Effective Rhizobium Strains Research Articles

Draft Genome Sequence of the Commercial Strain Rhizobium ruizarguesonis bv. viciae RCAM1022

Legume plants enter a symbiosis with soil nitrogen-fixing bacteria (rhizobia), thereby gaining access to assimilable atmospheric nitrogen. Since this symbiosis is important for agriculture, biofertilizers with effective strains of rhizobia are created for crop legumes to increase their yield and minimize the amounts of mineral fertilizers required. In this work, we sequenced and characterized the genome of Rhizobium ruizarguesonis bv. viciae strain RCAM1022, a component of the ‘Rhizotorfin’ biofertilizer produced in Russia and used for pea (Pisum sativum L.).

Enhancing Rhizobium–Legume Symbiosis and Reducing Nitrogen Fertilizer Use Are Potential Options for Mitigating Climate Change

This review article explores the impact of nitrogen fertilizers on the symbiotic relationship between Rhizobium bacteria and legume plants. Nitrogen fixation has the potential to address the global protein shortage by increasing nitrogen supply in agriculture. However, the excessive use of synthetic fertilizers has led to environmental consequences and high energy consumption. To promote sustainable agriculture, alternative approaches such as biofertilizers that utilize biological nitrogen fixation have been introduced to minimize ecological impact. Understanding the process of biological nitrogen fixation, where certain bacteria convert atmospheric nitrogen into ammonia, is crucial for sustainable agriculture. This knowledge helps reduce reliance on synthetic fertilizers and maintain soil fertility. The symbiotic relationship between Rhizobium bacteria and leguminous plants plays a vital role in sustainable agriculture by facilitating access to atmospheric nitrogen, improving soil fertility, and reducing the need for chemical fertilizers. To achieve optimal nitrogen fixation and plant growth, it is important to effectively manage nitrogen availability, soil conditions, and environmental stressors. Excessive nitrogen fertilization can negatively affect the symbiotic association between plants and rhizobia, resulting in reduced soil health, altered mutualistic relationships, and environmental concerns. Various techniques can be employed to enhance symbiotic efficiency by manipulating chemotaxis, which is the ability of rhizobia to move towards plant roots. Plant-specific metabolites called (iso)flavonoids play a crucial role in signaling and communication between legume plants and rhizobia bacteria, initiating the symbiotic relationship and enhancing nitrogen fixation and plant growth. Excessive nitrogen fertilizer application can disrupt the communication between rhizobia and legumes, impacting chemotaxis, root exudation patterns, nodulation, and the symbiotic relationship. High levels of nitrogen fertilizers can inhibit nitrogenase, a critical enzyme for plant growth, leading to reduced nitrogenase activity. Additionally, excessive nitrogen can compromise the energy demands of nitrogen fixation, resulting in decreased nitrogenase activity. This review discusses the disadvantages of using nitrogenous fertilizers and the role of symbiotic biological nitrogen fixation in reducing the need for these fertilizers. By using effective rhizobial strains with compatible legume cultivars, not only can the amounts of nitrogenous fertilizers be reduced, but also the energy inputs and greenhouse gas emissions associated with their manufacturing and application. This approach offers benefits in terms of reducing greenhouse gas emissions and saving energy. In conclusion, this paper provides a comprehensive overview of the current understanding of the impact of nitrogen fertilizers on the symbiotic relationship between Rhizobium and legume plants. It also discusses potential strategies for sustainable agricultural practices. By managing nitrogen fertilizers carefully and improving our understanding of the symbiotic relationship, we can contribute to sustainable agriculture and minimize environmental impact.

