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.     

A significant proportion of indigenous rhizobia from India associated with soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) distinctly belong to <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Ensifer</Emphasis> genera

The diversity among 269 rhizobia isolated from naturally occurring root nodules of soybean collected from two different agro-ecological regions of India, based on RFLP and sequences of the intergenic spacer (IGS) between the 16S and 23S rRNA genes, growth rate, and indole acetic acid production, revealed their significant, site-dependent genomic diversity. Among these bacteria, nine IGS genotypes were identified with two endonucleases. They were distributed into five divergent lineages by sequence analysis of each IGS representative strain, i.e., (1) comprising IGS genotypes I, II, III, and reference Bradyrhizobium yuanmingense; (2) with genotype IV and strains of unclassified bradyrhizobia genomic species; (3) including genotypes V, VI, and Bradyrhizobium liaoningense; (4) with IGS genotype VII and Bradyrhizobium elkanii strains; and (5) comprising IGS genotypes VIII, IX, and different Ensifer genus bacteria. Host-specificity test revealed that all rhizobia-nodulated soybean and cowpea and only part of them formed nodules on Arachis hypogeae and Cajanus cajan. The great diversity of soybean nodulators observed in this study emphasises that Indian soil is an important reservoir of nitrogen-fixing rhizobia.     

Effects of some Nigerian Bradyrhizobium inoculants on the performance of three cowpea cultivars (Vigna unguiculata) in two field plots

Summary We assessed the effectiveness of three locally made lignite, subbituminous coal and cowmanure-based cowpea Bradyrhizobium inoculants in comparison with a peat-based imported Bradyrhizobium incoculant in a two-field plot investigation. The local inoculants were prepared by incorporating three rhizobia strains (Ife CR9, Ife CR15 and Bradyrhizobium japonicum) into each of the above carrier materials and were used to inoculate three cowpea seed varieties: TVU 1190, IT 82E-60 and Ife brown. With lignite-based He CR9 inoculated into TVU 1190 seeds, total N content of the plants was 178.6 mg/plant compared with only 64.3 mg/plant for the uninoculated nitrate-free control plants. With Nigerian lignite, sub-bituminous coal and cow manure as carriers for cowpea rhizobia, the cowpea yield of the inoculated plants increased by 72%, 54% and 10%, respectively, compared with uninoculated plants, while the peat-based inoculant gave a 25% increase in cowpea yield. With lignite-based Ife CR9 inoculated into Ife brown seeds, total N content of the plants was 149.1 mg/plant, but with inoculation by lignite-based B. japonicum, total N content of the treated Ife brown plant was 132.4 mg/plant. Thus, the native Ife CR9 strain seems to be slightly better adapted to tropical conditions than the imported B. japonicum.     

Genetic variability of soybean bradyrhizobia populations under different soil managements

Due to their ecologic and economic importance, bradyrhizobia have been extensively studied in recent years. Since 1992, Bradyrhizobium elkanii SEMIA 587 and SEMIA 5019 and Bradyrhizobium japonicum SEMIA 5079 and SEMIA 5080 have been widely used in most Brazilian soybean fields. The objective of this work was to estimate the genetic variability of bradyrhizobial isolates recovered from soils under rhizobial inoculation and different soil managements. Only 25% of the isolates demonstrated high similarities to the original strains, and a strong correlation was obtained between the bradyrhizobial genetic variability and soil management. A high level of genetic diversity was observed both within isolates (H = 5.46) as well as among the different soil practices. Soil under no-tillage presented a higher bradyrhizobia diversity compared with bradyrhizobia isolated from soil under conventional tillage. Serological characterization also indicated that B. elkanii strains SEMIA 587 and SEMIA 5019 were more competitive and presented a higher nodular occupancy capacity than strains belonging to B. japonicum species in Southern Brazilian soils.     

Dual inoculation with strains of Bradyrhizobium japonicum and Azospirillum brasilense to improve growth and biological nitrogen fixation of soybean (Glycine max L.)

