Peanut Nodules Research Articles

Nodule INception-independent epidermal events lead to bacterial entry during nodule development in peanut (Arachis hypogaea).

Legumes can host nitrogen-fixing rhizobia inside root nodules. In model legumes, rhizobia enter via infection threads (ITs) and develop nodules in which the infection zone contains a mixture of infected and uninfected cells. Peanut (Arachis hypogaea) diversified from model legumes c. 50-55 million years ago. Rhizobia enter through 'cracks' to form nodules in peanut roots where cells of the infection zone are uniformly infected. Phylogenomic studies have indicated symbiosis as a labile trait in peanut. These atypical features prompted us to investigate the molecular mechanism of peanut nodule development. Combining cell biology, genetics and genomic tools, we visualized the status of hormonal signaling in peanut nodule primordia. Moreover, we dissected the signaling modules of Nodule INception (NIN), a master regulator of both epidermal infection and cortical organogenesis. Cytokinin signaling operates in a broad zone, from the epidermis to the pericycle inside nodule primordia, while auxin signaling is narrower and focused. Nodule INception is involved in nodule organogenesis, but not in crack entry. Nodulation Pectate Lyase, which remodels cell walls during IT formation, is not required. By contrast, Nodule enhanced Glycosyl Hydrolases (AhNGHs) are recruited for cell wall modification during crack entry. While hormonal regulation is conserved, the function of the NIN signaling modules is diversified in peanut.

Variation in rhizosphere microbial communities and its association with the nodulation ability of peanut

ABSTRACT Harnessing the rhizobia-legume symbiosis is a promising strategy to confront sustainability in agriculture. Agricultural research on legume nodulation has mostly focused on abiotic parameters, yet the importance of biotic variables on legumes nodulation is still ambiguous. We explored the variation of bacterial communities in peanut rhizosphere soil under different agricultural practices (peanut fallow rotation and Vicia villosa Roth-peanut rotation) using Illumina sequencing. Samples were collected in 2020 from a 9-year study (2012–2020) located at Dongxiang countryside (Jiangxi province, China). Compared with peanut fallow rotation (PFR), Vicia villosa Roth-peanut rotation (VPR) significantly (p< 0.05) decreased soil pH (by 0.4–3.9%) and number of peanut nodules (by 20–63.7%). By using the Random forest tool, we found that the difference in peanut nodulation was associated with specific microbiome alterations. The indicator taxa positively responded to the peanut nodulation was enriched in the PFR practice, while VPR led to increased abundance of indicator taxa which negatively responded to peanut nodulation. The path analysis indicated that soil pH has an indirect effect on peanut nodulation through its link with the abundance of indicator taxa. Our work provides evidence that the variation in peanut rhizosphere microbial communities was associated with peanut nodulation.

Do biofertilizers affect nodulation ability and pod production in peanut genotypes?

Peanuts are an important legume for the Northeastern Brazilian market, but their production in this region is low. The present study aimed to evaluate the effect of biofertilizer doses on peanut nodulation and production components, to define the best dose and genotype. The experiment was conducted in a completely randomized block design (DBC) in a 3 x 3 factorial arrangement, with three replications. The treatments consisted of two evaluation factors: 1) Fertilization via different doses of organic fertilizer applied to the substrate (D1 = 0mL; D2 = 500mL; and D3 = 1000mL); and 2) Peanut genotypes (BR-1, UNI43 and UNI08). The following traits were assessed: number of nodules per plant (NNP), number of mature pods (NVM), pod mass per plant (MVP), seed mass per plant (MSP) and root length (CR). Peanut production is affected by fertilization via bovine biofertilizer, mainly for the components NVM, MVP and MSP. Besides, nodulation is a likely conditioner. The 1000mL dose proved to be the best treatment for the traits analyzed, and UNI08 accession, the most responsive material.

