Hibridização in situ por fluorescência: análise histórica, inovações e perspectivas
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2022-09-05
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Universidade Federal de Goiás
Resumo
Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that is based on
the use of fluorescence-labeled probes to identify different regions in the genome. From the
improvement and development of new technologies, variations of the original technique
emerged making it possible to apply it to various issues, from identifying chromosomes
through descriptive cytogenetics to making evolutionary inferences or conducting applied
research in biotechnology and genetic improvement. Considering the great possibility of
applications of the FISH technique and its variations, the present work aims to detail the
methodological principles and applications, with emphasis on plants, and to investigate its use
in evolutionary studies. For this, the first chapter is a manuscript for the dissemination and
popularization of science that aims to discuss the various possibilities of using the FISH
technique, ranging from its methodological principles to applications in different areas. From
this study, it was verified that FISH can be applied to different biological materials and regions
of the genome and, despite being a technique developed for more than three decades, it can
generate impacts and contributions through its wide use until today. The second chapter is a
systematic review, whose main objective is to summarize and critically analyze the data
available in the literature on the applications of FISH for the cytogenetic study in plants, in
addition to detailing and discussing the applications of this technique for evolutionary studies
in plants. This study was conducted using the Preferred Reporting Items for Systematic
Reviews and Meta-Analyses (PRISMA) guidelines. The search was carried out in the Scopus
database and the keywords were directed to capture the breadth of the use of the FISH
technique aimed at the cytogenetic study of plants. In order to investigate the applications of
this technique, the articles found were grouped into different categories, called “Plant
Biotechnology”, “Evolutionary Studies”, “Descriptive”, “Techniques/Methodological” and “Gene
Expression”. A total of 4006 articles were selected that showed a trend of growth in the
number of publications between the years 1975 - 2020. China and the USA concentrated the
largest number of articles and Brazil was in 8th position. The analysis of the most used
keywords in the articles made it possible to trace a small history of the technique, ranging
from the beginning of indirect labeling in plant species, passing through the advances of FISH
and its variations, to the use of oligo-synthetic probes today. The categories “Plant
Biotechnology” and “Evolutionary Studies” represented more than 60% of the total works,
with emphasis on the use of GISH and FISH techniques with repetitive DNA probes and
indirect labeling. The Poaceae family stood out in the studies of "Plant Biotechnology" and
Leguminosae and Asteraceae were mostly found in the category of "Evolutionary Studies",
“Comparative analysis” and “Systems and cytotaxonomy.” Thus, the understanding of the
technique, the synthesis and measurement of data and the evolutionary path elaborated in
this study allowed the theoretical basis about the versatility of the technique and its use
today. demonstrate the main contributions of FISH directed to studies with plants,
highlighting the regions of research concentration, variations of the technique, main types of
studies and botanical families, as well as contributing to answering different evolutionary
questions.
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SILVA, T. F. Hibridização in situ por fluorescência: análise histórica, inovações e perspectivas. 2022. 84 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2022.