Effects of co-inoculation of Bradyrhizobium japonicum SAY3-7 and Streptomyces griseoflavus P4 on plant growth,nodulation, nitrogen fixation,nutrient uptake,and yield of soybean in a field condition

ABSTRACT

Co-inoculation of nitrogen-fixing bacteria with plant growth-promoting bacteria has become more popular than single inoculation of rhizobia or plant-growth-promoting bacteria because of the synergy of these bacteria in increasing soybean yield and nitrogen fixation. This study was conducted to investigate the effects of Bradyrhizobium japonicum SAY3-7 and Streptomyces griseoflavus P4 co-inoculation on plant growth, nodulation, nitrogen fixation, nutrient uptake, and seed yield of the ‘Yezin-6’ soybean cultivar. Nitrogen fixation was measured using the acetylene reduction assay and ureide methods. Uptake of major nutrients [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg)] was also measured. This study showed that single inoculation of SAY3-7 significantly increased shoot biomass; nodulation; Relative Ureide Index (RUI %), percent nitrogen derived from N fixation (% Ndfa); N, P, K, Ca, and Mg uptakes; during the later growth stages (R3.5 and R5.5), compared with control. These observations indicate that SAY3-7 is an effective N-fixing bacterium for the plant growth, nodulation, and nitrogen fixation with an ability to compete with native bradyrhizobia. Co-inoculation of SAY3-7 and P4 significantly improved nodule number; nodule dry weight; shoot and root biomass; N fixation; N, P, K, Ca, and Mg uptake; at various growth stages and seed yield in ‘Yezin-6’ soybean cultivar compared with the control, but not the single inoculation treatments. Significant differences in plant growth, nodulation, N fixation, nutrient uptake, and yield between co-inoculation and control, not between single inoculation and control, suggest that there is a synergetic effect due to co-inoculation of SAY3-7 and P4. Therefore, we conclude that Myanmar Bradyrhizobium strain SAY3-7 and P4 will be useful as effective inoculants in biofertilizer production in the future.     

Selection of type A and type B strains for improving symbiotic effectiveness on non-Rj and Rj4 genotype soybean varieties

The selection of effective rhizobia for higher efficiency nitrogen fixation is one of the most important steps for inoculant production. Therefore, this experiment was conducted to select the most effective type A and type B strains for specific Rj-gene harboring soybean varieties and to test the symbiotic effectiveness of selected strains on different Rj-gene harboring soybean varieties. Screening experiments using the specific soybean varieties were done with a completely randomized design and three replications in this study. Evaluation of the effective Myanmar Bradyrhizobium strains for plant growth, nodulation and N₂ fixation were studied in pot experiments using sterilized vermiculite in the Phytotron (controlled-environmental condition). Then, a pot experiment was conducted using Futsukaichi soil in the screen house (natural environmental condition). The N₂ fixation ability of soybean was evaluated by acetylene reduction activity (ARA) and the relative ureide index method. In the first screening experiment, type A and type B strains with higher nitrogen fixation and proper nodulation on their respective soybean cultivars were selected for the next screening. In the second screening, Bradyrhizobium elkanii AHY3-1 (type A), Bradyrhizobium japonicum SAY3-7 (type A), B. elkanii BLY3-8 (type B) and B. japonicum SAY3-10 (type B) isolates, which showed higher nitrogen fixation and nodulation in Yezin-3 (Rj₄) and Yezin-6 (non-Rj), were selected for the next experiment. In the third screening experiment, SAY3-7 and BLY3-8, which had higher nitrogen fixing potential and proper nodulation, were selected as effective isolates. These two isolates were compatible with non-Rj and Rj₄ soybean varieties for nodulation and nitrogen fixation. Based on the results of the screening experiment, these two strains were tested for their symbiotic efficacy in Futsukaichi soil. This study shows that inoculation treatment of SAY3-7 and BLY3-8 significantly increased plant growth, nodulation, and N₂ fixation at the V6, R3.5 and R8 stages in Yezin-3 (Rj₄) and/or Yezin-6 (non-Rj), and the seed yield at R8 stage, in Yezin-3 (Rj₄) and Yezin-6 (non-Rj) soybean varieties compared with the control treatment. It can be concluded that SAY3-7 and BLY3-8 are suitable for inoculant production because of their higher nitrogen fixation ability, proper nodulation and better productivity of Myanmar soybean cultivars.     