The effects of inoculation with Bradyrhizobium japonicum and Azospirillum brasilense strains on the growth of soybean were evaluated with regard to the estimation of N₂ fixation using the ¹⁵N isotope dilution technique. Inoculation, in general, increased the dry mass of soybean as well as nitrogen content. Dual inoculation with a mixture of B. japonicum and A. brasilense strains was superior over single inoculation with B. japonicum. Nitrogen fixed (Ndfa) varied according to inoculant and soil conditions. Percentages of nitrogen derived from air (% Ndfa) using a non-nodulating isoline were 72% and 76% for B. japonicum and B. japonicum plus A. brasilense, respectively, in non-sterile soil. A similar but higher trend was recorded in sterilized soil, in which the percentages of N₂ fixed were 81% and 86% for single and dual inoculation, respectively. The correlation coefficient between N₂ fixed and N uptake (r=0.94) and dry mass (r=0.89) was significant. Application of special bacterial inoculants in agricultural systems of Egypt seems to be a promising technology and could be used for improving soybean growth as well as soil fertility, thus minimizing environmental pollution. Received: 10 January 1996     

Potential of indigenous bradyrhizobia versus commercial inoculants to improve cowpea (Vigna unguiculata L. walp.) and green gram (Vigna radiata L. wilczek.) yields in Kenya

Limited information is available on reduced cowpea (Vigna unguiculata L. Walp.) and green gram (Vigna radiata L.Wilczek.) yields in Kenya. Declining soil fertility and absence or presence of ineffective indigenous rhizobia in soils are assumptions that have been formulated but still require to be demonstrated. In this study, soils were collected from legume growing areas of Western (Bungoma), Nyanza (Bondo), Eastern (Isiolo), Central (Meru) and Coast (Kilifi) provinces in Kenya to assess indigenous rhizobia in soils nodulating cowpea and green gram under greenhouse conditions. Our results showed that highest nodule fresh weights of 4.63 and 3.32?g plant^?1 for cowpea and green gram were observed in one soil from Isiolo and another from Kilifi, respectively, suggesting the presence of significant infective indigenous strains in both soils. On the other hand, the lowest nodule fresh weights of 2.17 and 0.72?g plant^?1 were observed in one soil from Bungoma for cowpea and green gram, respectively. Symbiotic nitrogen (N) fixation by cowpea and green gram was highest in Kilifi soil with values of 98% and 97%, respectively. A second greenhouse experiment was undertaken to evaluate the performance of commercial rhizobial inoculants with both legumes in Chonyi soil (also from Coast province) containing significant indigenous rhizobia [>13.5?×?10³ Colony Forming Units (CFU) g^?1]. Rhizobial inoculation did not significantly (P?相似文献   

Dual inoculation of peanut with Glomus sp. and Bradyrhizobium sp. enhanced the symbiotic nitrogen fixation as assessed by 15N-Technique

Abstract

The response of peanut (Arachis hypogaea L.) to inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi (Glomus etunicatum) and Bradyrhizobiurn sp. was studied in pots by the acetylene reduction activity (ARA) and ‘A-value’ methods. The soil used was a Light-coloured Andosol and the treatments consisted of the inoculation of VAM fungi only, inoculation of Bradyrhizobium only, dual inoculation of VAM fungi and Bradyrhizobium and control, under non-sterilized and sterilized soil conditions.

In the non-sterilized soil the ARA and nitrogen fixation determined by the ‘A-value’ method increased significantly only by dual inoculation of VAM fungi and Bradyrhizobium at 100 days after planting (DAP), but no significant difference was observed at 70 DAP. In the case of dual inoculation, 75% of the nitrogen of the plant was derived from fixation whereas the plants inoculated only with Bradyrhizobium derived 68% of their nitrogen from fixation and the control plants, 64%. Amount of P in plant increased significantly only by dual inoculation with VAM fungi and Bradyrhizobium.

In the sterilized soil a highly significant increase in the ARA was observed of the dual inoculation at all the sampling times. Nitrogen fixation determined by the A-value technique and N and P contents in plant also increased significantly by dual inoculation. Results obtained by the A-value method showed that plants with dual inoculation derived 68% of their nitrogen from fixation while the plants inoculated only with Bradyrhizobium, 38%.

From our this study we conclude that nitrogen fixation as well as N and P contents in peanut increased significantly only by dual inoculation with VAM fungi and Bradyrhizobium.     

Phosphate Solubilization by Several Genera of Saprophytic Fungi and Its Influence on Corn and Cowpea Growth

The aim of this study was to test the ability of several strains of fungi, which were isolated from the Brazilian Amazon, to solubilize calcium phosphate in vitro and to promote corn and cowpea growth under axenic conditions. Each plant species received six treatments: inoculation with strains with high solubilization index (SI) (Haematonectria ipomoeae CML 3249 and Pochonia chlamydosporia var. catenulate CML 3250) and control treatments: inoculation with strain that does not solubilize phosphate on Pikovskaya agar (PVK) (Acremonium polychromum FSA115), and non-inoculated treatments with high concentration of insoluble phosphate (HPins), high concentration of soluble phosphate (HPs) or low concentration of soluble phosphate (LPs). The fungi strains had SI between 1.07 cm and 2.03 cm including species without previous report in the literature of their capacity to solubilize calcium phosphate. The two phosphate-solubilizing strains promoted greater corn and cowpea root growth than the controls FSA115, HPins and LPs, to a level similar to the HPs control.     

