Nodulation of African yam bean (Sphenostylis stenocarpa) by Bradyrhizobium sp. isolated from Erythrina brucei

African yam bean (Sphenostylis stenocarpa), which is widely cultivated in Africa because of its growth capability on marginal soils, was nodulated by an endosymbiont (characterized and designed Bradyrhizobium sp. AUEB20) isolated from the Ethiopian tree Erythrina brucei with the formation of a small number of large, indeterminate N₂-fixing nodules. In contrast, 24 other isolates from Ethiopian woody legumes were ineffective. Strain AUEB20 promiscuously nodulated a number of tropical legumes, but none out of five European crop plants tested. Received: 17 September 1996     

Interactions between drought stress and vesicular-arbuscular mycorrhiza on the growth of Faidherbia albida (syn. Acacia albida) and Acacia nilotica in sterile and non-sterile soils

Summary Faidherbia albida (syn. Acacia albida) (Del.) A. Chev. and Acacia nilotica (L.) Willd. were grown for 18 weeks in sterile and non-sterile soils inoculated with Glomus clarum (Nicolson and Schenck). During this period, drought stress was imposed for the last 10 (F. albida) or 12 weeks (A. nilotica) at 2-week intervals. A greater number of leaves abscissed in drought-stressed mycorrhizal plants of A. nilotica than drought-stressed non-mycorrhizal and unstressed plants. In F. albida, the number of abscissed leaves was few and similar for all treatments. At the end of the drought stress, inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi in sterile soil increased the plant biomass of the two tree species compared to the control plants. In non-sterile soil, the mycorrhizal growth response of introduced G. clarum equalled the effect of indigenous VAM fungi. There were significant interactions between the mycorrhizal and drought stress treatments and between the mycorrhizal and soil treatments for plant biomass and P uptake in F. albida. The absence of these interactions except for that between the mycorrhizal and soil treatments in A. nilotica indicates that the increased plant biomass and nutrient uptake cannot be attributed directly to a mycorrhizal contribution to drought tolerance. F. albida tolerated the drought stress by producing long tap roots and similar weights of dry matter in shoots and roots. Whereas A. nilotica tolerated the drought stress by developing larger root systems able to explore a greater volume of soil, in addition to leaf abscission, for a favourable internal water status. The introduction of G. clarum increased nodulation by A. nilotica under unstressed conditions, but at the expense of a reduced P uptake in sterile soil.     

Co-introduction of exotic rhizobia to the rhizosphere of the invasive legume Acacia saligna,an intercontinental study

Invasive woody legumes have profound impacts in the nitrogen content and cycling of invaded ecosystems due to the ability to enter into symbiosis with nitrogen-fixing bacteria. In spite of the relevance of this symbiosis, the identity and origin of the symbionts involved in invasion are not well understood. We conducted a study to assess the diversity of symbiotic root-nodulating bacteria associated with the invasive Acacia saligna, in newly colonized areas in Portugal and Australia. BOX-PCR was used to discriminate the isolated bacteria and 16S rRNA and nifD genes were sequenced to identify the different isolates and their geographic origin. Bradyrhizobium and Mesorhizobium nodulated A. saligna in Australia while only Bradyrhizobium spp. were found in Portugal. The dominant strains nodulating A. saligna were related to Bradyrhizobium liaoningense and Bradyrhizobium canariense. Co-occurring Acacia longifolia and A. saligna in Australia harbor different rhizobial communities. As an example, we found Mesorhizobium sp. and Phyllobacterium trifolii in A. saligna and A. longifolia respectively, being this the first report for this association. The analysis of the phylogeographic marker nifD clustered most of the sequences obtained in this study with sequences of Australian origin, indicating that exotic bradyrhizobia might have been co-introduced with A. saligna in Portugal. This result highlights the risks of introducing exotic inoculants that might facilitate the invasion of new areas and alter native soil bacterial communities, hindering the recovery of ecosystems.     

