Rhizobium-legume Symbiosis Research Articles

ECONOMIC AND BIOENERGY EFFICIENCY OF CHICKPEA BACTERIZATION BY MESORHIZOBIUM CICERІ ND-64

Objective. Study the effect of chickpea seed inoculation with a suspension of Mesorhizobiumciceri ND-64 on crop yields, calculate the economic and bioenergy efficiency of seed bacterization.Methods. Microbiological, field experiment, economic, statistical. Results. It was shown that presowing inoculation of chickpea seeds with M. ciceri ND-64 during cultivation of this culture inPolissia of Ukraine conditions contributed to the increased efficiency of legume-rhizobial symbiosis. During three years of research there was an increase in structural parameters: the numberof beans per plant — by 15–42 %, seeds from the plant — by 12–32 %, weight of seeds from theplant — by 12–37 %, weight of 1000 seeds — by 6–9 % relative to positive (with inoculation usingreference strain of M. ciceri H-12) and absolute (without inoculation) controls, respectively. Theyield of Pamiat variety chickpeas increased by 12.8–26.5 %. The amount of profit when bacterialsuspension of M. ciceri ND-64 was applied for seed bacterization per 1 ha of sown area was UAH10,795 and the level of profitability was 106.1 %. The payback of each hryvnia of additional costswith additional income was UAH 19.43 after inoculation with M. ciceri H-12 and UAH 30.88 after M. ciceri ND-64. With anthropogenic energy consumption of 10,479 MJ in the variant with bacterization using M. ciceri ND-64 per 1 ha of sown area, the energy content of the obtained grain was26,181 MJ, respectively, the energy efficiency coefficient was 2.50. The additional energy costs associated with the use of M. ciceri ND-64 were 16.56 times paid back by the energy of the additionalcrop. Conclusion. The use of M. ciceri ND-64 for pre-sowing inoculation of chickpea seeds ofPamiat variety provided an increase in the number of seeds from the plant by 42 %, weight of seedsfrom the plant by 32 %, weight of 1000 seeds by 9 % and increase crop yields by 26.5 % relative tocontrol when the crop was cultivated in Polissia area during three years of research. All parameters of economic and energy efficiency of pre-sowing bacterization with M. ciceri ND-64 are moreeffective in comparison with inoculation using M. ciceri H-12.

EFFECT OF FUNGICIDES ON THE EFFICIENCY OF SOYBEAN INOCULATION WITH PESTICIDE-RESISTANT NODULE BACTERIA

Objective. Evaluate the efficiency of soybean inoculation with pesticide-resistant nodule bacteria Bradyrhizobium japonicum under the influence of pre-sowing seed dressing with Maxym XL,Standak Top and Fever and preventive spraying of plants with the fungicide Akanto Plus. Methods. Physiological, microbiological, gas chromatographic, vegetation experiment, statistical.Results. The number and weight of nodules formed on soybean roots during the growing seasonunder the action of the prothioconazole-based fungicide Fever were reported to be lower comparedwith the control and other variants of experiment. Pesticides Maxym XL and Standak Top had a lesspronounced toxic effect on the formation of legume-rhizobial symbiosis with the participation offungicide-resistant B. japonicum B78 Tn5 mutant compared to Fever. A decrease in the level of N2assimilation by 10.9–41.1 % in the three-leaf phase was found with the combined use of seed pesticides and nodule bacteria included in the study, relative to the variant with inoculation only. However, during the growing season, the negative effect of fungicidal preparations on the functioning oflegume-rhizobial symbiosis decreased. Conclusion. Under the conditions of the model vegetationexperiment with the concomitant use of Standak Top and Fever and bacterization, a decrease in individual plant productivity by 7.0 and 14.1 %, respectively, has been reported. The complex actionof the pesticide Maxym XL and inoculation with pesticide-resistant B. japonicum B78 transposonmutant, contributed to the maximum increase in yield by 27.2 %. The obtained data confirm the expediency of selection of fungicides and nitrogen-fixing microorganisms for concomitant use to ensure high economic and protective effects.

