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.     

N2 fixation in two Sesbania species and its transfer to rice (Oryza sativa L.) as revealed by 15N technology

Summary We used ¹⁵N technology to investigate N₂ fixation by Sesbania speciosa and Sesbania rostrata and its transfer to a lowland rice crop after incorporation of the Sesbania spp. into soil as green manure. During the first 50 days after establishment in November–December 1989, S. speciosa and S. rostrata produced 1126 and 923 kg dry matter ha^-1 respectively. They gathered 31 and 23 kg N ha^-1 respectively, of which 62%±5% and 55%±3% respectively, came from N₂ fixation. Both these species produced a greater biomass during September–October 1989, with S. rostrata producing more than S. speciosa. These results reflected differential responses by the plants to different day lengths at different times of the year. Furthermore, the dry matter yield and %N of ¹⁵N-labelled S. speciosa were smaller than those of the unlabelled plants, possibly due to inhibition of N₂ fixation in root nodules by the chemical N fertilizers added during labelling. These differences were not so pronounced in the stem-nodulated S. rostrata. The increased grain yield of rice fertilized with N in the form of chemical fertilizer or green manure was a result of an increased number of panicles per hill. The rice crop manured with S. speciosa produced a lower grain yield, with a lower grain weight than that manured with S. rostrata. This was due to a low uptake of soil N by rice manured with S. speciosa. Recovery of N from the green manure in rice straw with S. speciosa was significantly higher than from rice manured with S. rostrata, because of the higher applied N uptake by rice manured with the former.     

Biological nitrogen fixation in Azospirillum strain-maize genotype associations as evaluated by the 15N isotope dilution technique

Few studies of the inoculation of cereal crops with N₂-fixing bacteria have included more than one or two plant genotypes. In a recent study performed in Argentina using 12 different maize genotypes, it was found in 2 consecutive field experiments that several of them responded consistently, either negatively or positively, to inoculation with a mixture of strains of Azospirillum spp. The present study in post was performed to investigate the effect of inoculation of individual strains (and a mixture) of Azospirillum spp., and their nitrate reductase negative (NR-) mutants, on the growth of four of these maize genotypes. Two of these genotypes were grown in ¹⁵N-labelled soil with the aim of quantifying any contributions of biological N₂ fixation. Two genotypes (Morgan 318 and Dekalb 4D-70) produced similar increases in grain yield when they were inoculated with a mixture of Azospirillum spp. strains or fertilized with the equivalent of 100 kg N ha^-1. The other genotypes (Dekalb 2F-11 and CMS 22) showed little response to inoculation or N fertilization. The Morgan 318 and Dekalb 4D-70 genotypes showed a large increase in total N accumulation, suggesting that the response was due to increased N acquisition, but not due to bacterial nitrate reductase as the NR- mutants generally caused plant responses similar to those of the parent strains. Despite problems with the stabilization of the ¹⁵N enrichment in the soil, the ¹⁵N isotope dilution results indicated that there were very significant biological nitrogen fixation (BNF) contributions to the Dekalb 4D-70 and CMS 22 maize genotypes.Dedicated to Professor J.C.G. Ottow on the occasion of his 60th birthday     

