Direct amplification of nodD from community DNA reveals the genetic diversity of Rhizobium leguminosarum in soil

Sequences of nodD, a gene found only in rhizobia, were amplified from total community DNA isolated from a pasture soil. The polymerase chain reaction (PCR) primers used, Y5 and Y6, match nodD from Rhizobium leguminosarum biovar trifolii, R. leguminosarum biovar viciae and Sinorhizobium meliloti. The PCR product was cloned and yielded 68 clones that were identified by restriction pattern as derived from biovar trifolii [11 restriction fragment length polymorphism (RFLP) types] and 15 clones identified as viciae (seven RFLP types). These identifications were confirmed by sequencing. There were no clones related to S. meliloti nodD. For comparison, 122 strains were isolated from nodules of white clover (Trifolium repens) growing at the field site, and 134 from nodules on trap plants of T. repens inoculated with the soil. The nodule isolates were of four nodD RFLP types, with 77% being of a single type. All four of these patterns were also found among the clones from soil DNA, and the same type was the most abundant, although it made up only 34% of the trifolii-like clones. We conclude that clover selects specific genotypes from the available soil population, and that R. leguminosarum biovar trifolii was approximately five times more abundant than biovar viciae in this pasture soil, whereas S. meliloti was rare

The rhizobial microbiome from the tropical savannah zones in Northern Côte d’Ivoire

Over the past decade, many projects have been initiated worldwide to decipher the composition and function of the soil microbiome, including the African Soil Microbiome (AfSM) project that aims at providing new insights into the presence and distribution of key groups of soil bacteria from across the African continent. In this national study, carried out under the auspices of the AfSM project, we assessed the taxonomy, diversity and distribution of rhizobial genera in soils from the tropical savannah zones in Northern Côte d’Ivoire. Genomic DNA extracted from seven sampled soils was analyzed by sequencing the V4-V5 variable region of the 16S rDNA using Illumina’s MiSeq platform. Subsequent bioinformatic and phylogenetic analyses showed that these soils harbored 12 out of 18 genera of Proteobacteria harboring rhizobia species validly published to date and revealed for the first time that the Bradyrhizobium genus dominates in tropical savannah soils, together with Microvirga and Paraburkholderia. In silico comparisons of different 16S rRNA gene variable regions suggested that the V5-V7 region could be suitable for differentiating rhizobia at the genus level, possibly replacing the use of the V4-V5 region. These data could serve as indicators for future rhizobial microbiome explorations and for land-use decision-making.SUPPLEMENTARY MATERIAL : FIGURE S1: Covering area of each sampling site and its corresponding features, FIGURE S2: Rarefaction curve of the seven samples, indicated by the number of ASVs depending on the size of the sequence sample, FIGURE S3: 16S rDNA V4-V5 phylogenetic tree showing the relationship between all selected reference alpha and beta-rhizobia strains and the 77 ASVs detected in savannah soils of Northern Côte d’Ivoire, TABLE S1: Physico-chemical properties of samples soils, TABLE S2: Levels of similarity between the V4-V5 sequences of the 77 ASVs of rhizobia and all 18 alphaproteobacterial and betaproteobacterial genera harboring rhizobia strains, TABLE S3: Major characteristics of the 77 ASVs of rhizobia detected in savannah soils of Northern Côte d’Ivoire, TABLE S4: ASVs and their corresponding md5_hash identities, TABLE S5: ASVs richness, cumulative relative abundance of sequences and prevalence of ASVs per soil, TABLE S6: Measure of the richness and the alpha diversity per savannah zone.The US Agency for International Development (USAID)https://www.mdpi.com/journal/microorganismsam2022BiochemistryGeneticsMicrobiology and Plant Patholog

