Rhizobiophage effects on nodulation,nitrogen fixation,and yield of field-grown soybeans (Glycine max L. Merr.)

Summary Previous laboratory and greenhouse studies have shown that phages significantly reduce soil populations of homologous rhizobia. Reductions in nodulation and N₂ fixation have also been observed. The purpose of the current study was to examine the effect of a phage specific ofBradyrhizobium japonicum USDA 117 on nodulation, nodule occupancy, N₂ fixation and soybean growth and yield under field conditions. The phage was inoculated in combination withB. japonicum USDA 117 and/orB. japonicum USDA 110 (resistant strain) into a rhizobia-free sandy loam soil and planted toGlycine max (L.) Merr. Williams. When the phage was applied to soil inoculated withB. japonicum USDA 117 alone, significant reductions in nodule weight and number, shoot weight, foliar N, nitrogenase activity, and seed index were observed. When, however, the soil also contained the non-homologous strain,B. japonicum USDA 110, no significant effects on any of these parameters were found. Nodule occupancy by competing strains ofB. japonicum USDA 110 and USDA 117 was also affected by the phage. In soil which did not contain the phage, 46% and 44% of the identified nodules were occupied by USDA 110 and 117, respectively. When the phage was present in the soil, nodule occupancy byB. japonicum USDA 117 was reduced to 23%, while occupancy byB. japonicum USDA 110 was increased to 71%. These results suggest that nodulation by selected strains of rhizobia can be restricted and nodulation by more effective, inoculated strains can be increased through the introduction of a homologous phage to soils.     

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%.     

Effectiveness of foreign bradyrhizobia strains in enhancing nodulation, dry matter and seed yield of soybean (Glycine max L.) cultivars in Nigeria

 Field experiments were conducted to investigate the performance of three soybean cultivars with five foreign bradyrhizobia strains in different regions. The experiments at the two sites were designed with soybean (Glycine max L.) cultivars as the main factor and bradyrhizobia strains (USDA 136, TAL 122, USDA 6, TAL 377 and TAL 102) as the sub-factor. The experiments were arranged in randomised complete block design with four replications. Results show that nodule number, nodule dry weight and shoot dry weight, total N and seed yield were significantly increased when soybean cultivars were inoculated with foreign bradyrhizobia in two locations in the south east of Nigeria. At 63 days after planting the percentage increase in nodule number and dry weight after inoculation of soybean cultivars with bradyrhizobia strains ranged from 71 to 486% and from 0 to 200%, respectively. The percentage increase in shoot dry matter, %N and total N after bradyrhizobia inoculation ranged between 2–130%, 18–62% and 35–191%, respectively at Awka, and at the Igbariam site the percentage increase in shoot dry weight, %N and total N ranged between 3–76%, 0–43% and 19–125%, respectively. Seed yields after bradyrhizobia inoculation of soybean cultivar TGX 1485–1D at Igbariam ranged between 1.20 and 2.18 t ha^–1 against the uninoculated plants, which had seed yields of 1.05 t ha^–1. The poorest yield response after inoculation with bradyrhizobia strains was observed in soybean cultivar M-351, with a seed yield ranging from 0.60 to 0.98 t ha^–1. The fact that foreign bradyrhizobia strains were more effective than the indigenous strains for all the parameters studied suggests that there is a need to use bradyrhizobia inoculants for increased soybean production in Nigeria. The variations in the strain performance with the different soybean cultivars at the two sites, emphasises the need for careful Bradyrhizobium spp. strain selection. The fact that inoculation response was cultivar- and site-specific suggests that strategies for improving inoculation response in soybean cultivars should also consider the soil environment where the soybean is to be produced. Received: 25 May 1999     

Modelling of survival kinetics of some Bradyrhizobium japonicum strains in soils

Summary The existence of a balance survival rate of different B. japonicum strains in soils has been well established. Balance levels (M) were shown to be independent of inoculum size but differences between strains in a soil could not be discussed partly because of the instability of the recovery's efficiency in the area of the plateau. In this paper, a dynamic model, the Gompertz equation, is fitted to the data, to estimate the rate (A) at which equilibrium is reached. For the three strains in the three soils previously studied, this rate is also independent of inoculum size and the soil-strain interaction is highly significant. Some data from the literature are also well fitted by the model. We propose to characterize the saprophytic potential of a strain with the pair of parameters A-M. An optimal experimental procedure is proposed to obtain the most precise estimates for these parameters. Their biological significance is discussed, but the fact that they are not dependent upon inoculation level is of basic importance for agronomic practice.     