An Improvised Hairy Root Transformation Method for Efficient Gene Silencing in Roots and Nodules of Arachis hypogaea.

Peanut (Arachis hypogaea) is a major oilseed crop and is widely cultivated in tropical and subtropical climate zone worldwide. Peanut belongs to the Papilionoid family with an atypical nodule developmental program. In particular, rhizobia enter through developmental cracks and lead to the formation of aeschynomenoid subtype determinate nodules. Peanut nodules are efficient nitrogen-fixers and form swollen bacteroid containing symbiosomes. The allotetraploid genome and recalcitrance to stable transformation used to be the major bottleneck for peanut biologists. Recent genome sequencing of peanut cultivar Tifrunner has opened up a huge opportunity for molecular research. A composite plant contains transformed roots with a non-transformed shoot. The composite plant-based approach has already proven to be a tool of choice for high throughput studies in root biology. The available protocols failed to generate efficient hairy root transformation in the genome sequenced cultivar Tifrunner. Here we describe an efficient hairy root transformation and composite plant generation protocol for the peanut cultivar Tifrunner. Our protocol generated ~92% plant regeneration efficiency with between 21.8% and 58.6% co-transformed root regeneration. We also show that this protocol can be efficiently used for protein localization, promoter GUS analysis, monitoring hormone response, and RNAi mediated knockdown of the genes using genome sequenced cultivar Tifrunner.

BIOLOGICAL FEATURES OF GROUNDNUT MICROSYMBIONTS WIDESPREAD IN THE SOILS OF UKRAINE

Objective. Study the presence of nodule bacteria in the soils of Ukraine, capable of nodulatinggroundnut, isolate new strains of rhizobia from peanut nodules, study their morphological, culturaland serological properties and the ability to form symbiosis with different legumes. Methods. Microbiological (isolation of nodule bacteria from nodules and cultivation of microorganisms, studying biological properties of strains), serological (producing antisera to Bradyrhizobium lupini367a, studying rhizobia diversity in groundnut nodule populations, studying serological identity ofnew strains), vegetation experiment (studying formation and functioning of groundnut symbioticsystems with nodule bacteria, studying host specificity of new strains of groundnut rhizobia), gaschromatography (determining nitrogen-fixing activity of rhizobia in symbiosis with groundnut),mathematical and statistical. Results. Representatives of two species, B. lupini and B. japonicum,were found in nodule populations of rhizobia during the cultivation of groundnut plants on sodpodzolic soil and leached chernozem. The dominant microsymbionts of groundnut were lupine nodule bacteria of serogroup 367a (54.2 % and 45.8 % according to soils). Fewer nodules wereformed by intensive growing soybean rhizobia of serogroup КВ11 (16.7 % and 12.5 %). The shareof nodule bacteria not classified in the studied serogroups was 21.9 % and 41.7 %. Fifteen newstrains of nodule bacteria were isolated from groundnut nodules, which were morphologically, culturally and serologically identified as B. lupini serogroup 367a (7 of them), B. japonicumserogroup KB11 (4 of them) and Bradyrhizobium sp. (4 of them). New strains of B. lupini fromgroundnut nodules are able to infect white and yellow lupine but do not nodulate soybeans. Strainsidentified as B. japonicum form nodules on soybean roots but do not infect lupine. Serologically unidentified strain Bradyrhizobium sp. AR3, which is able to form a symbiosis with both lupine andsoybean (phenotypes Nod+Fix+) was obtained. Conclusion. For the first time it was established thatgroups of nodule bacteria capable of nodulating groundnut are present in the agrocenoses ofUkraine. Fifteen new strains of groundnut rhizobia were obtained, which were identified as B. lupini, B. japonicum and Bradyrhizobium sp.

Draft Genome Sequences of Nitrogen-Fixing Bradyrhizobia Isolated from Root Nodules of Peanut, Arachis hypogaea, Cultivated in Southern Tunisia.