The core bacteriobiome of Cote d’Ivoire soils across three vegetation zones

DATA AVAILABILITY STATEMENT : The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: https://www.ncbi.nlm.nih.gov/, PRJNA695288.The growing understanding that soil bacteria play a critical role in ecosystem servicing has led to a number of large-scale biogeographical surveys of soil microbial diversity. However, most of such studies have focused on northern hemisphere regions and little is known of either the detailed structure or function of soil microbiomes of sub-Saharan African countries. In this paper, we report the use of high-throughput amplicon sequencing analyses to investigate the biogeography of soil bacteria in soils of Côte d’Ivoire. 45 surface soil samples were collected from Côte d’Ivoire, representing all major biomes, and bacterial community composition was assessed by targeting the V4-V5 hypervariable region of the 16S ribosomal RNA gene. Causative relationships of both soil physicochemical properties and climatic data on bacterial community structure were infered. 48 phyla, 92 classes, 152 orders, 356 families, and 1,234 genera of bacteria were identified. The core bacteriobiome consisted of 10 genera ranked in the following order of total abundance: Gp6, Gaiella, Spartobacteria_ genera_incertae_sedis, WPS-1_genera_incertae_sedis, Gp4, Rhodoplanes, Pseudorhodoplanes, Bradyrhizobium, Subdivision3_genera_incertae_sedis, and Gp3. Some of these genera, including Gp4 and WPS-1_genera_incertae_sedis, were unequally distributed between forest and savannah areas while other taxa (Bradyrhizobium and Rhodoplanes) were consistently found in all biomes. The distribution of the core genera, together with the 10 major phyla, was influenced by several environmental factors, including latitude, pH, Al and K. The main pattern of distribution that was observed for the core bacteriobiome was the vegetation-independent distribution scheme. In terms of predicted functions, all core bacterial taxa were involved in assimilatory sulfate reduction, while atmospheric dinitrogen (N2) reduction was only associated with the genus Bradyrhizobium. This work, which is one of the first such study to be undertaken at this scale in Côte d’Ivoire, provides insights into the distribution of bacterial taxa in Côte d’Ivoire soils, and the findings may serve as biological indicator for land management in Côte d’Ivoire.The US Agency for International Development (USAID).http://www.frontiersin.org/Microbiologyam2024BiochemistryGeneticsMicrobiology and Plant PathologySDG-15:Life on lan

HyperEx: A Tool to Extract Hypervariable Regions from 16S rRNA Sequencing Data

ABSTRACTThe 16S ribosomal RNA gene is one of the most studied genes in biology. This 16S ribosomal RNA importance is due to its wide application in phylogenetics and taxonomic elucidation of bacteria and archaea. Indeed, 16S ribosomal RNA is present in almost all bacteria and archaea and has, among many other useful characteristics, a low mutation rate. The 16S ribosomal RNA is composed of nine hypervariable regions which are commonly targeted by high throughput sequencing technologies in identification or community studies like metabarcoding studies. Unfortunately, the hypervariable regions do not have the same taxonomic resolution among all bacteria taxa. This requires a preliminaryin silicoanalysis to determine the best hypervariable regions to target in a particular study. Nevertheless, to the best of our knowledge, no automated primer-based open-source tool exists to extract hypervariable regions from complete or near-complete 16S rRNA sequencing data. Here we present HyperEx which efficiently extracts the hypervariable region of interest based on embedded primers or user-given primers. HyperEx implements the Myers algorithm for the exact pairwise sequence alignment. HyperEx is freely available under the MIT license as an operating system independent Rust command-line tool athttps://github.com/Ebedthan/hyperexandhttps://crates.io.</jats:p

Increased Silicon Acquisition in Bananas Colonized by Rhizophagus irregularis MUCL 41833 Reduces the Incidence of Pseudocercospora fijiensis

