Combined effects of plant growth‐promoting rhizobacteria and genistein on nitrogen fixation in soybean at suboptimal root zone temperatures

Application of plant growth‐promoting rhizobacteria (PGPR) or the plant to bacteria signal molecule genistein has been shown to increase nodulation and nitrogen (N) fixation by soybean [Glycine max (L.) Merr.] over a range of root zone temperatures (RZTs) and, specifically, off‐sets at least some of the ill‐effects of low RZTs. Two sets of controlled‐environment experiments, one on a growth bench and the other in a greenhouse, were conducted to examine the combined ability of both PGPR and genistein to reduce the negative effects of low RZT on soybean nodulation and N fixation. Each of two the PGPR strains, Serratia proteamaculans 1–102 and Serratia liquefaciens 2–68 were co‐inoculated with Bradyrhizobium japonicum USDA 110 preincubated with 17.5 (somewhat inhibitory), and 15°C (very inhibitory). At RZTs of 25 and 17.5°C PGPR strains and genistein in combination increased the number of nodules and the amount of Nn fixed. The most stimulatory effect was observed at 17.5°C for the combination: S. proteamaculans 1–102 plus B. japonicum USDA 110 pre‐incubated in 15 μM genistein under greenhouse conditions. For most treatment combinations the stimulatory effects of PGPR and genistein were additive at RZTs of 17.5 and 25°C. Surprisingly, the combination of these two factors resulted in antagonism at the very inhibitory RZT of 15°C. The results suggest that the negative effects of certain low RZTs could be more effectively off‐set by combined treatments of PGPR plus geneistin pre‐incubation of rhizobial cultures than by their individual treatment.     

Nodulation competitiveness of Rhizobium leguminosarum bv. phaseoli and Rhizobium tropici strains measured by glucuronidase (gus) gene fusion

Summary The nodulation competitiveness of 17 Rhizobium leguminosarum bv. phaseoli and 3 R. tropici strains was analysed in growth pouches, at pH 5.2 and 6.4. All 20 strains were coinoculated with a gus ⁺ strain of R. leguminosarum bv. phaseoli strain KIM5s. The gus⁺ phenotype, carrying the glucuronidase gene, was used to type nodules directly in the growth pouches. Nodule occupancy ranged from 4% for the least competitive to 96% for the most competitive R. leguminosarum bv. phaseoli strain. The R. tropici strains showed low rates of nodule occupancy at pH 6.4 but their competitiveness improved significantly under acid conditions. CIAT 895 was the only R. leguminosarum bv. phaseoli strain that was less competitive (P<0.05) at the lower pH. The competitiveness of all the other R. leguminosarum bv. phaseoli strains was unaffected by pH. Various physiological and genetic properties of the strains were analysed in search of correlations with nodulation competitiveness. Hybridisation patterns with three different DNA probes (nif KDH, common nod genes, and hup genes) and the metabolism of 53 different C sources were compared. No general correlations were found between hybridisation or growth pattern and competitiveness. The less competitive R. tropici strains had a unique DNA hybridisation pattern and were not able to use shikimate, ferulate, coumarate, or asparagine as C sources. Most of the less competitive R. leguminosarum bv. phaseoli strains could not metabolize either ferulate or coumarate. This might indicate a relationship between nodulation competitiveness and the ability to degrade aromatic compounds.     