Adaptation of green manure legumes to adverse conditions in rice lowlands

Poor adoption of sustainable pre-rice green manure technology by lowland farmers is frequently associated with unreliable legume performance under adverse environmental conditions such as marginal soils, short photoperiod, and unfavorable hydrology. A series of field and microplot experiments were conducted at the International Rice Research Institute (IRRI) in 1991 and 1992 to screen and evaluate 12 promising flood-tolerant legumes for adaptation (N accumulation and biological N₂ fixation) to a range of environmental stresses, frequently encountered in rice lowlands. Legumes belonging to the genera Sesbania and Aeschynomene were grown for 8 weeks at 10×10 cm spacing: (1) in a fertile control soil and in four marginally productive irrigated lowland rice soils (sandy Entisol, P-deficient Inceptisol, acid Ultisol, and saline Mollisol); (2) during short- (11.7h) and long-day (12.3 h) seasons in a favorable irrigated lowland soil; and (3) in an aerobic soil (drought-prone rain-fed lowland) and a deep-flood-prone lowland soil (1 week seedling submergence). A large variability in N accumulation was obsersed among legume species and across different environments, ranging from less than 1 to over 70 mg N plant^-1. The nitrogen derived from the atmosphere (Ndfa) accounted on average for 82% of total N accumulation. Sesbania virgata was least affected by unfavorable soil conditions but its Ndfa was the lowest among the tested species (less than 60%). Stem nodule formation did not convey a significant advantage to legumes grown under adverse soil conditions. However, flooding reduced N₂ fixation less in stem-nodulating than in solely root-nodulating species. Most species drastically reduced N accumulation under short-day conditions. Aeschynomene afraspera and S. speciosa were least affected by photoperiod. The considerable genetic variability in the germplasm screened allows the selection of potentially appropriate legumes to most conditions studied, thus increasing N accumulation in green manures.     

Root-nodule bacteria from indigenous legumes in the north-west of Western Australia and their interaction with exotic legumes

Bacteria were isolated from root-nodules collected from indigenous legumes at 38 separate locations in the Gascoyne and Pilbara regions of Western Australia. Authentication of cultures resulted in 31 being ascribed status as root-nodule bacteria based upon their nodulation of at least one of eight indigenous legume species. The authenticated isolates originated from eight legume genera from 19 sites. Isolates were characterised on the basis of their growth and physiology; 20 isolates were fast-growing and 11 were slow-growing (visible growth within 3 and 7 d, respectively). Fast-growers were isolated from Acacia, Isotropis, Lotus and Swainsona, whilst slow-growers were from Muelleranthus, Rhynchosia and Tephrosia. Indigofera produced one fast-growing isolate and seven slow-growing isolates. Three indigenous legumes (Swainsona formosa, Swainsona maccullochiana and Swainsona pterostylis) nodulated with fast-growing isolates and four species (Acacia saligna, Indigofera brevidens, Kennedia coccinea and Kennedia prorepens) nodulated with both fast- and slow-growing isolates. Swainsona kingii did not form nodules with any isolates. Fast-growing isolates were predominantly acid-sensitive, alkaline- and salt-tolerant. All slow-growing isolates grew well at pH 9.0 whilst more than half grew at pH 5.0, but all were salt-sensitive. All isolates were able to grow at 37 °C. The fast-growing isolates utilised disaccharides, whereas the slow-growing isolates did not. Symbiotic interactions of the isolates were assessed on three annual, one biennial and nine perennial exotic legume species that have agricultural use, or potential use, in southern Australia. Argyrolobium uniflorum, Chamaecytisus proliferus, Macroptilium atropurpureum, Ononis natrix, Phaseolus vulgaris and Sutherlandia microphylla nodulated with one or more of the authenticated isolates. Hedysarum coronarium, Medicago sativa, Ornithopus sativus, Ornithopus compressus, Trifolium burchellianum, Trifolium polymorphum and Trifolium uniflorum did not form nodules. Investigation of the 31 authenticated isolates by polymerase chain reaction with three primers resulted in the RPO1 primer distinguishing 20 separate banding patterns, while ERIC and PucFor primers distinguished 26 separate banding patterns. Sequencing the 16S rRNA gene for four fast- and two slow-growing isolates produced the following phylogenetic associations; WSM1701 and WSM1715 (isolated from Lotus cruentus and S. pterostylis, respectively) displayed 99% homology with Sinorhizobium meliloti, WSM1707 and WSM1721 (isolated from Sinorhizobium leeana and Indigofera sp., respectively) displayed 99% homology with Sinorhizobium terangae, WSM1704 (isolated from Tephrosia gardneri) shared 99% sequence homology with Bradyrhizobium elkanii, and WSM1743 (isolated from Indigofera sp.) displayed 99% homology with Bradyrhizobium japonicum.     