CHARACTERIZATION AND COMPARATIVE RESPONSE OF RHIZOBIA ISOLATES FROM ROOT NOODLES OF SELECTED WEED SPECIES AND SOYBEAN (Glycine max) TO ENVIRONMENTAL FACTORS

Screening and selecting the most effective strains of rhizobium is important for biological nitrogen fixation. To determine the effects of environmental factors on rhizobia strains associated with some selected leguminous weed species, rhizobia strains isolated from root nodules of five weed species (Mimosa pudica, Crotolaria retusa, Desmodium triflorum, Canavalina ensiformis, Stylosanthes spp.) were subjected to varying temperature, salinity, and pH in the laboratory. Results indicated better growths at temperatures of 30 and 370C compared to the temperature of 250C. Results further showed that microbial growth was optimal at 1.5% salt concentration. Optimum growth was also obtained at the pH range of 5 and 8. The study further revealed by a series of morphological and biochemical tests that all the strains were gram-negative, rod-shaped, and mucous producing. Rhizobia strains from leguminous weeds were found to exhibit faster growth than the strain nodulating soybean in all the conditions to which they were subjected. A cross inoculation study was therefore recommended to access the compatibility of these rhizobial strains from the wild with cultivated leguminous plant species.

Genomic Diversity of Pigeon Pea (Cajanus cajan L. Millsp.) Endosymbionts in India and Selection of Potential Strains for Use as Agricultural Inoculants.

Pigeon pea (Cajanus cajan L. Millsp. ) is a legume crop resilient to climate change due to its tolerance to drought. It is grown by millions of resource-poor farmers in semiarid and tropical subregions of Asia and Africa and is a major contributor to their nutritional food security. Pigeon pea is the sixth most important legume in the world, with India contributing more than 70% of the total production and harbouring a wide variety of cultivars. Nevertheless, the low yield of pigeon pea grown under dry land conditions and its yield instability need to be improved. This may be done by enhancing crop nodulation and, hence, biological nitrogen fixation (BNF) by supplying effective symbiotic rhizobia through the application of “elite” inoculants. Therefore, the main aim in this study was the isolation and genomic analysis of effective rhizobial strains potentially adapted to drought conditions. Accordingly, pigeon pea endosymbionts were isolated from different soil types in Southern, Central, and Northern India. After functional characterisation of the isolated strains in terms of their ability to nodulate and promote the growth of pigeon pea, 19 were selected for full genome sequencing, along with eight commercial inoculant strains obtained from the ICRISAT culture collection. The phylogenomic analysis [Average nucleotide identity MUMmer (ANIm)] revealed that the pigeon pea endosymbionts were members of the genera Bradyrhizobium and Ensifer. Based on nodC phylogeny and nod cluster synteny, Bradyrhizobium yuanmingense was revealed as the most common endosymbiont, harbouring nod genes similar to those of Bradyrhizobium cajani and Bradyrhizobium zhanjiangense. This symbiont type (e.g., strain BRP05 from Madhya Pradesh) also outperformed all other strains tested on pigeon pea, with the notable exception of an Ensifer alkalisoli strain from North India (NBAIM29). The results provide the basis for the development of pigeon pea inoculants to increase the yield of this legume through the use of effective nitrogen-fixing rhizobia, tailored for the different agroclimatic regions of India.

Comparative genomic analysis of diverse rhizobia and effective nitrogen-fixing clover-nodulating Rhizobium strains adapted to Egyptian dry ecosystems

Rhizobium strains that nodulate Egyptian winter clover belong to two symbiotic species, Rhizobium aegyptiacum sv. trifolii and Rhizobium bangladeshense sv. trifolii. Rhizobia from Egyptian clover are often tolerant of extreme environmental conditions and are generally poorly characterized, despite the importance of this crop species. Here, we report on the comparative genomic analysis of four stress-tolerant, highly effective nitrogen-fixing, Egyptian-clover-nodulating rhizobia, R. aegyptiacum sv. trifolii strain Rhiz950 and Rhizobium bangladeshense sv. trifolii strains Rhiz1002, Rhiz1017 and Rhiz1024, as well as other diverse Rhizobium strains. While Rhizobium bangladeshense sv. trifolii strains Rhiz1002, Rhiz1017, and Rhiz1024 had similar genome sizes of around 6.6 Mb, the salt tolerant strain Rhiz950 had a slightly larger genome size of 7.4 Mb. All four strains lacked genes responsible for methylation and sulfation of nod factors (nodS and nodHPQ). They also lacked nodZ, and instead had the functionally-equivalent nod factor fucosylation gene nolK. No strains contained homocitrate synthase (encoded by nifV) that is essential for free-living nitrogen fixing by diazotrophs. A number of genes unique to the Egyptian clover strains and to other rhizobia are potential candidates for variation in stress tolerance and host-range.