氮磷钾肥用量对紫云英产量效应的研究

采用"3414"肥料效应试验设计方案对紫云英氮、磷、钾肥施用效应及养分的交互作用进行了研究,结果表明:与不施肥处理(CK)相比,13个施肥处理紫云英鲜草平均增产21.1 t·hm-2,平均产量为不施肥处理的2.35倍;分别固定磷(P2O5 60 kg·hm-2)、钾(K2O 60 kg·hm-2)肥,氮(N75 kg·hm-2)、钾(K2O 60 kg·hm-2)肥和氮(N 75 kg·hm-2)、磷(P2O5 60 kg·hm-2)肥用量,在施N 0～112.5 kg·hm-2,P2O5 0～90 kg·hm-2和K2O 0～90 kg·hm-2范围内,紫云英产量随相应肥料用量的增加而显著提高,N、P、K各养分施用的最高增产率分别为65.0%、27.8%和44.5%;从养分效率看,中量水平的氮(N 75.0 kg·hm-2)、磷(P2O5 60 kg·hm-2)和低量水平的钾(K2O 30 kg·hm-2)增产效果最好;氮、磷、钾肥之间存在一定的交互作用,互相影响肥效的发挥,中量水平的养分用量(N 75.0 kg·hm-2、P2O5 60 kg·hm-2和K2O 60 kg·hm-2)有利于各养分效果的发挥.结果说明,施肥对紫云英增产效果明显,氮、磷、钾肥用量和配比是影响紫云英产量的重要因素.     

不同施肥量对绿肥产量和养分积累的影响

大田试验条件下研究不同氮、磷、钾肥用量对绿肥[豆科绿肥紫云英(Astragalus sinicus L.)和禾本科绿肥看麦娘(Alopecurus aequalis Sobol.)]产量及养分积累量的影响。试验结果表明,施肥能显著增加绿肥产量,绿肥鲜草产量较不施肥处理增加42.8%~311.1%,与缺素处理相比,氮、磷、钾肥的增产率分别为57.1%~177.5%、27.8%~178.7%、14.2%~32.4%。两种绿肥的鲜草产量均随氮、磷、钾肥施用量的增加而增加,中、高水平的氮、磷肥用量下看麦娘的产量均高于紫云英,所有钾肥处理的看麦娘产量均高于紫云英。绿肥的养分积累量均随施肥量的增加而增加,中高水平的氮、磷肥用量下看麦娘的氮、磷积累量高于紫云英,不同钾水平下的看麦娘钾积累量均高于紫云英。氮、磷、钾肥用量分别为112.5 kg(N).hm?2、19.7 kg(P).hm?2、37.5 kg(K).hm?2时,两种绿肥总产量及碳、氮、磷、钾积累量最高,分别为49 424 kg.hm?2和3 212 kg(C).hm?2、151 kg(N).hm?2、19.8 kg(P).hm?2、156 kg(K).hm?2。此时看麦娘产量、碳、氮、磷、钾积累量分别占总量的59.0%、65.7%、66.3%和64.4%。紫云英田间自然洒种的看麦娘无成本投入且养分积累量高,不失为一种优质绿肥。     

Sesbania rostrata as a green manure for lowland rice: Growth,N2 fixation,Azorhizobium sp. inoculation,and effects on succeeding crop yields and nitrogen balance

Summary Root and stem nodulation, nitrogen fixation (acetylene-reducing activity), growth and N accumulation bySesbania rostrata as affected by season and inoculation were studied in a pot experiment. The effects ofS. rostrata as a green manure on succeeding wet-season and dry-season rice yields and total N balance were also studied.S. rostrata grown during the wet season showed better growth, nodulation, and greater acetylene-reducing activity than that grown during the dry season. Inoculation withAzorhizobium caulinodans ORS 571 StrSpc® (resistant to streptomycin and spectinomycin) on the stem alone or on both root and stem significantly increased N₂ fixation by the plants. Soil and seed inoculation yielded active root nodules under flooded conditions. Plants that were not inoculated on the stem did not develop stem nodules. The nitrogenase activity of the root nodules was greater than that of the stem nodules in about 50-day-oldS. rostrata. S. rostrata incorporation, irrespective of inoculation, significantly increased the grain yield and N uptake of the succeeding wet season and dry season rice crops. The inoculated treatments produced a significantly greater N gain (873 mg N pot^–1) than the noinoculation (712 mg N pot^–1) treatment. About 80% of the N gained was transferred to the succeeding rice crops and about 20% remained in the soil. The soil N in the flooded fallow-rice treatment significantly declined (–140 mg N pot^–1) but significantly increased in bothS. rostrata-rice treatments (159 and 151 mg N pot^–1 in uninoculated and inoculated treatments respectively). The N-balance data gave extrapolated values of N₂ fixed per hectare at about 303 kg N ha^–1 per two crops forS. rostrata (uninoculated)-rice and 383 forS. rostrata (inoculated)-rice.     