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.     

Hydrogen metabolism and nitrogen fixation of soybean (Glycine max. L.) inoculated with different strains ofRhizobium spp.

Summary Hydrogenase activities and N₂-fixing capacities of soybean nodules (Glycine max. cv. Hodgson), inoculated with strains ofBradyrhizobium japonicum andRhizobium fredii from different geographical regions, were measured after 35 days of culture under controlled conditions. Of the strains tested, 47% induced nodules with bacteroids which recycled H₂. The data obtained suggest that H₂-recycling ability is not a major factor influencing early N₂-fixation which depends essentially on the precocity and intensity of the initial nodulation.     

Classification and Characteristic of Maturity Groups of Chinese Landraces of Soybean [Glycine max (L.) Merr.]

To link Chinese soybean classification with the world soybean maturity group system suggested by American researchers, 264 soybean landraces from China and 48 varieties of 13 soybean maturity groups from the United States (US) were tested under the natural and extended day-length at Nanjing, China. Based on comparing Chinese with the US soybeans in days to maturity, Chinese soybean landraces were classified into maturity group (MG) 000, 00, 0, I, II,..., IX, which only MG X might not exist in China. Chinese soybean landraces in each of MG 0, I, II and III revealed large variation in days to flowering and were almost grown in all eco-regions in China. These four maturity groups were further divided into eight sub-groups according to the variation in days to flowering and geographical distribution of soybeans of the same maturity group. The geographic distribution of soybean maturity groups in China was also presented in this paper. This study should be significant for soybean researchers in the world to understand the whole feature of Chinese soybean landraces germplasm.     

AFLP Analysis of Genetic Diversity in Indian Soybean [Glycine max (L.) Merr.] Varieties

AFLP technique was used to assess genetic diversity in 72 soybean varieties under cultivation in India. Selected 12 AFLP primer pairs produced 1319 products of which 1257 were polymorphic (95%). Wide variations were observed for the number of amplification products, percent polymorphism and average polymorphism information content (PIC). The Jaccard's similarity indices (J) based on the AFLP profiles of the 72 soybean varieties were subjected to UPGMA cluster analysis. The dendrogram generated revealed four major clusters, which were strongly supported by the high bootstrap values obtained from analyses of 1000 bootstrap samples. In addition, the Mantel's test for cophenetic correlation with r = 0.955 indicated very good fit of the varieties to a group in the cluster analysis. Some correspondence between the clustering pattern and the pedigree, place of release or target area of the variety was observed. Overall moderately high genetic diversity was observed which appears to be due to the higher genetic diversity prevalent in 12 of the varieties included in three diverse clusters and was indicative of the need to include more diverse germplasm in the soybean improvement programs.     

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     

Variation of nitrogen accumulation and yield in response to phosphorus nutrition of soybean (Glycine max L. Merr.)

Phosphorus (P) is essential macronutrient for soybean [Glycine max (L.) Merr.] growth and function. The objective of this study was to determine effect of phosphorus nutrition (including phosphorus nutrition level and interruption of phosphorus supply) on nitrogen accumulation, nodule nitrogen fixation and yield of soybean plants by ¹⁵N labeling with sand culture. The results showed that they all presented a single peak curve with improvement of phosphorus nutrition level, when phosphorus concentration of nutrient solution was about 31 mg/L, they all reached the maximum and effect of phosphorus nutrition level on nodule nitrogen fixation was lower than that on yield formation level. Interruption of phosphorus supply during soybean growth period, nitrogen accumulation and nodule nitrogen fixation were seriously inhibited, and yield was decreased significantly when interruption of phosphorus supply during V₃-R₁ and R₁-R₅ period, while interruption of phosphorus supply during R₅-R₇ period had no significant effect on nitrogen accumulation, nodule nitrogen fixation and yield. So soybean nitrogen metabolism and yield were sensitive to phosphorus nutrition in the V₃-R₅ period, those were not sensitive to phosphorus nutrition after R₅ period.     

