Peanut Nodules Research Articles

Effects of Maize/Peanut Intercropping on Yield and Nitrogen Uptake and Utilization under Different Nitrogen Application Rates

The effects of maize/peanut intercropping on crop yields, peanut nodulation, biological nitrogen (N) fixation in peanuts, crop N uptake, and N use efficiency under different N application rates were studied. A long-term maize/peanut intercropping micro-plot experiment was started in 2015. The experiment included the following three planting patterns: maize sole crop (SM), peanut sole crop (SP), and maize and peanut intercropping (intercropping maize: IM; intercropping peanut: IP). Additionally, three N application rates were tested as follows: 0 kg·ha−1 (N0), 150 kg·ha−1 (N150), and 300 kg·ha−1 (N300). The results indicated that N fertilization significantly increased maize yield. Intercropping increased maize yield while decreasing peanut yield across different N application rates. Both N fertilization and intercropping significantly increased the maize harvest index (HI), whereas intercropping decreased the peanut HI under N300. The number and fresh weight of peanut nodules decreased with the increasing N application rate with reductions ranging from 31.15% to 45.23% and 39.60% to 46.67%, respectively. Intercropping increased the number of peanut nodules by an average of 62.56% under the N0 treatment. Intercropping significantly improved the N absorption capacity of the whole intercropping system, and the contribution of maize was higher than that of peanuts. Maize demonstrated a stronger competitive ability for N uptake compared with peanuts in the intercropping system. Intercropping significantly increased the N use efficiency for both maize and peanuts. However, the N use efficiency of maize increased with N application rates, while that of peanut decreased. Compared with sole crops, intercropping increased the partial factor productivity of maize by 55.2% but decreased that of peanuts by 56.3%. In conclusion, at an N application rate of 150 kg·ha−1, maize/peanut intercropping increased overall crop yield and improved the N absorption and use capacity of maize

Complete and circularized genome sequences of five nitrogen-fixing Bradyrhizobium sp. strains isolated from root nodules of peanut, Arachis hypogaea, cultivated in Tunisia.

This manuscript reports the complete and circularized Oxford Nanopore Technologies (ONT) long read-based genome sequences of five nitrogen-fixing symbionts belonging to the genus Bradyrhizobium, isolated from root nodules of peanut (Arachis hypogaea) grown on soil samples collected from Tunisia.

Maize/Peanut Intercropping Affects Legume Nodulation in Semi-Arid Conditions

Maize/peanut intercropping is practiced widely to increase land productivity and considered a sustainable way for using and saving resources through peanut’s complementary N source via biological N2 fixation. Our study aims to understand how maize/peanut intercropping affects the nodulation of peanuts under water-limiting conditions and different nitrogen inputs. A two-year micro-plot experiment in 2015–2016 and a two-year field experiment in 2017–2018 were conducted to quantify nodulation in maize/peanut intercropping and sole peanut cropping under four N fertilization rates (N-free, low, medium, and high N) in rain-fed water-limited conditions. In the micro-plot experiment, intercropped peanuts increased nodule biomass compared to sole peanuts. The nodule number of intercropped peanuts was 51.6% (p = 0.001) higher than that of sole cropped peanuts, while nodule weights did not differ at high N fertilization rates and were lower in the no-N fertilization control. However, the results were different in the field experiment. Both the nodule number and single weight of the sole cropped peanut were 48.7% (p = 0.020) and 58.9% (p = 0.014) higher than that of the intercropped peanut. The ratio of the nodule weight to aboveground dry matter at the beginning peg in the dry year of 2017 was lower in intercropping than sole cropping, especially at low N fertilization rates. The potential increase in nodulation found in a well-controlled micro-plot environment might be limited by strong water and light competitions in field conditions. The results could contribute to the understanding of interspecific interactions in cereal/legume intercropping.

Complete genome sequence of Bradyrhizobium sp. 62B, a native nitrogen-fixing rhizobium isolated from peanut nodules.

