ABSTRACT
Xylella fastidiosa genome sequencing has generated valuable data by identifying genes acting either on metabolic pathways or in associated pathogenicity and virulence. Based on available information on these genes, new strategies for studying their expression patterns, such as microarray technology, were employed. A total of 2,600 primer pairs were synthesized and then used to generate fragments using the PCR technique. The arrays were hybridized against cDNAs labeled during reverse transcription reactions and which were obtained from bacteria grown under two different conditions (liquid XDM2 and liquid BCYE). All data were statistically analyzed to verify which genes were differentially expressed. In addition to exploring conditions for X. fastidiosa genome-wide transcriptome analysis, the present work observed the differential expression of several classes of genes (energy, protein, amino acid and nucleotide metabolism, transport, degradation of substances, toxins and hypothetical proteins, among others). The understanding of expressed genes in these two different media will be useful in comprehending the metabolic characteristics of X. fastidiosa, and in evaluating how important certain genes are for the functioning and survival of these bacteria in plants.
ABSTRACT
Assimilation of nitrate and ammonium are vital procedures for plant development and growth. From these primary paths of inorganic nitrogen assimilation, this metabolism integrates diverse paths for biosynthesis of macromolecules, such as amino acids and nucleotides, and the central intermediate metabolism, like carbon metabolism and photorespiration. This paper reports research performed in the CitEST (Citrus Expressed Sequence Tag) database for the main genes involved in nitrogen metabolism and those previously described in other organisms. The results show that a complete cluster of genes involved in the assimilation of nitrogen and the metabolisms of glutamine, glutamate, aspartate and asparagine can be found in the CitEST data. The main enzymes found were nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthetase (GOGAT), glutamate dehydrogenase (GDH), aspartate aminotransferase (AspAT) and asparagine synthetase (AS). The different enzymes involved in this metabolism have been shown to be highly conserved among the Citrus and Poncirus species. This work serves as a guide for future functional analysis of these enzymes in citrus.
ABSTRACT
The mating system and genetic diversity were studied in a natural population of Myracrodruon urundeuva originating from 30 open-pollinated trees at the Paulo de Faria Ecological Station, SP, Brazil. The progenies were planted on the Teaching and Research Farm of the Ilha Solteira Engineering School, UNESP. Using the fAFLP molecular marker, eleven loci were selected to study the mating system. The mating system was analyzed using the multilocus mixed-mating model. The estimates of genetic divergence between pollen and ovule allele frequencies were significant for eight loci, suggesting nonrandom outcrossing. The estimates of the multilocus outcrossing rate revealed that M. urundeuva possesses a mating system with a predominance of outcrossing events (theta = 0.940 ± 0.086). The estimates of coancestry among plants within progenies (theta = 0.185) was higher than that expected for half-sib progenies (0.125) and the indirect estimate of the correlation of outcrossed paternity within progeny arrays (r p) was 0.403, suggesting that progenies have a high proportion of full-sibs. Result analysis suggests the need for the application of biometric models that take into account deviations from random outcrossing in the estimations of genetic parameters for quantitative traits and the need for retaining large sample sizes in order to preserve genetic variability.