Anthocyanin, a novel and user-friendly reporter for convenient, non-destructive, low cost, directly visual selection of transgenic hairy roots in the study of rhizobia-legume symbiosis

BackgroundAgrobacterium rhizogenes-mediated hairy root transformation provides a powerful tool for investigating the functions of plant genes involved in rhizobia-legume symbiosis. However, in the traditional identification methods of transgenic hairy roots based on reporter genes, an expensive chemical substrate or equipment is required.ResultsHere, we report a novel, low cost, and robust reporter for convenient, non-destructive, and directly visual selection of transgenic hairy roots by naked eye, which can be used in the study of rhizobia-legume symbiosis. The reporter gene AtMyb75 in Arabidopsis, encoding an R2R3 type MYB transcription factor, was ectopically expressed in hairy roots-mediated by A. rhizogenes, which induced purple/red colored anthocyanin accumulation in crop species like soybean (Glycine max (L.) Merr.) and two model legume species, Lotus japonicas and Medicago truncatula. Transgenic hairy roots of legumes containing anthocyanin can establish effective symbiosis with rhizobia. We also demonstrated the reliability of AtMyb75 as a reporter gene by CRISPR/Cas9-targeted mutagenesis of the soybean resistance to nodulation Rfg1 gene in the soybean PI377578 (Nod-) inoculated with Sinorhizobium fredii USDA193. Without exception, mature nitrogen-fixation nodules, were formed on purple transgenic hairy roots containing anthocyanin.ConclusionsAnthocyanin is a reliable, user-friendly, convenient, non-destructive, low cost, directly visual reporter for studying symbiotic nitrogen-fixing nodule development and could be widely applied in broad leguminous plants.

A Methionine Sulfoxide Reductase B Is Required for the Establishment of Astragalus sinicus-Mesorhizobium Symbiosis.

Methionine sulfoxide reductase B (MsrB) is involved in oxidative stress or defense responses in plants. However, little is known about its role in legume-rhizobium symbiosis. In this study, an MsrB gene was identified from Astragalus sinicus and its function in symbiosis was characterized. AsMsrB was induced under phosphorus starvation and displayed different expression patterns under symbiotic and nonsymbiotic conditions. Hydrogen peroxide or methyl viologen treatment enhanced the transcript level of AsMsrB in roots and nodules. Subcellular localization showed that AsMsrB was localized in the cytoplasm of onion epidermal cells and co-localized with rhizobia in nodules. Plants with AsMsrB-RNAi hairy roots exhibited significant decreases in nodule number, nodule nitrogenase activity and fresh weight of the aerial part, as well as an abnormal nodule and symbiosome development. Statistical analysis of infection events showed that plants with AsMsrB-RNAi hairy roots had significant decreases in the number of root hair curling events, infection threads and nodule primordia compared with the control. The content of hydrogen peroxide increased in AsMsrB-RNAi roots but decreased in AsMsrB overexpression roots at the early stage of infection. The transcriptome analysis showed synergistic modulations of the expression of genes involved in reactive oxygen species generation and scavenging, defense and pathogenesis and early nodulation. In addition, a candidate protein interacting with AsMsrB was identified and confirmed by bimolecular fluorescence complementation. Taken together, our results indicate that AsMsrB plays an essential role in nodule development and symbiotic nitrogen fixation by affecting the redox homeostasis in roots and nodules.