This work aimed to test the hypothesis that the combination of arbuscular mycorrhizal fungi (AMF) and accumulation of silicon (Si) in banana plants via its uptake and transport by the fungus reduces the incidence of Black Leaf Steak Disease (BLSD) caused by Pseudocercospora fijiensis. Methods: A pot experiment was conducted to compare BLSD symptoms on leaves of banana plants colonized or not by the AMF Rhizophagus irregularis MUCL 41833 and exposed or not to Si added to the growth substrate. Results: A marked increase in plant growth parameters (i.e., pseudostem diameter and height, leaf surface area, shoot, root and total dry weight) as well as accumulation of Si, P, and Ca were noticed in the AMF-colonized banana plants in presence as well as in absence of Si added to the growth substrate. Similarly Si addition to the substrate increased plant growth parameters. Leave symptoms caused by the pathogen were observed in all the treatments but were reduced in presence of AMF as well as in presence of Si added to the growth substrate. The more drastic reduction was noticed in the AMF-colonized plants with Si added to the growth substrate. The Severity Index as well as Area Under Disease Progress Curve were considerably decreased both at 21 (∼48% and 48%, respectively) and 35 days (∼21% and ∼32%, respectively) after inoculation of the pathogen as compared with non-AMF-colonized plants in absence of Si added to the substrate. Conclusion: Our findings revealed that AMF-colonized banana plants grown in a subs-trate supplemented with Si were less impacted by P. fijiensis than non-colonized plants grown without Si added to the growth substrate. The combination of AMF-colonized banana plants (during the weaning phase or in vitro) with the application of Si to soil seems thus a thoughtful option to mitigate the impact of BLSD in bananas, although such strategy needs first to be evaluated under field conditions to appraise its real potential

Two major clades of bradyrhizobia dominate symbiotic interactions with pigeonpea in fields of C&#244;te d&#39;Ivoire.

In smallholder farms of Côte d’Ivoire, particularly in the northeast of the country, Cajanus cajan (pigeonpea) has become an important crop because of its multiple beneficial facets. Pigeonpea seeds provide food to make ends meet, are sold on local markets and aerial parts serve as forage for animals. Since it fixes atmospheric nitrogen in symbiosis with soil bacteria collectively known as rhizobia, C. cajan also improves soil fertility and reduces fallow time. Yet, seed yields remain low mostly because farmers cannot afford chemical fertilizers. To identify local rhizobial strains susceptible to be used as bio-inoculants to foster pigeonpea growth, root nodules were collected in six fields of three geographically distant regions of Côte d’Ivoire. Nodule bacteria were isolated and characterised using various molecular techniques including matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) and DNA sequencing. These molecular analyses showed that 63 out of 85 nodule isolates belonged to two major clades of bradyrhizobia, one of which is known as the Bradyrhizobium elkanii super clade. Phylogenies of housekeeping (16S-ITS-23S, rpoB) and symbiotic (nifH) genes were not always congruent suggesting that lateral transfer of nitrogen fixation genes also contributed to define the genome of these bradyrhizobial isolates. Interestingly, no field-, plant- or cultivar-specific effect was found to shape the profiles of symbiotic strains. In addition, nodule isolates CI-1B, CI-36E and CI-41A that belong to distinct species, showed similar symbiotic efficiencies suggesting that any of these strains might serve as a proficient inoculant for C. cajan

Fluorescence and Reflectance Spectroscopy for Early Detection of Different Mycorrhized Plantain Plants

&lt;p class="1Body"&gt;Sustainable agriculture with use of Arbuscular Mycorrhizal Fungi (AMF) is an emerging farm management that improves crops nutrient and water use efficiency. Decision making on the effect of AMF is still dependent on agronomic diagnosis which is long, tedious, expensive and destructive. This study demonstrates the applicability of proximal fluorescence and reflectance spectroscopy for evaluating and detecting at early stage distinct types of mycorrhized plantain from two cultivars (&lt;em&gt;Musa paradisiaca&lt;/em&gt;).&lt;/p&gt;&lt;p class="1Body"&gt;Visible-near infrared (400-1000 nm) reflectance and fluorescence data were collected from control and three levels mycorrhized plants designed in randomized and complete block under greenhouse conditions. Two spectral measurements at a week interval were performed on plant leaves by using an USB spectrometer mounted with an Arduino-based LED driver clip.&lt;/p&gt;A new normalized reflectance water NWI5 index shows with Datt5 alone highly significant differences at P&amp;lt;0.001 respectively for Orishele and fhia21 cultivars. dNIRmin920_980, NDVI3 and GI reflectance index are significant at P&amp;lt;0.01. Seven other reflectance and 3 fluorescence indices ANTH, FRF_R and NBI_R are significant at P&amp;lt;0.05. The two first principal components for each cultivar spectral features explaining 94.1 % of variance were used to build predictive classification models. LogitBoost algorithm indicates accuracy of 90.27% on stratified cross-validation and 87.5% on test split. Our results confirm that fluorescence and reflectance spectroscopy is a valuable tool for early assessment of mycorrhization success rate evaluation and pattern recognition. They also show promise for the development of non-destructive and cost-effective detectors in monitoring crops under biofertilizers with arbuscular mycorrhizae.</jats:p