Quantification of nitrogen fixation by the peanut/Rhizobium symbiotic system in a virgin sandy soil

Nitrogen balance studies were conducted to quantify the nitrogen fixed by peanut/Rhizobium symbiotic system under field conditions in a sandy soil. Large scale inoculation with three NifTAL strains of cowpea rhizobia, 1000, 169, 1371 was done using two inoculation techniques: peat-based inoculant or injection of inoculant with irrigation water through an injection tank attached to the central pivot system. The results show nitrogen fixation amounting up to 186 kg N ha^?1 in peat-based inoculant and 171 kg N ha^?1 in liquid inoculant injected through the irrigation system. However, no significant differences in yield response were recorded between both inoculation techniques.     

Symbiotic performance of fast-growing rhizobia isolated from the coastal sand dune legumes of west coast of India

The symbiotic efficiency of coastal sand dune rhizobial isolates on four cultivated legumes, cowpea (Vigna unguiculata), green gram (Vigna radiata), black gram (Vigna mungo) and horse gram (Macrotyloma uniflorum), was assessed. Among the isolates of Someshwara (S1–S5), inoculation of S5 resulted in the highest increase of shoot biomass in cowpea (control vs experimental, 1:6), while inoculation of P1 among the Padubidri isolates (P1–P5) induced the highest shoot biomass in cowpea (1:14.4). Inoculation of the isolate P2 induced higher shoot biomass against uninoculated controls of horse gram (12.6:1), green gram (11.2:1) and black gram (6.1:1). One-way ANOVA revealed significant difference in the shoot biomass between uninoculated and inoculated cowpea plants with ten rhizobial isolates (P <0.05). Cultivation of surface-sterilized green gram seeds on unsterilized dune sand resulted in profuse flowering as well as nodules within 6 weeks indicating possibilities for isolating efficient rhizobial strains through cultivating edible legumes on coastal sand dune soils.     

Response of a wild-type and modern cowpea cultivars to arbuscular mycorrhizal inoculation in sterilized and non-sterilized soil

Cowpea is an important crop that serves as a legume and vegetable source to many smallholder farmers in sub-Saharan Africa. Soil fertility is a significant limitation to its production thus; inoculation with beneficial soil biota such as arbuscular mycorrhizal fungi (AMF) could improve its performance. However, plant–AMF interaction could vary based on crop cultivar hence affecting overall crop production. The present study aimed at determining the effect of AMF inoculation and soil sterilization on root colonization and growth of a wild-type and three modern cowpea cultivars grown by smallholder farmers in Kenya. Potted cowpea plants were inoculated with a commercial AMF inoculum comprising of Rhizophagus irregularis, Funneliformis mosseae, Glomus aggregatum and Glomus etunicatum and maintained in a greenhouse for 40 days. After harvesting, mycorrhizal colonization, nodule number and dry weight, root and shoot dry weights, nitrogen (N,) phosphorus (P) and potassium (K) content were determined. Interestingly, the modern cultivars showed significantly (p < 0.001) higher root colonization, nodulation, shoot P and N compared to the wild-type cultivar. Moreover, a strong positive correlation between AMF root colonization and shoot P (r² = 0.73, 0.90, p < 0.001), AMF root colonization and shoot N (r² = 0.78; 0.89, p < 0.001) was observed in both sterilized and non-sterilized soil, respectively. Soil sterilization affected root colonization and growth parameters with plants grown in non-sterilized soil performing better than those grown in sterilized soil. This study provides major evidence that modern cowpea cultivars are still responsive to mycorrhizal inoculation suggesting that modern breeding programs are not deleterious AMF symbiosis.     