Effects of symbiosis with Rhizobium fredii on transport of fixed nitrogen in the xylem of soybean plants

An experiment was conducted to identify the main nitrogenous compound transported in the xylem sap of soybean plants nodulated with Rhizobium fredii. Soybean (Glycine max L. Merr.) cultivars, wild type Bragg (nod⁺, fix⁺) and its nitrate tolerant, hypernodulating mutant ntsll16 (nod⁺⁺, fix⁺) were used for this experiment. These soybean plants were inoculated with a slowgrowing rhizobium, Bradyrhizobium japonicum USDAllO or fast-growing rhizobia consisting of a mixture of R. fredii USDA191, USDA193, and USDA-194 and grown in a phytotron under natural light and controlled temperature conditions. Xylem sap was collected from Bragg and ntsll16 plants at the flowering and pod elongation stages. Acetylene reduction activity per plant or per nodule weight was not different between soybean lines and inoculums. The composition of the nitrogenous compounds in the xylem sap was compared between the symbionts, with B. japonicum and R. fredii. At the flowering stage, ureide-N and amide-N accounted for 53 to 70% and 20 to 27% respectively of the total N in the sap collected from the plants inoculated either with B. japonicum or R. fredii. At the pod elongation stage, ureide-N and amide-N accounted for 74 to 85%, and 7 to 19% of total sap N. With the growth of the soybean plants, the ratio of ureide-N in the xylem sap increased. These results suggest that in the case of wild soybean and the hypernodulating mutant line nodulated by R. fredii, ureide is transported as the main nitrogenous compound of fixed nitrogen in the xylem sap in the same way as in plants nodulated with B. japonicum.     

Salinity effects on growth analysis and nutrient composition in four grain legumes‐rhizobium symbiosis

The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was investigated for four legumes: faba bean (Vicia faba L), pea (Pisum sativum L), soybean (Glycine max L), and common bean (Phaseolus vulgaris L). Plants were grown in a vermiculite culture system supplied with a N‐free nutrient solution with the addition of 0, 50, and 100 mM NaCl. Plants were harvested at the beginning of the flowering period and the dry weights of shoots and roots and acetylene reduction activity (ARA) were evaluated at the same time plant tissues were analysed for N, potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) contents.

The depressive effect of saline stress on ARA of nodules was directely related to the salt induced decline in dry weight and N content in shoots. Growth inhibition by NaCl treatments was greater for the pea than for other legumes, whereas the soybean was the most salt‐tolerant Saline stress also affected the N content in shoots and roots. In general the N content accumulated in the shoot and Na in the roots of the four legumes tested, while K accumulated both organs. The acquisition of other macronutrients differed according to the legume species. The legumes most sensitive were P. sativum and V. faba which accumulated Ca in shoot and Mg both in the shoot and the roots. On the contrary, in G. max and P. vulgaris, the two most salt tolerant legumes, accumulated Mg in the roots and Ca in both vegetative organs. Our results suggest a relationship between the salt‐tolerant range in legumes and the macronutrient accumulation in vegetative organs.     

Controlled ectomycorrhization of an exotic legume tree species Acacia holosericea affects the structure of root nodule bacteria community and their symbiotic effectiveness on Faidherbia albida, a native Sahelian Acacia