Antimicrobial resistance of Escherichia coli isolated from irrigation waters in selected urban fields around Yamoussoukro lakes in Côte d'Ivoire

This study examined the distribution of Escherichia coli (E. coli) resistant to eleven antibiotics and possible relationships among physico-chemical parameters and these resistances. It was carried out over two years during four climatic seasons in five lakes of the Yamoussoukro lake system in Côte d'Ivoire. The physicochemical parameters of the lakes studied are temperature, pH, dissolved oxygen (DO), dissolved organic carbon (DOC), turbidity, nitrates, orthophosphates and ammoniacal nitrogen. A total of 744 water samples were collected from five (5) lakes belonging to the lake system, over a two-year period, during four dry and four wet seasons. Escherichia coli loadings were evaluated after isolation on specific chromogenic medium COMPASS ECC Agar and the susceptibility to antibiotics by the agar diffusion method. Physico-chemical parameters were evaluated according to international standards. The physico-chemical results showed that the nutrient contents evolved generally from upstream to downstream of the lake system, independently of the sources of pollution of five lakes of system. However, the lake B, in upstream of the lake system, showed levels of nitrate, orthophosphate and ammoniacal nitrogen often similar to the downstream lakes. This demonstrates a source of chemical pollution in this lake, which flows to lakes D and E, which are themselves subject to other pollution. Bacterial loads and bacterial resistance to antibiotics have strictly evolved from upstream to downstream of the system. Most isolates were resistant to ampicillin (16.46%), tetracycline (12.87%), ciprofloxacin (12.86%) and sulfamethoxazole (10.14%). Antibiotic resistance patterns of E. coli isolates were similar in both years studied, but higher during rainy seasons (GRS and SRS). The spearman rank correlations and the principal component analysis (PCA) revealed significant correlations between bacterial resistance to antibiotics in lakes and water physico-chemical parameters. Turbidity and DOC can help to determine the main resistances in the lake D. Orthophosphates can help to understand the resistance of chloramphenicol and the nitrates can help to understand the resistance of ampicillin in the lake B. Resistance in the lakes A, B and C can be influenced by dissolved oxygen. Overall, this study provided baseline information on bacterial resistance in the lakes of Yamoussoukro in Côte d'Ivoire, showing that these lakes could be reservoirs of antibiotic resistance, potentially dangerous to public health.</jats:p

Hierarchical spatial sampling reveals factors influencing arbuscular mycorrhizal fungus diversity in Côte d’Ivoire cocoa plantations

AbstractWhile many molecular studies have documented arbuscular mycorrhizal fungi (AMF) communities in temperate ecosystems, very few studies exist in which molecular techniques have been used to study tropical AMF communities. Understanding the composition of AMF communities in tropical areas gains special relevance as crop productivity in typically low fertility tropical soils can be improved with the use of AMF. We used a hierarchical sampling approach in which we sampled soil from cocoa (Theobroma cacao L.) plantations nested in localities, and in which localities were nested within each of three regions of Côte d’Ivoire. This sampling strategy, combined with 18S rRNA gene sequencing and a dedicated de novo OTU-picking model, allowed us to study AMF community composition and how it is influenced at different geographical scales and across environmental gradients. Several factors, including pH, influenced overall AMF alpha diversity and differential abundance of specific taxa and families of the Glomeromycotina. Assemblages and diversity metrics at the local scale did not reliably predict those at regional scales. The amount of variation explained by soil, climate, and geography variables left a large proportion of the variance to be explained by other processes, likely happening at smaller scales than the ones considered in this study. Gaining a better understanding of processes involved in shaping tropical AMF community composition and AMF establishment are much needed and could allow for the development of sustainable, productive tropical agroecosystems.</jats:p