Análises in silico de genes associados à embriogênese somática em cana-de-açúcar
Resumo
Indirect somatic embryogenesis (ISE) is one of the most used regeneration pathways
to obtain sugarcane genetically modified plants. ISE regulatory genes have been
widely studied in monocotyledonous and dicotyledonous plants, and act as activators
of the expression of gene cascades or promoting cell growth and differentiation. In
contrast, in sugarcane, there are few reports in the literature with these genes involved
in ISE. Thus, the present study aimed to i) search for putative homologous genes
responsible for the regulation of ISE (families: BABYBOOM, WUSCHEL, LEAFY
COTYLEDON, VIVIPAROUS1, and AGAMOUS-LIKE) in sugarcane (SP80-3280 and
Saccharum spontaneum); ii) analyze the phylogeny between the sugarcane amino
acid sequences and their homologous in Arabidopsis thaliana and Poaceae species;
iii) design ideal primers for studying the expression of sugarcane ISE-related genes.
BLAST and BLASTp searches were carried out in the Phytozome, NCBI, UniProt, and
Gramene databases to obtain homologous sequences of the five gene families in corn,
sorghum, rice, and A. thaliana. Thus, phylogenetic analyzes of amino acid sequences
were performed with the aid of the software CLUSTALW and MEGAX, which showed
evolutionary proximity between the proteins of Zea mays, Sorghum bicolor, S.
spontaneum, and SP80-3280 in all the studied genes. In silico analyzes indicated that
they may have more than one gene from the BABYBOOM, WUSCHEL, and
VIVIPAROUS1 families regulating the pathways of ISE in sugarcane. In addition, with
the aid of the PRIMER3 software, 8 pairs of primers with ideal parameters were
obtained to be used in future analyzes of gene expression to understand more deeply
the mechanisms of ISE.
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