Root Nodules Of Plants Research Articles

Potential of novel Rhizobium strains as biofertilizer for improved groundnut (Arachis hypogaea L.) cultivation in Eastern India

ABSTRACT Comprehensive assessments were conducted on eight fast growing, novel Rhizobium isolates, short listed from a preliminary selection of 68 isolates obtained from root nodules of groundnut plants grown in diverse agro-climatic regions of eastern India. Additionally, two conventional rhizobia strains, viz. IGR6 and GN2, (collected from the Nodule Research Center, BCKV), were utilized for comparative analysis. Cluster analysis, based on carbon utilization and biochemical parameters, grouped all the test isolates with an average similarity level of 73%. The ability of the isolates to utilize a wide range of carbon sources, including hexoses, pentoses, and disaccharides, confirms their fast-growing nature. Further analysis through 16S rDNA extraction and nucleotide sequencing revealed that the promising fresh isolates exhibited 95–99% similarity to known Rhizobium species in public databases. When evaluated in pot culture experiments using new alluvial soil from the Gangetic Plains in eastern India, the native fresh isolate NRA1(PP355674) outperformed other isolates and conventional strains in terms of nodulation efficiency, plant dry matter production, and plant nitrogen uptake. This study highlights the significant potential of native Rhizobium isolates to surpass conventionally used strains in biological nitrogen fixation, supporting the development of sustainable, environmentally-friendly, and cost-effective agricultural practices.

Study of Plant Growth Promoting Abilities of Rhizobium Strains Isolated from Mungbean Root Nodules

Background: Plant Growth Promoting Rhizobacteria (PGPR) are a type of bacteria that use biofertilizers and other plant growth-promoting agents to increase plant output and growth. In the last several decades, there has been a global surge in the usage of beneficial soil microorganisms like PGPR for safe and sustainable agriculture due to the detrimental effects of artificial fertilizers on the environment and their rising prices. Plant-beneficial rhizobacteria have the potential to reduce the world's reliance on dangerous agricultural chemicals that upset agro-ecosystems. Today a large part of yield is lost due to the various stress mechanisms of plants. It could be classified into biotic and abiotic stress. These are generally a group of microorganisms that is found either in the rhizosphere or in the root nodules of plants. PGPR has beneficial impact, they colonize in the plant roots and improve crop yields and agricultural productivity and also produce some useful growth promoting growth hormones viz., IAA, Auxins, Cytokinin and Gibberellins. Methods: The bacterial isolates were screened for plant growth promoting traits such as phosphate solubilization, IAA production and siderophore production was tested qualitatively by using pikovskayas media, TSB (Tryptone Soy Broth) agar and Chrome Azurol Sulphate (CAS) blue agar respectively. Results: In present study out of the eight rhizobial isolates, five exhibited the phosphate solubilization zones, three shows negative result. However out of eight isolates five isolates produced IAA and three showed negative result. Among all isolates seven isolates were able to produce siderophore, while one showed negative result.

Microbiological mechanism of hydrogen fertilizer effect in soil.

For a long time, intercropping and rotation of leguminous with non-leguminous crops is widely used to reduce the application of nitrogen fertilizer and increase yield in agroecosystems. At present, most researchers considered that this management measure is helpful for reducing fertilizer consumption and increasing its efficiency, as it can improve nutrient supply of legumestonon-legumes, the spatial nutrient utilization efficiency by enhancing soil spatial heterogeneity, and improve soil structure and disease resistance. However, current theories cannot fully explain the positive effect of crop rotation and inter-cropping systems involving legumes. A large amount of hydrogen (H2) can be produced as an obligatory by-product of nitrogenase responsible for nitrogen (N2) fixation in the root nodules of leguminous plants. Despite of substantial amounts of H2 enriched in the rhizosphere of legumes, only a minor proportion of H2 is found to leak to soil surface. Increasing evidence showed that most H2 released in soil is immediately depleted in the surrounding of N2-fixing nodules by H2-oxidizing bacteria (HOB) thriving in soil. HOB can use H2 as an electron donor to assimilate and fix CO2 through redox reactions to synthesize cellular substances and consequently promote plant growth. To date, however, little is known about the biological mechanism and ecological process behind the "hydrogen fertilizer effect". Therefore, we review the H2-induced plant growth-promoting effects and its microbiological mechanisms. Our aims were to explore a new way for enhancing agroecosystem production, and to provide scientific basis for future utilization of H2 in agricultural production practices.