Ontogenic variation in nitrogen fixation and accumulation of nitrogen in mungbean,blackgram, cowpea,and groundnut

Ontogenic variations in N₂ fixation and accumulation of N by the mungbean (Vigna radiata L. Wilczek), blackgram (Vigna mungo L. Hepper), cowpea (Vigna unguiculata L. Walp.), and groundnut (Arachis hypogaea L.) were studied by a ¹⁵N-dilution technique. Pots filled with 7 kg of red yellow podzolic soil were used. Samples were taken 20, 40, 60, and 80 days after emergence which approximately corresponded to preflowering, flowering, early/mid-pod filling and late pod filling stages, respectively. During early growth (up to 40 days after emergence), the carryover of seed N accounted for a considerable fraction of the total plant N in the legumes, the highest being in the groundnut. With a correction for carryover, the groundnut derived over 45% of its N content from the atmosphere 20 days after emergence whereas the corresponding figures were 33% for the blackgram and about 28% for the cowpea and mungbean. Between flowering and early pod fill, there was a rapid increase in N₂ fixation in all legumes except in groundnut which showed highest fixation from 60 to 80 days after emergence. In the mungbean, N₂ fixation and uptake of soil N were insignificant 60 days after emergence while in other legumes these processes continued beyond this time. All legumes derived about 90% of their N from atmosphere by 80 days after emergence. However, due to considerable interspecific differences in total N yield the final amount of N₂ fixed showed an appreciable variation among legumes. It was highest in the groundnut (443 mg N plant^-1) followed by the cowpea (385), blackgram (273), and mungbean (145), respectively. The groundnut maintained nodules until the late pod filling stage while in other legumes, nodules senesced progressively following the mid-pod filling stage. During pod filling there was a net mobilization of N from vegetative tissues to developing pods in the mungbean, which amounted to about 20% of N in seeds. This mobilization was not evident in other legumes.     

Dual inoculation of Pisum sativum with Rhizobium leguminosarum and Penicillium bilaji

Summary To investigate the effect of single versus dual inoculation of peas (Pisum sativum L.) with Rhizobium leguminosarum biovar viceae and Penicillium bilaji (a soil fungi capable of solubilizing soil P) on N₂ fixation an experiment was carried out under controlled conditions. A sandy loam soil was selected which contained low levels of available N and P. P fertilizer [Ca(H₂PO₄)₂] and P. bilaji significantly increased dry matter production. Peas inoculated with R. leguminosarum showed only a small increase in dry matter, but the additional application of P significantly increased the yield. The total N accumulation was highly dependent on the presence of R. leguminosarum. Using the ¹⁵N method for estimating N₂ fixation, the highest level of N₂-fixing activity was observed in peas inoculated with R. leguminosarum and fertilized with inorganic P. Dual inoculation of peas with P. bilaji and R. leguminosarum significantly decreased the amount of N₂ fixed. Total P uptake was solely dependent on the P fertilizer.     

Inoculation withGlomus mosseae improves N2 fixation by field-grown soybeans

Summary A field study carried out in a sandy, relatively acid Senegalese soil with a low soluble P content (7 ppm) and low vesicular-arbuscular mycorrhizal (VAM) populations showed that soybean responded toGlomus mosseae inoculation when the soluble P level in the soil had been raised by the addition of 22 kg P ha^–1. In P-fertilized plots, N₂ fixation of soybean, assessed by the A value method, was 109 kg N₂ fixed hat when plants were inoculated withRhizobium alone and it reached 139 kg N₂ fixed ha^–1 when plants were dually inoculated withRhizobium andGlomus mosseae using an alginate bead inoculum. In addition to this N₂ fixation increase (+28%),Glomus mosseae inoculation significantly improved grain yield (+13%) and total N content of grains (+16%). This success was attributed mainly to the low infection potential of the native VAM populations in the experimental site. In treatments without solubleP or with rock phosphate, no effect of VAM inoculation was observed.     