Intensification of Symbiotic Performances of Grain Legumes and Rhizobia through Joint Use of Agro-inputs: Review

As it holds in sub-Saharan Africa, low grain legume crop productivity has been facing Ethiopia and is often associated with reduced N 2 -fixation. For low-input users, symbiotic nitrogen fixation (SNF) can be an affordable, feasible and sustainable option particularly when employed in appropriate inoculation of effective rhizobial strains to improve the productivity of legumes. Here, we review past research interventions on agro-inputs and Rhizobium -legume symbiosis interaction with a view of understanding the consequence on productivity of the symbiosis. First of all, information on farm-based need-to-inoculate and reinoculate legumes with rhizobia remain fundamental when benefit-oriented inoculation is thought. In addition, co-application of nutrients like phosphorus (P) or Sulphur (S), organic amendments, plant growth promoting microbes (PGPMs), arbuscular mycorrhizal fungi (AMF) and lime highly complemented rhizobia in enhancing symbiotic effectiveness. Sole lime application appeared no to improve yield of legumes, its co-application with rhizobia, acid tolerant variety and P fertilizer enhanced nitrogen (N) and grain yield of legumes. The reaction of rhizobia and symbiosis to seed-dresser pesticides may be detrimental or beneficial (indirectly) depending on the rate, and type of the pesticide and the legume. Thus, co-application of rhizobial strains and agro-inputs generally enhanced the productivity of rhizobial-legume symbiosis, but this should be done with prior refining through context-based scientific research. Key words : Agro-input, Inoculation, Nodulation, Rhizobia, Symbiotic Effectiveness DOI: 10.7176/JBAH/10-19-01 Publication date: October 31 st 2020

Towards sustainable yield improvement: field inoculation of soybean with Bradyrhizobium and co-inoculation with Azospirillum in Mozambique.

The effects of sole inoculation of soybean (Glycine max L. Merrill) with Bradyrhizobium and co-inoculation with Bradyrhizobium and Azospirillum on nodulation, plant growth and yields were investigated in the 2013/2014 and 2014/2015 cropping seasons under field conditions in Mozambique. The treatments included (1) Control (non-inoculated control, with symbiosis depending on indigenous rhizobia), (2) Urea (non-inoculated, receiving 200kgha-1 of N), (3) Sole inoculation with B. diazoefficiens strain USDA 110, and (4) Co-inoculation with B. diazoefficiens strain USDA 110 and A. brasilense strains Ab-V5 and Ab-V6, evaluated in a randomized complete block design with five replications. Nodule number and dry weight, shoot dry weight, biological and grain yields, grain dry weight, and harvest index were evaluated. In general, both sole inoculation and co-inoculation enhanced nodulation in relation to control. Sole inoculation increased grain yield by 22% (356kgha-1), the same enhancement magnitude attained under mineral N treatment, suggesting that Bradyrhizobium inoculation provides ecological and economic sustainability to the soybean crop in Mozambique or other countries with similar agro-climatic conditions. Co-inoculation did not increase grain yields in relation to neither the control nor sole inoculation, indicating that further research with adapted and high yielding soybean varieties along with effective rhizobial strains is required in Mozambique to attune the beneficial Azospirillum-plant cultivar-rhizobia interactions that have been reported in other countries for several legumes, including soybean.

Legume-Rhizobium Strain Specificity Enhances Nutrition and Nitrogen Fixation in Faba Bean (Vicia faba L.)