Searching for Myanmar indigenous Bradyrhizobium type C strains that best identify Rj4 genotypes in soybean

Bradyrhizobium species are symbiotic partners of soybean plants. However, some Bradyrhizobium bacteria do not form functional nodules on the roots of Rj₄ genotype soybean cultivars. Our objective was to identify the strains of Bradyrhizobium (i.e., type C strains) that are least competent to form nodules on the roots of this plant genotype. We checked (i) previously isolated type C strains of Myanmar Bradyrhizobium elkanii (MMY6-1, MMY6-2, and MMY6-5), (ii) previously isolated type C strains of Myanmarese Bradyrhizobium spp. (MMY3-5 and MMY3-7), and (iii) strain Is-34 of B. japonicum, for nodule formation when associated with Rj₄ and other Rj genotype soybeans. Strains in groups (i), (ii), and (iii) are known to be incompatible with Rj₄ soybean genotypes. MMY3-5 and MMY3-7 produced functional nodules when associated with Rj₄ and other Rj genotype soybean cultivars, except Hill (Rj₄) cultivar. The ratios of ineffective nodule numbers/total nodule numbers (I/T ratios) for MMY6-1, MMY6-2, MMY6-5, and Is-34 in association with Rj₄ soybean cultivars were > 0.5, demonstrating incompatibility between these bacterial strains and the Rj₄ genotype. Interestingly, the I/T ratios of MMY6-1 and MMY6-2 were higher than that of Is-34 in almost all Rj₄ soybean cultivars. Thus, the nodule-forming abilities of the B. elkanii strains MMY6-1 and MMY6-2 were strongly suppressed in Rj₄ soybean cultivars; these strains may therefore be useful to identify the Rj₄ genotype in soybean cultivars.     

Effects of root exudates of Rj2Rj3- and Rj4-genotype soybeans on growth and chemotaxis of Bradyrhizobium japonicum

Almost all of the soybean cultivars (Glycine max L. Merr.) form nodules on their roots by infection with Bradyrhizobium japonicum. However, it has been observed that the soybean cultivars harboring nodulation conditioning genes, for instance Rj ₂, Rj ₃, and Rj ₄, do not form effective nodules with some strains of B. japonicum. Ishizuka et al. (1991b) classified the rhizobia isolated from field-grown soybeans into nodulation-types A, B, and C based on the compatibility with these Rj-soybean cultivars. Nodulation-type B is incompatible with Rj ₂ Rj ₃-cultivars and type C is incompatible with Rj ₄-cultivars. Nodulation-type A rhizobia are compatible with both Rj ₂ Rj ₃ - and Rj ₄-cultivars. Furthermore, the Rj-cultivars were found to prefer appropriate nodulation-type rhizobia for nodulation, that is, non-Rj-, Rj ₂ Rj ₃ -, and Rj ₄-cultivars prefer nodulation-types A, C, and B rhizobia, respectively (Ishizuka et al. 1991a). In the present study, in order to examine the mechanisms of preference of Rj-cultivars for rhizobial strains for nodulation, root exudates from Rj-cultivars were examined for their effects on the growth rate and chemotaxis of various types of B. japonicum strains.     

