Paulo Ivan Fernandes-Júnior

Cowpea Nodules Harbor Non-rhizobial Bacterial Communities that Are Shaped by Soil Type Rather than Plant Genotype

Many studies have been pointing to a high diversity of bacteria associated to legume root nodules. Even though most of these bacteria do not form nodules with legumes themselves, it was shown that they might enter infection threads when co-inoculated with rhizobial strains. The aim of this work was to describe the diversity of bacterial communities associated with cowpea (Vigna unguiculata L. Walp) root nodules using 16S rRNA gene amplicon sequencing, regarding the factors plant genotype and soil type. As expected, Bradyrhizobium was the most abundant genus of the detected genera. Furthermore, we found a high bacterial diversity associated to cowpea nodules; OTUs related to the genera Enterobacter, Chryseobacterium, Sphingobacterium, and unclassified Enterobacteriacea were the most abundant. The presence of these groups was significantly influenced by the soil type and, to a lesser extent, plant genotype. Interestingly, OTUs assigned to Chryseobacterium were highly abundant, particularly in samples obtained from an Ultisol soil. We confirmed their presence in root nodules and assessed their diversity using a target isolation approach. Though their functional role still needs to be addressed, we postulate that Chryseobacterium strains might help cowpea plant to cope with salt stress in semi-arid regions.

Diversity and symbiotic performance of peanut rhizobia from Southeast region of Brazil

3Embrapa Semiarido, BR 428, km 152, Zona Rural, CEP 56302-970 Petrolina, PE. Brazil. 4 Embrapa Agrobiologia, BR 465, Km 07, Seropedica, RJ, CEP 23890-000. Brazil. Accepted 13 January, 2014 The use of peanut as a trap-plant for obtaining rhizobia may result in a high diversity of efficient isolates. This study aimed to evaluate the diversity and symbiotic efficiency of peanut rhizobia from the Southeastern region of Brazil. The bacteria were isolated from nodules of two genotypes in soils from the States of Sao Paulo and Rio de Janeiro, Brazil. The bacteria were authenticated and evaluated regarding to their symbiotic capacity. The rhizobial diversity was evaluated through their culture characteristics on yeast mannitol agar (YMA) culture media and by polymerase chain reaction Box(PCR). Two selected isolates were characterized phenotypically regarding their capacity to solubilize phosphate, to grow on media with different concentrations of NaCI, pHs and incubation temperatures. For these the 16S rRNA gene sequencing and symbiotic efficiency using non-autoclaved soils were conducted. The majority of the obtained isolates showed rapid growth and acidified the culture medium. Analysis of the isolates through the Box-PCR revealed low similarity to the reference strain indicated for the culture and a large genetic variability of the obtained isolates. The isolates AM 01 and AM 07, which clustered with Bradyrhizobium and Rhizobium genus, respectively, showed a potential for evaluations and a means to recommend new rhizobia strains for peanut.

Symbiotic and agronomic efficiency of new cowpea rhizobia from Brazilian Semi-Arid

Cowpea is a very important crop to Brazilian Semi-Arid mainly small family-based farmers. Rhizobia inoculation is a practice, easy to use, and cheap technology that increases cowpea productivity. The aim of this study was to evaluate the efficiency of two new rhizobia isolates in greenhouse and field as well as classify them taxonomically. To bacterial identification the 16S rRNA gene of ESA 17 and ESA 18 isolates were sequenced. The greenhouse test was conducted with pots containing 3 L of soil and the bacterial isolates evaluated were ESA 17, ESA 18,BR 3267 or BR 3262 strains. A field experiment was implemented on a Vertisol in Juazeiro, Bahia State, to evaluate the cowpea growth and productivity. In this experiment, the peat-based inoculants with ESA 17, ESA 18, BR 3267 or UFLA 3-84 were used in 2 cowpea cultivars. Both bacteria were identified as Bradyrhizobium, but related to different species. ESA 17 was related to B. japonicumand ESA 18 was closer to B. pachyrhizi. At greenhouse, both isolates increased cowpea nitrogen content in the shoots due to the presence of very efficient nodules. In the field, the isolate ESA 18 inoculated at BRS Pujante cultivar induced higher production than observed for the absolute control, and for BR 17 Gurgueia cultivar, he ESA 17 and BR 3267 stood out both by inducing high production and grain protein content. The results indicate that both isolates can be evaluated in network experiments aiming at official recommendation for new bacteria to cowpea inoculant in Brazil.

Phenotypic, genetic and symbiotic characterization of Erythrina velutina rhizobia from Caatinga dry forest.

Erythrina velutina (“mulungu”) is a legume tree from Caatinga that associates with rhizobia but the diversity and symbiotic ability of “mulungu” rhizobia are poorly understood. The aim of this study was to characterize “mulungu” rhizobia from Caatinga. Bacteria were obteined from Serra Talhada and Caruaru in Caatinga under natural regeneration. The bacteria were evaluated to the amplification of nifH and nodC and to metabolic characteristics. Ten selected bacteria identified by 16S rRNA sequences. They were tested in vitro to NaCl and temperature tolerance, auxin production and calcium phosphate solubilization. The symbiotic ability were assessed in an greenhouse experiment. A total of 32 bacteria were obtained and 17 amplified both symbiotic genes. The bacteria showed a high variable metabolic profile. Bradyrhizobium (6), Rhizobium (3) and Paraburkholderia (1) were identified, differing from their geographic origin. The isolates grew up to 45 °C to 0.51 mol L−1 of NaCl. Bacteria which produced more auxin in the medium with l-tryptophan and two Rhizobium and one Bradyrhizobium were phosphate solubilizers. All bacteria nodulated and ESA 90 (Rhizobium sp.) plus ESA 96 (Paraburkholderia sp.) were more efficient symbiotically. Diverse and efficient rhizobia inhabit the soils of Caatinga dry forests, with the bacterial differentiation by the sampling sites.