This study reports the effectiveness of some selected rhizobium strains in enhancing nitrogen fixation and nutrient uptake in Vicia faba L. Multi-location field experiments were conducted for two years (2016 and 2017) using a split-plot in randomized complete block design. Treatments comprised six rhizobium strains as the main plot factor and three varieties of Vicia faba as the sub-plot factor. Non-inoculated plants with or without N fertilizer served as +N and −N controls, respectively. Peat carrier-based inoculant of each strain was applied at the rate of 10 g kg−1 seed. Data on nodulation were taken at the late-flowering stage, whereas nitrogen and phosphorus concentrations in plant parts were analyzed at physiological maturity. The total nitrogen difference method was employed to quantify nitrogen fixation. Location x rhizobium strain x variety interaction had a significant effect on nodule dry weight plant−1. Rhizobium strains significantly enhanced nodulation, nitrogen fixation, nutrient uptake and soil nitrogen balance. Inoculation with NSFBR-12 and NSFBR-15 resulted in the highest nitrogen fixed, nutrient uptake and soil nitrogen balance. Vicia faba inoculated with the two top performing strains, NSFBR-12 and NSFBR-15 fixed respectively 87.7% and 85.5% of the total nitrogen uptake. Non-inoculated plants fulfilled proportionately more of the total nitrogen uptake through nitrogen derived from the soil rather than fixed nitrogen. Soil available phosphorus and pH had appreciable influences on nitrogen and phosphorus uptake of inoculated Vicia faba. Inoculation with competitive and effective rhizobium strains can improve soil nitrogen balance, nitrogen fixation and nutrient uptake of Vicia faba.

The promiscuity of Phaseolus vulgaris L. (common bean) for nodulation with rhizobia: a review.

Phaseolus vulgaris L. (common bean) is a legume indigenous to American countries currently cultivated in all continents, which is nodulated by different rhizobial species and symbiovars. Most of species able to nodulate this legume worldwide belong to the genus Rhizobium, followed by those belonging to the genera Ensifer (formerly Sinorhizobium) and Pararhizobium (formerly Rhizobium) and minority by species of the genus Bradyrhizobium. All these genera belong to the phylum alpha-Proteobacteria, but the nodulation of P. vulgaris has also been reported for some species belonging to Paraburkholderia and Cupriavidus from the beta-Proteobacteria. Several species nodulating P. vulgaris were originally isolated from nodules of this legume in American countries and are linked to the symbiovars phaseoli and tropici, which are currently present in other continents probably because they were spread in their soils together with the P. vulgaris seeds. In addition, this legume can be nodulated by species and symbiovars originally isolated from nodules of other legumes due its high promiscuity, a concept currently related with the ability of a legume to be nodulated by several symbiovars rather than by several species. In this article we review the species and symbiovars able to nodulate P. vulgaris in different countries and continents and the challenges on the study of the P. vulgaris endosymbionts diversity in those countries where they have not been studied yet, that will allow to select highly effective rhizobial strains in order to guarantee the success of P. vulgaris inoculation.

Isolation, characterization and symbiotic performance evaluation of soybean (Glycine max) nodulating rhizobia from different districts of Bangladesh

Rhizobia can meet up nitrogen requirement of legumes by biological nitrogen fixation through symbiosis. The main objective of this study was to find out the morpho-physiological diversity of indigenous soybean nodulating rhizobia and to find effective rhizobial strains for enhancing better soybean production at different environmental conditions. Fourteen rhizobial strains were isolated and studied their morph-physiological characteristics; evaluated their nodulation and symbiotic efficiency at potted soil. Among the strains, seven were slow growing, four were intermediate to slow growing and three were fast growing. The strains exposed creamy, milky-white and transparent single colonies between 1.17 and 3.67 mm in diameter after 3 to 7 days on growth medium. Twelve strains were alkali producers and only two strains were acid producers. Most of them tolerated both acidic pH (4.0 and 5.0) and alkaline pH (9.0 and 10.0) conditions. All strains tolerated to 1.0% NaCl but none of them survived at 5.0% salt stress. They retained their normal growth up to 37oC but most of them showed growth susceptibility at 45oC and growth was inhibited at 50oC. The strains were inoculated as treatments on soybean plants to compare their performance on growth of soybean along with urea and control treatment. Overall, rhizobial treatments significantly increased nodulation and growth of soybean plants over recommended dose of urea and negative control. Diverse rhizobial strains were associated with soybean root nodules in Bangladesh. The strains SB-27, SB-28 and SB-212 were very effective and produced almost double plant dry matter weight over some other strains, dose of urea and negative control. The physical stress tolerant and highly symbiotic strains deserve to be effective as bio-fertilizer for soybean crop production.