Many fast growing tree species have been introduced to promote biodiversity rehabilitation on degraded tropical lands. Although it has been shown that plant productivity and stability are dependent on the composition and functionalities of soil microbial communities, more particularly on the abundance and diversity of soil symbiotic micro-organisms (mycorrhizal fungi and rhizobia), the impact of tree introduction on soil microbiota has been scarcely studied. This research has been carried in a field plantation of Acacia holosericea (Australian Acacia species) inoculated or not with an ectomycorrhizal fungus isolate, Pisolithus albus IR100. After 7 year's plantation, the diversity and the symbiotic properties of Bradyrhizobia isolated from the plantation soil or from the surrounding area (Faidherbia albida (Del.) a. Chev. parkland) and able to nodulate F. albida, a native Sahelian Acacia species, have been studied. Results clearly showed that A. holosericea modified the structure of Bradyrhizobia populations and their effectiveness on F. albida growth. This negative effect was counterbalanced by the introduction of an ectomycorrhizal fungus, P. albus, on A. holosericea root systems.In conclusion, this study shows that exotic plant species can drastically affect genotypic and symbiotic effectiveness of native Bradyrhizobia populations that could limit the natural regeneration of endemic plant species such as F. albida. This effect could be counterbalanced by controlled ectomycorrhization with P. albus. These results have to be considered when exotic tree species are used in afforestation programs that target preservation of native plants and soil ecosystem rehabilitation.     

Characterization of local rhizobia in Thailand and distribution of malic enzymes

TWenty-six isolates were obtained from nodules of various legume plants (Glycine max, Vigna sinensis, Arachis hypogaea, Desmanthus virgatus, Acacia mangium, Centrosema pascuorum, Pterocarpus indicus, Xylia xylocarpa, and Sesbania rostrata) in Thailand. After confirming their nodulation and nitrogen-fixing abilities, they were identified by 16S rRNA gene analysis as Bradyrhizobium japonicum, Bradyrhizobium elkanii, Rhizobium leguminosarum, Rhizobium gallicum, and Rhizobium galegae. Using these local isolates, the distribution of the activities of both NAD⁺-dependent (DME: EC 1.1.1.39) and NADP⁺-dependent (TME: EC 1.1.1.40) malic enzymes was surveyed. The malic enzyme activities were present in all the isolated rhizobia and in other 17 local Bradyrhizobium strains in Thailand. In almost all the rhizobia, the DME activity predominated whereas the TME activity predominated only in the Rhizobium gallicum strains that were major symbionts of Sesbania rostrata. Southern hybridization analysis was performed to survey the distribution of the malic enzyme genes among the local rhizobia, which are similar to those of B. japonicum. DNA probes (ME1 for DME and ME2 for TME) were prepared by polymerase chain reaction (PCR) using degenerated primers from conserved regions of the protein sequences of bacterial malic enzymes. Southern blot analysis with ME1 as a probe showed a single band in about half of the isolates, especially in B. japonicum and R. leguminosarum strains, suggesting the wide distribution of such DME genes among local rhizobia. In contrast, Southern blot analysis with ME2 as a probe detected a single band only in five B. japonicum strains, suggesting that the TME genes, which are similar to those of B. japonicum, would be unique in a group of B. japonicum.     

Nodulation and Symbiotic Nitrogen Fixation of Cowpea Genotypes as Affected by Fertilizer Nitrogen

Abstract

Intercropping with legumes and non‐legumes is commonly practiced in many parts of the world to maximize productivity per unit area of land. In India, cowpea [Vigna unguiculate (L.) Walp] is a popular pulse legume component of intercropping farming systems. Often, however, potential production is compromised, particularly in high fertilizer input systems, because legume component competes with the non‐legume component of the system for nitrogen (N) in the soil. An experiment was conducted in order to identify lines of cowpea that could obtain the majority of their nitrogen requirements from symbiotic fixation of atmospheric nitrogen rather than from uptake of soil nitrogen. Twenty‐nine genotypes of cowpea were screened for tolerance to (applied) nitrogen in soil in field condition. The parameters used to appraise tolerance were extent of root nodulation, the amount of nitrogen fixed, nitrate reductase activity (NR) in roots and nodules, and nitrite content of roots and nodules. There were two nitrogen treatments applied as urea, 40 kg N per ha (N40), and 120 kg N per ha (N120). There were three genotypes whose nitrogen‐fixing effectiveness was apparently unimpaired by applications of nitrogen to the soil. Genotype EC‐170442‐3 nodulated and fixed atmospheric nitrogen satisfactorily at higher levels of applied nitrogen. At N40, genotypes EC‐244390 and EC‐240900 formed a great abundance of large nodules effective in nitrogen fixation; even at N120, EC‐240900 had better symbioses than the majority of the 29 cowpea lines originally screened. These three genotypes are deemed worthy of further examination for their suitability for intercropping systems. How this might be achieved is discussed.     

