The pea ( Pisum sativum L.) genes sym33 and sym40 control infection thread formation and root nodule function

Two novel non-allelic mutants that were unable to fix nitrogen (Fix⁻) were obtained after EMS (ethyl methyl sulfonate) mutagenesis of pea (Pisum sativum L.). Both mutants, SGEFix⁻–1 and SGEFix⁻–2, form two types of nodules: SGEFix⁻–1 forms numerous white and some pink nodules, while mutant SGEFix⁻–2 forms white nodules with a dark pit at the distal end and also some pinkish nodules. Both mutations are monogenic and recessive. In both lines the manifestation of the mutant phenotype is associated with the root genotype. White nodules of SGEFix⁻–1 are characterised by hypertrophied infection threads and infection droplets, mass endocytosis of bacteria, abnormal morphological differentiation of bacteroids, and premature degradation of nodule symbiotic structures. The structure of the pink nodules of SGEFix⁻–1 does not differ from that of the parental line, SGE. White nodules of SGEFix⁻–2 are characterised by “locked” infection threads surrounded with abnormally thick plant cell walls. In these nodules there is no endocytosis of bacteria into host-cell cytoplasm. The pinkish nodules of SGEFix⁻–2 are characterised by virtually undifferentiated bacteroids and premature degradation of nodule tissues. Thus, the novel pea symbiotic genes, sym40 and sym33, identified after complementation analysis in SGEFix⁻–1 and SGEFix⁻–2 lines, respectively, control early nodule developmental stages connected with infection thread formation and function. Received: 12 June 1998 / Accepted: 25 June 1998     

Effect of rhizobia and rock biofertilizers with <Emphasis Type="Italic">Acidithiobacillus</Emphasis> on cowpea nodulation and nutrients uptake in a tableland soil

Chemical fertilizers have been used in the cultivation of plants due to their high solubility and effect on crops yield. Biofertilizers with phosphate rock (PR) and potash rock (KR) plus sulfur inoculated with Acidithiobacillus may improve plant growth and contribute to addition of available P and K in soil. The effectiveness of biofertilizers from phosphate and potash rocks mixed with sulfur and Acidithiobacillus was studied in a Typic Fragiuldult soil of the Brazilian Northeast Tableland. Cowpea (cv. “IPA 206”) was grown with and without rhizobia inoculation. Treatments were: (a) phosphate rock (1000 kg ha⁻¹); (b) Biofertilizers-B_P (250 and 500 kg ha⁻¹); (c) triple superphosphate-TSP (250 kg ha⁻¹); (d) potash rock (1000 kg ha⁻¹); (e) biofertilizer-B_K (250; 500 and 750 kg ha⁻¹); (f) potassium chloride-KCl (250 kg K₂0 ha⁻¹); (g) control without P or K fertilization (P₀K₀). The soil was maintained under water submersion covered with black plastic (solarization process) for a period of 30 days. Biofertilizers (B_p and B_K) and soluble fertilizers increased plant growth and NPK uptake. Biofertilizers reduced soil pH, especially when applied in highest rates. Biofertilizers and TSP+KCl showed the best values of available P and K in soil. Rhizobial inoculation was effective on cowpea, but no nodules were formed by bacteria native from the soil, probably due to the effect of the solarization process. From obtained PK biofertilizers could be used as alternative for cowpea fertilization in Tableland soils.     

Studies on the efficacy of a phytohormone producing phosphate solubilizingBacillus firmus in augmenting paddy yield in acid soils of Nagaland