We present the complete genome sequence of Bradyrhizobium sp. 62B, a strain isolated from the root nodules of peanut plants that grow in central Argentina. The genome consists of 8.15 Mbp, distributed into a chromosome of 7.29 Mbp and a plasmid of 0.86 Mbp.

Effect of Three Different Nitrogen Rates and Three Rhizosphere N2- Fixing Bacteria on Growth, Yield and Quality of Peanuts

Raising prices of organic fertilizers, environmental relation of their usage and need for global foodstuff, placed a topic of universal interest in cultivation of legume plants for human nutrients and soil fertility supplementation. Three rates of N fertilizer were studied in the field: 0, 20 and 40 kg N/ha. Three bacterial strains were identified and inoculated in the experiment: Enterobacter asburiae (E. asburiae), Klebsiella quasipneumoniae (K. quasipneumoniae) and Enterobacter cloacae (E. cloacae). The soil samples were taken before and after experiment for chemical attributes analysis. The effect of N2 fertilizer and 3 bacterial strains was on soil chemical properties, yield components and yield of peanut plant. The results of present study showed that application of different N2 fertilizer rates was a little influence on the N2-fixing ability of rhizosphere bacteria, but high effect was significant at the peanut nodule number. In the 3 N fertilizer rates and 3 bacteria, which only had N2 fertilizer rate of 40 kg N/ha and E. asburiae significantly raised the maximum yield component and yield of peanut, while there was the lowest yield component and yield of peanut was observed in N fertilizer rate application of 0 kg N/ha and E. cloacae inoculation. The differences in rhizosphere bacterium strains between different N fertilizer rates was better than that caused by the nitrogen fertilizer amount. This could be due to the substantial improvement in soil chemical attributes at different soils, especially in, pH, OM and total N, as these significantly impacted the soil structure. HIGHLIGHTS Increasing pH, organic matter and total nitrogen of soil were statistically significantly different by 3 N2 fertilizer rates. There is a positive interaction between nitrogen rates and RNFB increasing yield of peanut. Number and weight of peanut nodules obtained the maximum value without applying N2 application compare to other treatments. The maximum yield components of peanut were inoculated with asburiae to compare with K. quasipneumoniae and E. cloacae and applied weight of 40 kg N/ha. The maximum yield of peanut was inoculated with asburiae to compare with K. quasipneumoniae and E. cloacae. GRAPHICAL ABSTRACT

Exchangeable Al, peanut growth, and nodulation on Ultisol Bogor as affected by ameliorant proportion of lime and organic matter

Ultisols are characterized as acid soils with high exchangeable Al as the primary constraint for plant growth. This study aimed to investigate the effect of the proportion of lime and organic matter in decreasing exchangeable Al on Ultisol Bogor and continued to observe the impact on the growth and nodulation of peanuts. The pot experiment used a completely randomized design with a single treatment of the addition of ameliorant consisting of 12 levels of the treatment of lime (calcite and dolomite), organic matter (dry cow dung and rice straw compost), and their combinations with three replications. The first step was applying the ameliorant and incubating for eight weeks, then continuing the cultivation of peanuts. Results showed that during incubation, the decrease in exchangeable Al levels varied between treatments, along with an increase in soil pH and available P. The treatments of calcite 100% (T1) and dolomite 100% (T2) showed the highest increase in soil pH and available P and the highest decrease in exchangeable Al. The result of plant growth showed that the highest plant dry weight, N-uptake, and P-uptake were obtained by the treatment of dolomite 25% + rice straw compost 25% + dry cow dung 25% + NPK 25% (T10), whereas the highest number and total weight of nodules were obtained by the rice straw compost 50% + dry cow dung 50% (T7) and dolomite 25% + rice straw compost 75% (T8).

Isolation and Characterization Identification of Edophytic Nitrogen-Fixing Bacteria from Peanut Nodules.