Use of image analysis in the evaluation of radicular nodules in chickpeas

The use of advanced computational systems facilitates the application of a wide range of statistical techniques, available in open-source format, to perform analyses of substantial complexity in biological systems, particularly in the field of plant biology. The aim of this study is to demonstrate the application of image analysis in the evaluation of root nodules in Cicer arietinum plants. The research was conducted in the field, where roots were collected, subjected to rigorous cleaning protocols and subsequently photographed under precisely controlled studio conditions. This photographic process was performed using equipment configured with meticulous parameter settings. Subsequently, image analyses were performed using the statistical software package, R. Parameters of interest were quantified, including metrics such as root area, nodule area, the proportional representation of nodules in relation to the total root mass and the absolute count of nodules present. A critical comparison between the proposed analytical methodology and conventional approaches evidenced a substantial improvement in efficiency, thus highlighting the robustness and validity of this analytical framework for the outlined research objectives. Therefore, the developed methodology has significant potential to facilitate accurate and comprehensive analyses of nodules and root systems, thus making a notable contribution to fostering a deeper understanding and exploration of plant-microorganism interactions. Furthermore, this methodological advancement has the potential to contribute to reducing operational costs and optimizing the time spent on such meticulous evaluative undertakings.

Adaptation of Rhizobium leguminosarum sv. trifolii strains to low temperature stress in both free-living stage and during symbiosis with clover

Legume-rhizobial symbiosis plays an important role in agriculture and ecological restoration. This process occurs within special new structures, called nodules, formed mainly on legume roots. Soil bacteria, commonly known as rhizobia, fix atmospheric dinitrogen, converting it into a form that can be assimilated by plants. Various environmental factors, including a low temperature, have an impact on the symbiotic efficiency. Nevertheless, the effect of temperature on the phenotypic and symbiotic traits of rhizobia has not been determined in detail to date. Therefore, in this study, the influence of temperature on different cell surface and symbiotic properties of rhizobia was estimated. In total, 31 Rhizobium leguminosarum sv. trifolii strains isolated from root nodules of red clover plants growing in the subpolar and temperate climate regions, which essentially differ in year and day temperature profiles, were chosen for this analysis. Our results showed that temperature has a significant effect on several surface properties of rhizobial cells, such as hydrophobicity, aggregation, and motility. Low temperature also stimulated EPS synthesis and biofilm formation in R. leguminosarum sv. trifolii. This extracellular polysaccharide is known to play an important protective role against different environmental stresses. The strains produced large amounts of EPS under tested temperature conditions that facilitated adherence of rhizobial cells to different surfaces. The high adaptability of these strains to cold stress was also confirmed during symbiosis. Irrespective of their climatic origin, the strains proved to be highly effective in attachment to legume roots and were efficient microsymbionts of clover plants. However, some diversity in the response to low temperature stress was found among the strains. Among them, M16 and R137 proved to be highly competitive and efficient in nodule occupancy and biomass production; thus, they can be potential yield-enhancing inoculants of legumes.

Root nodules of red alder (Alnus rubra) and Sitka alder (Alnus viridis ssp. sinuata) are inhabited by taxonomically diverse cultivable microbial endophytes