Nitrogen fixation and CO2 exchange in soybeans (Glycine max L.) inoculated with mixed cultures of different microorganisms

N₂ fixation, photosynthesis of whole plants and yield increases in soybeans inoculated with mixed cultures of Bradyrhizobium japonicum 110 and Pseudomonas fluorescens 20 or P. fluorescens 21 as well as Glomus mosseae were found in pot experiments in gray forest soil carried out in a growth chamber. The effects of pseudomonads and vesicular-arbuscular (VA) mycorrhizal fungus on these parameters were found to be the same. Dual inoculation of soybeans with mixed cultures of microorganisms stimulated nodulation, nitrogenase activity of nodules and enhanced the amount of biological nitrogen in plants as determined by the ¹⁵N dilution method in comparison to soybeans inoculated with nodule bacteria alone. An increased leaf area in dually infected soybeans was estimated to be the major factor increasing photosynthesis. P. fluorescens and G. mosseae stimulated plant growth, photosynthesis and nodulation probably due to the production of plant growth-promoting substances. Increasing phosphorus fertilizer rates within the range of 5–40 mg P 100 g^-1 1:1 (v/v) soil: sand in a greenhouse experiment led to a subsequent improvement in nodulation, and an enhancement of N₂ fixation and yield in soybeans dually inoculated with B. japonicum 110 and P. fluorescens 21. These indexes were considerably higher in P-treated plants inoculated with mixed bacterial culture than in plants inoculated with nodule bacteria alone.     

The fate of markerAzospirillum lipoferum inoculated into rice and its effect on growth,yield and N2 fixation of plants studied by acetylene reduction,15N2 feeding and15N dilution techniques

Summary A spontaneous mutant ofAzospirillum lipoferum, resistant to streptomycin and rifampicin, was inoculated into the soil immediately before and 10 days after transplanting of rice (Oryza sativa L.). Two rice varieties with high and low nitrogen-fixing supporting traits, Hua-chou-chi-mo-mor (Hua) and OS4, were used for the plant bacterial interaction study. The effect of inoculation on growth and grain and dry matter yields was evaluated in relation to nitrogen fixation, by in situ acetylene reduction assay,¹⁵N₂ feeding and¹⁵N dilution techniques. A survey of the population of marker bacteria at maximum tillering, booting and heading revealed poor effectivety. The population of nativeAzospirillum followed no definite pattern. Acetylene-reducing activity (ARA) did not differ due to inoculation at two early stages but decreased in the inoculated plants at heading. In contrast, inoculation increased tiller number, plant height of Hua and early reproductive growth of both varieties. Grain yield of both varieties significantly increased along with the dry matter. Total N also increased in inoculated plants, which was less compared with dry matter increase.¹⁵N₂ feeding of OS4 at heading showed more¹⁵N₂ incorporation in the control than in the inoculated plants. The ARA,¹⁵N and N balance studies did not provide clear evidence that the promotion of growth and nitrogen uptake was due to higher N₂ fixation.     

Phosphorus requirements and nitrogen accumulation by N2-fixing and non-N2-fixing leguminous trees growing in low P soils