ABSTRACTHere, we report the draft genome sequences of two nitrogen-fixing symbionts, Bradyrhizobium sp. strain sGM-13 and Bradyrhizobium sp. strain sBnM-33, isolated from root nodules of peanut grown on soil samples collected from two regions in South Tunisia. The draft genome sizes of these two strains are 8.31 × 106 bp and 8.97 × 106 bp, respectively.

Root endophyte-enhanced peanut-rhizobia interaction is associated with regulation of root exudates

Root exudates play a crucial role in the symbiosis between leguminous plants and rhizobia. Our previous studies have shown that a fungal endophyte Phomopsis liquidambaris promotes peanut-rhizobia nodulation and nitrogen fixation, but the underlying mechanism are largely unknown. Here, we explore the role of peanut root exudates in Ph. liquidambaris-mediated nodulation enhancement. We first collected root exudates from Ph. liquidambaris-inoculated and un-inoculated peanuts and determined their effects on rhizobial growth, biofilm formation, chemotaxis, nodC gene expression, and peanut nodulation. Our results found a positive effect of Ph. liquidambaris-inoculated root exudates on these characteristics of rhizobia. Next, we compared the root exudates profile of Ph. liquidambaris-inoculated and un-inoculated plants and found that Ph. liquidambaris altered the concentrations of phenolic acids, flavonoids, organic acids and amino acids in root exudates. Furthermore, the rhizobial chemotaxis, growth and biofilm formation in response to the changed compounds at different concentrations showed that all of the test compounds induced rhizobial chemotactic behavior, and organic acids (citric acid and oxalic acid) and amino acid (glutamate, glycine and glutamine) at higher concentrations increased rhizobial growth and biofilm formation. Collectively, our results suggest that root exudates alterations contribute to Ph. liquidambaris-mediated peanut-rhizobia nodulation enhancement.

Novel digital assessment technique used to describe peanut nodulation life history

Biological nitrogen fixation (BNF) in legume root nodules is an important source of N globally. Typically, the quantification of BNF is an estimate of N fixation through techniques such as an acetylene reduction assay or isotopic signatures. However, these methods have large logistical limitations, and do not allow for assessments of the nodule morphology or the nodulation process itself, knowledge that is important to fully understand the BNF process. To provide an updated method of nodulation assessments, peanut (Arachis hypogaea L.) nodules were sampled from a two-year field trial and quantified using scanned images of excised nodules, allowing digital assessment of nodule number and size. Further, a digital color analysis method was developed to assess internal nodule color (a known proxy for nodule BNF activity and senescence). To apply and evaluate the new method, peanut nodulation trait data was used for an exploration of the relationship between nodulation traits, plant physiological processes, and nitrogen status in the plant, followed by a nodulation life history analysis. The nodulation assessment method was confirmed by strong correlation with measures of peanut tissue N, biomass, photosynthetic function, and yield. Nodulation life history data suggest novel findings regarding differences in the development of nodules between taproots and lateral roots, contrary to the previous understanding of peanut nodulation life history. This new nodulation assessment method can provide further insight into the BNF process by allowing the collection of detailed data in a relatively simplified manner in field conditions.

Effects of maize/peanut intercropping system on nodulation of peanut in northwest Liaoning

The maize/peanut intercropping system was considered as a technique, which can both reduce drought risk of sole maize and wind erosion of sole peanut in northwest Liaoning. To clarify the nodulation characteristic of peanut in maize/peanut intercropping system of semi-arid area in northwest Liaoning, a field experiment was conducted at National Agricultural Experimental Station for Agricultural Environment, in northwest Liaoning, China. Including three cropping system (M2P4, M4P4 and Sole peanut) and three densities of maize (2, 3, 4 plants m−2 for M2P4, 3, 4.5, 6 plants m−2 for M4P4) based on the configuration. The results showed that Intercropped with maize significantly promote the nodulation of peanut. The nodule number and nodule size of peanut in M4P4 was significantly more than that of intercrops M2P4 and monoculture. Increasing the proportion of maize in row promotes the nodulation of peanut in maize/peanut intercropping system. The middle density of maize (3 plants m−2) is the best density for peanut nodulation in M2P4 intercropping system. 3 plants m-1 density of maize is better for nodulation of peanut in earlier stage of peanut growth in M4P4 system, while the high maize density (6 plants m−2) is more suitable for peanut nodulation in middle and later periods.