Effects of Kocide 101® on the bean (Phaseolus vulgaris L.)‐Rhizobium symbiosis

A glasshouse study was undertaken to investigate the effects of the copper fungicide Kocide 101 and its residues in soil on the growth, nodulation and nitrogen fixation of beans (Phaseolus vulgaris L.). The soil used was a sandy clay loam classified as Typic Rhodustalf. The bean variety SUA 90 was used as test crop. The bean rhizobia strains CIAT 899, PV, and a local isolate were used. Kocide 101 applied at the recommended rate (equivalent to 1.7 mg kg^‐1 soil) had no significant negative effects on the growth, nodulation or nitrogen fixation of bean plants. Higher levels of Kocide 101 significantly (P < 0.05) reduced plant growth, nodulation and nitrogen fixation. The bean plants inoculated with the “local isolate”; rhizobia had the highest dry matter weights, nodule numbers and nodule dry weights, and also had more N fixation. They were followed by those inoculated with the PV, strain and, lastly, those inoculated with CIAT 899. The growth and nodulation of bean plants were still curtailed by the Kocide 101 residues four months after the fungicide was first applied to the soil. Therefore, occurrence of high levels of Kocide 101 in soils can have long‐term effects on the performance of the bean‐rhizobia symbiosis.     

Manganese modulates the responses of nitrogen-supplied and Rhizobium-nodulated Phaseolus vulgaris L. to inoculation with arbuscular mycorrhizal fungi

In Venezuela, low yields of black bean crops are attributed, in part, to the low manganese (Mn) and phosphorus (P) contents in the Quartzipsamment soils where this crop is usually sown. To test this hypothesis, black bean plants were grown in sterilized sand to simulate soil physical properties, were fertilized with increasing Mn concentrations (0.1-20 μM) and inoculated with a commercial mixture of Rhizobium leguminosarum bv phaseoli strains 127K44, 127K89, 127K105 (+Rh), in combination with arbuscular mycorrhizal fungi Scutellospora heterogama and Entrophospora colombiana (+AMF). Non-inoculated plants fertilized with 6 mM NO₃ and 2 mM P served as controls. Plants were harvested at 18, 25, 33, and 40 days after emergence. At all harvests, the greatest growth and highest P and iron (Fe) leaf concentrations occurred in control plants grown in 5 μM Mn. The growth of +AMF plants was promoted at 0.1 μM Mn and inhibited at higher than 1 μM Mn. Whereas, concentrations of 5-10 μM Mn enhanced the growth and the Mn concentrations in leaves of +Rh plants 40 days after emergence. The tripartite symbiosis (+Rh+AMF) decreased growth, nodulation and leaf ureide and chlorophyll concentrations in plants grown in less than 20 μM Mn, imputed to severe ultrastructural alterations in the leaf and nodule tissues. Only +Rh+AMF plants grown in 20 μM Mn were effectively nodulated, AMF colonized and reached the flowering stage, although with diminished growth and low chlorophyll concentrations. Results confirm the high Mn requirement of +Rh plants for growth and nodulation and question the implementation of the tripartite symbiosis to improve yields in early flowering black bean varieties planted in soils deficient in Mn and P.     

Competition and persistence of Rhizobium tropici and Rhizobium etli in tropical soil during successive bean (Phaseolus vulgaris L.) cultures

Strains of Rhizobium tropici IIB, CIAT899 and F98.5, both showing good N₂ fixation, and a R. etli strain W16.3SB were introduced into a field which had no history of bean culture. Plant dilution estimates showed that in the presence of its host (Phaseolus vulgaris cv. Carioca) during the cropping seasons and the subsequent fallow summer periods, the bean rhizobial populations increased from less than 30 to 10³ g^–1 dry soil after 1 year and to 10⁴ g^–1 dry soil after 2 years. In the 1st year crop, the inoculated strains occupied most of the nodules, which resulted in a higher nodulation and C₂H₂ reduction activity. Without reinoculation for the second and third crops, however, little R. tropici IIB was recovered from the nodules and the bean population consisted mainly of R. etli, R. leguminosarum bv. phaseoli, and R. tropici IIA. Reinoculation with our superior R. tropici IIB strains before the second crop resulted in R. tropici IIB occupying the main part of the nodules and a positive effect on nodulation and C₂H₂ reduction activity, but reintroduction of the inoculant strain in the third season did not have any effect.     