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     

Iron requirement for soybean [Glycine max (L.) Merrill] inoculated with selected exotic and native isolates of Bradyrhizobium sp. under irrigated conditions

A field and greenhouse experiments were conducted to determine the requirement of Fe nutrient supplied through foliar and soil application in soybean inoculated with different selected isolates of exotic and native Bradyrhizobium spp. in saline soils. Six soybean genotypes and three Bradyrhizobium spp. were used for the greenhouse experiments, whereas only two soybean genotypes, namely TGx-1336424 and GIZA, were selected for further study under field conditions. Two levels of FeSO₄ (0 and 4 mg Fe kg^?1 soil) directly supplied to the soil and three levels of Fe-ethylenediaminetetraacetic acid (0–2% of Fe) through foliar application were used for greenhouse and field experiments, respectively. The results of the greenhouse experiment indicated a non-significant effect of Fe application on nodulation and shoot biomass in soybean. Fe application did not improve the grain yield and total biomass yield in soybean inoculated with UK isolate and local isolate but showed remarkable improvement with TAL-379. High soil native N might be the cause for insignificant effect of Fe applied at 2% in highly effective inoculated plants. Therefore, it can be concluded that the symbiotic effectiveness of Bradyrhizobium sp. and the native soil N would affect the soybean Fe requirement supplied through foliar application.     

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.     

Effect of high temperature on plasmid curing of Rhizobium spp. in relation to nodulation of pigeonpea [Cajanus cajan (L.) Millsp.]

Summary Fifty-six isolates of Rhizobium and Bradyrhizobium spp. (Cajanus) were studied for their plasmid profile and N₂-fixation efficacy. One to three plasmids were reproducibly detected in all the Rhizobium spp. strains but no plasmid was detected in the Bradyrhizobium spp. strains. Rhizobium sp. strain P-1 was mutagenized by Tn5 and three nod^– and six nod⁺fix^– were screened for symbiotic parameters. Neomycin-sensitive mutants were isolated by elevated temperatrue (40°C) from tranconjugants carrying Tn5 insertions. The high temperature cured these mutants from the single large plasmid present in the parent strain P-1. All these cured mutants were nod^–, indicating that the genes for nodulation were present on this plasmid, which is readily cured at a high temperature (40°C). The high temperature in the semi-arid zones of Haryana could be responsible for the low nodulation of pigeonpea because the plasmid carrying the nodulation genes is cured at 40°–45°C giving rise to non-nodulating mutants.     

Effect of harvesting and drying conditions on chlorophyll levels of soybean (Glycine max L. Merr)

Chlorophyll in soybean represents a downgrading factor for the crops. Five Brazilian cultivars were harvested between R(6) and R(8) stage of development (Fehr & Caviness scale) and dried at 25 degrees and 40 degrees C. The effect of maturity stages and two drying conditions after harvest were studied to achieve reduction of moisture and chlorophylls to acceptable levels. When seeds were dried at 25 degrees C, even harvesting at early stages of development such as R(6), the green pigments were almost degraded, and 16 ppm of chlorophyll were found at maximum, accompanied by loss of moisture. Moisture and chlorophyll declines as seed matures, but at intermediary stages (R(6)-R(7)), chlorophyll degrades first, so the rate of moisture loss should not be used to predict chlorophyll contents. At 40 degrees C, complete degradation of chlorophyll pigments is only achieved when seeds are swathed from R(7) stage up, otherwise the seed quality could be compromised. Slow drying allows almost complete removal of green pigments, even when seeds are swathed a few days before the physiological maturity stage.     

Effect of deep placement of calcium cyanamide,coated urea,and urea on soybean (Glycine max (L.) Merr.) seed yield in relation to different inoculation methods

Abstract

The main objective of this study was to increase the productivity of soybean [Glycine max (L.) Merr. cv. Enrei] seed by deep placement of controlled release nitrogen fertilizers and by the application of different methods of inoculation of bradyrhizobia. Ten days old seedlings in an inoculated paper pot (IPP), in a non-inoculated paper pot (NIPP), and those grown in a vermiculite bed without paper pot (DT) were transplanted to an upland field converted from a drained paddy field in Nagaoka. In addition to the basal application of 16 kg N ha^?1 in the surface layer (Control), deep placement of 100 kg N ha^?1 of urea (Urea), 100-day type coated urea (CU-100), and calcium cyanamide (CaCN₂) treatments were applied at the depth of 20 cm. In the IPP method, a significantly higher seed yield was obtained with the deep placement of CaCN₂ and CU-100 compared with the Urea and Control treatments. A similar tendency was observed for the DT and NIPP methods. Among the same N fertilizer treatments, the seed yield for IPP and DT tended to exceed that for NIPP, although the NIPP roots also showed nodulation probably due to infection with indigenous bradyrhizobia. The percentage of nitrogen derived from atmospheric N₂ estimated by the simple relative ureide method was higher in the plants with CU-100 and CaCN₂ compared with those with the Urea and Control treatments at the RI stage, suggesting that the basal deep placement of CaCN₂ or CU-100 for soybean cultivation enabled the supply of N without concomitant depression of N₂ fixation. Thus the deep placement of cheaper CaCN₂ was found to be as effective as that of CU-100 for enhancing the soybean seed yield.     