Symbiotic dinitrogen fixation - its dependence on plant nutrition and its ecophysiological impact

The cultivation of symbiotic legumes and the consequent small uptake of nitrate by the host plant depresses soil pH and impairs subsequent legume growth. The rate of soil pH decline depends on the soil H⁺ buffer power, climatic conditions, soil permeability, and on the kind of legume cropping. It is shown for some leguminous crop species from the temperate climate that in many cases the N₂ fixation performance of Rhizobium/Bradyrhizobium symbiosis is more dependent on the growth conditions of the host plant than on the nitrogenase potential of the bacteroid. This is true for the supply of the host plant with phosphate, potassium and water as well as for light intensity. Nitrogen deficiency of the host plant because of insufficient nitrogenase activity was observed at low soil pH where nodulation and the development of the nitrogenase activity were delayed. Nodulation and nitrogenase activity are suppressed by high levels of available nitrogen in the soil. There are indications that the nitrogenase activity is very flexible and adjusts to the demand of the host plant. The mechanism of this regulation is not yet understood.     

Influence of reference trees on N2-fixation estimates inLeucaena leucocephala andAcacia albida using15N-labelling techniques

Summary We examined the suitability of four reference crops, i.e., two non-fixing trees,Cassia siamea andEucalyptus grandis, and two uninoculated fixing trees,Leucaena leucocephala andAcacia albida, for measuring fixed N₂ fixed in inoculatedL. leucocephala andA. albida grown for 36 weeks in pots. The¹⁵N isotope-dilution (involving the addition of equal amounts of labelled N fertilizer to the non-fixing and the fixing plants) and theA-value (with different amounts of labelled N fertilizer added to the fixing and the non-fixing crops) methods were used. The isotope dilution approach gave several large negative estimates of fixed N₂ inA. albida. Positive and similar values of fixed N₂ were measured in all four reference crops using theA-value approach. ForL. leucocephala the isotope-dilution approach gave different estimates of fixed N₂, with the different reference crops; the uninoculated N₂-fixing crops indicated significantly less fixed N₂ than the non-fixing reference crops. Similar values for N₂ fixed inL. leucocephala were obtained using the two non-fixing trees, either by the isotope-dilution or theA-value method. On average,A. albida derived about twice as much N from fertilizer asL. leucocephala. In both species, the atom %¹⁵N excess declined by about 50% in successive harvests.     

Nodulation status of native woody legumes and phenotypic characteristics of associated rhizobia in soils of southern Ethiopia

The nodulation of provenances of Acacia seyal, Acacia tortilis and Faidherbia albida, and other indigenous multipurpose tree species were tested in 14 different soil samples collected from diverse agro-ecological zones in southern Ethiopia. Associated rhizobia were isolated from these and from excavated nodules of field standing mature trees, and phenotypically characterized. Indigenous rhizobia capable of eliciting nodules on at least one or more of the woody legume species tested were present in most of the soils. Tree species were markedly different in nodulation in the different site soils. Sesbania sesban and Acacia abyssinica showed higher nodulation ability across the different sites indicating widespread occurrence of compatible rhizobia in the soils. The nodulation patterns of the different provenances of Acacia spp. suggested the existence of intraspecific provenance variations in rhizobial affinity which can be exploited to improve N fixation through tree selection. Altogether, 241 isolates were recovered from the root nodules of trap host species and from excavated nodules. Isolates were differentiated by growth rate and colony morphology and there were very fast-, fast-, slow-, and very slow-growing rhizobia. The bulk of them (68.5%) were fast-growing acid-producing rhizobia while 25.3% were slow-growing alkali-producing types. Fast-growing alkali-producing (2.9%) and slow-growing acid-producing strains (3.3%) were isolated from trap host species and excavated nodules, respectively. All isolates fell into four colony types: watery translucent, white translucent, dull glistering and milky (curdled) type. The diversity of indigenous rhizobia in growth rate and colony morphology suggested that the collection probably includes several rhizobial genera.     