Summary A super strain ofBacillus firmus (NCIM-2636) producing a phytohormone, indole-3-acetic acid, in addition to its high ability to solubilize insoluble inorganic phosphates were applied in acid soils of Nagaland, India. Rice (Oryza sativa L.) variety Jaya and IR-8 were grown in kharif season in two successive years 1980 and 1981. After proper manuring the soils received single super phosphate (S.S.P.) and Mussoorie Rock phosphate (R.P.) separately at different doses. Yield of crop in both the years increased significantly due to bacterial inoculation. Maximum grain yield was recorded in Jaya variety under S.S.P. and R.P. when treatments were at the dose of 43.75 and 17.5 kg P ha⁻¹ respectively while the same in IR-8 variety under S.S.P. and R.P. treatments were at the dose of 35 and 17.5 kg P ha⁻¹ respectively. Maximum straw yield was produced by Jaya variety when 35 and 43.75 kg P ha⁻¹ in the form of S.S.P. and R.P. respectively were applied. Highest straw yield of IR-8 variety was obtained after the application of 17.5 kg P ha⁻¹ (S.S.P. and R.P.) in combination with phosphate solubilizing bacteria. Bacterial inoculation decreased the phosphorus availability in 1 st year but increased the same in 2nd year. Phosphorus content in grains was significantly enhanced in both the trials. Maximum uptake of phosphorus by grains was noted in Jaya variety at the dose of 47.5 kg P ha⁻¹ and in IR-8 variety at the dose of 52.5 kg P ha⁻¹ under S.S.P. treatment, while 8.75 and 35 kg P ha⁻¹ in the form of R.P. yielded similar results in Jaya and IR-8 varieties respectively. Phosphorus at the dose of 35 kg ha⁻¹ was found to cause more P-uptake by straw in both S.S.P. and R.P. treatments. The various data from the experiment conclusively proved that the bacterium in combination with R.P. produced the desired effect more prominently than when bacterium applied in combination with S.S.P.     

Improvement of growth,under field conditions,of wheat inoculated with <Emphasis Type="Italic">Pseudomonas chlororaphis</Emphasis> subsp.<Emphasis Type="Italic"> aurantiaca</Emphasis> SR1

Effects of inoculation of wheat (Triticum aestivum L.) with the rhizobacterium Pseudomonas chlororaphis subsp. aurantiaca strain SR1 (termed SR1) were studied at an experimental field site in Río Cuarto, Argentina. Treatments involved SR1 inoculation with or without nitrogen/phosphorus fertilization. Inoculation produced a significant increase in plant height and root length in early growth stages. Inoculation plus fertilization with 40 kg ha⁻¹ urea/30 kg ha⁻¹ diamonic phosphate (“50% dose”) gave a yield increase of 636 kg ha⁻¹ relative to control, and an increase of 472 kg ha⁻¹ relative to fertilization with 80 kg ha⁻¹ urea/60 kg ha⁻¹ phosphate without inoculation. SR1 inoculation without fertilization, compared to control, produced increases of 6% in weight of 1,000 grains, 13% in number of spikes per plant, and 30% in number of grains per spike. Inoculation plus 50% dose fertilization also improved these parameters. Results of the study indicate that inoculation of wheat with SR1 improves various growth and yield parameters, and allows reduced dosage of nitrogen/phosphorus fertilizers in the field.     

Phosphorus fertilization and tillage effect on dinitrogen fixation in soybeans

Summary Tillage has been shown to affect the uptake of phosphorus (P) and yield of soybeans, [Glycine max (L.) Merr.], but there is little information concerning the effects of P fertilization on nitrogen (N₂) fixation in soybeans under no-tillage. Two field experiments were conducted in 1980 and 1981 to determine the effects of soil P on N₂ fixation under no-tillage and to study the interaction of P fertilization and tillage of N₂ fixation, nutrient uptake, and yield of soybeans. In Exp. I, P was applied in 1977 at five rates up to 384 kg P ha⁻¹ and the effects of residual soil P were evaluated in 1980 and 1981 under no-tillage management. Nitrogen fixation rates, as measured by acetylene reduction assay, were significantly affected by soil P in Exp. I, but the assay proved to be a poor technique for estimating total plant N in these tests. Acetylene reduction rates and plant P increased rapidly as soil P increased from 2 to 20 mg kg⁻¹, with little additional increase above 20 mg P kg⁻¹. In Exp. II, rates (0, 32, 64, and 128 kg P ha⁻¹) and time (fall, spring and fall plus spring) of P application were compared under conventional tillage and no tillage. However, plant P increased with increasing levels of applied P. Applied P had no affect on acetylene reduction rates but rates were greater for no-tillage than conventional tillage at the V9 and R5 stages of growth in 1981. Plant uptake of P was more efficient under no-tillage than under conventional tillage in 1980 and 1981. Application of 64 kg P ha⁻¹ under no-tillage resulted in equivalent plant P levels as the 128 kg P ha⁻¹ applied under conventional tillage.     