Lotus japonicus alters in planta fitness of Mesorhizobium loti dependent on symbiotic nitrogen fixation.

Rhizobial bacteria are known for their capacity to fix nitrogen for legume hosts. However ineffective rhizobial genotypes exist and can trigger the formation of nodules but fix little if any nitrogen for hosts. Legumes must employ mechanisms to minimize exploitation by the ineffective rhizobial genotypes to limit fitness costs and stabilize the symbiosis. Here we address two key questions about these host mechanisms. What stages of the interaction are controlled by the host, and can hosts detect subtle differences in nitrogen fixation? We provide the first explicit evidence for adaptive host control in the interaction between Lotus japonicus and Mesorhizobium loti. In both single inoculation and co-inoculation experiments, less effective rhizobial strains exhibited reduced in planta fitness relative to the wildtype M. loti. We uncovered evidence of host control during nodule formation and during post-infection proliferation of symbionts within nodules. We found a linear relationship between rhizobial fitness and symbiotic effectiveness. Our results suggest that L. japonicus can adaptively modulate the fitness of symbionts as a continuous response to symbiotic nitrogen fixation.

Differential growth costs and nitrogen fixation in Cytisus multiflorus (L'Hér.) Sweet and Cytisus scoparius (L.) Link are mediated by sources of inorganic N.

Shrubby legumes in Mediterranean-type ecosystems face strong nutrient limitations that worsen in summer, when water is absent. Nitrogen-fixing legumes are likely to be able to switch between soil N and atmospheric N (N2 ) sources to adjust the C costs of N acquisition in different seasons. We investigated the utilisation of different inorganic N sources by two indigenous shrubby legumes (Cytisus multiflorus and Cytisus scoparius). Plant performance in terms of photosynthesis and biomass production was also analysed. Plants were cultivated in sterile river sand supplied with Hoagland nutrient solution, grown in N-free nutrient solution and inoculated with effective rhizobial strains from nodules of adult plants of the same species. A second treatment consisted of plants given 500μm NH4 NO3 added into the nutrient solution. In a third treatment, plants were watered with another source of N (500μm NH4 NO3 ) as well as being inoculated with effective rhizobial strains. The application of NH4 NO3 to the legumes resulted in a larger increase in plant dry matter. Carbon construction costs were higher in plants supplied with mineral and symbiotic N sources and always higher in the endemic C.multiflorus. Differences in photosynthesis rates were only observed between species, regardless of the N source. Non-fertilised inoculated plants had more effective root nodules and a clear dependence on N2 fixation. We propose that the ability of C.scoparius to change N source makes it a plastic species, which would account for its broader distribution in nature.

Isolation, characterization and selection of indigenous Bradyrhizobium strains with outstanding symbiotic performance to increase soybean yields in Mozambique.