Biological effects of native and exotic plant residues on plant growth, microbial biomass and N availability under controlled conditions

The leaf litter of six tropical tree species (Acacia holosericea, Acacia tortilis, Azadirachta indica, Casuarina equisetifolia, Cordyla pinnata and Faidherbia albida) frequently used in agroforestry plantations in Sahelian and Soudano-Sahelian areas were tested for their influence on soil nitrogen content, microbial biomass and plant growth under controlled greenhouse conditions. Half of the soil was planted with onion (Allium cepa L.) seedlings and the other half was not. Two herbaceous species, Andropogon gayanus and Eragrostis tremula, were also studied. Co-inertia analysis (CIA) and one-way analysis of variance (ANOVA) analysis showed that C. pinnata and F. albida leaf powder amendment induced the highest plant growth, whereas leaf powder of E. tremula is associated to higher microbial biomass and NH₄⁺ content. Higher onion seedlings growth is associated with higher concentration of nitrogen and lignin in leaf powders. Conversely, lower plant growth is associated to higher rates of cellulose, hemicellulose and phenols in leaves. Higher rates of cellulose and hemicellulose are associated with higher microbial biomass and NH₄⁺, whereas phenols are associated to lower microbial biomass. The results showed that amendment of A. holosericea leaf powder (high concentrations of phenol) to the soil resulted in a lower microbial biomass and lower onion seedlings growth. Data showed that the plant residue quality index (PRQI) could be a useful tool to predict the effects of litter materials on root growth in glasshouse conditions. The highest values on soil and plant parameters were recorded with C. pinnata litter. While powdered leaf material increased the accessibility of substrates to microbes, more research with C. pinnata leaf litter (under a wider range of ecological conditions) is needed. It could add deeper on its agronomic impact in the tropics.     

Response of Vigna Unguiculata Grown Under Different Soil Moisture Regimes to the Dual Inoculation with Nitrogen-Fixing Bacteria and Arbuscular Mycorrhizal Fungi

The interaction between legumes, rhizobial and arbuscular mycorrhizal (AM) partners benefits plant nutrition and improves plant tolerance to water stress. The present research evaluated the effectiveness of symbioses between cowpea plants (Vigna unguiculata (L.) Walp.), AM fungi (Glomus intraradices) and two strains of Bradyrhizobium japonicum on the mycorrhization, acid phosphatase activity (APase), enzymes related to nitrogen fixation and assimilation, and biomass accumulation at three soil moisture levels. The results revealed that the soil moisture optimal for the formation of active symbiotrophic associations in cowpea cultivation was about 60% water-holding capacity (WHC), where both Bradyrhizobium strains and AM fungi function well with respect to mycorrhization, nitrogen and phosphorus uptake, nitrogen fixation and plant biomass production. Under conditions of reduced water supply, the symbiotic association between Br. japonicum-273 and Gl. intraradices was better for cowpea cultivation, while in elevated soil moisture association between Br. japonicum-269 and Gl. intraradices was more appropriate.     

Partitioning of above and below ground costs during phosphate stress in Medicago truncatula

Phosphate (P) availability for plant uptake can limit the yield of natural and agricultural systems. Under P limited conditions, the P-requirement of symbiotic nodules of legumes may exacerbate the P stress of host plants. Adaptations to survive under P stress may vary between different functioning tissues. This study investigated the physiological adaptations to P stress in above and below ground organs of nodulated Medicago truncatula Gaertner. Seedlings were inoculated with Sinorhizobium meliloti in quartz sand and fed nutrient solution with either 0.01?mM or 0.5?mM P concentrations. P-stressed nodulated plants showed compromised photosynthetic responses. Alternative growth allocation during stressed conditions was observed between different organs. The concentration of inorganic P, carbon, and nitrogen were lower during stressed conditions. The above ground tissues scavenged P and lowered their dependence on adenosine-triphosphate required for metabolism. Whereas the below ground tissues recycled phosphate from phosphate monoesters in the cell.     