This work was carried out to isolate and perform molecular identification and selection of endophytic nitrogen-fixing bacteria (ENFB) to be utilized as biofertilizer. In this research, nodulous samples of peanuts were collected from inside dyke areas, namely, Phuoc Hung of An Phu, An Giang, Vietnam. Ten colonies were isolated from nutrient agar plates containing YMA's medium. All isolates were rod shaped, Gram negative, and no spore creation. Biochemical tests indicated that they were obligate aerobes, catalase, oxidase, urea hydrolysis, well motile ability, and no nitrate reduction. The salt tolerance observed that most survived at 0.5% and 2% salinity (except Enterobacter cloacae subsp. dissolvens strain LMG 2683), while at 4%, only 3 isolates (Bacillus aryabhattai strain CM44, Enterobacter asburiae strain IIWM-JS-07L, and Bacillus songklensis strain KCa6) and at 5% only, 2 isolates survived, namely, Enterobacter asburiae strain IIWM-JS-07L and Bacillus songklensis strain KCa6. The result showed that most of ten ENFB strains could adapt to the range of 25°C and 45°C (except Enterobacter cloacae subsp. dissolvens strain LMG 2683 and Enterobacter mori strain cjy13 at 25°C). Out of ten isolates, three were finally selected for the next studies, which potentially have N-fixing ability and are utilized as biofertilizer in agricultural cultivation.

Optimizing Initial Nitrogen Application Rates to Improve Peanut (Arachis hypogaea L.) Biological Nitrogen Fixation

The application of nitrogen fertilizer is crucial to the growth and biological nitrogen fixation of peanut, especially in the seedling stage where nodules have not yet formed. However, it is still uncertain how much initial nitrogen fertilizer should be applied to promote peanut root growth, nodule formation, and biological nitrogen fixation (BNF). There, a 2-year pot experiment was conducted using Huayu 22 (HY22, large-grain cultivar) and Huayu 39 (HY39, small-grain cultivar) as experimental materials to research the effects of different initial nitrogen fertilizer application rates on peanut root growth (root weight, root length, root mean diameter, root activity) and biological nitrogen fixation capacity (nodule number, nodule weight, biological nitrogen fixation, and nitrogen fixation potential per plant). N0, as control, four initial nitrogen fertilizer application rates were established: 15 kg·hm−2 (N15), 30 kg·hm−2 (N30), 45 kg·hm−2 (N45), and 60 kg·hm−2 (N60). The present results showed that the nodule number, nodule dry weight, nitrogenase activity, and biological nitrogen fixation of the HY22 cultivar under the N15 treatment were higher compared to those under other treatments over the two growing seasons. In addition, the cultivar of HY39 treated with the N15 treatment also increased the nitrogen fixation potential per plant and BNF relative to other treatments. Although the application of 60 kg·hm−2 nitrogen increased the root surface area and root volume, it decreased the nitrogenase activity, nodule dry weight, and nitrogen fixation potential per plant of HY22 and HY39 varieties in both growing seasons. Above all, an initial nitrogen application of 15 kg·hm−2 may be the optimal treatment for promoting peanut nodule formation and biological nitrogen fixation.

Effect of Reducing Nitrogen Fertilization and Adding Organic Fertilizer on Net Photosynthetic Rate, Root Nodules and Yield in Peanut.

Long-term excessive application of chemical fertilizers can cause many problems, such as soil degradation and environmental pollution. Therefore, we reduced conventional nitrogen fertilization and added organic fertilizers in some cases to investigate the response of photosynthetic characteristics, root nodules and yield on reduced nitrogen fertilization. Compared to conventional nitrogen fertilization, the 25% and 35% nitrogen reduction treatments reduced the leaf area index, net photosynthetic rate, 100-fruit weight, 100-kernel weight and the yield of peanut, but had no significant effect on the kernel rate. With constant N fertilizer, adding organic fertilization alone increased leaf area index, chlorophyll, net photosynthetic rate and yield of peanut. In compounded treatments of nitrogen and organic fertilizer, the highest yields were achieved in the 25% N reduction with the 3000 kg/hm-2 organic fertilizer treatment (T3) and the 4500 kg/hm-2 organic fertilizer treatment (T4); furthermore, the net photosynthetic rate, leaf area index, yield and fertilizer contribution were significantly higher in these two treatments than in the conventional fertilizer treatments. Nitrogen fertilizer had significant effects on the quantity and fresh weight of root nodules. Concretely, nitrogen reduction increased the quantity and fresh weight of root nodules of peanut in the early stage of fertility but decreased them in the harvest stage. Nitrogen reduction with an additional organic fertilizer in the late stage of fertility increased the quantity and fresh weight of root nodules of peanut. Considering the property of root nodules was significantly positively correlated with net photosynthetic rate and yield, the arguments above may be the mechanism of the highest yields found in T3 and T4. This work can provide empirical and instructional support for a balanced fertilization strategy in peanut agriculture and high-yielding and efficient cultivation of peanut.