СИМБІОТИЧНА ПРОДУКТИВНІСТЬ ГОРОХУ ПОСІВНОГО ЗАЛЕЖНО ВІД ВПЛИВУ МІНЕРАЛЬНИХ ДОБРИВ ТА РЕГУЛЯТОРІВ РОСТУ В УМОВАХ ЛІСОСТЕПУ ЗАХІДНОГО

У статті представлено результати досліджень із вивчення залежності формування симбіотичної продуктивності гороху посівного від рівня удобрення різними дозами мінеральних добрив та регуляторів росту в умовах Лісостепу Західного. Експерементальну частину проводили впродовж 2016-2018 рр. на дослідному полі Навчально-виробничого центру «Поділля» ПДАТУ, на чорноземі типовому, глибокому, малогумусному важкосуглинкову на лесовидних суглинках в умовах польового досліду. У ході наших досліджень, встановлено, що продуктивність гороху посівного в значній мірі залежала від ефективної взаємодії рослини-господаря та бульбочкових бактерій в оптимальних умовах. Внесення фосфорно-калійних добрив та невеликих доз азотних добрив покращували біологічну фіксацію азоту, а обприскування рослин регуляторами росту у фазі бутонізації – цвітіння продовжило функціонування червоного пігменту синтезованого леггемоглобіну у бульбочках. В результаті проведених обрахунків встановлено, позитивний вплив дії мінеральних добрив та регуляторів росту на показники загального та активного симбіотичних потенціалів рослин гороху посівного. Загальний симбіотичний потенціал характеризує стан бобоворизобіального симбіозу за вегетаційний період. Тривалість активного функціонування бульбочок на коренях гороху та здатність їх фіксувати біологічний азот протягом вегетаційного періоду є величина активного симбіотичного потенціалу. Всього за вегетаційний період показники загального симбіотичного потенціалу були найвищими у сорту гороху Чекбек, і становили - 13,1 тис. кг*діб/га на варіанті удобрення N30P30K45 у поєднанні з регулятором росту Вимпел. Для сорту Готівський на цьому ж варіанті живлення – 10,6 тис. кг*діб/га, у сорту Фаргус - 10,0 тис. кг*діб/га.

Unexplored Arsenals of Legume Peptides With Potential for Their Applications in Medicine and Agriculture.

During endosymbiosis, bacteria live intracellularly in the symbiotic organ of their host. The host controls the proliferation of endosymbionts and prevents their spread to other tissues and organs. In Rhizobium-legume symbiosis the major host effectors are secreted nodule-specific cysteine-rich (NCR) peptides, produced exclusively in the symbiotic cells. NCRs have evolved in the Inverted Repeat Lacking Clade (IRLC) of the Leguminosae family. They are secreted peptides that mediate terminal differentiation of the endosymbionts, forming polyploid, non-cultivable cells with increased membrane permeability. NCRs form an extremely large family of peptides, which have four or six conserved cysteines but otherwise highly diverse amino acid sequences, resulting in a wide variety of anionic, neutral and cationic peptides. In vitro, many synthetic NCRs have strong antimicrobial activities against both Gram-negative and Gram-positive bacteria, including the ESKAPE strains and pathogenic fungi. The spectra and minimal bactericidal and anti-fungal concentrations of NCRs differ, indicating that, in addition to their charge, the amino acid composition and sequence also play important roles in their antimicrobial activity. NCRs attack the bacteria and fungi at the cell envelope and membrane as well as intracellularly, forming interactions with multiple essential cellular machineries. NCR-like peptides with similar symbiotic functions as the NCRs also exist in other branches of the Leguminosae family. Thus, legumes provide countless and so far unexplored sources of symbiotic peptides representing an enormous resource of pharmacologically interesting molecules.

The Lotus japonicus Ubiquitin Ligase SIE3 Interacts With the Transcription Factor SIP1 and Forms a Homodimer.