Effects of resident rhizobial communities and soil type on the effective nodulation of pulse legumes

Communities of resident rhizobia capable of effective nodulation of pulse crops were found to vary considerably over a range of soil environments. These populations from soils at 50 sites in Southern Australia were evaluated for nitrogen fixing effectiveness in association with Pisum sativum, Vicia faba, Lens culinaris, Vicia sativa, Cicer arietinum and Lupinus angustifolius. The values for nitrogen fixing effectiveness could be related to soil pH as determined by soil type and location. It was found that 33% of paddocks had sufficient resident populations of Rhizobium leguminosarum bv viciae for effective nodulation of faba bean, 54% for lentils, 55% for field pea and 66% for the effective nodulation of the vetch host plant. Mesorhizobium cicer populations were very low with only 7% of paddocks surveyed having sufficient resident populations for effective nodulation. Low resident rhizobial populations (<10 rhizobia g⁻¹ soil) of R. leguminosarum bv viciae and M. cicer were found in acid soil conditions. In contrast, Bradyrhizobium populations increased as soil pH decreased. Inoculation increased faba bean yields from 0.34 to 4.4 t ha⁻¹ and from 0.47 to 2.37 t ha⁻¹ for chickpeas on acid soils. On alkaline soils, where resident populations were large there was no consistent response to inoculation. Observations at experimental field sites confirmed the findings from the survey data, stressing the importance of rhizobial inoculation, especially on the acid soils in south-eastern Australia.     

Changes in Phosphate Fractions Extracted From Different Organs of Phosphorus Starved Nitrogen Fixing Pea Plants

The present work investigates the impact of phosphorus (P) starvation on plant growth, symbiotic nitrogen fixation, and internal P status (determined as extracted P fractions) of leaves, roots, and nodules of 27-days–old pea (Pisum sativum L) plants inoculated with Rhizobium leguminosarum bv viciae strain D293. The procedure of separation of organic and inorganic P compounds in 10% perchloric acid (HCLO₄) and the absorption of nucleotides in active charcoal gave several fractions, containing different phosphorus compounds, which were extracted and determined as inorganic phosphate after combustion. These are acid soluble and insoluble P, sugar P, nucleotide P, and inorganic P. The P starvation of plants inhibited significantly plant dry mass accumulation, nodulation rate and specific nitrogenase activity of nodules. These results were accompanied with lower quantities of total P per plant, acid soluble and acid non-soluble P fractions in all plant organs. The inhibited accumulation of P in the acid soluble P fraction was associated with decrease of sugar, nucleotide and inorganic P in all plant organs. The most negatively affected were all P fractions extracted from nodules and leaves. The low content of inorganic P in the stressed plant tissues was regarded as primary reason for induced alterations in the content of analyzed P fractions.     

Enhanced dissipation of chrysene in planted soil: the impact of a rhizobial inoculum

Results from an innovative approach to improve remediation in the rhizosphere by encouraging healthy plant growth and thus enhancing microbial activity are reported. Mixed grass-legume systems, together with microbial inoculants, were used to remediate a polycyclic aromatic hydrocarbon (chrysene) spiked agricultural soil. Inoculants were symbiotic rhizobia, which may play an important role in rhizoremediation by increasing plant and root growth. An inoculum of an isolate of Rhizobium leguminosarum bv. trifolii, selected for PAH tolerance, was produced using a peat carrier. The inoculum and white clover (Trifolium repens L.), were planted into soils with ryegrass (Lolium perenne L.). The soils spiked with chrysene (500 mg kg⁻¹) then aged for 4 weeks. Shoot- and root-biomass of plants, and the amount of root nodulation, were determined. Rhizobial populations, soil pH and soil nitrogen were also monitored throughout the trial. In addition, the ability of the inoculated rhizobial strain to utilise chrysene as a sole carbon source was assessed. Direct uptake and/or degradation of chrysene by the clover and ryegrass did not occur to a significant degree. Enhanced losses of chrysene were seen in planted, non-sterile soils that contained a rhizobial inoculum. No direct degradation of chrysene by R. leguminosarum bv. trifolii was observed and no enhanced losses of PAHs were detected in sterile soils after inoculation with rhizobia. Results suggest that the enhanced dissipation of chrysene, observed in the non-sterile planted inoculated pots, was not a result of degradation of chrysene by R. leguminosarum bv. trifolii. The symbiotic association with R. leguminosarum bv. trifolii improved plant vigour and growth in inoculated planted treatments. This may have stimulated the rhizospheric microflora to degrade chrysene.     