Interactive effects of co-inoculation of Bradyrhizobium japonicum strains and mycorrhiza species on soybean growth and nutrient contents in plant

The main objective of this study was to investigate the effects of co-inoculation with different strains of Bradyrhizobium japonicum (i.e. Helinitro, Rizoking, and Nitragin) and arbuscular mycorrhizal fungi (AMF) species (i.e. Glomus fasciculatum, Glomus versiforme, Glomus intraradices, Glomus mosseae, and Glomus etunicatum) on soybean growth, fungal root colonization, and nutrient uptake of nitrogen (N), phosphorus (P), zinc (Zn), iron (Fe), and copper (Cu). Co-inoculation with various AMF species and rhizobia significantly (p<0.01) increased the soybean biomass production as compared to the non-inoculated controls. Furthermore, AMF colonization of roots of soybean plants increased by 79, 70.1, 67, 63, 57.5, and 50.1% in the presence of G. fasciculatum (GF), G. versiforme (GV), G. intraradices (GI), G. mosseae (GM), and G. etunicatum (GE), and Gmix (a mixed culture of fungi), respectively. Higher nutrient contents were observed in plants co-inoculated with Helinitro and GF. More insight into these results will enable optimization of the effective use of AM fungi in combination with their bacterial partners as a tool for increasing soybean yields in Iran; however, its general analytical framework could be applied to other parts of the world.     

Evaluation of effective Myanmar Bradyrhizobium strains isolated from Myanmar soybean and effects of coinoculation with Streptomyces griseoflavus P4 for nitrogen fixation

In order to substitute the use of chemical fertilizers in legume production, there is a need for the production of rhizobial inoculants which are capable of being used as biofertilizers. To achieve this, an effective symbiotic nitrogen (N) fixation between legumes and root nodule bacteria will be essential. Evaluation of effective Myanmar Bradyrhizobium (Jordan 1982) strains isolated from Myanmar soybean (Glycine max L. Merr.) and effects of coinoculation with Streptomyces griseoflavus Krainsky 1914 P4 for N fixation were studied in pot experiments using sterilized vermiculite and Hoagland solution in the Phytotron (25°C and 70% relative humidity) with completely randomized design and three replicates. N fixation ability of soybean was evaluated by acetylene reduction activity (ARA) by gas chromatography. It was found that MAS23 showed a relatively high degree of stability and a high level of ARA per plant on both Yezin-3 and Yezin-6 soybean varieties. In the symbiotic relationship between Bradyrhizobium strains and P4 experiments, the treatments consisted of six Bradyrhizobium strains (MAS23, MAS33, MAS34, MAS43, MAS48 and USDA110) and Streptomyces griseoflavus P4 were evaluated with four Myanmar soybean varieties (Yezin-3, Yezin-6, Hinthada and Shan Sein). In the Yezin-3 soybean variety, the best treatment for ARA per plant was found in the dual inoculation of P4 and MAS34. In the Yezin-6 soybean variety, the highest nodule dry weight was found in dual inoculation of P4 with MAS34 but the highest ARA per plant was observed in the dual inoculation of P4 and MAS23. On the other hand, single inoculation of MAS43 and coinoculation of P4 with MAS48 were significantly higher in N fixation of Hinthada, and coinoculation of P4 with MAS33 was significant improvement of ARA per plant (P < 0.05) in Shan Sein soybean.     

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%.     

Copper,nitrogen, and Rhizobium japonicum relationships in determinate soybean

A relationship among Cu, N, and Rhizobium japonicum was hypothesized because previous research had shown that (a) 35% or more legumes in the Atlantic Coastal Plain have Cu concentrations of 6 mg kg^‐1 or less, (b) Cu influences N fixation in some legumes, and (c) irrigated soybean (Glycine max L. Merr.) can accumulate most of its N through fixation. Soybean were grown on a Cu‐deficient Norfolk (fine‐loamy, siliceous, thermic Typic Paleudult) loamy sand with 3 fertilizer sources of Cu, 2 strains of R. japonicum, and with or without 336 kg ha^‐1 of N fertilizer. Application of Cu significantly increased the number of pods plant^‐1 suggesting pod abortion in determinate soybean may be caused by low Cu, but seed yield was not increased. Fertilization with N increased vegetative growth, but not total biomass or seed yield. Inoculation with R. japonicum strain 110 significantly increased seed yield by 0.3 Mg ha^‐1 compared to strain 587. The yield increase was similar with or without fertilizer N application indicating strain response was not totally caused by improved N efficiency. There was no relationship between seed yield and nodule occupancy as measured by the ELISA technique.     