Soybean inoculation with effective rhizobial strains makes unnecessary the use of N-fertilizers in the tropics. A frequently reported problem is the failure of the inoculant strains to overcome the competition imposed by indigenous rhizobial populations. The screening of indigenous rhizobia, already adapted to local conditions, searching for highly effective strains for use as inoculants represents a promising strategy in overcoming inoculation failure. The objective of this study was to isolate and characterize indigenous rhizobia and to identify strains that hold potential to be included in inoculant formulations for soybean production, with both promiscuous and non-promiscuous soybean cultivars, in Mozambican agro-climatic conditions. A total of 105 isolates obtained from nodules of promiscuous soybean grown at 15 sites were screened for N2-fixation effectiveness in the greenhouse along with five commercial strains. Eighty-seven isolates confirmed the ability to form effective nodules on soybean and were used for genetic characterization by rep-PCR (BOX) and sequencing of the 16S rRNA gene, and also for symbiotic effectiveness. BOX-PCR fingerprinting revealed remarkable genetic diversity, with 41 clusters formed, considering a similarity level of 65%. The 16S rRNA analysis assigned the isolates to the genera Bradyrhizobium (75%) and Agrobacterium/Rhizobium (25%). Great variability in symbiotic effectiveness was detected among the indigenous rhizobia from Mozambique, with ten isolates performing better than the commercial strain B. diazoefficiens USDA 110, the best reference strain, and 51 isolates with lower performance than all reference strains. Thirteen of the best isolates from the first greenhouse trial were evaluated, along with the five commercial strains, in two promiscuous (TGx 1963-3F and TGx 1835-10E) and one non-promiscuous (BRS 284) soybean cultivars in a second greenhouse trial. In general the promiscous soybeans responded better to inoculation. The 13 isolates were also characterized for tolerance to acidity and alkalinity (pH 3.5 and 9.0, respectively), salinity (0.1, 0.3 and 0.5molL-1 of NaCl) and high temperatures (35, 40 and 45°C) in vitro. Five isolates, three (Moz 4, Moz 19 and Moz 22) belonging to the superclade B. elkanii and two (Moz 27 and Moz 61) assigned to the superclade B. japonicum, consistently showed high symbiotic effectiveness, suggesting that the inoculation with indigenous rhizobia adapted to local conditions represents a possible strategy for increasing soybean yields in Mozambique. Phylogenetic position of the five elite isolates was confirmed by the MLSA with four protein-coding housekeeping genes, dnaK, glnII, gyrB and recA.

Improvement of Faba Bean Yield Using Rhizobium/Agrobacterium Inoculant in Low-Fertility Sandy Soil

Soil fertility is one of the major limiting factors for crop’s productivity in Egypt and the world in general. Biological nitrogen fixation (BNF) has a great importance as a non-polluting and a cost-effective way to improve soil fertility through supplying N to different agricultural systems. Faba bean (Vicia faba L.) is one of the most efficient nitrogen-fixing legumes that can meet all of their N needs through BNF. Therefore, understanding the impact of rhizobial inoculation and contrasting soil rhizobia on nodulation and N2 fixation in faba bean is crucial to optimize the crop yield, particularly under low fertility soil conditions. This study investigated the symbiotic effectiveness of 17 Rhizobium/Agrobacterium strains previously isolated from different Egyptian governorates in improving the nodulation and N2 fixation in faba bean cv. Giza 843 under controlled greenhouse conditions. Five strains that had a high nitrogen-fixing capacity under greenhouse conditions were subsequently tested in field trials as faba bean inoculants at Ismaillia Governorate in northeast Egypt in comparison with the chemical N-fertilization treatment (96 kg N·ha−1). A starter N-dose (48 kg N·ha−1) was applied in combination with different Rhizobium inoculants. The field experiments were established at sites without a background of inoculation under low fertility sandy soil conditions over two successive winter growing seasons, 2012/2013 and 2013/2014. Under greenhouse conditions, inoculated plants produced significantly higher nodules dry weight, plant biomass, and shoot N-uptake than non-inoculated ones. In the first season (2012/2013), inoculation of field-grown faba bean showed significant improvements in seed yield (3.73–4.36 ton·ha−1) and seed N-yield (138–153 Kg N·ha−1), which were higher than the uninoculated control (48 kg N·ha−1) that produced 2.97 Kg·ha−1 and 95 kg N·ha−1, respectively. Similarly, in the second season (2013/2014), inoculation significantly improved seed yield (3.16–4.68 ton·ha−1) and seed N-yield (98–155 Kg N·ha−1) relative to the uninoculated control (48 kg N·ha−1), which recorded 2.58 Kg·ha−1 and 80 kg N·ha−1, respectively. Interestingly, faba bean inoculated with strain Rlv NGB-FR 126 showed significant increments in seed yield (35%–48%) and seed N-yield (34%–49%) compared to the inorganic N fertilizers treatment (96 kg N·ha−1) over the two cropping seasons, respectively. These results indicate that inoculation of faba bean with effective rhizobial strains can reduce the need for inorganic N fertilization to achieve higher crop yield under low fertility soil conditions.