Effect of seed phosphorus concentration on nodulation and growth of three common bean cultivars

Abstract

An experiment was conducted in the greenhouse to evaluate the effects of seed phosphorus (P) concentration on growth, nodulation, and nitrogen (N) and P accumulation of three common bean (Phaseolus vulgaris L.) cultivars. Seeds were produced under low or high soil P levels, and soaked, or not, in 200 mM KH₂PO₄ solution. The experiment had a 3×3×2×2 factorial block design: three cultivars (ICA Col 10103, Carioca and Honduras 35), three levels of applied P (15, 30 and 45 mg P kg^?1 soil), two native seed P concentrations, and two seed soaking treatments. Plants were harvested at flowering. Soaked seeds increased the number, dry mass and P content of nodules, but did not affect plant growth. Plants originated from seeds with high native P concentration presented higher shoot dry mass and nodule number and mass at every soil P level, and were less responsive to increased soil P supply, than plants from low seed P. In plants from seeds with high P, soil P levels did not alter significantly root dry mass, while in plants from seeds with low P bean cultivars expressed wider differences in root dry mass. The genotypic variability of nodulation was influenced by soil P levels and seed P concentration. Both higher soil or seed P supply enhanced N and P accumulation in shoots. These results indicate that a high seed P concentration produces plants less dependent on soil P supply, and can enhance nodulation and N₂ fixation of common bean. Seed P supply affected the cultivar performance, and should be considered in evaluation of bean genotypes.     

Genistein addition to the rooting medium of soybean at the onset of nitrogen fixation increases nodulation

In the legume‐(Brady)Rhizobium symbiosis, signal exchange between the host‐plant and the symbiotic bacterium is an essential step in nodule formation. Genistein is the most effective plant‐to‐bacterium signal in the soybean [Glycine max (L.) Merr.] N₂‐fixing symbiosis. Its concentration in soybean root system increases with seedling development, and decreases immediately after the onset of N₂ fixation. This study was conducted to determine whether addition of genistein to the rooting medium at the onset of N₂ fixation would increase nodulation thereafter. The results indicated that watering soybean plants with a solution containing genistein beginning at the onset of N₂‐fixation increased nodule size, nodule number and nodule weight per plant. Shoot nitrogen (N) concentration was also increased. Soybean cultivar AC Bravor was more sensitive to genistein addition than Maple Glen.     

Competitiveness of Rhizobium leguminosarum bv. phaseoli strains in relation to environmental stress and plant defense mechanisms

Summary Six Rhizobium leguminosarum bv. phaseoli strains (Ciat 151, Ciat 895, Ciat 899, CE3, H2C, Kim5s) were tested for nodule occupancy in different bean cultivars at two field sites (one fertile, one acid tropical soil) and in the greenhouse. The effects of several environmental factors such as low pH, high temperature, Al and Mn toxicity, iron deficiency, bean tannins, and bean phytoalexins were tested in vitro. Strain Kim5s was competitive under all tested conditions while strains CE3 and H2C had consistently low nodule occupancy levels. Strain Ciat 151 was superior to the other inoculant strains in the acid soil but competed poorly in the fertile soil. Strain Ciat 895 was more competitive in the fertile soil. There was a decline in nodule occupancy for all strains tested from the first trifoliate leaf stage to the pod-filling stage. No plant genotype effect on nodule occupancy was observed. There were significant (P<0.05) plant genotype and location effects, but no significant strain effect on acetylene reduction activity, plant dry weight, and nodule number. The greenhouse experiments confirmed, at least partially, the results from the field trials. In Leonard jars with an acid soil, strains Ciat 151 and Kim5s were amongst the most competitive strains. In coinoculation experiments, Kim5s was the most competitive strain, followed by Ciat 899 and Ciat 895. The competitiveness of a given strain was affected by the coinoculant strain. Tolerance in vitro to low pH, high growth temperature, Al or Mn toxicity, or Fe limitation was not related to competitiveness of the inoculum strains. The sensitivity of the strains towards bean tannins or bean phytoalexins also was not correlated with their competitiveness.     