The application of CRISPR/Cas9 in hairy roots to explore the functions of AhNFR1 and AhNFR5 genes during peanut nodulation

BackgroundPeanut is an important legume crop growing worldwide. With the published allotetraploid genomes, further functional studies of the genes in peanut are very critical for crop improvement. CRISPR/Cas9 system is emerging as a robust tool for gene functional study and crop improvement, which haven’t been extensively utilized in peanut yet. Peanut plant forms root nodules to fix nitrogen through a symbiotic relationship with rhizobia. In model legumes, the response of plants to rhizobia is initiated by Nod factor receptors (NFRs). However, information about the function of NFRs in peanut is still limited. In this study, we applied the CRISPR/Cas9 tool in peanut hairy root transformation system to explore the function of NFR genes.ResultsWe firstly identified four AhNFR1 genes and two AhNFR5 genes in cultivated peanut (Tifrunner). The gene expression analysis showed that the two AhNFR1 and two AhNFR5 genes had high expression levels in nodulating (Nod+) line E5 compared with non-nodulating (Nod-) line E4 during the process of nodule formation, suggesting their roles in peanut nodulation. To further explore their functions in peanut nodulation, we applied CRISPR technology to create knock-out mutants of AhNFR1 and AhNFR5 genes using hairy root transformation system. The sequencing of these genes in transgenic hairy roots showed that the selected AhNFR1 and AhNFR5 genes were successfully edited by the CRISPR system, demonstrating its efficacy for targeted mutation in allotetraploid peanut. The mutants with editing in the two AhNFR5 genes showed Nod- phenotype, whereas mutants with editing in the two selected AhNFR1 genes could still form nodules after rhizobia inoculation.ConclusionsThis study showed that CRISPR-Cas9 could be used in peanut hairy root transformation system for peanut functional genomic studies, specifically on the gene function in roots. By using CRISPR-Cas9 targeting peanut AhNFR genes in hairy root transformation system, we validated the function of AhNFR5 genes in nodule formation in peanut.

Identification of miRNAs linked to peanut nodule functional processes

microRNAs (miRNAs) are non-coding small RNAs that regulate gene expression at post-transcriptional level. Thousands of miRNAs have been identified in legumes, but studies about miRNAs linked to peanut nodule functionality are scarce. In this work we analyzed transcriptional changes in peanut nodules to identify miRNAs involved in functional processes of these organs. We found 32 miRNAs precursors differentially expressed in nodules compared with roots, and predicted the potential targets of their corresponding mature miRNAs. Among them, 20 belong to 14 conserved miRNAs families and 12 are Arachis hypogaea-specific miRNAs. Expression levels of 3 miRNAs (ahy-miR399, ahy-miR159 and ahy-miR3508) were confirmed experimentally by qPCR. We also demonstrated that the expression of these miRNAs was not affected by inoculation of a biocontrol bacterium or a fungal pathogen. The catalogue of differentially expressed miRNA precursors and the expression of the corresponding mature miRNA potential targets in the nodules of A. hypogaea obtained in this work is a database of strong candidates, including A. hypogaea-specific miRNAs, for the regulation of the nodule functionality. The analysis of their role in this process will certainly lead to the characterization of essential regulators in these particular aeschynomenoid nodules.

Comparison of SNP Calling Pipelines and NGS Platforms to Predict the Genomic Regions Harboring Candidate Genes for Nodulation in Cultivated Peanut.