Reversibility of phosphorus sorption by ferruginous nodules

Ferruginous nodules sorb significant amounts of available soil and fertiliser phosphate. The effect of this sorption on phosphorus availability of an agricultural soil was tested by sequential extraction and by exhaustive cropping with millet (Pennisetum typhoides) in a greenhouse trial following fertilisation of the original soil containing 70% nodules and of prepared samples containing various mixes of separated soil fines and nodules. Phosphorus sorption maxima by the soil fines and nodules were 190 mg kg⁻¹ and 380 mg kg⁻¹ respectively. Samples of fines and nodules which had sorbed 110 and 194 mg kg⁻¹ were submitted to 8 successive extractions with 0.01 M KCl, after which P desorption amounted to 117 mg kg⁻¹ and 103 mg kg⁻¹ respectively. Hysteresis between sorption and desorption was negligible for the soil fines and increased with increasing nodule content of the samples. In the greenhouse experiment, P uptake at the first cropping was highest in the soil fines at all levels of phosphorus applied. Subsequent croppings, however, showed higher P uptake in the concretionary soils. These results indicate a higher initial P release from the soil fines with cropping followed by an earlier exhaustion of phosphorus. At the end of the greenhouse experiment, yields were low in spite of the large quantities of P still remaining in the soils. Phosphorus fractionation showed that, of the P left in the soil after cropping 20% was in labile, 29% in Fe or Al-associated, and 51% in low-availability forms.     

Effect of phosphorus deficiency on acid phosphatase and phytase activities in common bean (Phaseolus vulgaris L.) under symbiotic nitrogen fixation

Changes in growth, symbiotic nitrogen fixation (SNF), acid phosphatase (ACP), and phytase activities to phosphorus availability (15 and 60 μmol KH₂PO₄ plant⁻¹ week⁻¹) were compared in two recombinant lines (115 and 147) of common bean. Plant growth, nodulation and SNF were genotype and P level-dependent. 147 was more affected by P shortage (15 μmol P) than 115. Four ACP types were revealed in the nodules of both lines, ACP1 exhibiting a higher specific activity under P shortage as compared to the 60 μmol P treatment, especially in 115. A single phytase was revealed for the nodules of both lines and was significantly enhanced by P deficiency. Three ACP types were found in roots and leaves, showing increasing activity under P deficiency, especially in 115. Regardless of P supply, leaf ACP specific activity was higher than that of nodules and roots in the both lines. Interestingly, phosphorus use efficiency for N₂ fixation significantly correlated to nodule ACP activity under P shortage in the both lines. The relatively better performance of 115 as compared to 147 under P deficiency could be partly ascribed to the ability of 115 to maintain higher ACP activity. This enzyme might be involved in the remobilization of the plant Pi and its utilization for SNF.     

Role of the polychaete <Emphasis Type="Italic">Neanthes succinea</Emphasis> in phosphorus regeneration from sediments in the Salton Sea,California

The Salton Sea currently suffers from several well-documented water quality problems associated with high nutrient loading. However, the importance of phosphorus regeneration from sediments has not been established. Sediment phosphorus regeneration rates may be affected by benthic macroinvertebrate activity (e.g. bioturbation and excretion). The polychaete Neanthes succinea (Frey and Leuckart) is the dominant benthic macroinvertebrate in the Salton Sea. It is widely distributed during periods of mixing (winter and spring), and inhabits only shallow water areas following development of anoxia in summer. The contribution of N. succinea to sediment phosphorus regeneration was investigated using laboratory incubations of cores under lake temperatures and dissolved oxygen concentrations typical of the Salton Sea. Regeneration rates of soluble reactive phosphorus (SRP) were lowest (−0.23–1.03 mg P m⁻² day⁻¹) under saturated oxygen conditions, and highest (1.23–4.67 mg P m⁻² day⁻¹) under reduced oxygen levels. N. succinea most likely stimulated phosphorus regeneration under reduced oxygen levels via increased burrow ventilation rates. Phosphorus excretion rates by N. succinea were 60–70% more rapid under reduced oxygen levels than under saturated or hypoxic conditions. SRP accounted for 71–80% of the dissolved phosphorus excreted under all conditions. Whole-lake SRP regeneration rates predicted from N. succinea biomass densities are highest in early spring, when the lake is mixing frequently and mid-lake phytoplankton populations are maximal. Thus, any additional phosphorus regenerated from the sediments at that time has potential for contributing to the overall production of the lake. Guest Editor: John M. Melack Saline Water and their Biota     