Influence of Pseudomonas syringae R25 and P. putida R105 on the growth and N2 fixation (acetylene reduction activity) of pea (Pisum sativum L.) and field bean (Phaseolus vulgaris L.)

The effects of inoculating field peas (Pisum sativum L.) with Rhizobium leguminosarum and field beans (Phaseolus vulgaris L.) with R. phaseoli, alone or in combination with Pseudomonas syringae R25 and/or P. putida R105, were assessed under gnotobiotic conditions in growth pouches and in potted soil in a growth chamber. Inoculation of peas with P. syringae R25 or P. putida R105 alone had no effect on plant growth in pouches. In soil, however, the isolate R25 inhibited nitrogenase activity (as assessed by acetylene reduction assay) of nodules formed by indigenous rhizobia; strain R105 stimulated pea seedling emergence and nodulation. P. syringae R25 inhibited the growth of beans in either plant-growth system. P. putida R105, however, had no effects on beans in pouches, but reduced plant root biomass and nodulation by indigenous rhizobia in soil. Coinoculation of pea seeds with R. leguminosarum and either of the pseudomonads significantly (P<0.01) increased shoot, root, and total plant weight in growth pouches, but had no effect in soil. Co-inoculation of field beans with R. phaseoli and P. putida R105 had no effects on plant biomass in growth pouches or in soil, but the number of nodules and the acetylene reduction activity was significantly (P<0.01) increased in the soil. In contrast, co-inoculation of beans with rhizobia and P. syringae R25 had severe, deleterious effects on seedling mergence, plant biomass, and nodulation in soil and growth pouches. Isolate R25 was responsible for the deleterious effects observed. Although plant growth-promoting rhizobacteria may interact synergistically with root-nodulating rhizobia, the PGPR selected for one crop should be assessed for potential hazardous effects on other crops before being used as inoculants.     

Cropping history affects nodulation and symbiotic efficiency of distinct hairy vetch (Vicia villosa Roth.) genotypes with resident soil rhizobia

Compatible rhizobia strains are essential for nodulation and biological nitrogen fixation (BNF) of hairy vetch (Vicia villosa Roth, HV). We evaluated how past HV cultivation affected nodulation and BNF across host genotypes. Five groups of similar HV genotypes were inoculated with soil dilutions from six paired fields, three with 10-year HV cultivation history (HV+) and three with no history (HV?), and used to determine efficiency of rhizobia nodulation and BNF. Nodulation was equated to nodule number and mass, BNF to plant N and Rhizobium leguminosarum biovar viceae (Rlv) soil cell counts using qPCR to generate an amplicon of targeted Rlv nodD genes. Both HV cultivation history and genotype affected BNF parameters. Plants inoculated with HV+ soil dilutions averaged 60 and 70 % greater nodule number and mass, respectively. Such plants also had greater biomass and tissue N than those inoculated with HV? soil. Plant biomass and tissue N were strongly correlated to nodule mass (r ²?=?0.80 and 0.50, respectively), while correlations to nodule number were low (r ²?=?0.50 and 0.31, respectively). Although hairy vetch rhizobia occur naturally in soils, past cultivation of HV was shown in this study to enhance nodulation gene-carrying Rlv population size and/or efficiency of rhizobia capable of nodulation and N fixation.     