Effects of hairy vetch foliage application on nodulation and nitrogen fixation in soybean cultivated in three soil types

We investigated the effects of applying hairy vetch foliage on nodulation and atmospheric nitrogen (N₂) fixation in soybean cultivated in three soil types in pot experiments. Soybean plants were grown in Gley Lowland soil (GLS), Non-allophanic Andosol (NAS), and Sand-dune Regosol (SDR) with hairy vetch foliage application in a greenhouse for 45 days. In GLS, the nodule number was not influenced by the application, however, nodule dry weight and N₂ fixation activity tended to increase. In NAS and SDR, nodule formation was depressed by foliage application. Soybean plant growth was promoted in GLS and SDR but not in NAS. These promotive effects of hairy vetch foliage application on soybean plant growth in GLS were considered to be mainly caused by the increase in N₂ fixation activity of the nodules, whereas it was considered to be mainly caused by the increase in nitrogen uptake activity of the roots in SDR. The varying effects of hairy vetch foliage application on soybean nodulation may be due to soil chemical properties such as pH and cation exchange capacity, which are related to soil texture. Therefore, we conclude that it is important to use hairy vetch for soybean cultivation based on the different effects of hairy vetch on soybean plant growth in different soil types.     

Nitrogen fixation and soil N level during maturation affect the contents of storage compounds of soybean seeds

We compared the concentrations and contents of protein and oil in mature seeds from nodulated and non-nodulated soybean plants grown on soils with four different N levels during maturation. We observed a positive correlation between the contents of protein and oil in seeds from nodulated plants. Seeds from nodulated plants grown on urea-treated soil showed higher protein and lower oil contents than those from plants grown on soil treated with coated slow release N fertilizer (LP-100). Contents of these compounds in seeds from nodulated plants grown on LP-100 soil were almost the same as those from non-nodulated plants on the same soil. These observations indicated that N economy in roots during seed maturation affects the contents of storage compounds. We suggested that the control of the N₂ fixation activity of soybean plants and management of soil N level during seed maturation are important to determine the contents of protein and oil in seeds.     

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.     

Alleviation of drought stress in the common bean (Phaseolus vulgaris L.) by co-inoculation with Paenibacillus polymyxa and Rhizobium tropici

A greenhouse experiment was performed to evaluate the influence of Rhizobium when co-inoculated with each of two Paenibacillus polymyxa strains, singly and in mixture on growth, nitrogen content, phytohormone levels and nodulation of the common bean (Phaseolus vulgaris L.) under three levels of drought stress. Stress was applied continuously by the control of matric potential (ψ_m) through a porous cup. Bean plants cv. Tenderlake were grown in pots with Fluvic Neosol eutrophic soil under three different ψ_m (S₁ −7.0; S₂ −70.0 and S₃ < −85 kPa). The seeds were inoculated with Rhizobium tropici (CIAT 899) and each of P. polymyxa (DSM 36) and P. polymyxa Loutit (L) singly and in mixture (CIAT 899 + DSM36 + Loutit). Co-inoculation of bean with Rhizobium and both Paenibacillus strains resulted increased plant growth, nitrogen content and nodulation compared to inoculation with Rhizobium alone. This was particularly evident at the most negative ψ_m (S₃ < −85 kPa) we used. Drought stress triggered a change in phytohormonal balance, including an increase in leaf abscisic acid (ABA) content, a small decline in indole acetic acid (IAA) and gibberellic acid (GA₃) and a sharp fall in zeatin content in bean leaves. The content of endogenous Cks decreased under water stress, possibly amplifying the response of shoots to increasing ABA content. We hypothesize that co-inoculation of bean with R. tropici (CIAT 899) and P. polymyxa strains (DSM 36) and Loutit (L) mitigates some of the negative effects of drought stress on bean.     