COWPEA (VIGNA UNGUICULATA) CROPS IN AFRICA CAN RESPOND TO INOCULATION WITH RHIZOBIUM

SUMMARYCowpea (Vigna unguiculata L. Walp) is the most important food grain legume in Africa. Cowpea is nodulated by rhizobium bacteria in almost all soils of the tropics, but studies performed in the 1970s and 1980s in Nigeria suggested only modest responses of grain yield in the field to inoculation of selected rhizobium strains. More recently, experiments performed in Brazil have shown that cowpea responded to inoculation of rhizobium selected locally and grain yields increased by up to 30%. We tested some of the Brazilian strains on cowpea at a site in northern Mozambique and at several sites in Northern Ghana. At all sites phosphorus fertilizer (26 kg P ha−1) was added to all plots. At the site in Mozambique despite considerable damage to the crop by the parasitic yellow witchweed (Alectra vogelii), grain yields were more than doubled by inoculation of one of the Brazilian strains and reached 1.4 Mg ha−1. In on-station experiments conducted in 2012 in June and August in northern Ghana using the local cowpea variety Padi-Tuya as the test crop, nodule weight at 35 days after planting (dap) tripled with rhizobium strain BR 3299 (530 mg plant−1) in August with the other inoculants (BR 3267 and a mixture of BR 3267 and BR 3299) also increased nodule weight to over 300 mg plant−1. In the first on-station experiment, grain yields were doubled by the inoculation of any of the three rhizobium strains, and in the second experiment, significant increases in grain yield ranged from 39% to 57% and reached over 2.0 Mg ha−1. Similar increases in nodulation and grain yield due to inoculation were observed in 22 on-farm trials. Nitrogen fertilizer application promoted vegetative growth but did not increase grain yield and nodulation. Inoculating cowpea with highly effective rhizobium strains can therefore enhance grain yield of smallholder farmers in Africa.

Patterns of Growth Costs and Nitrogen Acquisition in Cytisus striatus (Hill) Rothm. and Cytisus balansae (Boiss.) Ball are Mediated by Sources of Inorganic N.

Nitrogen-fixing shrubby legumes in the Mediterranean area partly overcome nutrient limitations by making use of soil N and atmospheric N2 sources. Their ability to switch between different sources lets them adjust to the carbon costs pertaining to N acquisition throughout the year. We investigated the utilization of different inorganic N sources by Cytisus balansae and Cytisus striatus, shrubby legumes under low and a sufficient (5 and 500 µM P, respectively) levels of P. Plants grew in sterile sand, supplied with N-free nutrient solution and inoculated with effective Bradyrhizobium strains; other treatments consisted of plants treated with (i) 500 µM NH4NO3; and (ii) 500 µM NH4NO3 and inoculation with effective rhizobial strains. The application of NH4NO3 always resulted in greater dry biomass production. Carbon construction costs were higher in plants that were supplied with mineral and symbiotic N sources and always greater in the endemic C. striatus. Photosynthetic rates were similar in plants treated with different sources of N although differences were observed between the two species. Non-fertilized inoculated plants showed a neat dependence on N2 fixation and had more effective root nodules. Results accounted for the distribution of the two species with regards to their ability to use different N sources.

Effect of plant population and N fertilizer on the growth and yield of bambara groundnut (<i>Vigna subterranean</i> (L.) Verdc.)

Responses of bambara groundnut (Vigna subterranea (L.) Verdc) to 3 levels of fertilizer N (0, 50, and 100 kg N/ha) and seven plant populations (55555, 63492, 74074, 88888, 111111, 148148 and 222222 plants/ha) were studied under field conditions in Nsukka, Nigeria. The experimental design was a randomized complete block and there were three replications. Seed yield progressively increased with increasing plant population, being significantly highest at 222,222 plants/ha. The application of 100 kg N/ha increased the total dry matter per plant compared with no application or with 50 kg N/ha, although seed yield benefits were not readily apparent. Nodulation was not statistically influenced by N treatment although zero or- low N application showed a slight increase in nodulation, suggesting a lack of adequate population of the effective rhizobium in the location. Innoculation of bambara groundnut seeds with effective rhizobium strain before planting becomes necessary for good nodulation and production of the crop under Nsukka agro-ecological conditions.Key words: Vigna subterranea, nodulation, N-fertilizer, plant population, seed yield, drymatter yield.