AbstractThe root nodules of actinorhizal plants are home to nitrogen‐fixing bacterial symbionts, known as Frankia, along with a small percentage of other microorganisms. These include fungal endophytes and non‐Frankia bacteria. The taxonomic and functional diversity of the microbial consortia within these root nodules is not well understood. In this study, we surveyed and analyzed the cultivable, non‐Frankia fungal and bacterial endophytes of root nodules from red and Sitka alder trees that grow together. We examined their taxonomic diversity, co‐occurrence, differences between hosts, and potential functional roles. For the first time, we are reporting numerous fungal endophytes of alder root nodules. These include Sporothrix guttuliformis, Fontanospora sp., Cadophora melinii, an unclassified Cadophora, Ilyonectria destructans, an unclassified Gibberella, Nectria ramulariae, an unclassified Trichoderma, Mycosphaerella tassiana, an unclassified Talaromyces, Coniochaeta sp., and Sistotrema brinkmanii. We are also reporting several bacterial genera for the first time: Collimonas, Psychrobacillus, and Phyllobacterium. Additionally, we are reporting the genus Serratia for the second time, with the first report having been recently published in 2023. Pseudomonas was the most frequently isolated bacterial genus and was found to co‐inhabit individual nodules with both fungi and bacteria. We found that the communities of fungal endophytes differed by host species, while the communities of bacterial endophytes did not.

In Vitro Screening of Endophytic Micromonospora Strains Associated with White Clover for Antimicrobial Activity against Phytopathogenic Fungi and Promotion of Plant Growth

Bacteria belonging to the genus Micromonospora are recognized as microorganisms with the potential to be used in biotechnology processes, given their beneficial influence on plant growth and the biocontrol of phytopathogens. In this study, nineteen Micromonospora isolates originating from the root nodules of white clover plants were taxonomically assigned based on the phylogenetic analysis of the 16S rRNA gene and four housekeeping genes. The antifungal properties of the bacteria against phytopathogenic Botrytis cinerea, Fusarium oxysporum, Fusarium equiseti, Sclerotinia sclerotiorum, and Verticillium albo-atrum were tested with the agar plug test and the dual culture test. The ability to produce various metallophores was determined with the agar plug diffusion test on modified chrome azurol S (CAS) agar medium. International Streptomyces Project-2 medium (ISP2) broth amended with 0.2% L-tryptophan was used to indicate the bacterial ability to produce auxins. The strains belonging to M. tulbaghiae, M. inaquosa, and M. violae showed in vitro potential as antimicrobial agents against the tested fungi. M. inaquosa strain 152, M. violae strain 126, M. violae strain 66, and M. violae strain 45 were recognized as the most efficient metallophore producers. M. alfalfae strain 55 and M. lupini strain 5052 were identified as the most promising auxin compound producers and, therefore, show potential as plant-growth-promoting bacteria.

The Effectiveness of Plant Growth Promoting Rhizobacteria (PGPR) on the Growth of Root Nodules of Peanut Genotypes under Water Deficit Condition

The deficiency of water sources and soil fertility are obstacles to producing peanuts in dry land. One of effort to increase production on dry land is the utilization of PGPR biofertilizers. This study aimed to determine the growth of plants and root nodules of several peanut genotypes applied with PGPR, to understand the interaction between several peanut genotypes and PGPR on plant growth and root nodules, and to examine the relationship between water deficit conditions and the formation of root nodules. This experiment used a Complete Randomized Design-split split-plot design with the main plots being D0 = optimal conditions (no water deficit) and D1 = water deficit. The subplots were P0 = no PGPR and P1 = with PGPR. The sub-subplots consisted of five peanut genotypes: V1 = Hypoma-I, V2 = Domba, V3 = Talam, V4 = Bison, and V5 = G300-II. The observations showed that the addition of PGPR resulted in higher plant growth compared to those without PGPR and had more effective root nodules in nitrogen fixation compared to treatments without PGPR. The application of PGPR and genotype did not show significant interaction on plant growth and root nodules under water deficit conditions. Water deficit significantly affected all parameters of plant growth and root nodules. PGPR significantly affected the parameters of leaf number rate, nodule number, and nodule weight. Genotype significantly affected the plant height rate. The interaction of water deficit and genotype significantly affected the leaf number rate, while the interaction of water deficit and PGPR, and the interaction of PGPR and genotype did not significantly affect all observation parameters.

