Validação temporal e tissular de genes envolvidos no desenvolvimento de botão floral de algodoeiro

Abstract

A growing number of studies reports the identification and characterization of genes associated with flower bud and the isolation of its regulatory regions. These strands are helping to unravel a number of issues concerning the regulation and cellular interactions, thus generating a great impact on improving various plant species. In cotton (Gossypium hirsutum) these studies have contributed substantially in obtaining of genetically modified GM plants, holders of genes for resistance to herbicides and lepidopteran, and are becoming the possibility of increasing the control efficiency against target pests that act directly in floral structures and / or even enhance the expression of the coloring effect of the fibers. Different techniques are available for functional genomics analysis, among the most adopted is the construction of expressed sequence tags (ESTs) libraries that allows to identify which RNAs are expressed in an organism at a given time. Using this methodology in a previous study, were characterized by in silico analyzes genes involved in reproductive functions of cotton, some of whom expressed in male and female gametophytes. At that moment, from RT-PCR assays and qRT-PCR, six genes (GhASH, GhFIB010, GhGLUC, GhMYB, GhOVU and GhUDP) were investigated for the temporal expression during ontogeny flower bud, at stages 2-4 6-8, 10, 12, 14-16, and 18-20 mm. All of the genes investigated were active in floral buds of different sizes and in different expression levels, however, the highest level observed was 10 mm, except for gene GhASH with the highest expression in 10 to 16 mm. In these stages the cotton bud is in intense cellular activity including reduced cell wall formation of the complete embryonic sac, appearance and degeneration of the antipodes suggesting that these genes that may be involved in the formation and development or ovules, pollen, pollen tube, as well as on fibers. The gene was GhASH which also had the highest level of expression in all stages compared to other genes. Due to the regulation of these genes involved precursor steps, prior to anthesis and possibly runs through the final process of maturation, we proceeded further a study of tissue expression in organs flowering cotton (bracts, sepals, petals, ovules, anthers and fibers with 8, 10 and 18 DPA) to understand the interrelationships of the genes involved in the reproductive phase and its performance during development. It was found that all genes showed equivalent expression with its features, described in the databases. Analyses of gene activity for genes GhASH, GhOVU and GhGLUC proposed performances in training and / or development of all floral organs especially in pollen grains, pollen tube and anthers. For genes involved in fiber GhMYB, GhFIB010, GhUDP expression was observed for all studied growth stage, suggesting that are directly involved in elongation phase of the fibers, this phase has been widely investigated since the length of the cotton fiber is a key feature in determining the quality and yield. Additional studies to functional characterization of promising genes and isolation of their regions upstream become necessary in order to contribute to the progress and molecular breeding of this oilseed.