Effects of temperature, and availability of nitrogen and phosphorus on the abundance of Anabaena and Microcystis in Lake Biwa, Japan: an experimental approach

Under optimal nutrient conditions, both Microcystis sp. and Anabaena sp. isolated from Lake Biwa grew optimally at 28–32°C but differed in maximal growth rates, phosphate uptake kinetics, maximal phosphorus quotas, and growth responses to nitrogen and phosphorus limitation. The maximal growth rates of Microcystis and Anabaena were 1.6 and 1.25 divisions day⁻¹, respectively. With phosphate and nitrate in the growth-limiting range, the growth of Microcystis was optimal at an N : P ratio of 100 : 1 (by weight) and declined at lower (nitrogen limitation) and higher (phosphorus limitation) ratios. In contrast, Anabaena growth rates did not change at N : P ratios from 1000 : 1 to 10 : 1. Starting with cells containing the maximal phosphorus quota, Microcystis growth in minus-phosphorus medium ceased in 7–9 days, compared with 12–13 days for Anabaena. The phosphate turnover time in cultures starved to their minimum cell quotas was 7.9 min for Microcystis and 0.6 min for Anabaena. Microcystis had a higher K _s (0.12 μg P l⁻¹ 10⁻⁶ cells) and lower V _max (9.63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), than Anabaena (K _s 0.02 μg P l⁻¹ h⁻¹ 10⁻⁶ cells; V _max 46.25 63 μg P l⁻¹ h⁻¹ 10⁻⁶ cells), suggesting that Microcystis would not be able to grow well in phosphorus-limited waters. We conclude that in spite of the higher growth rate under ideal conditions, Microcystis does not usually bloom in the North Basin because of low availability of phosphorus and nitrogen. Although Anabaena has an efficient phosphorus-uptake system, its main strategy for growth in low-phosphorus environments may depend on storage of phosphorus during periods of abundant phosphorus supply, which are rare in the North Basin. Received: July 31, 2000 / Accepted: October 18, 2000     

The root surface phosphatases ofEriophorum vaginatum: Effects of temperature,pH, substrate concentration and inorganic phosphorus

Eriophorum vaginatum L. subsp.spissum (Fern.) Hult., a dominant plant in arctic tundra ecosystems, has acid phosphatase activity evenly distributed along its root surface from the root tip to a distance at least 16 cm from the tip. These root surface phosphatases have optimal activity from pH 3.5 to 4.0; mean soil pH for soil samples collected with roots was 3.69. Apparent energy of activation and Q₁₀ values (14.0 kcal mol⁻¹ and 2.2, respectively) do not provide evidence for temperature acclimation, but substantial phosphatase activity was measured at 1°C. Kinetic parameters determined for this root surface phosphatase were as follows: Km=9.23 mM, Vmax=1.61×10⁻³ μmoles mm⁻²h⁻¹. The presence of inorganic phosphorus in the assay medium did not inhibit root surface phosphatase activity except at very high concentrations (100 mM); even then, only slight inhibition was detected (7 to 19%). A comparison of hydrolysis rates with inorganic phosphate assimilation rates measured forE. vaginatum indicates that organic phosphate hydrolysis may occur at approximately one third the rate of inorganic phosphate absorption. Calculations show that inorganic phosphate produced by root surface phosphatase activity may satisfy 65% of the annual phosphate demand ofE. vaginatum. Since arctic tundra soils are typically higher in dissolved organic phosphorus compounds than in inorganic phosphate, root surface phosphatase activity may make a considerable contribution to the phosphate nutrition of this widespread and abundant arctic plant.     