Plant-associated bacteria and enzymes support Canavalia rosea growth in coastal hypersaline soils

Canavalia rosea is an extremophilic legume that grows in hypersaline and nutrient-deficient ecosystems. The extremophilic nature of C. rosea may be attributed to its ability to establish symbiotic associations with nutrient mineralizing and plant growth promoting (PGP) bacteria housed in the nodules. This study examined legume-microbe symbiosis and plant nutrition of C. rosea growing in subtropical coastal zone in KwaZulu-Natal province, South Africa. Canavalia rosea adult plants of the same age from Westbrook, Scottburgh and Durban were collected for plant biomass and plant nutrition and root nodules were used for bacterial extraction and identification. Rhizosphere soils sampled from the three localities were used for bacterial extraction and identification, extracellular enzyme assays and soil characteristics (pH, nutrient concentrations, total cation, and exchange acidity). Westbrook, Scottburgh and Durban soils were nutrient-deficient with varying total cations, acid saturation and a pH range of 7.3–7.6. Soil nutrient mineralizing extracellular enzyme activities varied across study sites. The culturable bacterial strains isolated from the sampled soils belonged to the Pseudomonas, Pantoea and Flavobacterium genera. Canavalia rosea root nodules were nodulated by Pseudomonas guariconensis, Pseudomonas fulva, Pseudomonas fluorescens, Pseudomonas chlororaphis and Pseudomonas chlororaphis subsp. aurantiaca. Plants growing in Westbrook soils had a significantly higher total plant biomass compared to Scottburgh and Durban plants. Plant P concentration did not vary significantly between sites while plant N and C concentrations varied significantly. Plant-associated and soil bacteria with phosphorus (P) solubilising, nitrogen (N) cycling, and N fixing functions and associated enzymes seem to facilitate the mobilization of nutrients enabling C. rosea to thrive in hypersaline and low-nutrient environments.

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.

Expression and mutagenesis studies in the Medicago truncatula iron transporter MtVTL8 confirm its role in symbiotic nitrogen fixation and reveal amino acids essential for transport.

The model legume Medicago truncatula establishes a symbiosis with soil bacteria (rhizobia) that carry out symbiotic nitrogen fixation (SNF) in plant root nodules. SNF requires the exchange of nutrients between the plant and rhizobia in the nodule that occurs across a plant-derived symbiosome membrane. One iron transporter, belonging to the Vacuolar iron Transporter-Like (VTL) family, MtVTL8, has been identified as essential for bacteria survival and therefore SNF. In this work we investigated the spatial expression of MtVTL8 in nodules and addressed whether it could be functionally interchangeable with a similar nodule-expressed iron transporter, MtVTL4. Using a structural model for MtVTL8 and the previously hypothesized mechanism for iron transport in a phylogenetically-related Vacuolar Iron Transporter (VIT), EgVIT1 with known crystal structure, we identified critical amino acids and obtained their mutants. Mutants were tested in planta for complementation of an SNF defective line and in an iron sensitive mutant yeast strain. An extended phylogenetic assessment of VTLs and VITs showed that amino acids critical for function are conserved differently in VTLs vs. VITs. Our studies showed that some amino acids are essential for iron transport leading us to suggest a model for MtVTL8 function, one that is different for other iron transporters (VITs) studied so far. This study extends the understanding of iron transport mechanisms in VTLs as well as those used in SNF.

Newly isolated Brevundimonas naejangsanensis as a biocontrol agent againstFusarium redolens the causal of Fusarium yellows ofchickpea.

