Austropuccinia psidii, the causal agent of myrtle rust, is a biotrophic pathogen whose growth and development depends on the host tissues. The uredospores of A. psidii infect Eucalyptus by engaging in close contact with the host surface and interacting with the leaf cuticle that provides important chemical and physical signals to trigger the infection process. In this study, the cuticular waxes of Eucalyptus spp. were analyzed to determine their composition or structure and correlation with susceptibility/resistance to A. psidii. Twenty-one Eucalyptus spp. in the field were classified as resistant or susceptible. The resistance/susceptibility level of six Eucalyptus spp. were validated in controlled conditions using qPCR, revealing that the pathogen can germinate on the eucalyptus surface of some species without multiplying in the host. CG-TOF-MS analysis detected 26 compounds in the Eucalyptus spp. cuticle and led to the discovery of the role of hexadecanoic acid in the susceptibility of Eucalyptus grandis and Eucalyptus phaeotricha to A. psidii. We characterized the epicuticular wax morphology of the six previously selected Eucalyptus spp. using scanning electron microscopy and observed different behavior in A. psidii germination during host infection. It was found a correlation of epicuticular morphology on the resistance to A. psidii. However, in this study, we provide the first report of considerable interspecific variation in Eucalyptus spp. on the susceptibility to A. psidii and its correlation with cuticular waxes chemical compounds that seem to play a synergistic role as a preformed defense mechanism.
Pasture degradation can cause changes in diazotrophic bacterial communities. Thus, this study aimed to evaluate the culturable and total diazotrophic bacterial community, associated with regions of the rhizosphere and roots of Brachiaria decumbens Stapf. pastures in different stages of degradation. Samples of roots and rhizospheric soil were collected from slightly, partially, and highly degraded pastures. McCrady's table was used to obtain the Most Probable Number (MPN) of bacteria per gram of sample, in order to determine population density and calculate the Shannon-Weaver diversity index. The diversity of total diazotrophic bacterial community was determined by the technique of Denaturing Gradient Gel Electrophoresis (DGGE) of the nif H gene, while the diversity of the culturable diazotrophic bacteria was determined by the Polymerase Chain Reaction (BOX-PCR) technique. The increase in the degradation stage of the B. decumbens Stapf. pasture did not reduce the population density of the cultivated diazotrophic bacterial community, suggesting that the degradation at any degree of severity was highly harmful to the bacteria. The structure of the total diazotrophic bacterial community associated with B. decumbens Stapf. was altered by the pasture degradation stage, suggesting a high adaptive capacity of the bacteria to altered environments.
Plant growth-promoting bacteria (PGPB) have received great interest in recent decades. However, PGPB mechanisms remain poorly understood in forage species. We aimed to evaluate roots endophytic and rhizospheric bacteria strains from Brachiaria humidicola and Brachiaria decumbens. The strains were evaluated for biological nitrogen-fi xing in saline stress (0 to 10.0 g L -1 of NaCl), N-acyl homoserine lactones and indole-like compounds (ILC) production, the activity of hydrolytic enzymes, and inorganic phosphate solubilization (IPS) under different C sources. The diversity of strains was assessed by BOX-PCR. About 58% of strains were positive for BNF. High salinity levels reduced the growth and BNF. About 58% produced N-acyl homoserine lactones. The ILC was present in 39% of strains. Cellulase, polygalacturonase, pectate lyase, and amylase production were observed in 77, 14, 22, and 25% of strains, respectively. The IPS was observed in 44, 81, and 87% of isolates when glucose, mannitol and sucrose were used, respectively. Comparing two plant species and niches, the strains associated with B. humidicola and root endophytic presented more PGPB mechanisms than others. We found high strain diversity, of which 64% showed similarity lower than 70%. These results can be supporting the bioproducts development to increase forage grasses production in tropical soils.
2018. Diazotrophic bacteria isolated from Brachiaria spp.: genetic and physiological diversity. Cien. Inv. Agr. 45(3): 277-289.Grass from the genus Brachiaria spp. predominates in pastures with low fertile soils. This scenario highlights the importance of the association with microorganisms to foster plant growth, which becomes essential to the successful establishment of this forage in such environments. This study aimed to evaluate the genetic variability and identify the mechanisms of plant growth promotion, in vitro, of bacteria associated with Brachiaria decumbens Stapf. and Brachiaria humidicola (Rendle.) Schweickerdt in Pernambuco, Brazil. We evaluated 20 isolates of diazotrophic bacteria obtained from the endophyte or rhizosphere communities. The genetic characteristics were determined via sequencing the 16S rRNA region, which allowed us to identify ten different bacterial genera: Bacillus sp., Burkholderia sp., Enterobacter sp., Klebsiella sp., Microbacterium sp., Pantoea sp., Ralstonia sp., Rhizobium sp., Sinomonas sp., and Sphingomonas sp., with a specificity of the genus Rhizobium sp. to Brachiaria decumbens Stapf.. The phenotypic and functional characteristics revealed that 100% of the bacterial strains produced indol-3-acetic acid (IAA) with the addition of L-tryptophan, and 60% presented IAA production independent of the L-tryptophan pathway. We also detected that 70% of the isolated bacteria possessed the capacity to solubilize phosphorus. The analysis of the enzymatic output revealed that 30% of the bacterial isolates produced cellulase, 60% produced pectate lyase, 15% produced polygalacturonase, and 30% produced amylase. We also detected the production of N-acyl homoserine lactones in 65% of bacterial strains. In summary, our results showed that plants of B. decumbens Stapf. and B. humidicola (Rendle.) Schweickerdt interacted with different bacterial genera capable of promoting plant growth.
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