The variation in P uptake and use efficiency and N accumulation by Gliricidia sepium (N₂-fixing tree), Senna siamea and S. spectabilis (leguminous non-N₂-fixing trees) were examined in the field at Fashola (savanna zone), southwestern Nigeria, using four P rates, 0, 20, 40 and 80 kg P ha^-1. Growth of G. sepium and S. spectabilis responded to P application at 24 weeks after planting (WAP) and average yield increases of 58% and 145% were observed by the application of 40 kg P ha^-1 for the two species, respectively. Such a P response was not found in S. siamea at 24 WAP and for any of the species at 48 WAP. G. sepium accumulated more P (on average 162%) than S. siamea and S. spectabilis at 24 WAP and had greater root length and a higher percentage of mycorrhizal infection. However, at 48 WAP S. siamea had 2.5 times more P than G. sepium. Differences in the physiological P use efficiency (PPUE) between G. sepium and the non-N₂-fixing trees were significant at the 0 P level, being higher for S. siamea (average, 0.61 g shoot mg^-1 P) than for G. sepium (0.27 g shoot mg^-1 P). G. sepium had a consistently lower atom % ¹⁵N than S. spectabilis, while that of S. siamea for most of the time did not differ from that of G. sepium. The reference plant affected N₂ fixation extimates, with negative values and a higher variability (CV 60%) associated with S. siamea than with S. spectabilis (CV<20%). Consequently, S. spectabilis was selected as a better reference plant for measuring N₂ fixation in G. sepium. G. sepium fixed on average 35% and 54% of its N at 24 and 48 WAP, respectively. Except at the lowest P rate, percentage and amount of N fixed were not generally enhanced by P application.     

Effects of inoculation with N2-fixing cyanobacteria on the nitrogenase activity in soil and rhizosphere of wetland rice (Oryza sativa L.)

Summary A greenhouse experiment was conducted with wetland rice (Oryza sativa cv. IR-50) in a clay-loam soil (Fluventic Eutrochrept) to study the effect of cyanobacterial inoculation a mixed culture of Aulosira fertilissima, Nostoc muscorum, Nostoc spp., and Anabaena spp., applied at the rate of 0.15 g (dry weight pot^-1 or 43 kg ha^-1) on acetylene reduction activity in soil and the root system (excised root), and the grain and straw yield. The effects of applying P (40 kg ha^-1), N (60 kg ha^-1), and P+N to the soil were also evaluated. Cyanbacterial inoculation significantly increased (more than 200% on average) photo-dependent acetylene reduction activity in soils, particularly where the indigenous activity was considerably low, i.e. under unfertilized and N-fertilized conditions. The effect of inoculation was prominent at the maximum tillering and grain formation stages of the crop. This inoculation benefit was, however, marginal in P-applied soils (P and P+N), where the indigenous activity was stimulated more than threefold. The inoculation led to a remarkable increase in root-associative acetylene reduction activity after the maximum tillering stage of the crop, particularly with applied N but for other treatments this inoculation effect was not significant. Cyanobacterial inoculation also increased the grain and straw yield of the crop when N was not applied. The grain and straw yield was significantly correlated with the acetylene reduction activity in flooded soils and in the root system during the tillering and maximum tillering stages of rice growth, respectively.     

Ammonia volatilization from nitrogen fertilizers surface-applied to corn (Zea mays) and grass pasture (Dactylis glomerata)

Summary The major agronomic concern with NH₃ loss from urea-containing fertilizers is the effect of these losses on crop yields and N fertilizer efficiency. In this 2-year study, NH₃ volatilization from surface-applied N fertilizers was measured in the field, and the effects of the NH₃ losses detected on corn (Zea mays L.) and orchardgrass (Dactylis glomerata L.) yield and N uptake were determined. For corn, NH₄NO₃ (AN), a urea-AN solution (UAN), or urea, were surface-broadcast at rates of 0, 56 and 112 kg N ha^–1 on a Plano silt loam (Typic Argiudoll) and on a Fayette silt loam (Typic Hapludalf). Urea and AN (0 and 67 kg N ha^–1) were surface-applied to grass pasture on the Fayette silt loam. Significant NH₃ losses from urea-containing N sources were detected in one of four corn experiments (12%–16% of applied N) and in both experiments with grass pasture (9%–19% of applied N). When these losses occurred, corn grain yields with UAN and urea were 1.0 and 1.5 Mg ha^–1, respectively, lower than yields with AN, and orchardgrass dry matter yields with urea were 0.27 to 0.74 Mg ha^–1 lower than with AN. Significant differences in crop N uptake between N sources were detected, but apparent NH₃ loss based on N uptake differences was not equal to field measurements of NH₃ loss. Rainfall following N application markedly influenced NH₃ volatilization. In corn experiments, NH₃ loss was low and yields with all N sources were similar when at least 2.5 mm of rainfall occurred within 4 days after N application. Rainfall within 3 days after N application did not prevent significant yield reductions due to NH₃ loss from urea in grass pasture experiments.     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.     