Three endophytic bacteria, namely BvV, BvP and BvL, were newly isolated from the root nodules of bean, pea and lentil plants respectively cultivated in Mascara the northwest of Algeria, and identified by 16S ribosomal RNA gene sequencing as Brevundimonas naejangsanensis. These strains were able to produce hydrolytic enzymes and hydrogen cyanide. All strains produced a growth-promoting hormone, indole acetic acid, varying in concentration from 83.2 to 171.7 µg/mL. The phosphate solubilizing activity of BvV, BvP and BvL varied from 25.5 to 42.02 µg/mL for tricalcium phosphate. The three antagonistic Brevundimonas spp. showed in vitro the most inhibitory effect on mycelial growth of Fusarium redolens FRC (from 78.33 to 85.55%). Strain BvV, BvP and BvL produced also volatile metabolites which inhibited mycelial FRC growth up to 39.2%. All strains showed significant disease reduction in pot experiments. Chickpea Fusarium yellows severity caused by FRC was reduced significantly from 89.3 to 96.6% in the susceptible cultivar ILC 482 treated with antagonistic B. naejangsanensis. The maximum stimulatory effect on chickpea plants growth was observed by inoculation of strain BvV. This treatment resulted in a 7.40-26.21% increase in shoot height as compared to the control plants. It is concluded that the endophytic bacterial strains of B. naejangsanensis having different plant growth promoting (PGP) activities can be considered as beneficial microbes for sustainable agriculture. To our knowledge, this is the first report to use B. naejangsanensis strains as a new biocontrol agent against F. redolens, a new pathogen of chickpea plants causing Fusarium yellows disease in Algeria.

Nodulation Experiment by Cross-Inoculation of Nitrogen-Fixing Bacteria Isolated from Root Nodules of Several Leguminous Plants.

Root-nodule nitrogen-fixing bacteria are known for being specific to particular legumes. This study isolated the endophytic root-nodule bacteria from the nodules of legumes and examined them to determine whether they could be used to promote the formation of nodules in other legumes. Forty-six isolates were collected from five leguminous plants and screened for housekeeping (16S rRNA), nitrogen fixation (nifH), and nodulation (nodC) genes. Based on the 16S rRNA gene sequencing and phylogenetic analysis, the bacterial isolates WC15, WC16, WC24, and GM5 were identified as Rhizobium, Sphingomonas, Methylobacterium, and Bradyrhizobium, respectively. The four isolates were found to have the nifH gene, and the study confirmed that one isolate (GM5) had both the nifH and nodC genes. The Salkowski method was used to measure the isolated bacteria for their capacity to produce phytohormone indole acetic acid (IAA). Additional experiments were performed to examine the effect of the isolated bacteria on root morphology and nodulation. Among the four tested isolates, both WC24 and GM5 induced nodulation in Glycine max. The gene expression studies revealed that GM5 had a higher expression of the nifH gene. The existence and expression of the nitrogen-fixing genes implied that the tested strain had the ability to fix the atmospheric nitrogen. These findings demonstrated that a nitrogen-fixing bacterium, Methylobacterium (WC24), isolated from a Trifolium repens, induced the formation of root nodules in non-host leguminous plants (Glycine max). This suggested the potential application of these rhizobia as biofertilizer. Further studies are required to verify the N2-fixing efficiency of the isolates.

COMPLEX INOCULATION OF SOYBEANS WITH NODULE BACTERIA BRADYRHIZOBIUM JAPONICUM AS A MEASURE TO OPTIMIZE SYMBIOTIC NITROGEN FIXATION