Isolation and Characterization of Bacterial Endophytes from Small Nodules of Field-Grown Peanut.

It is evident that legume root nodules can accommodate rhizobial and non-rhizobial bacterial endophytes. Our recent nodule microbiome study in peanuts described that small nodules can harbor diverse bacterial endophytes. To understand their functional role, we isolated 87 indigenous endophytes from small nodules of field-grown peanut roots and characterized them at molecular, biochemical, and physiological levels. The amplified 16S rRNA genes and phylogenetic analysis of these isolates revealed a wide variety of microorganisms related to the genera Bacillus, Burkholderia, Enterobacter, Herbaspirillum, Mistsuaria, Pantoea, Pseudomonas, and Rhizobia. It was observed that 37% (100% identity) and 56% (>99% identity) of the isolates matched with the amplified sequence variants (ASVs) from our previous microbiome study. All of these isolates were tested for stress tolerance (high temperature, salinity, acidic pH) and phosphate (P) solubilization along with ammonia (NH3), indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate deaminase (ACCD), and siderophore production. The majority (78%) of the isolates were found to be halotolerant, thermotolerant, and acidophilic, and a few of them showed a significant positive response to the production of IAA, NH3, siderophore, ACCD, and P-solubilization. To evaluate the plant growth promotion (PGP) activity, plant and nodulation assays were performed in the growth chamber conditions for the selected isolates from both the non-rhizobial and rhizobial groups. However, these isolates appeared to be non-nodulating in the tested conditions. Nonetheless, the isolates 2 (Pantoea), 17 (Burkholderia), 21 (Herbaspirillum), 33o (Pseudomonas), and 77 (Rhizobium sp.) showed significant PGP activity in terms of biomass production. Our findings indicate that these isolates have potential for future biotechnological applications through the development of biologicals for sustainable crop improvement.

Response of peanut quality and yield to chicken manure combined with Rhizobium inoculation in sandy soil

Chemical fertilizers, which contain a nitrogen (N) element, has been intensively used to increase the peanut productivity. However, the unstable and high cost of N fertilizer, and the great demand for N fertilizer sources have strongly increased the strategical plan of nitrogen fixation (NF). Therefore, the field research was carried out to appraise the ability of Rhizobium sp. trains and chicken manure (CM) on the quality and yield of peanuts. This research has four ratios, which valued from 0.0, 2.0, 4.0 to 6.0 t CM per ha in the combination with the Rhizobium sp. inoculum, expect control treatment (without CM and Rhizobium sp.). Different rates of CM combined with Rhizobium sp. inoculation was added by using 6.0 tons CM/ ha, which had number of the highest peanut nodules. Research results observed that the inoculant of Rhizobium sp. strain combined with CM remarkably increased the yield components per plant such as biomass, number of nodules, weight of dry nodules, weight of fill and empty pods and fresh yield of groundnut. The highest yield and quality of peanut (7.60 t/ha), oil % (50.6%), seed protein percentage (26.8%), as well as NPK content in seed (4.32, 0.912 and 0.999%, respectively) were obtained under the application of NPK+6.0 t CM/ha+ Rhizobium sp. inoculation. Co-application of 6t CM/ha and Rhizobium sp. inoculation increased by 20.5% when compared without CM application and no Rhizobium sp. inoculation. The study showed that both possibility of nitrogen fixation of peanut and nitrogen uptake of the sandy soil were raised by field inoculant with effectiveness of Rhizobium sp. with animal manure application. In really, Rhizobium sp. inoculation and CM proved a great method to increase soil nutrients for subsequent crops and it helped to enhance the taking of nitrogen from the air into the crop soil.