The fungicides thiram and captan affect the phenotypic characteristics of Rhizobium leguminosarum strain C1 as determined by FAME and Biolog analyses

 We assessed the effects of the fungicides captan and thiram on the survival and phenotypic characteristics of Rhizobium leguminosarum bv. viceae, strain C1. Fungicide was applied to pea seed at rates of 0.25–2 g a.i. kg^–1 seed, and treated seed was inoculated with strain C1. After 24 h, rhizobia were extracted from treated seed and viable numbers determined by plating onto yeast extract mannitol agar. Phenotypic characteristics were assessed using FAME (fatty acid methyl ester) profiles and Biolog substrate utilization patterns. Captan and thiram significantly reduced the numbers of rhizobia recovered from seed and altered the FAME and Biolog profiles of recovered rhizobia. However, only the highest concentrations of captan affected nodulation and plant growth. Contact with some seed-applied fungicides can significantly alter phenotypic characteristics of rhizobia, but these changes might be offset by the presence of host plants. Received: 20 February 1999     

PLANT GROWTH PROMOTING RHIZOBACTERIA INCREASE GROWTH,YIELD AND NITROGEN FIXATION IN PHASEOLUS VULGARIS

Nitrogen (N) fixation by legume-Rhizobium symbiosis is important to agricultural productivity and is therefore of great economic interest. Growing evidence indicates that soil beneficial bacteria can positively affect symbiotic performance of rhizobia. The effect of co-inoculation with plant growth-promoting rhizobacteria (PGPR) and Rhizobium, on nodulation, nitrogen fixation, and yield of common bean (Phaseolus vulgaris L.) cultivars was investigated in two consecutive years under field conditions. The PGPR strains Pseudomonas fluorescens P-93 and Azospirillum lipoferum S-21 as well as two highly effective Rhizobium strains were used in this study. Common bean seeds of three cultivars were inoculated with Rhizobium singly or in a combination with PGPR to evaluate their effect on nodulation and nitrogen fixation. A significant variation of plant growth in response to inoculation with Rhizobium strains was observed. Treatment with PGPR significantly increased nodule number and dry weight, shoot dry weight, amount of nitrogen fixed as well as seed yield and protein content. Co-inoculation with Rhizobium and PGPR demonstrated a significant increase in the proportion of nitrogen derived from atmosphere. These results indicate that PGPR strains have potential to enhance the symbiotic potential of rhizobia.     

Nodulation and N2 fixation of faba bean (Vicia faba L.) after soil amendment with crop residues

The effect of prior soil amendment with different N sources at 50 mg N (kg soil)^—1 on nodulation and N₂ fixation of faba bean (Vicia faba L. cv. Troy) using wheat (Triticum aestivum L. cv. Star) as reference crop was assessed in a pot experiment. Four treatments viz legume manure (LEGM) as clover shoots, cereal manure (CEREM) as barley straw, N fertilizer (FERT‐N) as Ca(NO₃)₂, and no‐manure control (NOMAN) were investigated consecutively at 45, 70, and 90 days after sowing (DAS). Faba bean nodulated profusely, with an increase on average from 629 nodules per pot at 45 DAS to nearly 2.3‐ and 3.3‐fold at 70 and 90 DAS, respectively. Low nodule numbers and nodule dry matter occurred under FERT‐N and CEREM, whereas high values were found for NOMAN and LEGM. Soil amendment affected percent N₂ fixation in relation to N source and plant age. Highest percent N₂ fixation (≥ 90 %) was found under the lowest N‐supplying amendments, no‐manure, and cereal manure, respectively. FERT‐N depressed N₂ fixation particularly at 45 DAS when N₂ fixation was reduced to as low as 23 %. The rise in N₂ fixation thereafter suggests that faba bean adjusted after depletion of mineral N in the soil. N₂ fixation was also decreased after cereal straw application, even though N concentration in faba bean plants was high. The results indicate that plant residues, both with high and low N concentration, applied to soil to raise its fertility may interfere with N₂ fixation of faba bean.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