Objective. Establish effective mixed inoculants for soybeans based on new active strains of Bradyrhizobium japonicum with different intensity and dynamics of nitrogen-fixing activity to improve nodulation processes and functional activity of legume-rhizobia symbiosis. Methods. Microbiological, physiological, statistical, gas chromatography. Legume-rhizobia systems based on Glycine max L. (Merill) soybean, variety Samorodok, were studied using recombinant strains of B. japonicum: B199, B201, Д47, Д48, Д52, Д60, Д63. Monoinoculation of soybean with B. japonicum 634б served as the control. Results. Mixed inoculation of B. japonicum ensured positive ontogenetic dynamics of root nodule formation in soybean. The indicator of the number of root nodules in experimental plants was 1.2–1.9 times higher versus the control plants from Day 18 to 45 of soybean vegetation. When soybeans were inoculated with B. japonicum 634б + B201 and 634б + Д 52, the mass of nodules at Day 45 of vegetation increased by 30.0–35.3 % versus the control and was the highest (1.10–1.15 g/plant) among the studied variants, as a result of the strengthening effect of complex bacterization. A positive synergistic effect of the combined use of strains was obtained — intensification of the nitrogen-fixing activity of soybean root nodules. In the phase of budding/beginning of flowering and full flowering under combined inoculation with active strains of B. japonicum 634б + Д47, 634б + Д52 and 634б + В201, nitrogen fixation of soybean nodules increased by 10.7–25.5 % (at Day 32), by 13.2–30.9 % (at Day 38), and was also 2.3–2.8 times higher in the phase of full flowering (Day 45 of soybean vegetation) versus the similar indicator of control plants. In the phase of full flowering of plants (Day 45 of vegetation), under mixed inoculation with strains of B. japonicum 634б + Д47, 634б + В201, 634б + Д52 and В201 + Д52, the aboveground mass of plants exceeded that of control plants by 8.3–17.2 % variant, which was probably provided by improved conditions of their nitrogen nutrition, due to enhanced assimilation of N2 by root nodules. The most developed root system was in soybeans inoculated with strains B. japonicum 634б and B. japonicum 634б + Д52, B. japonicum 634б + В201 and B. japonicum Д48 + Д52. Conclusion. Symbiotic features of active recombinant strains of B. japonicum and their variability in functional activity at different stages of soybean ontogenesis should be taken into account when creating complex nitrogen-fixing bacterial preparations for soybeans with the effect of microbiological synergy.

Viability of Nodule Bacteria Bradyrhizobium japonicummon Soybean Seeds by Treatment With Fungicides During Extended Storage

Background. With the increase in soybean cultivation areas, inoculants are becoming increasingly sought after. They are not only compatible with the original preparations for seed treatment of soybean but also offer the possibility of applying them for several days or even months before sowing into the soil. Objective. The viability of new strains of nodule bacteria Bradyrhizobium japonicum (strain PC07 and strain B78) was investigated on the surface of soybean seeds treated with fungicides Fever and Maxim XL during the extended storage of inoculated seeds. Additionally, their ability for nitrogen fixation under symbiotic conditions was evaluated. Methods. Сultivation, serial dilution method, determination of bacterial titer of inoculants, quantification of colony forming units (CFU), gas chromatography. Results. In laboratory conditions, on the soybean seeds of Almaz and Vasylkivska varieties, the viability of the nodule bacteria B. japonicum PC07 and B78, which are more resistant to the active substances of these fungicides in pure culture, decreased. This reduction was dependent on the storage period, the fungicide used, and the strain of inoculant. One day after inoculation on the seeds without the use of fungicide, 68.6–75.4% CFU of the initial number was retained. Seeds treated with the fungicide Fever contained 36.5–38.3% CFU, and those treated with Maxim XL contained 26.2–27.0% CFU. The use of inoculants based on fungicide-resistant strains of B. japonicum PC07 and B78 also provided high cell viability – 2.41–2.8×106 (on the seeds without treatment), 1.40–1.70×106 (with Fever) and 0.8–1.17×106 (with Maxim XL) CFU/seed after 5 days of storage. The nitrogen-fixing activity of root nodules in soybean plants of Almaz and Vasylkivska varieties, whose seeds were treated with Fever, decreased on both varieties by 18.4–22.4% and 32.1–39.5%, respectively and for treatment with Maxim XL, the reduction was 24.5–33.7% and 47.7–75.2%, respectively, during the storage of seeds for 5 and 7 days before sowing, compared to control variants (seeds without fungicide treatment). Conclusions. The utilization of fungicide-resistant strains of B. japonicum PC07 and B78 for bacterization of soybean seeds treated with fungicides Fever and Maxim XL provides a high inoculation titer during 5 days of storage. Viability of microbial cells on seeds treated with fungicides significantly diminishes during storage for more than 7–14 days. Insufficient titer of CFU of B. japonicum strains PC07 and B78 on the seed surface after 14 days of storage can considerably impede the efficacy of biopreparations. Therefore, it is crucial to seek substances that can enhance the resistance of rhizobia on seeds to the adverse effects of fungicide seed treatment, ensuring a longer period of their viability.