Effect of nodulation with Bradyrhizobium japonicum and Shinorhizobium fredii on xylem sap composition of Peking (Glycine max L. Merr )

Five barley cultivars were grown together in complete, low-P·low-pH and high-Al medium containing only NO₃, only NH₄ or both NO₃ and NH₄ as N sources, respectively using an automatic control system of pH for water culture, and the relationship between the differential Al tolerance and the plant-induced pH change of medium among the barley cultivars was investigated.

The pH of the medium containing only NO₃ as N source tended to increase, whereas the pH of the other media containing only NH₄ or both NO₃ and NH₄ as N sources tended to decrease, but the fluctuations of the medium pH could be maintained within the value of 0.2 pH in the complete medium and within the value of 0.1 pH in the high-Al medium.

Barley cultivars still differed in their Al tolerance in the medium which was continuously stirred and circulated at a constant pH. The pattern of Al tolerance was not affected by the N sources in the medium. The plant-induced pH change of medium for each cultivar was influenced by the N sources in the medium, and was not correlated positively with Al tolerance. The contents of Al and Ca or other nutrient cations in roots were positively correlated with Al tolerance and positive correlations were recognized also between the contents of Al and Ca or some other nutrient cations in the roots.

In conclusion, the following mechanisms are proposed. Al tolerant barley cultivars exclude Al actively outside the plasmalemma of the root cells, and the excluded Al may polymerize and or react with P to form Al precipitates. Consequently, in the Al tolerant barley cultivars the Al content may be low in the root protoplasts, high in the whole root tissues and the contents of Ca or other nutrients may be high in the roots. The plant-induced pH change of medium is not considered to be the cause of the differential Al tolerance among barley cultivars.     

Rapid and reversible nitrate inhibition of nodule growth and N2 fixation activity in soybean (Glycine max (L.) Merr.)

Nodulated soybean (Glycine max. (L) Merr. cv. Williams) plants were hydroponically cultured, and various combinations of 1-week culture with 5 or 0 mm nitrate were applied using 13-d-old soybean seedlings during three successive weeks. The treatments were designated as 0-0-0, 5-5-5, 5-5-0, 5-0-0, 5-0-5, 0-5-5, and 0-0-5, where the three sequential numbers denote the nitrate concentration (mm) applied in the first-second-third weeks. The size of the individual nodule was measured periodically using a slide caliper. All the plants were harvested after measurement of the acetylene reduction activity (ARA) at the end of the treatments. In the 0-0-0 treatment, the nodules grew continuously during the treatment period. Individual nodule growth was immediately suppressed after 5 mm nitrate supply. However, the nodule growth rapidly recovered by changing the 5 mm nitrate solution to a 0 mm nitrate solution in the 5-0-0 and 5-5-0 treatments. In the 5-0-5 treatment, nodule growth was completely inhibited in the first and the third weeks with 5 mm nitrate, but the nodule growth was enhanced in the second week with 0 mm nitrate. The nodule growth response to 5 mm nitrate was similar between small and large size nodules. After the 5-5-5, 5-0-5, 0-0-5, and 0-5-5 treatments, where the plants were cultured with 5 mm nitrate in the last third week, the ARA per plant was significantly lower compared with the 0-0-0 treatment. On the other hand, the ARA after the 5-0-0 and 5-5-0 treatments was relatively higher than that after the 0-0-0 treatment, possibly due to the higher photosynthate supply associated with the vigorous vegetative growth of the plants supplemented with nitrate nitrogen. It is concluded that both soybean nodule growth and N₂ fixation activity sensitively responded to the external nitrate level, and that these parameters were reversibly regulated by the current status of nitrate in the culture solution, possibly through sensing of the nitrate concentration in roots and / or nodules.     