Cultivated peanut (Arachis hypogaea L.) forms root nodules to enable a symbiotic relationship with rhizobia for biological nitrogen fixation. To understand the genetic factors of peanut nodulation, it is fundamental to genetically map and clone the genes involved in nodulation. For genetic mapping, high throughput genotyping with a large number of polymorphic markers is critical. In this study, two sets of sister recombinant inbred lines (RILs), each containing a nodulating (Nod+) and non-nodulating (Nod-) line, and their Nod+ parental lines were extensively genotyped. Several next generation sequencing (NGS) methods including target enrichment sequencing (TES), RNA-sequencing (RNA-seq), genotyping by sequencing (GBS), and the 48K Axiom Arachis2 SNP array, and various analysis pipelines were applied to identify single nucleotide polymorphisms (SNP) among the two sets of RILs and their parents. TES revealed the largest number of homozygous SNPs (15,947) between the original parental lines, followed by the Axiom Arachis2 SNP array (1,887), RNA-seq (1,633), and GBS (312). Among the five SNP analysis pipelines applied, the alignment to A/B genome followed by HAPLOSWEEP revealed the largest number of homozygous SNPs and highest concordance rate (79%) with the array. A total of 222 and 1,200 homozygous SNPs were polymorphic between the Nod+ and Nod− sister RILs and between their parents, respectively. A graphical genotype map of the sister RILs was constructed with these SNPs, which demonstrated the candidate genomic regions harboring genes controlling nodulation across the whole genome. Results of this study mainly provide the pros and cons of NGS and SNP genotyping platforms for genetic mapping in peanut, and also provide potential genetic resources to narrow down the genomic regions controlling peanut nodulation, which would lay the foundation for gene cloning and improvement of nitrogen fixation in peanut.

Isolation, Characterization, and Complete Genome Sequence of a Bradyrhizobium Strain Lb8 From Nodules of Peanut Utilizing Crack Entry Infection.

In many legumes, the colonization of roots by rhizobia is via “root hair entry” and its molecular mechanisms have been extensively studied. However, the nodulation of peanuts (Arachis hypogaea L.) by Bradyrhizobium strains requires an intercellular colonization process called “crack entry,” which is understudied. To understand the intercellular crack entry process, it is critical to develop the tools and resources related to the rhizobium in addition to focus on investigating the mechanisms of the plant host. In this study, we isolated a Bradyrhizobium sp. strain, Lb8 from peanut root nodules and sequenced it using PacBio long reads. The complete genome sequence was a circular chromosome of 8,718,147 base-pair (bp) with an average GC content of 63.14%. No plasmid sequence was detected in the sequenced DNA sample. A total of 8,433 potential protein-encoding genes, one rRNA cluster, and 51 tRNA genes were annotated. Fifty-eight percent of the predicted genes showed similarity to genes of known functions and were classified into 27 subsystems representing various biological processes. The genome shared 92% of the gene families with B. diazoefficens USDA 110T. A presumptive symbiosis island of 778 Kb was detected, which included two clusters of nif and nod genes. A total of 711 putative protein-encoding genes were in this region, among which 455 genes have potential functions related to symbiotic nitrogen fixation and DNA transmission. Of 21 genes annotated as transposase, 16 were located in the symbiosis island. Lb8 possessed both Type III and Type IV protein secretion systems, and our work elucidated the association of flagellar Type III secretion systems in bradyrhizobia. These observations suggested that complex rearrangement, such as horizontal transfer and insertion of different DNA elements, might be responsible for the plasticity of the Bradyrhizobium genome.