Effectiveness of halo-tolerant, auxin producing Pseudomonas and Rhizobium strains to improve osmotic stress tolerance in mung bean (Vigna radiata L.)

Halo-tolerant, auxin producing bacteria could be used to induce salt tolerance in plants. A number of Rhizobium and auxin producing rhizobacterial strains were assessed for their ability to tolerate salt stress by conducting osmoadaptation assay. The selected strains were further screened for their ability to induce osmotic stress tolerance in mung bean seedlings under salt-stressed axenic conditions in growth pouch/jar trials. Three most effective strains of Rhizobium and Pseudomonas containing ACC-deaminase were evaluated in combination, for their ability to induce osmotic stress tolerance in mung bean at original, 4, and 6 dS m(-1) under axenic conditions. Results showed that sole inoculation of Rhizobium and Pseudomonas strains improved the total dry matter up to 1.4, and 1.9 fold, respectively, while the increase in salt tolerance index was improved up to 1.3 and 2.0 fold by the Rhizobium and Pseudomonas strains, respectively. However, up to 2.2 fold increase in total dry matter and salt tolerance index was observed due to combined inoculation of Rhizobium and Pseudomonas strains. So, combined application of Rhizobium and Pseudomonas strains could be explored as an effective strategy to induce osmotic stress tolerance in mung bean.

<b>Cowpea nodulation, biomass yield and nutrient uptake, as affected by biofertilizers and rhizobia, in a sodic soil amended with <i>Acidithiobacillus</i></b> - doi: 10.4025/actasciagron.v35i4.16994

Sodic soils require application of amendments as gypsum and organic matter. Many types of compost have been tested in sodic soils reclamation; however, these materials often do not provide satisfactory pH reduction. A recent study reported effective effects applying mixture of gypsum and sulfur inoculated with Acidithiobacillus in sodic soils with high pH and exchangeable sodium, though the effects on plant parameters were not evaluated. The present study was conducted to verify the effects of BPK rock biofertilizers on nodulation, biomass yield and nutrient uptake in cowpea compared with mineral fertilizer after sodic soil amendment. The BPK biofertilizers and PK mineral fertilizer were applied at different rates, and plants were inoculated with effective rhizobia strains. A control that did not receive PK fertilization was included. The results indicated that gypsum and sulfur with Acidithiobacillus reduced the soil’s pH and the amount of soil exchangeable sodium. BPK rock biofertilizer increased cowpea nodulation, biomass yield and nutrient uptake. The native rhizobia in the soil exhibited effectiveness in cowpea growth; displaying similar results compared with the rhizobia inoculated plants. BPK biofertilizers may be used as alternative to mineral PK fertilizers in sodic soils after the application of gypsum and sulfur inoculated with Acidithiobacillus.

SYMBIOTIC NITROGEN-FIXING ACTIVITY AND YIELD POTENTIAL OF INOCULATED HEDYSARUM CORONARIUM IN ISRAEL

ABSTRACT Hedysarum coronarium L. has recently been introduced into Israel for evaluation as a “new” winter forage crop. The lack of a native rhizobial population suitable for this legume and the need for inoculation were assessed. Field inoculation experiments with effective Rhizobium strains, previously selected under greenhouse conditions, proved the benefits of symbiotically fixed N2 to yield potential and forage quality of H. coronarium. Seasonal changes in nodulation, plant dry matter accumulation and N2 (C2 H2)-fixation activity under field conditions were followed. The effect of defoliation on root nodule development and nitrogen fixation rate during the vegetative regrowth period are briefly discussed.