The symbiosis receptor kinase SymRK plays an essential role in symbiotic signal transduction and nodule organogenesis. Several proteins bind to SymRK, but how the symbiosis signals are transduced from SymRK to downstream components remains elusive. We previously demonstrated that both SymRK interacting protein 1 (SIP1, an ARID-type DNA-binding protein) and SymRK interacting E3 ligase [SIE3, a RING (Really Interesting New Gene)-containing E3 ligase] interact with SymRK to regulate downstream cellular responses in Lotus japonicus during the legume-rhizobia symbiosis. Here, we show that SIE3 interacts with SIP1 in both yeast cells and Nicotiana benthamiana. SIE3 associated with itself and formed a homodimer. The cysteine 266 residue was found to be essential for SIE3 dimerization and for promoting nodulation in transgenic hairy roots of L. japonicus. Our findings provide a foundation for further investigating the regulatory mechanisms of the SymRK-mediated signaling pathway, as well as the biological function of E3 ligase dimerization in nodule organogenesis.

In Vivo Metabolic Regulation of Alternative Oxidase under Nutrient Deficiency-Interaction with Arbuscular Mycorrhizal Fungi and Rhizobium Bacteria.

The interaction of the alternative oxidase (AOX) pathway with nutrient metabolism is important for understanding how respiration modulates ATP synthesis and carbon economy in plants under nutrient deficiency. Although AOX activity reduces the energy yield of respiration, this enzymatic activity is upregulated under stress conditions to maintain the functioning of primary metabolism. The in vivo metabolic regulation of AOX activity by phosphorus (P) and nitrogen (N) and during plant symbioses with Arbuscular mycorrhizal fungi (AMF) and Rhizobium bacteria is still not fully understood. We highlight several findings and open questions concerning the in vivo regulation of AOX activity and its impact on plant metabolism during P deficiency and symbiosis with AMF. We also highlight the need for the identification of which metabolic regulatory factors of AOX activity are related to N availability and nitrogen-fixing legume-rhizobia symbiosis in order to improve our understanding of N assimilation and biological nitrogen fixation.

The Role of Nitric Oxide in Nitrogen Fixation by Legumes.

The legume-rhizobia symbiosis is an important process in agriculture because it allows the biological nitrogen fixation (BNF) which contributes to increasing the levels of nitrogen in the soil. Nitric oxide (⋅NO) is a small free radical molecule having diverse signaling roles in plants. Here we present and discuss evidence showing the role of ⋅NO during different stages of the legume-rhizobia interaction such as recognition, infection, nodule development, and nodule senescence. Although the mechanisms by which ⋅NO modulates this interaction are not fully understood, we discuss potential mechanisms including its interaction with cytokinin, auxin, and abscisic acid signaling pathways. In matures nodules, a more active metabolism of ⋅NO has been reported and both the plant and rhizobia participate in ⋅NO production and scavenging. Although ⋅NO has been shown to induce the expression of genes coding for NITROGENASE, controlling the levels of ⋅NO in mature nodules seems to be crucial as ⋅NO was shown to be a potent inhibitor of NITROGENASE activity, to induce nodule senescence, and reduce nitrogen assimilation. In this sense, LEGHEMOGLOBINS (Lbs) were shown to play an important role in the scavenging of ⋅NO and reactive nitrogen species (RNS), potentially more relevant in senescent nodules. Even though ⋅NO can reduce NITROGENASE activity, most reports have linked ⋅NO to positive effects on BNF. This can relate mainly to the regulation of the spatiotemporal distribution of ⋅NO which favors some effects over others. Another plausible explanation for this observation is that the negative effect of ⋅NO requires its direct interaction with NITROGENASE, whereas the positive effect of ⋅NO is related to its signaling function, which results in an amplifier effect. In the near future, it would be interesting to explore the role of environmental stress-induced ⋅NO in BNF.

Transcriptional regulation of NIN expression by IPN2 is required for root nodule symbiosis in Lotus japonicus.