Allelic expression of AhNSP2-B07 due to parent of origin affects peanut nodulation.

Legumes are well-known for establishing a symbiotic relationship with rhizobia in root nodules to fix nitrogen from the atmosphere. The nodulation signaling pathway 2 (NSP2) gene plays a critical role in the symbiotic signaling pathway. In cultivated peanut, an allotetraploid (2n = 4x = 40, AABB) legume crop, natural polymorphisms in a pair of NSP2 homoeologs (Na and Nb) located on chromosomes A08 and B07, respectively, can cause loss of nodulation. Interestingly, some heterozygous (NBnb) progeny produced nodules, while some others do not, suggesting non-Mendelian inheritance in the segregating population at the Nb locus. In this study, we investigated the non-Mendelian inheritance at the NB locus. Selfing populations were developed to validate the genotypical and phenotypical segregating ratios. Allelic expression was detected in roots, ovaries, and pollens of heterozygous plants. Bisulfite PCR and sequencing of the Nb gene in gametic tissue were performed to detect the DNA methylation variations of this gene in different gametic tissues. The results showed that only one allele at the Nb locus expressed in peanut roots during symbiosis. In the heterozygous (Nbnb) plants, if dominant allele expressed, the plants produced nodules, if recessive allele expressed, then no nodules were produced. qRT-PCR experiments revealed that the expression of Nb gene in the ovary was extremely low, about seven times lower than that in pollen, regardless of genotypes or phenotypes of the plants at this locus. The results indicated that Nb gene expression in peanut depends on the parent of origin and is imprinted in female gametes. However, no significant differences of DNA methylation level were detected between these two gametic tissues by bisulfite PCR and sequencing. The results suggested that the remarkable low expression of Nb in female gametes may not be caused by DNA methylation. This study provided a unique genetic basis of a key gene involved in peanut symbiosis, which could facilitate understanding the regulation of gene expression in symbiosis in polyploid legumes.

Transcriptomic and Metabolomic Analyses Reveal the Roles of Flavonoids and Auxin on Peanut Nodulation.

Rhizobia form symbiotic relationships with legumes, fixing atmospheric nitrogen into a plant-accessible form within their root nodules. Nitrogen fixation is vital for sustainable soil improvements in agriculture. Peanut (Arachis hypogaea) is a leguminous crop whose nodulation mechanism requires further elucidation. In this study, comprehensive transcriptomic and metabolomic analyses were conducted to assess the differences between a non-nodulating peanut variety and a nodulating peanut variety. Total RNA was extracted from peanut roots, then first-strand and second-strand cDNA were synthesized and purified. After sequencing adaptors were added to the fragments, the cDNA libraries were sequenced. Our transcriptomic analysis identified 3362 differentially expressed genes (DEGs) between the two varieties. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that the DEGs were mainly involved in metabolic pathways, hormone signal transduction, secondary metabolic biosynthesis, phenylpropanoid biosynthesis, or ABC transport. Further analyses indicated that the biosynthesis of flavonoids, such as isoflavones, flavonols, and flavonoids, was important for peanut nodulation. A lack of flavonoid transport into the rhizosphere (soil) could prevent rhizobial chemotaxis and the activation of their nodulation genes. The downregulation of AUXIN-RESPONSE FACTOR (ARF) genes and lower auxin content could reduce rhizobia's invasion of peanut roots, ultimately reducing nodule formation. Auxin is the major hormone that influences the cell-cycle initiation and progression required for nodule initiation and accumulates during different stages of nodule development. These findings lay the foundation for subsequent research into the nitrogen-fixation efficiency of peanut nodules.

