Symbiotic nitrogen-fixing bacterial populations trapped from soils under agroforestry systems in the Western Amazon

Cowpea (Vigna unguiculata) is an important grain-producing legume that can forego nitrogen fertilization by establishing an efficient symbiosis with nitrogen-fixing bacteria. Although inoculating strains have already been selected for this species, little is known about the genotypic and symbiotic diversity of native rhizobia. Recently, Bradyrhizobium has been shown to be the genus most frequently trapped by cowpea in agricultural soils of the Amazon region. We investigated the genetic and symbiotic diversity of 148 bacterial strains with different phenotypic and cultural properties isolated from the nodules of the trap species cowpea, which was inoculated with samples from soils under agroforestry systems from the western Amazon. Sixty non-nodulating strains indicated a high frequency of endophytic strains in the nodules. The 88 authenticated strains had varying symbiotic efficiency. The SPAD (Soil Plant Analysis Development) index (indirect measurement of chlorophyll content) was more efficient at evaluating the contribution of symbiotic N2-fixation than shoot dry matter under axenic conditions. Cowpea-nodulating bacteria exhibited a high level of genetic diversity, with 68 genotypes identified by BOX-PCR. Sequencing of the 16S rRNA gene showed a predominance of the genus Bradyrhizobium, which accounted for 70 % of all strains sequenced. Other genera identified were Rhizobium, Ochrobactrum, Paenibacillus, Bosea, Bacillus, Enterobacter, and Stenotrophomonas. These results support the promiscuity of cowpea and demonstrate the high genetic and symbiotic diversity of rhizobia in soils under agroforestry systems, with some strains exhibiting potential for use as inoculants. The predominance of Bradyrhizobium in land uses with different plant communities and soil characteristics reflects the adaptation of this genus to the Amazon region

Diversity and Efficiency of Rhizobia Communities from Iron Mining Areas Using Cowpea as a Trap Plant

<div><p>ABSTRACT Mining is an important economic activity. However, its impact on environment must be accessed, mainly on relevant processes for their sustainability. The objective of this study was to evaluate the diversity and efficiency of symbiotic nitrogen fixing bacterial communities in soils under different types of vegetation in the Quadrilátero Ferrífero: ironstone outcrops, Atlantic Forest, neotropical savanna, and a rehabilitated area revegetated with grass. Suspensions of soil samples collected under each type of vegetation were made in a saline solution to capture rhizobia communities that were then inoculated on cowpea [Vigna unguiculata (L.) Walp.], which was used as a trap plant. The symbiotic efficiency of the communities was evaluated in a greenhouse experiment and the data obtained were correlated to the chemical and physical properties of the soils under each type of vegetation. At the end of the experiment, the bacteria present in the nodules were isolated to evaluate their diversity. The highest numbers of nodules occurred in the treatment inoculated with soil samples from rehabilitated area revegetated with grass and neotropical savanna vegetation, and the lowest numbers were observed in the treatment inoculated with soil samples from ironstone outcrops and Atlantic Forest. In relation to root dry matter, the treatment inoculated with soil samples from Neotropical savanah was superior to those inoculated with soil samples from the other areas; already, in relation to the shoot dry matter, no significant difference among the treatments was observed. The soil properties with the greatest influence on the microbial communities were Al3+ content, considered as high in the Atlantic Forest and neotropical savanna vegetation, as intermediate in the iron outcrops, and as very low in the rehabilitated area revegetated with grass; organic matter, considered as very high in the ironstone outcrops and neotropical savanna, as high in the Atlantic Forest, and as low in the rehabilitated area revegetated with grass; and the pH, with intermediate acidity level in the rehabilitated area revegetated with grass, high level of acidity in the iron outcrops and neotropical savanna, and very high acidity in the Atlantic Forest. After isolation of the nodules, 380 bacterial strains were obtained and separated into 27 groups by cultural characterization analysis. Genetic diversity was evaluated by the 16S rRNA gene partial sequencing of 156 strains, which identified some bacteria belonging to nitrogen-fixing Leguminosae nodulating bacterial genera (Rhizobium, Bradyrhizobium, Burkholderia, and Cupriavidus), some representative of associative bacteria (Bacillus, Paenibacillus, Herbaspirillum, Pseudomonas, and Agrobacterium), and other genera (Brevibacillus, Novosphingobium, Chitinophaga, Dyella, Acinetobacter, and Stenotrophomonas). The highest genetic diversity of bacteria was found in the rehabilitated area revegetated with grass indicated that it was effective in soil rehabilitation</p></div

Symbiotic Efficiency and Genotypic Characterization of Variants of Bradyrhizobium spp. in Commercial Inoculants for Soybeans

<div><p>ABSTRACT High yields obtained from soybean [Glycine max (L.) Merrill], and consequent profits, are related to efficient symbiosis of soybean with nitrogen fixing bacteria of the Bradyrhizobium genus. Inoculation with strains belonging to the Bradyrhizobium japonicum, B. elkanii, and B. diazofficiens species, approved by the Brazilian Ministry of Agriculture (MAPA), is a common practice for this crop. However, variations in the composition of inoculants can change the quality of the product. The aim of this study was to genetically identify and to evaluate the symbiotic efficiency of isolates with cultural characteristics different from those of strains approved by MAPA and which are present in two commercial inoculants for soybean. Isolates obtained from inoculants and the strains SEMIA 5079 (CPAC 15), SEMIA 5080 (CPAC 7), SEMIA 587 (BR 96), and SEMIA 5019 (BR 29), indicated in the formulation of these inoculants, were tested for symbiotic efficiency in soybeans (cultivar: COODETEC 5G 830 RR) in a greenhouse. Identification by partial sequencing of the 16S rRNA gene, together with BOX-PCR analysis, showed that variant isolates were not similar to the inoculant strains indicated in the commercial products, despite belonging to the same genus and/or species. No loss was observed in the nodulation ability of the isolates; however, significant differences were reported in the symbiotic efficiency of the isolates compared to the inoculant strains approved by MAPA. Nevertheless, the presence of morphological variants did not influence the efficiency of inoculant products.</p></div