Effect of placement of urea and coated urea fertilizers on yield and quality of soybean (Glycine max (L.) Merr.) seeds

The objective of the present study was to record the seed yield and to examine visually the quality of soybean seeds cultivated under different types and placements of urea fertilizers. In addition to the conventional fertilizer application (including ammonium sulfate 16 kg N ha^-1 broadcasting (100 kg N ha^-1 of urea (0B) and X00-d type coated urea CU-100 (CUB), and deep placement (100 kg N ha^-1) of urea (UD) and 100-d type coated urea CU-100 (CUD) was conducted in separate plots in a paddy field converted to an upland field located at Shindori Experimental Station of Niigata University. Soybean plant growth was periodically analyzed and the quality of harvested seeds was also visually examined (hereafter referred to as “visual quality”). It was found that the deep placement treatments were more conducive 1o nitrogen (N₂)fixation, based on the relative mreide N concentration in the xylem sap, which is a good indicator of N~fixation by soybean. Also the total seed yield was the highest in CUD (82 g plant^-1) and 0D (81 g plant^-1), compared to the control (62 g plant^-1), UB (68 g plant^-1), and CUB (68 g plant^-1). The visual quality of harvested seeds showed that CUD enhanced the quality of seeds compared to the other treatments, in which the percentage of good quality seeds, hereafter referred to as "good seeds," based on the dry weight was 51 (control), 65 (K3B), 61 (CUB), 61 (0D), and 6696 (CUD). In terms of diseased seeds, the percentage of turtle wrinkle and broken seed coats was found to decrease by N application compared to the control. Thus, it is suggested that N fertilization management is important for maximum yield of soybean as well as for the enhancement of seed quality.     

Effects of CO2 concentration in rhizosphere on nodulation and N2 fixation of soybean and cowpea

Soybean (Glycine max L. Merr.) cvs. Akisengoku and Peking, and cowpea (Vigna unguiculata Walp.) cv. Kegonnotaki were inoculated with Bradyrhizobium japonicum AlO17, Shinorhizobium fredii USDAI93, and B. sp. Vigna MAFF03-03063, respectively and were cultured hydroponically with supply of CO₂-free air, 3dm³ m^-3 CO₂ air, or 25 dm³ m^-3 CO₂ air to study the effects of the CO₂ concentration in the rhizosphere on plant growth, nodulation, and nitrogen fixation. Increase of the CO₂ concentration in the rhizosphere led to the increase of the plant dry weight in the symbiosis between Peking and USDAI93, and that between Kegonnotaki and MAFF03-03063. On the other hand, dry matter accumulation in the symbiosis between Akisengoku and AI017 decreased under the supply of 25 dm³ m^-3 CO₂ air aimed at increasing the CO₂ concentration in the rhizosphere beyond the optimum CO₂ concentration for growth. Nodule mass and nodule number per plant were highest in Akisengoku, followed by Kegonnotaki and lowest in Peking. Also the increase of the CO₂ concentration in the rhizosphere led to the increase of the nodule mass and number in Kegonnotaki, while no changes were observed in Akisengoku and Peking. Biological nitrogen fixation (BNF) was highest in Akisengoku, followed by Kegonnotaki, and lowest or near zero in Peking. BNF in Akisengoku and Kegonnotaki showed a similar tendency to that of dry matter accumulation. BNF of Peking was especially low under the supply of CO₂-free air, and it increased with the increase of the CO₂ concentration in the rhizosphere. For the symbiosis of Bradyrhizobium strains with soybean and cowpea, the most suitable CO₂ concentration for N₂ fixation and plant growth was estimated to be about 10 dm³ m^-3, while for the symbiosis of S. fredii with soybean, the value was estimated to be above 30 dm³ m^-3.     

本质素对土壤N、P转化及玉米产量的影响

研究造纸黑液中提取的木质素对土壤N、P转化及其对玉米生长和产量的影响结果表明,木质素可减缓NH4+向NO3氧化,且随其施用量的增加效果更显著.木质素与磷酸二铵混合施用效果最佳,其次为硫酸铵＞尿素.在30℃温度下培养27d,施用量为2%和5%的木质素可分别减少施尿素土壤N2O释放83%和96%;而施磷酸二铵的土壤则分别减少83%和93%.施用木质素可促进难溶性P的溶解,对作物生长极为有利.玉米盆栽试验中施用木质素的根系较发达、粗壮,平均株高、地上部和地下部的鲜物质量和干物质量均高于不施木质素的处理.木质素用量为50μg/g和200μg/g时玉米籽粒产量分别提高4.2%和18.8%.     