Influence of arbuscular mycorrhizae and phosphorus fertilization on growth, nodulation and N2 fixation (15N) inMedicago sativa at four salinity levels

The rose of an isolate of the arbuscular mycorrhizal (AM) fungusGlomus mosseae in the protection ofMedicago sativa (+Rhizobium meliloti) against salt stress induced by the addition of increasing levels of soluble salts was studied. The interactions between soluble P in soil (four levels), mycorrhizal inoculum and degree of salinity in relation to plant growth, nutrition and infective parameters were evaluated. Salt stress was induced by sequential irrigation with saline water having four concentrations of three salts (NaCl, CaCl₂, and MgCl₂).¹⁵N-labelled ammonium sulphate was added to provide a quantitative estimate of N₂ fixation under moderate to high salinity levels. N and P concentration and nodule formation increased with the amount of plant-available P or mycorrhizal inoculum in the soil and generally declined as the salinity in the solution culture increased from a moderate to a high level. The mycorrhizal inoculation protected the plants from salt stress more efficiently than any amount of plant-available P in soil, particularly at the highest salinity level applied (43.5 dS m^–1). Mycorrhizal inoculation matched the effect on dry matter and nutrition of the addition in the soil of 150 mg P kg^–1. Nevertheless the highest saline solution assayed (43.5 dS m^–1) affected more severely plants supplemented with phosphorus than those with the addition of mycorrhizal inoculum. Such a saline-depressing effect was 1.5 (biomass), 1.4 (N) and 1.5 (P) times higher in plants supplied with soluble phosphate than with AM inoculum. Mechanisms beyond those mediated by P must be involved in the AM-protectioe effect against salinity. The¹⁵N methodology used allowed the determination of N₂ fixation as influenced by different P applications compared to mycorrhizal inoculation. A lack of correlation between nodule formation and function (N₂ fixation) was evidenced in mycorrhizal-inoculated plants. In spite of the reduced activity per nodule in mycorrhizal-inoculated In spite of the reduced activity per nodule in mycorrhizal-inoculated plants, the N contents determined indicated the highest acquisition of N occurred in plants with the symbiotic status. Moreover, N and P uptake increased while Ca and Mg decreased in AM-inoculated plants. Thus P/Ca ratios and cation/anion balance in general were altered in mycorrhizal treatments. This study therefore confirms previous findings that AM-colonized plants have optional and alternative mechanisms available to satisfy their nutritive requirements and to maintain their physiological status in stress situations and in disturbed ecosystems.     

Use of 15N isotope dilution for quantification of N2 fixation associated with roots of kallar grass (Leptochloa fusca (L.))

Summary Leptochloa fusca (L.) Kunth (kallar grass) has previously been found to exhibit high rates of nitrogen fixation. A series of experiments to determine the level of biological nitrogen fixation using ¹⁵N isotopic dilution were carried out in nutrient solution and saline soil. In the nutrient solution, E. coli inoculated plants were taken as non-nitrogen-fixing control. It was observed that nearly 60%–80% of the plant N was derived from atmospheric fixation. Estimations based on the N difference method gave much lower values (18%–35%). In experiments with saline soil which was initially sterilized with chloroform fumigation, a mixed culture of N₂-fixing rhizospheric isolates from kallar grass roots was inoculated and planted to kallar grass. Uninoculated treatments were regarded as controls. The soil was previously labelled with ¹⁵N by adding cellulose and (¹⁵NH₄)₂SO₄. The results of these studies showed fixation values of 6%–32% when estimated by ¹⁵N dilution, whereas by the N difference method 54% of the plant N was estimated to be derived from fixation. This discrepancy is due to the increase in root proliferation due to inoculation, which results in greater uptake of soil N. The distribution of ¹⁵N in different fractions of the soil-N indicted isotopic dilution due to bacterial fixation of atmospheric N₂.     