The unique root-nodule symbiosis between Rhizobium and the aquatic legume,Neptunia natans (L. f.) Druce

We examined the development of the aquatic N₂-fixing symbiosis between Rhizobium sp. (itNeptunia) and roots of Neptunia natans L. f. (Druce) (previously N. oleracea Lour.) under natural and laboratory conditions. When grown in its native marsh habitat, this unusual aquatic legume does not develop root hairs, the primary sites of rhizobial infection for most temperate legumes. Under natural conditions, the aquatic plant floats and develops nitrogen-fixing nodules at emergence of lateral roots on the primary root and on adventitious roots at stem nodes, but not from the stem itself. Cytological studies using various microscopies revealed that the mode of root infection involved an intercellular route of entry followed by an intracellular route of dissemination within nodule cells. After colonizing the root surface, the bacteria entered the primary root cortex through natural wounds caused by splitting of the epidermis and emergence of young lateral roots, and then stimulated early development of nodules at the base of such roots. The bacteria entered the nodule through pockets between separated host cells, then spread deeper in the nodule through a narrower intercellular route, and eventually evoked the formation of infection threads that penetrated host cells and spread throughout the nodule tissue. Bacteria were released from infection droplets at unwalled ends of infection threads, became enveloped by peribacteroid membranes, and transformed into enlarged bacteroids within symbiosomes. In older nodules, the bacteria within symbiosomes were embedded in an unusual, extensive fibrillar matrix. Cross-inoculation tests of 18 isolates of rhizobia from nodules of N. natans revealed a host specificity enabling effective nodulation of this aquatic legume, with lesser affinity for Medicago sativa and Ornithopus sp., and an inability to nodulate several other crop legume species. Acetylene reduction (N₂ fixation) activity was detected in nodules of N. natans growing in aquatic habitats under natural conditions in Southern India. These studies indicate that a specific group of Rhizobium sp. (Neptunia) occupies a unique ecological niche in aquatic environments by entering into a N₂-fixing root-nodule symbiosis with Neptunia natans.We thank J. Whallon for technical assistance, G. Truchet, J. Vasse, S. Wagener, J. Beaman, F. DeBruijn, F. Ewers, and A. Squartini for helpful comments, and N.N. Prasad and G. Birla for assistance in conducting field observations. This work was supported by the Michigan Agricultural Experiment Station and National Science Foundation grants DIR-8809640 and BIR-9120006 awarded to the MSU Center for Microbial Ecology. This study is dedicated to the memory of Dr. Joseph C. Burton, a friend and colleague who made many contributions to the study of the Rhizobiumlegume symbiosis.     

Phosphorus dynamics and bioavailability in sediments of the Penzé Estuary (NW France): in relation to annual P-fluxes and occurrences of Alexandrium Minutum

The macrotidal estuary of Penzé (Brittany, Western part of the Channel, France) has been subjected to recurrent annual toxic blooms of Alexandrium minutum since 1988. This study aims to specify the phosphorus dynamics and bioavailability in sediments in order to improve our understanding of Alexandrium occurrences. Sediment-P pools and diffusive phosphate fluxes were studied under similar hydrodynamic conditions, in the intermediate estuary in May, June and July 2003 and along the salinity gradient from August 2004 to June 2005. The results highlight a decrease in bioavailable phosphorus (iron and organic bound) from the inner part of the estuary seaward. The ratio of iron-bound phosphorus to iron-oxyhydroxides is lower in the inner and intermediate estuaries (5–8) than in the outer site (15), suggesting a saturation of sorption sites and greater phosphorus bioavailability in this area. Pools of bioavailable phosphorus in surficial sediments are about eight times higher than the annual net-export of P (7 ton year⁻¹). Phosphate releases from sediments are always lower than 5 μmol m⁻² d⁻¹ in March. The highest supplies occur in June and August in the intermediate area (up to 400 μmol m⁻² d⁻¹) where they represent up to 50% of river loadings. These results further suggest that phosphate pulses coincide with occurrences of Alexandrium reported in June.     