Expression of Nodule Inception (NIN) is essential for initiation of legume-rhizobial symbiosis. An existing model regarding the regulation of NIN expression involves two GRAS transcription factors - NSP1 (Nodulation Signaling Pathway 1) and NSP2. NSP2 forms a complex with NSP1 to directly bind to NIN promoter. However, rhizobial treatment-induced NIN expression could still be detected inthe nsp1 mutant plants, suggesting that other proteins must be involved inthe regulation of NIN expression. A combination of molecular, biochemical and genetic analyses was used to investigate the molecular basis of IPN2 in regulating root development and NIN expression in Lotus japonicus. In this study, we identified that IPN2 is a close homolog of Arabidopsis APL (ALTERED PHLOEM DEVELOPMENT) with essential function in root development. However, Lotus IPN2 has a different expression pattern compared with the Arabidopsis APL gene. IPN2 binds to the IPN2-responsive cis element (IPN2-RE) of NIN promoter and activates NIN expression. IPN2, NSP1 and NSP2 form a protein complex to directly target NIN promoter and activate NIN expression in the legume-rhizobial symbiosis. Our data refine the regulatory model of NIN expression that NSP2 works together with NSP1 and IPN2 to activate the NIN gene allowing nodulation in L.japonicus.

QTL Mapping and Data Mining to Identify Genes Associated With the Sinorhizobium fredii HH103 T3SS Effector NopD in Soybean.

In some legume–rhizobium symbioses, host specificity is influenced by rhizobial type III effectors-nodulation outer proteins (Nops). However, the genes encoding host proteins that interact with Nops remain unknown. In this study, we aimed to identify candidate soybean genes associated with NopD, one of the type III effectors of Sinorhizobium fredii HH103. The results showed that the expression pattern of NopD was analyzed in rhizobia induced by genistein. We also found NopD can be induced by TtsI, and NopD as a toxic effector can induce tobacco leaf death. In 10 soybean germplasms, NopD played a positively effect on nodule number (NN) and nodule dry weight (NDW) in nine germplasms, but not in Kenjian28. Significant phenotype of NN and NDW were identified between Dongnong594 and Charleston, Suinong14 and ZYD00006, respectively. To map the quantitative trait locus (QTL) associated with NopD, a recombinant inbred line (RIL) population derived from the cross between Dongnong594 and Charleston, and chromosome segment substitution lines (CSSLs) derived from Suinong14 and ZYD00006 were used. Two overlapping conditional QTL associated with NopD on chromosome 19 were identified. Two candidate genes were identified in the confident region of QTL, we found that NopD could influence the expression of Glyma.19g068600 (FBD/LRR) and expression of Glyma.19g069200 (PP2C) after HH103 infection. Haplotype analysis showed that different types of Glyma.19g069200 haplotypes could cause significant nodule phenotypic differences, but Glyma.19g068600 (FBD/LRR) was not. These results suggest that NopD promotes S. fredii HH103 infection via directly or indirectly regulating Glyma.19g068600 and Glyma.19g069200 expression during the establishment of symbiosis between rhizobia and soybean plants.

Flavonoids repress the production of antifungal 2,4-DAPG but potentially facilitate root colonization of the rhizobacterium Pseudomonas fluorescens.

In the well-known legume-rhizobia symbiosis, flavonoids released by legume roots induce expression of the Nod factors and trigger early plant responses involved in root nodulation. However, it remains largely unknown how the plant-derived flavonoids influence the physiology of non-symbiotic beneficial rhizobacteria. In this work, we demonstrated that the flavonoids apigenin and/or phloretin enhanced the swarming motility and production of cellulose and curli in Pseudomonas fluorescens 2P24, both traits of which are essential for root colonization. Using a label-free quantitative proteomics approach, we showed that apigenin and phloretin significantly reduced the biosynthesis of the antifungal metabolite 2,4-DAPG and further identified a novel flavonoid-sensing TetR regulator PhlH, which was shown to modulate 2,4-DAPG production by regulating the expression of 2,4-DAPG hydrolase PhlG. Although having similar structures, apigenin and phloretin could also influence different physiological characteristics of P. fluorescens 2P24, with apigenin decreasing the biofilm formation and phloretin inducing expression of proteins involved in the denitrification and arginine fermentation processes. Taken together, our results suggest that plant-derived flavonoids could be sensed by the TetR regulator PhlH in P. fluorescens 2P24 and acts as important signalling molecules that strengthen mutually beneficial interactions between plants and non-symbiotic beneficial rhizobacteria.