GFP labeling of a Bradyrhizobium strain and an attempt to track the crack entry process during symbiosis with peanuts.

Compared to the well-studied model legumes, where symbiosis is established via root hair entry, the peanut is infected by Bradyrhizobium through the crack entry, which is less common and not fully understood. Crack entry is, however, considered a primitive symbiotic infection pathway, which could be potentially utilized for engineering non-legume species with nitrogen fixation ability. We utilized a fluorescence-labeled Bradyrhizobium strain to help in understanding the crack entry process at the cellular level. A modified plasmid pRJPaph-bjGFP, harboring the codon-optimized GFP gene and tetracycline resistance gene, was created and conjugated into Bradyrhizobium strain Lb8, an isolate from peanut nodules, through tri-parental mating. Microscopic observation and peanut inoculation assays confirmed the successful GFP tagging of Lb8, which is capable of generating root nodules. A marking system for peanut root potential infection sites and an optimized sample preparation protocol for cryostat sectioning was developed. The feasibility of using the GFP-tagged Lb8 for observing crack entry was examined. GFP signal was detected at the nodule primordial stage and the following nodule developmental stages with robust GFP signals observed in infected cells in the mature nodules. Spherical bacteroids in the root tissue were visualized at the nodules' inner cortex under higher magnification, reflecting the trace along the rhizobial infection path. The GFP labeled Lb8 can serve as an essential tool for plant-microbe studies between the cultivated peanut and Bradyrhizobium, which could facilitate further study of the crack entry process during the legume-rhizobia symbiosis.

Isolation of Peanut (<i>Arachis hypogaea</i> L.) Nodulating Rhizobia and Assessment of Their Phosphate Solubilizing Activity

Phosphate-solubilizing bacteria can enhance the dissolution of insoluble phosphorus in the soil, promoting the availability of soluble phosphorus. Thus, their application can reduce the use of chemical fertilizers and ease sustainable agriculture. The study aimed at isolating and assessing the phosphate solubilizing activity of peanut rhizobia isolated from root nodules of peanut in three Cameroonian soils (Douala, Bafoussam and Ebolowa). Rhizobia were trapped by seeding peanut in plastic pots containing different soils. 45 days later, at the flowering time, root nodules were harvested. From these nodules, rhizobia were isolated, purified on Yeast Extract Mannitol Agar (YEMA) medium and authenticated by inoculating them on sterile sand containing peanut plants and watered with a nutrient solution without nitrogen. The phosphate solubilizing activity of the rhizobia isolates was then assessed in Modified Mineral Salt Medium (MMSM) containing bromocresol green and where the soluble phosphate was replaced by insoluble inorganic phosphates from Cameroon, Algeria, Senegal and tricalcium phosphate in both solid and broth media. The results were analysed statistically by one-way ANOVA using SPSS. A collection of twenty-five isolates was constituted among which 04 from Douala, 12 from Bafoussam and 09 from Ebolowa. All isolates were distinct morphologically. There was significant solubilization of inorganic phosphate in both solid and broth media (p<0.05). Isolated bacteria were characterized as being phosphate solubilizers with values ranging from 1.75 (AhBf1 on Algeria rock phosphate) to 18.9 mg/L (AhDa3 on Algeria rock phosphate). The Algeria rock phosphate was the most solubilized by peanut nodulating bacteria, followed by the tricalcium phosphate, the Senegal rock phosphate and the Cameroon rock phosphate. The solubilizing activity of inorganic phosphates in the broth medium was associated with a global decrease in the pH of the culture medium. The peanut nodulating rhizobia isolates that show the best solubilizing capacity could alleviate the problem of phosphorus availability in agricultural soils.

Mapping of QTLs Associated with Biological Nitrogen Fixation Traits in Peanuts (Arachis hypogaea L.) Using an Interspecific Population Derived from the Cross between the Cultivated Species and Its Wild Ancestors.