Pengaruh Pemberian Biochar Sekam Padi dan Molybdenum Terhadap Pertumbuhan dan Hasil Tanaman Kedelai (GlycineMax. L.Merill) Pada Ultisol

Soybeans are a very important food crop commodity because they are the main source of vegetable protein. In jambi province, the ultisol soil type is dominated. Ultisols has the potential to be used as agricultural land. Improving proper soil conditions such as ultisols can be done by adding natural ingridients such as biuochar. Molybdenum plays a role in activating the nitrogenase enzyme to form root nodules in legume plants. The aim of thisn research to concentrate on the impact of biochar and Mo components on the devolment and yield of soybean plant in ultisols. This research was carried out at the Teaching and research farm, faculty of agriculture, jambi university for approximately 4 month starting fromseptember to december 2022. This research used a randomized block design (RAK) with a factorial pattern consisting of 2 factors, namely rice husk biochar and Molybdenum. Factor I biochar ( B) which consists of three dose levels, namely without biochar as a control (b0), biochar 6 (b1), biochar 12 (b2), Factor II Molybdenum consists of three dose levels, namely without molybdenum as a control (m0), 75 (m1), and 150 (m2). Thus three are 9 treatment combinationts. Each treatment was repeated 3 times that there were 27 exxperiental plots. The variables observed were plant height, leaf area, total dry weight, number of roor nodules, number of branches, number of pods, weight of 100 dry seeds, number of seeds per plant. To detemine the effect of treatment, statistical analisys was carried out using ANOVA, if the treatment shows a real difference to the observed variables, it is continued with the DMRT at the α = 5% levels. The results of the research show that each treatment has a single factor effect on the observed variables. In this study, there was no interraction between rich husk biochar and molybdenum on all the variables observed, but asw a single factor, rice husk biochar influenced the height and leaf area variables. Meanwhile, molybdenum effects leafarea, total dry weight, number of root nodules, number of pods, and weight poer plant. The rice husk biochar provided in this tusdy was not able to increase the Mo element in the soil, as well as nitrate reductas activity.

Detection of Rhizobium leguminosarum bv. trifolii and Rhizobium leguminosarum bv. phaseoli isolated from root nodules of leguminous plants by multiplex-PCR

The genetic diversity of 4 R. leguminosarum bv. trifolii and R. leguminosarum bv. Phaseoli that was extracted from root nodes of Trifolium spp and Phaseolus vulgaris L cultivars circulated in Mosul soils were investigated. The bacteria strains under study show similar tolerance levels against some external factors. In the current study, using classical diagnosis methods, similarity and distance indices and DNA polymorphism were examined with the randomly amplified polymorphic DNA. The isolates were distributed through genetic diversification into two main classes, including two main branches, Rl2 and the other sub-group was divided into Rl3, and the neighboring group involved Rl1 and Rl4. It is noted that individual plants and abiotic aspects were less affected than the Genetic factor on bacterial diversity. On the other hand, in multiplex-PCR reaction, Our outcomes revealed specific amplified for R. leguminosarum bv. trifolii with amplification products 419 bp and R. leguminosarum bv. Phaseoli with amplification products 362 bp. Keywords: R. leguminosarum bv. phaseoli, R. leguminosarum bv. trifolii , nodules, multiplex-PCR, RAPD-PCR .

Silicon supplementation and jasmonate activation synergistically increase phenolic defences against a legume herbivore