Characterization of phosphobacteria isolated from eutrophic aquatic ecosystems

Phosphobacteria are able to enhance phosphorus availability in soil and improve crop yields. To develop such biofertilizers, 14 predominant phosphobacteria were isolated from eutrophic aquatic ecosystems. Molecular identification and phylogenetic analysis revealed three groups among the nine isolates of inorganic phosphate-solubilizing bacteria (IPSB): IPSBl and IPSB2 belonged to the actinobacteria and flavobacteria, respectively, and the other seven belonged to the γ-proteobacteria. Among five isolates of organic phosphorus-mineralizing bacteria (OPMB), two groups were present: OPMB1 and OPMB3 belonged to the β-proteobacteria, while the other three belonged to the γ-proteobacteria. The IPSB isolates released 62.8–66.7 mg P 1⁻¹ from tricalcium phosphate under shaking conditions, and 26.8 to 43.7 mg P 1⁻¹ under static conditions; the OPMB strains released 23.5–30.2 mg P 1⁻¹ from lecithin under shaking conditions, and 16.7–27.6 mg P 1⁻¹ under static conditions. To the best of our knowledge, this is the first report indicating that IPSBl (designated Aureobacterium resistents) as a tricalcium phosphate-solubilizing bacterium and OPMB1 and OPMB3 (designated Acidovorax temperans and Achromobacter xylosoxidans, respectively) are lecithin-mineralizing bacteria. This investigation demonstrated that a eutrophic aquatic ecosystem is a selective source of phosphobacteria and the screened phosphobacteria are a potential alternative to the development of biofertilizers.     

Phosphate solubilization activity of rhizobia native to Iranian soils

Agricultural soils in Iran are predominantly calcareous with very low plant available phosphorus (P) content. In addition to their beneficial N₂-fixing activity with legumes, rhizobia can improve plant P nutrition by mobilizing inorganic and organic P. Isolates from different cross-inoculation groups of rhizobia, obtained from Iranian soils were tested for their ability to dissolve inorganic and organic phosphate. From a total of 446 rhizobial isolates tested for P solubilization by the formation of visible dissolution halos on agar plates, 198 (44%) and 341(76%) of the isolates, solubilized Ca₃(PO₄)₂ (TCP) and inositol hexaphosphate (IHP), respectively. In the liquid Sperber TCP medium, phosphate-solubilizing bacteria (Bacillus sp. and Pseudomonas fluorescens) used as positive controls released an average of 268.6 mg L⁻¹ of P after 360 h incubation. This amount was significantly (P < 0.05) higher than those observed with all rhizobia tested. The group of Rhizobium leguminosarum bv. viciae mobilized in liquid TCP Sperber medium significantly (P < 0.05) more P (197.1 mg L⁻¹ in 360 h) than other rhizobia tested,. This group also showed the highest dissolution halo on the TCP solid Sperber medium. The release of soluble P was significantly correlated with a drop in the pH of the culture filtrates indicating the importance of acid production in the mobilization process. None of the 70 bradyrhizobial isolates tested was able to solubilize TCP. These results indicate that many rhizobia isolated from soils in Iran are able to mobilize P from organic and inorganic sources and this beneficial effect should be tested with crops grown in Iran.     

<Emphasis Type="Italic">Spartina maritima</Emphasis> influence on the dynamics of the phosphorus sedimentary cycle in a warm temperate estuary (Mondego estuary,Portugal)

During the last decades the Mondego estuary has been under severe ecological stress mainly caused by eutrophication. In this salt march system, Spartina maritima covers about 10.5 ha of the intertidal areas. The objective of the present study was to evaluate the effect of Spartina maritima marshes on the dynamics of phosphorus (P) binding in the surface sediment. We compare phosphate and oxygen fluxes, P-adsorption capacity, phosphate concentrations and total amount, and the extractable P forms in the upper 20 cm of sediment in vegetated sediment with adjacent mudflats without vegetation. Sediment pore-water profiles followed a clear trend, with lower P concentrations in more superficial layers, and increasing with depth. The vegetated mudflats presented lower concentrations of dissolved inorganic phosphorus than adjacent bare bottom mudflats, lower phosphate total amount, as well as higher P-adsorption capacity. Results from the extraction procedure show that the superficial layers are the most important for estuarine phosphorus dynamics, since maximum concentrations of labile P pools are present here. In contrast, higher proportions of refractory P pool are found in deeper layers. Spartina marsh sediments had less total P, less iron bound P, and less exchangeable P than adjacent bare bottom mudflats. Also the pool of loosely sorbed P is lower in the Spartina marsh. Phosphate regeneration from the sediment to the overlying water was only 11.8 kg ha⁻¹ year⁻¹ in vegetated sediment while 25.8 kg ha⁻¹ year⁻¹ in the bare mud flat. Plant uptake for growth combined with an enhanced P-adsorption capacity of the sediment, may explain these differences. Therefore, Spartina marshes are very important agents in the sedimentary P cycle worldwide, and can be considered a useful management tool in estuarine ecosystem recovery efforts.     