Microsymbionts of forage peanut under different soil and climate conditions belong to a specific group of Bradyrhizobium strains

The use of mixed pastures of grasses with forage peanut (Arachis pintoi) is growing and offers promising results. This leguminous forage plant has a stoloniferous growth habit and tolerates more intense defoliation and shading. However, there are few studies on its symbiosis with nitrogen (N2) fixing bacteria and no studies on genetic identification of its native microsymbionts. The aim of this study was to evaluate phenotypic and genetic diversity and the symbiotic efficiency of bacterial strains isolated from nodules of forage peanut intercropped with grasses in pasture areas. Root nodules were collected from the Belomonte cultivar in Itabela, Bahia, and BRS Mandobi cultivar in Lavras, Minas Gerais, Brazil. The areas of sample collection had contrasting physical and chemical characteristics. Samples of 10 plants per area and 5 nodules per plant were taken. The strains isolated from the nodules were analyzed for culture characteristics (growth period and change in pH of the culture medium), their 16S rRNA, gyrB and nifH genes were partially sequenced, and authentication and analysis of symbiotic efficiency in forage peanut plants were carried out. Both the authentication and symbiotic efficiency experiments were carried out in a greenhouse under axenic conditions, using A. pintoi cv. BRS Mandobi in a completely randomized design with four replications. Authentication was performed in long neck bottles (500 mL) filled with nutrient solution, and symbiotic efficiency was performed in polypropylene seedling plug pots containing a mixture of sand and vermiculite with a nutrient solution. A total of 74 bacterial strains were obtained from isolation of the forage peanut nodules, 40 from Itabela, and 34 from Lavras. Partial sequencing of the 16S rRNA gene was obtained for 48 strains, 30 from Itabela, and 18 from Lavras. The strains isolated from nodules of forage peanut in pasture areas of Itabela, were identified in the following genera: Bradyrhizobium (21), Rhizobium (5), Burkholderia (2), Kocuria (1), and Paenibacillus (1). The strains coming from soils of Lavras, were identified in the following genera: Bradyrhizobium (13), Rhizobium (2), Burkholderia (1), Mucilaginibacter (1), and Enterobacter (1). In the authentication experiment, 17 strains from Itabela, and 13 strains from Lavras, exhibited nodulation. From these authenticated strains, 18 were identified by 16S rRNA sequencing as belonging to the Bradyrhizobium genus. Of these strains, 11 from the two areas exhibited high symbiotic efficiency in A. pintoi, and their results were better than those of the strains currently approved as inoculants for this crop. The non-nodulating strains are probably nodule endophytes. Sequencing of the gyrB housekeeping gene, performed in 13 strains, confirms phylogenetic groups of potential new species among the strains isolated from A. pintoi, which include strains able to nodulate this species, belonging to a specific phylogenetic group of Bradyrhizobium. Sequences of nifH gene of efficient strains belonged to a group different from that formed by non-nodulating strains.

Endophytic Fungus Drives Nodulation and N2 Fixation Attributable to Specific Root Exudates.