Effects of Rhizobium and vesicular-arbuscular mycorrhiza inoculations on nodulation,symbiotic nitrogen fixation and soybean yield in subtropical-tropical fields

Summary Field experiments were carried out to determine the effects of single and mixed inoculations with Rhizobium and vesicular-arbuscular mycorrhiza (VAM) on nodulation, symbiotic N₂ fixation and yield of soybeans in six Taiwan subtropical-tropical sites. Inoculation with Rhizobium alone significantly increased nodulation, nodule weight and nitrogenase activity of nodules in three out of six experimental fields, and affected soybean yields in the range –13% to + 134%. Inoculation with VAM fungi alone did not have a significant effect on nodulation and nitrogenase activity. Mycorrhiza inoculation affected soybean yields in the range –13% to + 65%, but only the yield increases at one out of six sites with N application were statistically significant. Mixed inoculation with Rhizobium and mycorrhiza affected yields in the range –8% to + 145% A synergistic effect from mixed inoculation of Rhizobium-mycorrhiza on soybean yields was found in one out of six experimental fields. The yield response to N application (40 kg N ha^–1) in these six paddy-field trials was not significant. These results suggest that single or mixed inoculation of rhizobia can greatly assist soybean grain yields and can replace N fertilizers.     

Survival and competitiveness of Bradyrhizobium sp. in the rhizosphere of pigeonpea (Cajanus cajan)

Summary The survival of Bradyrhizobium Cajanus strain CC1021 in the soil and rhizosphere of pigeonpea [Cajanus cajan (L.) Millsp] was determined by most probable number and fluorescent antibody techniques. The survival of strain CC1021 was poor under field compared with pot-culture conditions. Although the rhizosphere of pigeonpea promoted the growth of native pigeonpea rhizobia, although no increase in the number of rhizobia was observed with the inoculant strain. Under similar conditions of rhizosphere colonization the competitiveness of strain CC1021 with cv UPAS 120 and ICPL 312 was 10% and 66016 during the first year and 8.4% and 33.3%, respectively, during the second year.     

Effects of deep placement of controlled-release nitrogen fertilizer on soybean growth and yield under sulfur deficiency

Sulfur (S) and Nitrogen (N) metabolisms in plants are interacted and it is known that S deficiency decrease N absorption and metabolism. In leguminous plants S deficiency also decreases N₂ fixation by rhizobia in the nodules. Deep placement of a controlled-release N fertilizer is a good method to provide nitrogen to soybean without inhibiting N₂ fixation; thus, it was hypothesized that this method is able to provide nitrogen effectively to sulfur-deficient soybean plants. In this study effects of deep placement of coated urea on S-N physicological interaction, growth and productivity in soybean plants were examined using pot experiments. Soybean plants were grown with sulfate concentrations of 30, 100, or 1000 μM, with or without deep placement of coated urea. Shoot weights at the developing stage were not affected by S deficiency. SPAD values of leaves during the flowering stage decreased with S deficiency and increased with the deep placement of coated urea. S deficiency decreased seed weight per plant at the harvesting stage, but this decrease was attenuated by the deep placement of coated urea. N and S content in shoots at the developing stage increased with the deep placement of coated urea, whereas in seeds, only the N content increased. N₂ fixation activity based on the relative ureide-N content in xylem sap indicated that the deep placement of coated urea did not inhibit N₂ fixation activity at the early flowering stage. Without deep placement of coated urea, the relative ureide-N content decreased under S deficiency at the seed filling stage. These results suggest that the deep placement of coated urea is an efficient method to supply N to support soybean yield under S deficiency.

Abbreviations: Deep+: with deep placement of coated urea; Deep–: without deep placement of coated urea