Polyploid plants obtain greater fitness benefits from a nutrient acquisition mutualism.

Polyploidy is a key driver of ecological and evolutionary processes in plants, yet little is known about its effects on biotic interactions. This gap in knowledge is especially profound for nutrient acquisition mutualisms, despite the fact that they regulate global nutrient cycles and structure ecosystems. Generalism in mutualistic interactions depends on the range of potential partners (niche breadth), the benefits obtained and ability to maintain benefits across a variety of partners (fitness plasticity). Here, we determine how each of these is influenced by polyploidy in the legume-rhizobium mutualism. We inoculated a broad geographic sample of natural diploid and autotetraploid alfalfa (Medicago sativa) lineages with a diverse panel of Sinorhizobium bacterial symbionts. To analyze the extent and mechanism of generalism, we measured host growth benefits and functional traits. Autotetraploid plants obtained greater fitness enhancement from mutualistic interactions and were better able to maintain this across diverse rhizobial partners (i.e. low plasticity in fitness) relative to diploids. These benefits were not attributed to increases in niche breadth, but instead reflect increased rewards from investment in the mutualism. Polyploid plants displayed greater generalization in bacterial mutualisms relative to diploids, illustrating another axis of advantage for polyploids over diploids.

Exogenous application of signaling molecules to enhance the resistance of legume-rhizobium symbiosis in Pb/Cd-contaminated soils

Being signaling molecules, nitric oxide (NO) and hydrogen sulfide (H2S) can mediate a wide range of physiological processes caused by plant metal toxicity. Moreover, legume-rhizobium symbiosis has gained increasing attention in mitigating heavy metal stress. However, systematic regulatory mechanisms used for the exogenous application of signaling molecules to alter the resistance of legume-rhizobium symbiosis under metal stress are currently unknown. In this study, we examined the exogenous effects of sodium nitroprusside (SNP) as an NO donor additive and sodium hydrosulfide (NaHS) as a H2S donor additive on the phytotoxicity and soil quality of alfalfa (Medicago sativa)-rhizobium symbiosis in lead/cadmium (Pb/Cd)-contaminated soils. Results showed that rhizobia inoculation markedly promoted alfalfa growth by increasing chlorophyll content, fresh weight, and plant height and biomass. Compared to the inoculated rhizobia treatment alone, the addition of NO and H2S significantly reduced the bioaccumulation of Pb and Cd in alfalfa-rhizobium symbiosis, respectively, thus avoiding the phytotoxicity caused by the excessive presence of metals. The addition of signaling molecules also alleviated metal-induced phytotoxicity by increasing antioxidant enzyme activity and inhibiting the level of lipid peroxidation and reactive oxygen species (ROS) in legume-rhizobium symbiosis. Also, signaling molecules improved soil nutrient cycling, increased soil enzyme activities, and promoted rhizosphere bacterial community diversity. Both partial least squares path modeling (PLS-PM) and variation partitioning analysis (VPA) identified that using signaling molecules can improve plant growth by regulating major controlling variables (i.e., soil enzymes, soil nutrients, and microbial diversity/plant oxidative damage) in legume-rhizobium symbiosis. This study offers integrated insight that confirms that the exogenous application of signaling molecules can enhance the resistance of legume-rhizobium symbiosis under metal toxicity by regulating the biochemical response of the plant-soil system, thereby minimizing potential health risks.