Endophytic fungi play important roles in the modification of ecosystem productivity; however, the underlying mechanisms are only partly understood. A 2-year field plot experiment verified that the endophytic fungus Phomopsis liquidambaris increased peanut (Arachis hypogaea L.) yields and significantly increased nodulation and N2 fixation regardless of whether N fertilizers were added. Root exudates collected from P. liquidambaris-colonized plants significantly improved nodulation and N2 fixation. Rhizosphere stimulation experiments further showed that colonized root exudates had significantly decreased soil nitrate (NO3-) concentrations, with decreased abundance and diversity of ammonia oxidizing archaea (AOA). In contrast, the abundance and diversity of diazotrophs significantly increased, and most diazotrophs identified were peanut nodulation-related strains (Bradyrhizobium sp.). P. liquidambaris symbiosis increased the expression of phenolic and flavonoid synthesis-related genes, and the derived phenolics and flavonoids could effectively increase the chemotaxis, biofilm formation, and nodC gene expression (nodulation-related biological processes) of the Bradyrhizobium strain. Metabolic pattern analysis showed that phenolics and flavonoids are more likely to accumulate to higher levels in the rhizosphere soil of peanuts colonized with P. liquidambaris Finally, a synthetic root exudate experiment further confirmed the underlying mechanisms for the P. liquidambaris-induced improvement in nodulation and N2 fixation, i.e., that the specific root exudates derived from P. liquidambaris colonization decrease nitrate concentration and increase the population and biological activities of peanut nodulation-related Bradyrhizobium species, which beneficially enhance peanut-Bradyrhizobium interactions. Therefore, this study is the first to provide new insight into a positive relationship between an exotic endophytic fungus, crop nodulation, and N2 fixation increase.IMPORTANCE Endophytic fungi play an important role in balancing the ecosystem and boosting host growth; however, the underpinning mechanisms remain poorly understood. Here, we found that endophytic fungal colonization with P. liquidambaris significantly increased the productivity, nodulation, and N2 fixation of peanuts through the secretion of specific root exudates. We provide a reasonable mechanism explaining how P. liquidambaris promotes peanut nodulation and N2 fixation, whereby the specific root exudates produced by P. liquidambaris colonization decrease rhizosphere soil nitrate (NO3-) and increase the population and biological activities of peanut-nodulating-related Bradyrhizobium strains, which is beneficial to enhancing the peanut-Bradyrhizobium symbiotic interaction. Our study provides reliable empirical evidence to show the mechanism of how an exotic endophytic fungus drives an increase in nodulation and N2 fixation, which will be helpful in erecting a resource-efficient and sustainable agricultural system.

Phylogenetic diversity and cross-inoculation of indigenous isolated Bradyrhizobium from nodules of peanut in Liaoning province of China.

Arachis hypogaea. L is a legume of economic importance, which is nodulated by Bradyrhizobium, a slow-growing bacteria. However there is no well characterization of this rhizobia in many areas of China. In the present study, cross-inoculation experiments were performed in cowpea and soybean. The isolated bacteria strains were characterized physiologyically, biochemically and identified through 16S rDNA sequence analysis showing that it belongs to Bradyrhizobium japonicum. The genetic diversity of the seventeen isolated strains were assessed through PCR-RFLP of 16S rDNA and 16S-23S rDNA IGS region. Cross inoculation test indicated that isolates could nodulate cowpea but not soybean. The cluster analysis based on physiological and biochemical characteristics showed the lower correlation between isolates and sites. The isolates were grouped into four clusters based on 16S rDNA gene sequence analysis. Thirteen polymorphisms were variable across all observations in 16S rDNA RFLP and six different IGS types from isolates. The results implies that there was some association between geographical factor and phylogenetic diversity of indigenous Bradyrhizobium isolates.

The Alleopathic Compound Luteolin from Peanut Litter Affects Peanut Nodule Formation and the Rhizosphere Microbial Community

Core Ideas Flavonoid luteolin can accumulate in consecutively monocultured peanut fields. Luteolin treatment changes the rhizosphere microbial community of peanut. Flavonoid luteolin in peanut residues may cause continuous cropping obstacles. The flavonoid luteolin is a common flavone released from peanut (Arachis hypogaea L.) residues. Luteolin accumulates in consecutively monocultured soil and affects the growth and nodulation of continuously cropped peanut. A microcosm experiment was performed to determine the effects of luteolin released from peanut residue on peanut nodule formation and on the rhizosphere soil microbial community. Luteolin reached a maximum concentration of 0.42 μg g−1 soil in litterbag experiments, and the half‐life of luteolin at 2.5 to 40 μg g−1 soil varied from 1.13 to 2.91 d. Results from pot experiments indicate that soil with luteolin treatment inhibited peanut seedling nodulation and growth, soil dehydrogenase activity, microbial biomass carbon content, biofilm formation, and chemotactic response of Rhizobium sp. and changed the microbial community structures and abundances at concentration of luteolin as low as 0.42 μg g−1 in soil. This might explain why peanut performs poorly with continuous consecutive monocropping.