Abstract The accumulation of silicon (Si) is widely reported to have anti‐herbivore defensive properties in grasses. There is emerging, but fragmentary, evidence that Si could play a similar role in legumes. Here, we sought to understand the effects of Si supplementation on anti‐herbivore defensive properties in lucerne (Medicago sativa), especially in relation to other potential defences (i.e. phenolics) and the phytohormone that regulates anti‐herbivore defences, jasmonic acid or jasmonate (JA), which is also linked to Si accumulation. We determined how growth, root nodulation and chemistry (carbon, nitrogen and phenolic concentrations) of four genotypes of lucerne responded to Si supplementation, with and without the application of JA, and we used feeding assays to determine the subsequent effects on the feeding success of adult Sitona discoideus weevils. Si supplementation increased plant mass and root nodulation of M. sativa by 61% and 227%, respectively, and reduced relative consumption (RC) and frass production by S. discoideus by 38% and 30% respectively. Si supplementation had no effect on foliar nitrogen concentrations, most likely due to the dilution effects of increased plant growth and foliar carbon. Phenolic concentrations were negatively correlated with leaf RC; RC also decreased by 34% when JA was applied to plants. When Si was combined with JA application, phenolics were significantly enhanced, demonstrating the potential to stimulate multiple anti‐herbivore properties in M. sativa. Overall, the Si‐ and JA‐induced phytochemical and herbivore feeding responses were consistent between the four lucerne genotypes tested. Synthesis. The novel findings suggest that Si accumulation may play a more important role in legume resistance to herbivorous animals than previously thought. The ubiquity of soil Si and its emerging functional role in plant biology, including plant–animal interactions, suggest that these patterns could be common among legumes.

Bambara Nut Root-Nodules Bacteria from a Semi-Arid Region of South Africa and Their Plant Growth-Promoting Traits.

Rhizobial nitrogen-fixing bacteria are the main inhabitants of the root nodules of legume plants. Studying the bacterial community of legume nodules is important in understanding plant growth and nutrient requirements. Culture-based technique was used to examine the bacterial community of these underground organs from Vigna subterranea L. Verdc (Bambara nut), an underutilized legume in Africa, for plant growth-promoting traits. In this study, Bambara nuts were planted to trap root-nodule bacteria, and the bacteria were morphologically, biochemically, and molecularly characterized. Five selected isolates were screened in vitro for their plant growth-promoting traits and exhibited differences in their phenotypic traits. The polymerase chain reaction (PCR) products were subjected to partial 16S rRNA gene sequencing for phylogenetic analysis. Based on 16S rRNA gene sequence, the isolates were identified as BA1 (Stenotrophomonas maltophilia), BA2 (Chryseobacterium sp.), BA3 (Pseudomonas alcaligenes), BA4 (Pseudomonas plecoglossicida), and BA5 (Pseudomonas hibiscicola). Results showed that four of the five isolates could produce IAA. The capability to solubilize phosphate in Pikovskaya's agar plates was positively shown by four isolates (BA2, BA3, BA4, and BA5). Three isolates could produce hydrogen cyanide while isolates BA1, BA3, BA4, and BA5 were found to have ammonia-production traits. The results suggest that these plant growth-promoting isolates can be used as inoculants for plant growth and productivity.

Isolation and Characterization of Thermotolerant Mungbean (Vigna Radiata L.) Rhizobial Isolate As Biofertilizer

Rhizobia act as primary nitrogen fixing microorganisms in the root of leguminous plants. Efficient rhizobia are helpful in increasing productivity of legumes. In the present investigation a total of twenty rhizobial isolates were retrieved from root nodules of mungbean plants growing in field during summer season. On evaluation of plant growth promoting traits of all rhizobial isolates it was found that rhizobial isolate HSR1 had maximum indole acetic acid (IAA) production (58.83 μg/ml), ammonia excretion (4.78 μg/ml) and phosphate solubilization potential. Other plant growth promoting traits like aminocyclopropane- 1-carboxylic acid (ACC) deaminase activity, siderophore production and hydrogen cyanide (HCN) production were also exhibited by isolate HSR1. This isolate was able to survive 45ºC and exhibited plant growth promoting traits at high temperature. Genomic DNA extracted from isolate HSR1 was subjected to amplification of 16S rRNA gene for identification. The similarity searching of the sequence obtained after 16S rRNA gene sequencing showed 99% similarity with Sinorhizobium sp. T25, which was confirmed by morphological and biochemical tests. Presence of good plant growth promoting traits makes HSR1 a promising strain to be developed as biofertilizer to improve soil fertility and plant growth. Bangladesh J. Bot. 52(2): 217-224, 2023 (June)