Peanuts (Arachis hypogaea L.) are an allotetraploid grain legume mainly cultivated by poor farmers in Africa, in degraded soil and with low input systems. Further understanding nodulation genetic mechanisms could be a relevant option to facilitate the improvement of yield and lift up soil without synthetic fertilizers. We used a subset of 83 chromosome segment substitution lines (CSSLs) derived from the cross between a wild synthetic tetraploid AiAd (Arachis ipaensis × Arachis duranensis)4× and the cultivated variety Fleur11, and evaluated them for traits related to BNF under shade-house conditions. Three treatments were tested: without nitrogen; with nitrogen; and without nitrogen, but with added0 Bradyrhizobium vignae strain ISRA400. The leaf chlorophyll content and total biomass were used as surrogate traits for BNF. We found significant variations for both traits specially linked to BNF, and four QTLs (quantitative trait loci) were consistently mapped. At all QTLs, the wild alleles decreased the value of the trait, indicating a negative effect on BNF. A detailed characterization of the lines carrying those QTLs in controlled conditions showed that the QTLs affected the nitrogen fixation efficiency, nodule colonization, and development. Our results provide new insights into peanut nodulation mechanisms and could be used to target BNF traits in peanut breeding programs.

Microbiome analysis revealed distinct microbial communities occupying different sized nodules in field-grown peanut.

Legume nodulation is the powerhouse of biological nitrogen fixation (BNF) where host-specific rhizobia dominate the nodule microbiome. However, other rhizobial or non-rhizobial inhabitants can also colonize legume nodules, and it is unclear how these bacteria interact, compete, or combinedly function in the nodule microbiome. Under such context, to test this hypothesis, we conducted 16S-rRNA based nodule microbiome sequencing to characterize microbial communities in two distinct sized nodules from field-grown peanuts inoculated with a commercial inoculum. We found that microbial communities diverged drastically in the two types of peanut nodules (big and small). Core microbial analysis revealed that the big nodules were inhabited by Bradyrhizobium, which dominated composition (>99%) throughout the plant life cycle. Surprisingly, we observed that in addition to Bradyrhizobium, the small nodules harbored a diverse set of bacteria (~31%) that were not present in big nodules. Notably, these initially less dominant bacteria gradually dominated in small nodules during the later plant growth phases, which suggested that native microbial communities competed with the commercial inoculum in the small nodules only. Conversely, negligible or no competition was observed in the big nodules. Based on the prediction of KEGG pathway analysis for N and P cycling genes and the presence of diverse genera in the small nodules, we foresee great potential of future studies of these microbial communities which may be crucial for peanut growth and development and/or protecting host plants from various biotic and abiotic stresses.

The Relationship between Core Rhizosphere Taxa and Peanut Nodulation Capacity under Different Cover Crop Amendments

Adequate exploitation of legume–rhizobia symbiosis for nitrogen fixation may help to alleviate the overuse of chemical nitrogen fertilizer and aid in sustainable agricultural development. However, controlling this beneficial interaction requires thorough characterization of the effects of soil rhizosphere microorganisms, especially core taxa, on the legume–rhizobia symbiosis. Here, we used Illumina sequencing to investigate the effects of cover crop (Raphanus sativus L. and Lolium perenne L.) residue on the rhizosphere soil microbial community and peanut nodulation ability. The results indicated that Raphanus sativus L. amendment (RS) significantly increased soil available phosphorus (AP) content and peanut nodulation ability, while the Lolium perenne L. amendment (LP) had no noticeable impact on peanut nodulation. LP and RS significantly elevated bacterial and rhizobial diversity, reduced fungal diversity, and shifted microbial community structure (bacteria, 14.7%, p = 0.001; rhizobia, 21.7%, p = 0.001; fungi, 25.5%, p = 0.001). Random forest analysis found that the core rhizosphere taxa, sharing similar ecological preferences, were the primary drivers of peanut nodulation. By least squares regression, soil AP content was found to be positively correlated with the relative abundance of key ecological clusters. Furthermore, RS was found to promote peanut nodulation by increasing the relative abundance of critical rhizosphere taxa. Overall, our findings emphasize that core microbial taxa might play an essential function in the modulation of legume nodulation and provide scientific evidence for the effective management of the plant microbiome.