Competition between inoculated and indigenous Rhizobium/Bradyrhizobium spp. strains for nodulation of grain and fodder legumes in Pakistan

Summary The competitive ability of inoculated and indigenous Rhizobium/Bradyrhizobium spp. to nodulate and fix N₂ in grain legumes (Glycine max, Vigna unguiculata, Phaseolus vulgaris) and fodder legumes (Vicia sativa, Medicago sativa, and Trifolium subterraneum) was studied in pots with two local soils collected from two different fields on the basis of cropping history. The native population was estimated by a most-probable-number plant infectivity test in growth pouches and culture tubes. The indigenous rhizobial/bradyrhizobial population ranged from 3 to 2×10⁴ and 0 to 4.4×10³ cells g^-1 in the two soils (the first with, the second without a history of legume cropping). Inoculated G. max, P. vulgaris, and T. subterraneum plants had significantly more nodules with a greater nodule mass than uninoculated plants, but N₂ fixation was increased only in G. max and P. vulgaris. A significant response to inoculation was observed in the grain legume P. vulgaris in the soil not previously used to grow legumes, even in the presence of higher indigenous population (>10³ cells g^-1 soil of Rhizobium leguminosarum bv phaseoli). No difference in yield was observed with the fodder legumes in response to inoculation, even with the indigenous Rhizobium sp. as low as <14 cells g^-1 soil and although the number and weight of nodules were significantly increased by the inoculation in T. subterraneum. Overall recovery of the inoculated strains was 38–100%, as determined by a fluorescent antibody technique. In general, the inoculation increased N₂ fixation only in 3 out of 12 legume species-soil combinations in the presence of an indigenous population of rhizobial/bradyrhizobial strains.     

Foliar application of molybdenum in common bean. III. Effect on nodulation

Two field experiments were conducted, one in Viçosa, Brazil (high fertility soil), and the other in Coimbra, Brazil (low fertility soil) to verify the effect molybdenum (Mo), nitrogen (N), or Rhizobium on growth of common bean (Phaseolus vulgaris). Plants were either given a foliar application of 40 g Mo ha^‐1, or an addition of N fertilizer (20 kg ha^‐1 at planting and/or 30 kg ha^‐1 as a side dressing), or inoculated with two selected Rhizobium strains. Nodulation parameters of common bean (cv. Ouro Negro 1992) were measured. At both sites, Mo application at 25 days after plant emergence decreased nodule number per plant, while nodule weight was not affected in Viçosa and was increased in Coimbra at 46 days after plant emergence. However, N application at planting decreased the number and the weight of nodules at both sites, while N side dressed did not affect nodule number or weight. Therefore, the main effect of Mo fertilization was an increase in the size of nodules. The main effect of Mo on nodulation appears to be the avoidance of nodule senescence, and so maintaining a longer period of effective N₂ fixation.     

Effect of Root Zone Temperature on Oilseed Rape (Brassica napus) Response to Boron

Abstract

Oilseed rape (Brassica napus) is sensitive to low boron (B) supply, and its growth response to B may be influenced by soil temperature. To test the relationship between B and temperature, oilseed rape (cv. Hyola 42) seedlings were grown at 10°C (low) root zone temperature (RZT) with B supply from deficient to adequate B levels until growth of low B plants just began to slow down. Half of the pots were then transferred to 20°C (warm) RZT for 11 days before they were moved back to 10°C RZT for the final 4 days. Both plant dry mass and B uptake increased after plants were exposed to warm RZT. However, plant B deficiency was exacerbated by warm RZT in low B plants because of increased relative growth rate and shoot–root ratio without a commensurate increase in B uptake rate. It is concluded that RZT above the critical threshold for chilling injury in oilseed rape can nevertheless affect the incidence of B deficiency by altering shoot–root ratio and hence the balance between shoot B demand and B uptake.