage genome sequencing to identify 45S rDNA in Passiflora L. and
5. Conclusions
Amplification of 26S rDNA via PCR and FISH chromosomal mapping was successful in Passiflora and in other plant species. The amplification of the probe by PCR makes this methodology more advantageous when compared to the maintenance of the probe in vectors, which makes this technique expensive, laborious and passive of cumulative errors in function of the several steps of bacterial replication, extraction, quantification and marking of
57
plasmids. The probe presented in this work will be useful for studies on karyotype variation, cytotaxonomy and evolution in Passiflora.
Information on the composition of the GC content of the Passiflora genome corroborates the relationship between CMA3+/DAPI- bands and 45S rDNA sites observed in different species, and shows that ribosomal genes themselves are rich in GC. The presence of CMA3
bands related to 45S rDNA sites shows the usefulness of this fluorochrome in the identification of these sites in species of this genus.
Supplementary Materials: All datasets supporting the conclusions of this article are included within the article.
Acknowledgments: Thanks to UESC, CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) and FAPESB (Fundação de Amparo à Pesquisa do Estado da Bahia) and CNPq for the financial support for research. FAPESB for the scholarships granted to the first author. Centro de Biologia Computacional e Gestão de informações Biotecnológicas (NBCGIB) by bioinformatic support.
Author Contributions: Experiments conception and design: Gonçalo S. Silva, Margarete M.
Souza. Sequencing data: Gonçalo S. Silva, Fabienne Micheli, Cláusio A. F. de Melo, Vanessa de C. C. Pamponet. RepeatExplorer analysis: Gonçalo S. Silva, Vanessa de C. C. Pamponet, Sarah G. de Oliveira. Cytogenetic experiments: Gonçalo S. Silva, Cláusio A. F. de Melo.
Paper writing: Gonçalo S. Silva, Margarete M. Souza.
Conflicts of Interest: The authors declare no conflict of interest.
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CAPÍTULO 3: Caracterização citogenômica do DNA repetitivo e mapeamento