Abstract
ABSTRACT Tree genomes have been sequenced in recent years providing a source of basic information on multigenic family characterization. Comparative genomics based on those complete genome sequences available in public database is an important tool providing useful information to progress on functional gene characterization. In this work, we focus on gene encoding for Thioredoxins (Trxs) in Eucalyptus grandis genome, which are oxidoreductase enzymes, involved in significant biochemical processes, above all the maintenance of cellular homeostasis. Here we investigate the diversity, structure and expression of these genes in eucalyptus. For this purpose, bioinformatics tools were employed, using public platforms data, to identify coding sequences and validate gene expression. Specific softwares were employed to characterize gene structure and expression. RT-PCR assays were carried out to specifically verify the expression of 4 cytoplasmic thioredoxin genes, observed in silico from leaf, phloem, xylem and apical meristem tissues. Twenty-two Trxs with characteristic and canonic active sites were identified, confirming the presence of all types of the three main groups already defined as plastidial (m, f, x, y, z) cytoplasmatic (h) and mitochondrial (o). However, differences in the number of genes per group were observed when compared with other tree genomes. The expression of these thioredoxin genes compared to some homologous genes presented divergent expression patterns compared to Arabidopis thaliana suggesting a functional specificity in eucalyptus, such as in the case of Eucgr.F01604 gene encoding an h1 cytoplasmic Trx, which presents a strong expression in conductor tissues.
Highlights
Thioredoxins (Trxs) are small ubiquitous enzymes – approximately 14kDa - present in all organisms, and responsible for maintaining the redox state in cells (Geigenberger et al, 2017)
We focus on gene encoding for Thioredoxins (Trxs) in Eucalyptus grandis genome, which are oxidoreductase enzymes, involved in significant biochemical processes, above all the maintenance of cellular homeostasis
The expression of these thioredoxin genes compared to some homologous genes presented divergent expression patterns compared to Arabidopis thaliana suggesting a functional specificity in eucalyptus, such as in the case of Eucgr.F01604 gene encoding an h1 cytoplasmic Trx, which presents a strong expression in conductor tissues
Summary
Thioredoxins (Trxs) are small ubiquitous enzymes – approximately 14kDa - present in all organisms, and responsible for maintaining the redox state in cells (Geigenberger et al, 2017). Thioredoxins present a surprising complexity compared to animals in terms of number of genes per genome, and functional diversity They were identified as responsible for the activation of two important Calvin-Benson cycle enzymes, FBPase-Fructose 1,6-bisphosphatase - Malate dehydrogenase - MDH (Jacquot and Buchanan, 1981). The complexity and abundance of Trxs in plants was confirmed by the Arabidopsis thaliana genome-sequencing project (Arabidopsis Genome Initiative, 2000) They were classified according to their cell compartimentalization, as chloroplastic, cytoplasmic (Reichheld et al, 2002), mitochondrial (Laloi et al, 2001), and, more recently, nuclear (Reichheld et al, 2007). Rivera-Madrid et al (1995) clearly characterized cytoplasmic h Trxs (heterotrophic)
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