Occurrence and nature of mixed infections in nodules of field-grown soybeans (Glycine max)

Summary Mixed infections of Bradyrhizobium japonicum strains in early and late nodules of four soybean cultivars were studied in a field soil. Nodule occupants were identified by immunofluorescence using serogroup specific antibodies prepared against B. japonicum strains USDA 110, USDA 123, and USDA 138. Double infection was determined directly by combined examination of the same microscopic field by fluorescence and phase contrast microscopy. Double strain occupancy was observed consistently, and its occurrence did not differ substantially in pouch, soil pot, and field experiments, ranging in incidence from 12% to 32%. No significant differences in the incidence or nature of double infection could be attributed to cultivar, seed inoculation, or plant maturity. Strains reactive to strain USDA 123-fluorescent antibody were dominant in both singly and doubly infected nodules irrespective of cultivar, plant age, or seed inoculation with strain USDA 110.Paper no. 15092 in the Scientific Journal Series of the Minnesota Agricultural Experiment Station, St. Paul     

接种bradyrhizobium sp.对豇豆生物量及根瘤的影响

Vigna unguiculata(L.)Walp (cowpea)is a tropical legume of very high nutrive and economic values.A laboratory experiment.was conducted to evaluate the BNF (biological nitrogen fixation) effciency of 4 strains of Bradyrhizobium sp.(VUXYI,VUIE1,VUMDS1 and VUID1).From the results obtained these strains were classified into efficiency groups,with VUID1 being the most efficient and VUXY1 the least efficient.the effect of inoculation was tested on 2 cowpea varieties in a field experiment.Remarkable increases in nodulation biomass and crop yield were observed.An increase from 58% ti 81% in dry seed was obtained for the two varieties.A significance test revealed a signifecantly positive correlation between nodulation and biomass.     

土壤中大豆根瘤菌之间竞争结瘤的研究——Ⅲ.接种菌量对大豆生长的影响

本文研究了大豆根瘸菌PRC005的接菌量对大豆生长的影响.田间试验结果表明:接菌量在播种后40天和60天没有显著增加根瘤数、根瘤干重、地上部植株干重和植株含氮量.施氮肥处理和较高接菌量处理之间的大豆种子产量差异不显著,与不接菌对照处理相比,施氮肥和较高接菌量两个处理的种子产量显著增加.施氮肥处理没有使植株含氮量增加,并且还妨碍了大豆的结瘸作用.室内盆栽试验结果表明:只有当接菌量高于土著菌数1200倍时,才能显著地提高大豆的结瘤数和植株干重.     

DEHP污染的两种土壤中接种丛枝菌根对豇豆生长和DEHP降解的影响

A 60-day pot experiment was carried out using di-(2-ethylhexyl) phthalate (DEHP) as a typical organic pollutant phthalic ester and cowpea (Vigna sinensis) as the host plant to determine the effect of arbuscular mycorrhizal inoculation on plant growth and degradation of DEHP in two contaminated soils, a yellow-brown soil and a red soil. The air-dried soils were uniformly sprayed with different concentrations of DEHP, inoculated or left uninoculated with an arbuscular mycorrhizal (AM) fungus, and planted with cowpea seeds. After 60 days the positive impact of AM inoculation on the growth of cowpea was more pronounced in the red soil than in the yellow-brown soil, with significantly higher (P < 0.01) mycorrhizal colonization rate, shoot dry weight and total P content in shoot tissues for the red soil. Both in the yellow-brown and red soils, AM inoculation significantly (P < 0.01) reduced shoot DEHP content, implying that AM inoculation could inhibit the uptake and translocation of DEHP from roots to the aboveground parts. However, with AM inoculation no positive contribution to the degradation of DEHP was found.     

Field performance of three Bradyrhizobium japonicum strains with two soybean (Glycine max. L.) cultivars

Summary A field experiment was condutced in a clay loam soil to study the performance of three Bradyrhizobium japonicum strains; USDA 110, USDA 138 and TAL 379, in relation to their N₂-fixing potential and competitiveness on two soybean cultivars (Clark and Calland). Inoculation of soybean cultivars with these strains, either singly or in combination, induced significant increases in plant dry weight, N₂ fixation and seed yields. However, no significant differences were found between the rhizobial strains and/or their mixtures in N₂ fixation and increased seed yield for both cultivars. The two soybean cultivars gave similar responses to inoculation. No significant differences in seed yield were observed between Clark and Calland cultivars. The interaction between inoculant strain and soybean cultivar was not significant. The competition between strains for nodulation was assessed. Strain USDA 110 was the most competitive, followed by USDA 138. Strain TAL 379 was always less competitive on both cultivars. The incidence of double-strain occupancy of nodules varied from 8% to 40%.     

