Random amplified polymorphic DNA (RAPD) markers were used to analyze the genetic structure of Eufriesea violacea populations in three fragments (85.47, 832.58 and 2800 ha) of Atlantic rainforest located in the north of the Brazilian state of Paraná. A total of twelve primers produced 206 loci, of which 129 were polymorphic (95% criterion). The proportions of polymorphic loci in each population ranged from 57.28% to 59.2%, revealing very similar levels of genetic variability in the groups of bees from each fragment. Unbiased genetic distances between groups ranged from 0.0171 to 0.0284, the smallest genetic distance occurring between bees from the two larger fragments. These results suggest that the E. violacea populations from the three fragments have maintained themselves genetically similar to native populations of this species originally present in northern Paraná.

In this study, random amplified polymorphic DNA (RAPD) markers were applied to analyze the genetic diversity of samples of the Neotropical catfish Hypostomus ancistroides, collected from four sites (S1, S2, S3 and S4) along an urban stream in Southern Brazil. The 11 primers used in RAPD analysis amplified 147 loci, 76 (51.7%) of which were polymorphic. The proportions of polymorphic loci observed in the four samples were: 29.93% (S1), 31.97% (S2), 23.81% (S3) and 38.77% (S4). The average heterozygosity within sampling localities ranged from 0.1230 to 0.1526 and unbiased genetic distances ranged from 0.0253 to 0.0445. AMOVA partitioned 90.85% of the total variation within samples and 9.15% among samples. Excepting for the sample pair S1-S2 (φ ST = 0.02784; p > 0.05), all others pairwise φ ST values were significantly greater than zero, indicating moderate genetic differentiation among catfish samples from four localities. The relative low levels of genetic variation detected in all groups studied could be related to different factors, including the sedentary habit of these fish, which can be eroding the genetic variation of H. ancistroides from each locality.

BackgroundHerbaspirillum rubrisubalbicans was first identified as a bacterial plant pathogen, causing the mottled stripe disease in sugarcane. H. rubrisubalbicans can also associate with various plants of economic interest in a non pathogenic manner.ResultsA 21 kb DNA region of the H. rubrisubalbicans genome contains a cluster of 26 hrp/hrc genes encoding for the type three secretion system (T3SS) proteins. To investigate the contribution of T3SS to the plant-bacterial interaction process we generated mutant strains of H. rubrisubalbicans M1 carrying a Tn5 insertion in both the hrcN and hrpE genes. H. rubrisulbalbicans hrpE and hrcN mutant strains of the T3SS system failed to cause the mottled stripe disease in the sugarcane susceptible variety B-4362. These mutant strains also did not produce lesions on Vigna unguiculata leaves. Oryza sativa and Zea mays colonization experiments showed that mutations in hrpE and hrcN genes reduced the capacity of H. rubrisulbalbicans to colonize these plants, suggesting that hrpE and hrcN genes are involved in the endophytic colonization.ConclusionsOur results indicate that the T3SS of H. rubrisubalbicans is necessary for the development of the mottled stripe disease and endophytic colonization of rice.

Ocotea catharinensis is a rare tree species indigenous to the Atlantic rainforest of South America. In spite of its value as a hardwood species, it is in danger of extinction. The species erratically produces seeds showing irregular flowering and slow growth. Therefore, plants are not easily replaced. Tissue culture-based techniques are commonly used for obtaining living material for tree propagation and in vitro preservation. Therefore, a high-frequency somatic embryogenic system was developed for the species. In the present work, the genetic fidelity of cell aggregates and somatic embryos at various stages of in vitro development of O. catharinensis was investigated using RAPD and AFLP markers. Both analyses confirmed the absence of genetic variation in all developmental stages of O. catharinensis embryogenic cultures, verifying that the in vitro system is genetically stable. The cultures were also analyzed for their methylation profiles at 5′-CCGG-3′ sites by identifying methylation-sensitive amplification polymorphisms. Some of these markers differentiated cell aggregates from embryo bodies. The sequencing of ten MSAP markers revealed that four sequences showed significant similarity to genes encoding plant proteins. Particularly, the predicted amino acid sequence of the fragment designated as OcEaggHMttc155 was similar to the enzyme 1-aminocyclopropane-1-carboxylate oxidase (ACO), which is involved in the biosynthesis of ethylene, and its expression was reported to occur from the beginning to the intermediate stages of plant embryo development. Here, we suggest that this enzyme is possibly involved in the control of the earliest stages of somatic embryogenesis of O. catharinensis, and an approach to study ACO expression during somatic embryogenesis is proposed.