Bacteroid Development, Transcriptome, and Symbiotic Nitrogen-Fixing Comparison of Bradyrhizobium arachidis in Nodules of Peanut (Arachis hypogaea) and Medicinal Legume Sophora flavescens.

Bradyrhizobium arachidis strain CCBAU 051107 could differentiate into swollen and nonswollen bacteroids in determinate root nodules of peanut (Arachis hypogaea) and indeterminate nodules of Sophora flavescens, respectively, with different N2 fixation efficiencies. To reveal the mechanism of bacteroid differentiation and symbiosis efficiency in association with different hosts, morphologies, transcriptomes, and nitrogen fixation efficiencies of the root nodules induced by strain CCBAU 051107 on these two plants were compared. Our results indicated that the nitrogenase activity of peanut nodules was 3 times higher than that of S. flavescens nodules, demonstrating the effects of rhizobium-host interaction on symbiotic effectiveness. With transcriptome comparisons, genes involved in biological nitrogen fixation (BNF) and energy metabolism were upregulated, while those involved in DNA replication, bacterial chemotaxis, and flagellar assembly were significantly downregulated in both types of bacteroids compared with those in free-living cells. However, expression levels of genes involved in BNF, the tricarboxylic acid (TCA) cycle, the pentose phosphate pathway, hydrogenase synthesis, poly-β-hydroxybutyrate (PHB) degradation, and peptidoglycan biosynthesis were significantly greater in the swollen bacteroids of peanut than those in the nonswollen bacteroids of S. flavescens, while contrasting situations were found in expression of genes involved in urea degradation, PHB synthesis, and nitrogen assimilation. Especially higher expression of ureABEF and aspB genes in bacteroids of S. flavescens might imply that the BNF product and nitrogen transport pathway were different from those in peanut. Our study revealed the first differences in bacteroid differentiation and metabolism of these two hosts and will be helpful for us to explore higher-efficiency symbiosis between rhizobia and legumes. IMPORTANCE Rhizobial differentiation into bacteroids in leguminous nodules attracts scientists to investigate its different aspects. The development of bacteroids in the nodule of the important oil crop peanut was first investigated and compared to the status in the nodule of the extremely promiscuous medicinal legume Sophora flavescens by using just a single rhizobial strain of Bradyrhizobium arachidis, CCBAU 051107. This strain differentiates into swollen bacteroids in peanut nodules and nonswollen bacteroids in S. flavescens nodules. The N2-fixing efficiency of the peanut nodules is three times higher than that of S. flavescens. By comparing the transcriptomes of their bacteroids, we found that they have similar gene expression spectra, such as nitrogen fixation and motivity, but different spectra in terms of urease activity and peptidoglycan biosynthesis. Those altered levels of gene expression might be related to their functions and differentiation in respective nodules. Our studies provided novel insight into the rhizobial differentiation and metabolic alteration in different hosts.

Root Nodule Categorization and Their Relation with Plant Growth in Peanut Crop Grown in Alfisols

To investigate the relationship between nodule category and plant growth characteristics in peanut, nodules were classified into three categories: large size nodule (LSN) ( greater than 2 mm), medium size nodule (MSN) (1-2 mm) and small size nodule (SSN) ( less than 1 mm) and their position on roots (primary and lateral roots). The number of LSNs on primary roots was found to be much higher than on lateral roots. The number of MSN developed on primary and lateral roots was comparable, whereas, lateral roots had a higher SSN number. In addition, when comparing the LSN and MSN numbers to SSN number, the plant growth parameters showed a high positive correlation. Multivariate regression analysis yielded similar results. This nodule classification technique aids in the identification of peanut nodules and their role in quantification of plant growth characteristics. Further, this technique can be used in identifying/screening of efficient nodulating peanut genotypes.