Effect of plant growth-promoting bacteria and soil compaction on barley seedling growth, nutrient uptake, soil properties and rhizosphere microflora

Inoculants are of great importance in sustainable and/or organic agriculture. In the present study, plant growth of barley (Hordeum vulgare) has been studied in sterile soil inoculated with four plant growth-promoting bacteria and mineral fertilizers at three different soil bulk densities and in three harvests of plants. Three bacterial species were isolated from the rhizosphere of barley and wheat. These bacteria fixed N₂, dissolved P and significantly increased growth of barley seedlings. Available phosphate in soil was significantly increased by seed inoculation of Bacillus M-13 and Bacillus RC01. Total culturable bacteria, fungi and P-solubilizing bacteria count increased with time. Data suggest that seed inoculation of barley with Bacillus RC01, Bacillus RC02, Bacillus RC03 and Bacillus M-13 increased root weight by 16.7, 12.5, 8.9 and 12.5% as compared to the control (without bacteria inoculation and mineral fertilizers) and shoot weight by 34.7, 34.7, 28.6 and 32.7%, respectively. Bacterial inoculation gave increases of 20.3–25.7% over the control as compared with 18.9 and 35.1% total biomass weight increases by P and NP application. The concentration of N and P in soil was decreased by increasing soil compaction. In contrast to macronutrients, the concentration of Fe, Cu and Mn was lower in plants grown in the loosest soil. Soil compaction induced a limitation in root and shoot growth that was reflected by a decrease in the microbial population and activity. Our results show that bacterial population was stimulated by the decrease in soil bulk density. The results suggest that the N₂-fixing and P-solubilizing bacterial strains tested have a potential on plant growth activity of barley.     

Nitrogen and phosphorus fertilization leads to soil arbuscular mycorrhizal fungal diversity changes and rainfed crop yield increase on the Loess Plateau of China: A 37-year study

More than 80% of plants form mutualistic symbiotic relationships with arbuscular mycorrhizal fungi (AMF), and the application of fertilizers, such as nitrogen (N) and phosphorus (P) fertilizers, is a common agricultural management practice to improve crop yield and quality. However, the potential effects of long-term N and P fertilization on the AMF community in the rainfed agricultural system of the Loess Plateau of China are still not well understood. In this study, a long-term field experiment was conducted based on orthogonal design, with three N levels (0, 90, and 180 kg ha^-1 year^-1) and three P levels (0, 90, and 180 kg ha^-1 year^-1) for wheat fertilization. Changes in AMF community and correlations between AMF community composition, soil environmental factors, and wheat yield component traits were analyzed using traditional biochemical methods and high-throughput sequencing technology. The results showed that long-term N and P addition had a significant effect on the AMF community structure and composition. Nitrogen application alone significantly reduced the richness and diversity of AMF community, whereas the combined application of N and P significantly increased the richness and diversity of AMF community. The AMF community was driven mainly by soil available P, total P, and pH. There was a significant positive correlation between Glomus abundance and wheat yield and a significant negative correlation between Paraglomus abundance and wheat yield. Long-term N and P addition directly increased crop yield and affected yield indirectly by influencing soil chemical properties and the AMF community. Combined application of N and P both at 90 kg ha^-1 year^-1 could improve the ecological and physiological functions of the AMF community and benefit the sustainable development of rainfed agriculture.     

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.