Variation in rhizosphere microbial communities and its association with the symbiotic efficiency of rhizobia in soybean

Rhizobia–legume symbiosis is an important type of plant–microbe mutualism; however, the establishment of this association is complicated and can be affected by many factors. The soybean rhizosphere has a specific microbial community, yet whether these organisms affect rhizobial nodulation has not been well investigated. Here, we analyzed the compositions and relationships of soybean rhizocompartment microbiota in three types of soil. First, we found that the rhizosphere community composition of soybean varied significantly in different soils, and the association network between rhizobia and other rhizosphere bacteria was examined. Second, we found that some rhizosphere microbes were correlated with the composition of bradyrhizobia and sinorhizobia in nodules. We cultivated 278 candidate Bacillus isolates from alkaline soil. Finally, interaction and nodulation assays showed that the Bacillus cereus group specifically promotes and suppresses the growth of sinorhizobia and bradyrhizobia, respectively, and alleviates the effects of saline–alkali conditions on the nodulation of sinorhizobia as well as affecting its colonization in nodules. Our findings demonstrate a crucial role of the bacterial microbiota in shaping rhizobia–host interactions in soybean, and provide a framework for improving the symbiotic efficiency of this system of mutualism through the use of synthetic bacterial communities.

The impact of the rhizobia-legume symbiosis on host root system architecture.

Legumes form symbioses with rhizobia to fix N2 in root nodules to supplement their nitrogen (N) requirements. Many studies have shown how symbioses affect the shoot, but far less is understood about how they modify root development and root system architecture (RSA). RSA is the distribution of roots in space and over time. RSA reflects host resource allocation into below-ground organs and patterns of host resource foraging underpinning its resource acquisition capacity. Recent studies have revealed a more comprehensive relationship between hosts and symbionts: the latter can affect host resource acquisition for phosphate and iron, and the symbiont's production of plant growth regulators can enhance host resource flux and abundance. We review the current understanding of the effects of rhizobia-legume symbioses on legume root systems. We focus on resource acquisition and allocation within the host to conceptualize the effect of symbioses on RSA, and highlight opportunities for new directions of research.

Optimizing Rhizobium-legume symbioses by simultaneous measurement of rhizobial competitiveness and N2 fixation in nodules

Legumes tend to be nodulated by competitive rhizobia that do not maximize nitrogen (N2) fixation, resulting in suboptimal yields. Rhizobial nodulation competitiveness and effectiveness at N2 fixation are independent traits, making their measurement extremely time-consuming with low experimental throughput. To transform the experimental assessment of rhizobial competitiveness and effectiveness, we have used synthetic biology to develop reporter plasmids that allow simultaneous high-throughput measurement of N2 fixation in individual nodules using green fluorescent protein (GFP) and barcode strain identification (Plasmid ID) through next generation sequencing (NGS). In a proof-of-concept experiment using this technology in an agricultural soil, we simultaneously monitored 84 different Rhizobium leguminosarum strains, identifying a supercompetitive and highly effective rhizobial symbiont for peas. We also observed a remarkable frequency of nodule coinfection by rhizobia, with mixed occupancy identified in ∼20% of nodules, containing up to six different strains. Critically, this process can be adapted to multiple Rhizobium-legume symbioses, soil types, and environmental conditions to permit easy identification of optimal rhizobial inoculants for field testing to maximize agricultural yield.

Native Bradyrhizobium Strains From Ghana Can Enhance Grain Yields of Field-Grown Cowpea and Groundnut

The existence of large population of ineffective native rhizobia and inconsistent performance of exotic strains in Ghanaian soils necessitate the need to identify effective and locally adapted elite strains for enhanced legume-rhizobium symbiosis. This study was designed to test the suitability of two previously selected potential elite Bradyrhizobium strains for use as inoculants on cowpea (Vigna unguiculata L. Walp) and groundnut (Arachis hypogaea L.) in multi-location experiments. Field experiments were set up at 26 locations (12 planted with cowpea and 14 planted with groundnut) in the Northern region of Ghana. Four treatments were applied at each location: inoculation with Bradyrhizobium strains KNUST 1002 and KNUST 1006, a positive nitrogen (+N) control and a negative control (without nitrogen or inoculation) arranged in randomized complete blocks with four replications. The results showed that inoculation with strains KNUST 1002 and KNUST 1006 promoted significant increases in grain yields of both cowpea and groundnut. On average, inoculating cowpea with strains KNUST 1002 and KNUST 1006 resulted in 63 and 52% increases in grain yield when compared to the negative control. Pod yields of groundnut on the other hand, were significantly (p< 0.05) increased at 57% of the study location with one or both test strains. Responses to inoculation were, however, highly variable across the different study locations (i.e. significant Treatment ⸼ Location Interaction, TLI). A detailed analysis of this significant TLI based on the genotype main effect (G) plus genotype-by-environment (GE) interaction (GGE) biplot analysis revealed that location contributed 71 and 88% of the variation observed in cowpea and groundnut, respectively, and grouped the locations into mega-environments. These results demonstrate that native elite Bradyrhizobium strains KNUST 1002 and KNUST 1006 have potential for use as inoculants to increase cowpea and groundnut production in Northern Ghana.