The effect of acidity on the distribution and symbiotic efficiency of rhizobia in Lithuanian soils

The distribution and symbiotic efficiency of nodule bacteria Rhizobium leguminosarum_bv. trifolii F., Sinorhizobium meliloti D., Rhizobium galegae L., and Rhizobium leguminosarum bv. viciae F. in Lithuanian soils as dependent on the soil acidity were studied in the long-term field, pot, and laboratory experiments. The critical and optimal pH values controlling the distribution of rhizobia and the symbiotic nitrogen fixation were determined for every bacterial species. The relationship was found between the soil pH and the nitrogen-fixing capacity of rhizobia. A positive effect of liming of acid soils in combination with inoculation of legumes on the efficiency of symbiotic nitrogen fixation was demonstrated.     

Rhizobium strain,a banana (Musa spp.)-associated bacterium with a high potential as biofertilizer

A diazotrophic bacterial strain denominated 11B isolated from the rhizosphere of a banana plant (Musa spp.) was characterized morphologically, biochemically, and phylogenetically, whereas the symbiotic potential of the strain was assessed through tests of host range, interstrain nodulation competitiveness, and capacity to synthesize indole-3-acetic acid (IAA) and solubilize phosphate. Based on morphological and physiological–biochemical properties, as well as 16S ribosomal deoxyribonucleic acid (rDNA) sequence and phylogenetic analysis, the strain 11B belonged to the genus Rhizobium with 100.0% sequence similarity with Rhizobium tropici CAF440. The optimum growth temperature and pH for strain 11B are 28°C and 7.2, respectively. This strain was able to produce IAA, solubilize phosphate, and fix large amounts of nitrogen (N₂) and form effective nodules on the legumes Acaciella angustissima, Gliricidia sepium, Leucaena leucocephala, Lysiloma acapulcensis, and Phaseolus vulgaris. The rhizobial strain 11B was used successfully as a biofertilizer in agriculturally important legumes, forest trees, and agroindustrial plants.     

Enhanced soybean (Glycine max L.) plant growth and nodulation by Bradyrhizobium japonicum-SB1 in presence of Bacillus thuringiensis-KR1

Abstract

Nodulation and subsequent nitrogen fixation are important factors that determine the productivity of soybean (Glycine max L.). The beneficial effects of nodulation can be enhanced when rhizobial inoculation is combined with plant-growth-promoting bacteria (PGPB). The PGPB strain Bacillus thuringiensis-KR1, originally isolated from the nodules of Kudzu vine (Pueraria thunbergiana), was found to promote growth of soybean plants (variety VL Soya 2) under Jensen's tube and growth pouch conditions, when co-inoculated with Bradyrhizobium japonicum-SB1. Co-inoculation with Bacillus thuringiensis-KR1 (at a cell density of 10 cfu) provided the highest and most consistent increase in nodule number, shoot weight, root weight, root volume, and total biomass, over rhizobial inoculation and control, under both conditions. The results demonstrate the potential benefits of using nonrhizobial nodule occupants of wild legumes for the co-inoculation of soybean, with Bradyrhizobium japonicum-SB1, in order to achieve plant-growth promotion and increased nodulation.     

Proton release by nodulated roots varies among common bean genotypes (Phaseolus vulgaris) under phosphorus deficiency

Responses of proton release to phosphorus (P) availability by nodulated roots of common bean (Phaseolus vulgaris L.) were investigated for lines BAT 477 and CocoT, inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic culture under glasshouse conditions. Phosphorus was supplied as KH₂PO₄ at 15 and 60 μmol plant^–1 week^–1 (15P and 60P). Proton release was higher for BAT 477 than for CocoT under both P supplies. However, it was higher for 60P than 15P, whatever the line. The ratio of proton release per unit biomass of nodulated root was higher for BAT 477 than for CocoT, independent of P deficiency. Proton release was correlated with the nodulated‐root respiration for both genotypes and with the nodule respiration linked with nitrogen fixation for CocoT. Thus, the nodulation was more limited by 15P than root and shoot growth and more in CocoT than in BAT 477. It is concluded that independent of symbiotic N₂ fixation, proton release was higher in BAT 477 than in CocoT and that the nodulated legume releases a substantial amount of protons into its rhizosphere that is correlated with its nitrogen fixation that eventually depends upon the nodule permeability to O₂ diffusion.