Genetic diversity and symbiotic efficiency of rhizobial strains isolated from nodules of peanut (Arachis hypogaea L.) in Senegal

• This is the first report on the genetic and phenotypic diversity of rhizobia associated with Peanut in Senegal. • We characterize a new collection of Bradyrhizobia strain obtained from peanut nodules in Senegal. • We compared strains from the peanut center of origin (South America) to strains obtained in Senegal. • We identify effective strains with potential for use as peanut inoculants. • Peanut was probably introduced in Senegal without its original strains, or those strains were lost or transformed.

Morphological and Genetic Characterization of Non‐Nodulating Peanut Recombinant Inbred Lines

Peanut (Arachis hypogaea L.) nodulation is relatively diverged from other legumes. Further understanding of nodulation mechanisms would facilitate improvement of biological N fixation and yield in peanut. The objective of this study was to characterize the non‐nodulating (Nod−) peanut recombinant inbred lines (RILs) in comparison to their nodulating (Nod+) sister RILs morphologically and genetically. Two pairs of Nod− and Nod+ RILs were inoculated with rhizobia for morphological observation. Nodules and root hairs were absent in Nod− lines and present in Nod+ lines. The Nod− lines had a much smaller canopy with yellow leaves and fewer pods than Nod+ lines. Simple‐sequence repeat (SSR) marker genotyping of the two pairs of RILs and their parental lines revealed several chromosome regions differing between Nod− and Nod+ sister lines, which potentially harbor the genes controlling nodulation. Two F2 populations segregating for nodulation were constructed by crossing the two Nod− lines with their Nod+ sister lines. The segregation ratios of Nod−/Nod+ of the two populations followed 7:9 and 7:57, respectively, indicating that multiple genes were involved in controlling peanut nodulation. The results from this study provided important morphological and genetic information related to peanut nodulation, which paved the road for further mapping and cloning of peanut nodulation genes and for eventually developing peanut cultivars with improved N fixation efficiency.

Involvement of Root Hair during &amp;lt;i&amp;gt;Rhizobial&amp;lt;/i&amp;gt; Invasion in Cultivated Peanut (&amp;lt;i&amp;gt;Arachis hypogaea&amp;lt;/i&amp;gt; L.)

Peanut root invasion by Bradyrhizobia is through a crack entry, which is different from many other legumes applying an infection thread entry in root hair. Understanding the role of root hair in the crack entry of Bradyrhizobia invasion of peanut root and subsequent peanut nodulation would facilitate improvement of biological nitrogen fixation in cultivated peanut. The objective of this study was to investigate the involvement of root hair in Bradyrhizobial invasion of peanut. Seedling roots of a nodulating peanut cultivar were observed for root hair emergence, its life span, and nodule formation at the base of the lateral roots with and without rhizobia inoculation for 14 days after germination (DAG). Scanning electron microscopy (SEM) was utilized to observe rhizobia accumulation at lateral roots at 24 hours after inoculation (HAI) before the emergence of root hair. Root hair emerged at 7 DAG with or without rhizobia inoculation. Two variations of rosette hair (RoH) were observed, the transient-thin RoH had life span of 3 days after root hair emergence and the thick and densely distributed RoH type stayed till the time of nodule emergence (9 days after inoculation). The lateral root devoid of root hair at the top 2 cm region was found to produce nodules. The SEM observation of seedling roots at 24 HAI showed that Bradyrhizobia invaded the roots at epidermis, protoplasm of cortical cell, and cortical cells of the main root near the newly emerged lateral root in the absence of RoH. The observations validated that root hair is not required in the Bradyrhizobia invasion of peanut root in the crack entry mode. Results from this study provided important morphological information for the hypothesis of close relationship between RoH and peanut nodulation for further genetic study of crack entry mechanism and signaling pathway of symbiosis between Bradyrhizobia and peanut.