Is it time to include legumes in plant silicon research?

Abstract To date, the functional role of plant silicon has mostly been investigated in grasses (Poaceae). This potentially overlooks the importance of silicon in other plant functional groups such as legumes (Fabaceae). Legumes form a symbiotic relationship with nitrogen‐fixing bacteria (rhizobia) inside the root nodules for fixing atmospheric nitrogen. A small, but growing number of studies suggest that silicon promotes this symbiotic relationship. We consider how legumes may take up and deposit silicon relative to what is known about these processes in grasses. We synthesize information about how silicon affects legume growth and function in the context of environmental stresses and the legume–rhizobia symbiosis. The available literature indicates that silicon is broadly beneficial to legumes, alleviating the effects of stresses including metal toxicity, salinity, alkalinity and pathogens. Crucially, there is also evidence that silicon promotes the legume–rhizobia interaction including increased root nodulation, numbers of bacteroids and nitrogen fixation across several legume species. We propose a model for how silicon may benefit the legume–rhizobia interaction. We hypothesize that silicification in the tissues may reduce the high metabolic cost of carbon‐based compounds in cell wall construction, optimize solute transport and gas exchange in root nodules and/or promote protection against environmental stresses. We therefore propose a hypothetical framework to better understanding the impacts of silicon on legume–rhizobia relationships. We also suggest potential research priorities that would help us to better understand the functional role of silicon in nitrogen‐fixing legumes. These research priorities focus on characterizing how silicon affects the chemical dialogues between the host plant and its rhizobial partner, how silicon is deposited in legume roots and how resources are exchanged by the two. Given the growing importance of legumes at a global scale, silicon could play a vital role in improving legume health and productivity with manifold environmental benefits. A free Plain Language Summary can be found within the Supporting Information of this article.

Rhizobial infection triggers systemic transport of endogenous RNAs between shoots and roots in soybean.

Legumes have evolved a symbiotic relationship with rhizobial bacteria and their roots form unique nitrogen-fixing organs called nodules. Studies have shown that abiotic and biotic stresses alter the profile of gene expression and transcript mobility in plants. However, little is known about the systemic transport of RNA between roots and shoots in response to rhizobial infection on a genome-wide scale during the formation of legume-rhizobia symbiosis. In our study, we found that two soybean (Glycine max) cultivars, Peking and Williams, show a high frequency of single nucleotide polymorphisms; this allowed us to characterize the origin and mobility of transcripts in hetero-grafts of these two cultivars. We identified 4,552 genes that produce mobile RNAs in soybean, and found that rhizobial infection triggers mass transport of mRNAs between shoots and roots at the early stage of nodulation. The majority of these mRNAs are of relatively low abundance and their transport occurs in a selective manner in soybean plants. Notably, the mRNAs that moved from shoots to roots at the early stage of nodulation were enriched in many nodule-related responsive processes. Moreover, the transcripts of many known symbiosis-related genes that are induced by rhizobial infection can move between shoots and roots. Our findings provide a deeper understanding of endogenous RNA transport in legume-rhizobia symbiotic processes.