Physical mapping and candidate gene prediction of branch number on the main stem in soybean [Glycine max (L.) Merr.]

Genetic Resources and Crop Evolution - Branch number on the main stem (BNMS) is an important factor that affects crop plant architecture and yield in soybean [Glycine max (L.) Merr.]. With the aim...     

Effect of temperature on flavonoid production in pigeonpea [Cajanus cajan (L) Millsp] in relation to nodulation

To ascertain the reasons for poor nodulation of pigeonpea, we studied the effect of high temperature on the production of flavonoids by the pigeonpea host. A high temperature affected flavonoid production by pigeonpea and mungbean. At 37°C pigeonpea root exudates contained four flavonoids and the root extract contained five. The proportion of the second flavonoid in the pigeonpea and the mungbean was higher and the proportion of the third flavonoid was lower at 37°C compared to 30°C. At the higher temperature the flavonoids exuded from pigeonpea roots were same those in the root homogenate.     

Influence of lime and phosphorus on soybean (Glycine max (L.) Merrill) yield,leaf and seed composition on a paleudult soil

Abstract

This research was undertaken on a paleudult soil in southern Brazil, 30° south latitude, to quantify lime and P effect upon soybean (Glycine max (L.) Merrill). A lime x P factorial experience with lime treatments of 0, 0.5, 1, and 2 times SMP interpretation to pH 6.5, and 0, 44, 88, 132, and 176 kg P/ha with 3 replications were installed. The experiment was conducted for 2 years (1973–74, 1974–75), with leaf‐N, P, and K; yield; seed‐N, P, and K; Bray P₂ (0.03N NH₄F + 0.1N HC1) avail‐able‐P and soil pH measurements completed each year. Data was evaluated with linear, quadratic, logarithmic, polynomial, segmented line, and multiple regression using the coefficient of determination as goodness of fit.

The best model fit between P treatment and Bray P₂ available‐P was a quadratic equation; the model between relative yield and Bray P_2‐P with 54% of the relative yield attributed to Bray P₂ available‐P, a sigmented line. This model indicated point of maximum yield (91% relative yield) was obtained at 7.4 ppm‐P, with no increase in relative yield with increasing levels of soil available‐P. To calculate the P fertilizer necessary to increase available soil‐P to the level of maximum yield of equation Y_p = [1639(7.4 ‐ x_s)]^1/2, where Y_p = kg P/ha fertilizer needed; and x_s = initial Bray P₂ soil available‐P in ppm's. The lime effect upon soil pH was best described as a linear relationship. Yield increase with lime at this site was not significant at the 5% level.

The leaf‐N, P, and K increased significantly with soil available‐P levels. A second degree polynomial with logarithmic function best defined these relationships. The calculated DRIS indices and sum proved useful to evaluate the plant‐N, P, and K balance of each treatment.

Only seed‐P level was directly related to soil available‐P. Both seed‐N and seed‐K were highly correlated with indirect effects of soil available‐P levels.

Results from this study suggest the segmented line model would best interpret soybean yield response to Bray P₂ available‐P for this soil. To obtain maximum yield using this model rather than the second degree polynomial would require less fertilizer P. Foliar analyses interpretation confirmed adequate plant‐P level would be supplied for maximum yield at this level of fertilization.     

Effects of black soybean [Glycine max (L.) Merr.] seed coats and its anthocyanidins on colonic inflammation and cell proliferation in vitro and in vivo

Anti-inflammatory and antiproliferative activities of anthocyanidins and anthocyanin-rich black soybean seed coats were studied in HT-29 human colon adenocarcinoma cells and carcinogen-treated F344 rats, respectively. Cyanidin and delphinidin significantly inhibited cell growth at concentrations of >or=1 microM. Anthocyanidins suppressed cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) mRNAs in TPA-stimulated HT-29 cells. Both yellow and black soybean seed coat supplementation (10%, w/w) did not significantly reduce the number of aberrant crypt foci (ACF), although a modest decrease in the number of ACF was observed in animals fed soybean seed coats. The colonic COX-2 mRNA level was suppressed in rats fed both soybean seed coat diet. The plasma prostaglandin E 2 (PGE 2) level was reduced only in rats fed black soybean seed coats. No difference was observed in either colonic iNOS mRNA or plasma nitric oxide level. These results indicate that anthocyanidins are possible anti-inflammatory agents; however, further studies are required to determine required intake levels in vivo to exert antitumor effect.