Diversity,phylogeny and host specificity of soybean and peanut bradyrhizobia

A study on the diversity, phylogeny, and host specificity of soybean (Glycine max L.) and peanut (Arachis hypogaea L.) bradyrhizobia was conducted based on the 16S ribosomal RNA (rRNA) restriction fragment length polymorphisms (RFLPs), 16S rRNA sequencing, and 16S–23S rRNA intergenetic spacer (IGS) RFLP assays. Based on 16S rRNA RFLP assay, tested bradyrhizobia were divided into five genotypes, which could be further clustered into five groups by IGS RFLP assays. According to the 16S rRNA sequencing, strains of IGS-II, IV, and V were phylogenetically related to Bradyrhizobium liaoningense, Bradyrhizobium japonicum, and Bradyrhizobium elkanii, while strains of IGS-Ic and IGS-III related to Bradyrhizobium yuanmingense and Bradyrhizobium canariense, respectively. All isolates could crossly nodulate Phaseolus vulgaris, forming small white nodules. Strains of IGS-II originally isolated from peanut could efficiently nodulate Glycine soja, and two strains isolated from soybean could also nodulate peanut.     

Co-selection of compatible rhizobia and vesicular-arbuscular mycorrhizal fungi for cowpea in sterilized and non-sterilized soils

Summary We selected two isolates of Rhizobium for cowpea (Vigna unguiculata) with sterilized soil tests and two different isolates by non-sterilized soil testing. The four rhizobia were then paired individually with either Glomus pallidum, Glomus aggregatum, or Sclerocystis microcarpa in separate, sterilized, or non-sterilized soil experiments. The purpose of the experiments was to determine the effect of soil sterilization on the selection of effective cowpea rhizobia, and to see whether these rhizobia differed in their effects on cowpea growth when paired with various vesicular-arbuscular mycorrhizal (VAM) fungi. Our experiments showed that the rhizobia selected in sterilized soil tests produced few growth responses in the cowpea compared to the other introduced rhizobia, irrespective of pairing with VAM fungi in sterilized or non-sterilized soil. In contrast, the two rhizobia initially selected by non-sterilized soil testing significantly improved cowpea growth in non-sterilized soil, especially when paired with G. pallidum. Our results suggest that it is important to select for effective rhizobia in non-sterilized soil, and that pairing these rhizobia with specific, coselected VAM fungi can significantly improve the legume growth response.     

Effect of soil physical properties on soybean nodulation and N2 fixation at the early growth stage in heavy soil field in Hachirougata Polder,Japan

We studied the effect of the soil physical properties on soybean nodulation and N₂ fixation in the heavy soil of an upland field (UF) and an upland field converted from a paddy field (UCPF) in the Hachirougata polder, Japan. Seeds of the soybean cultivar Ryuho were sown in each field with or without inoculation of Bradyrhizobium japonicum A1017. The soybean plants were sampled at 35 (V3) and 65 (Rl) d after sowing (DAS), and then nodulation and the percentage of N derived from N₂ fixation in the xylem sap were determined. The soil physical properties were different between UF and UCPF, especially the air permeability and soil water regime. Nodule growth was restricted in UCPF irrespective of rhizobial inoculation, though rhizobial infection was not inhibited by the unfavorable soil physical conditions. Soybean plant growth was closely related to the nodule mass and N₂ fixation activity, and the inoculation of a superior rhizobium strain was effective only at 35 DAS. These results indicate that soybean nodulation and N₂ fixation was considerably affected by the physical properties of heavy soil, and that it is important to maintain the N₂ fixation activity and inoculate the soybean plants with a superior rhizobium strain at a later growth stage in order to increase soybean production in heavy soil fields.     

Survival and potential denitrifying activity of Azospirillum lipoferum and Bradyrhizobium japonicum inoculated into sterilized soil

Summary Potential denitrifying activity and population dynamics of Azospirillum lipoferum (137C) and Bradyrhizobium japonicum (G2sp) inoculated into a -sterilized soil were studied for a period of 3 weeks. The denitrifying enzyme potential of soil inoculated independently with each bacterial species was strongly stimulated by the presence of a plant (Zea mays L.). Simultaneous inoculation of both bacteria also produced a higher denitrifying enzyme potential than simple inoculation. Even with double inoculation, the presence of a plant did not modify the evolution of the activity. The response of the population dynamics to these treatments followed a different pattern. The population dynamics of A. lipoferum was not affected by the presence of the plant or by the presence of B. japonicum. In contrast, the presence of both a plant and of A. lipoferum seemed to promote the growth of B. japonicum.