Influence of drought on competition between selected Rhizobium meliloti strains and naturalized soil rhizobia in alfalfa

Drought is an important environmental factor that can affect rhizobial competition and N₂ fixation. Three alfalfa (Medicago sativa L. and M. falcata L.) accessions were grown in pots containing soil from an irrigated (Soil 1) and a dryland (Soil 2) alfalfa field in northern Utah, USA. Mutants of three strains of Rhizobium meliloti Dang. from Pakistan (UL 136, UL 210, and UL 222) and a commercial rhizobial strain 102F51a were developed with various levels of resistance to streptomycin. Seeds inoculated with these individual streptomycin-resistant mutants were sown in the two soils containing naturalized rhizobial populations. Soils in the pots were maintained at −0.03, −0.5, and −1.0 MPa. After 10 weeks, plants were harvested and nodule isolates were cultured on agar medium with and without streptomycin to determine nodule occupancy (proportion of the nodules occupied by introduced rhizobial strains). Number of nodules, nodule occupancy, total plant dry weight, and shoot N were higher for Soil 1 than Soil 2. Number of nodules, plant dry weight, and shoot N decreased as drought increased from −0.03 to −1.0 MPa in the three alfalfa accessions. Rhizobial strains UL 136 and UL 222 were competitive with naturalized alfalfa rhizobia and were effective at symbiotic N₂ fixation under drought. These results suggest that nodulation, growth, and N₂ fixation in alfalfa can be improved by inoculation with competitive and drought-tolerant rhizobia and may be one economically feasible way to increase alfalfa production in water-limited environments. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931. Joint contribution from USDA-ARS and the Utah Agric. Exp. Sta., Utah State Univ., Logan, UT 84322-4810, USA. Journal Paper No. 4931.     

Inoculation of forage grasses with N2-fixing Enterobacteriaceae

Summary Previous investigations indicated some forage grass roots in Texas are heavily colonized with N₂-fixing bacteria. The most numerous N₂-fixing bacteria were in the genera Klebsiella and Enterobacter. In the present investigation inoculation experiments were conducted using 18 isolates of these bacteria to determine if a N₂-fixing association could be established between the bacteria and the grassesCynodon dactylon andPanicum coloratum. Plants were grown in soil for approximately 5 months in a greenhouse and were measured periodically for dry matter, nitrogen accumulation, and acetylene reduction activity. Results of the investigation indicated that 25% of the plant-soil systems were active in acetylene reduction and the activity was high enough to indicate agronomically significant quantities of N₂ were being fixed (>8kg N ha⁻¹). However, plant systems extrapolated to fix>8 kg N ha⁻¹ contained less nitrogen and accumulated less dry matter than plants less active in acetylene reduction. Inocula could not be re-isolated from healthy grass roots indicating that the N₂-fixing activity may have not have been closely assiciated with plant roots. Future research is needed to determine factors limiting colonization of grass roots.     

Different nutrient use strategies of expansive grasses <Emphasis Type="Italic">Calamagrostis epigejos</Emphasis> and <Emphasis Type="Italic">Arrhenatherum elatius</Emphasis>

Enhanced nitrogen (N) levels accelerate expansion of Calamagrostis epigejos and Arrhenatherum elatius, highly aggressive expanders displacing original dry acidophilous grassland vegetation in the Podyjí National Park (Czech Republic). We compared the capability of Calamagrostis and Arrhenatherum under control and N enhanced treatments to (i) accumulate N and phosphorus (P) in plant tissues, (ii) remove N and P from above-ground biomass during senescence and (iii) release N and P from plant material during decomposition of fresh formed litter. In control treatment, significantly higher amounts of total biomass and fresh aboveground litter were observed in Calamagrostis than in Arrhenatherum. Contrariwise, nutrient concentrations were significantly higher (11.6–14.3 mg N g⁻¹ and 2.3 mg P g⁻¹) in Arrhenatherum peak aboveground biomass than in Calamagrostis (8.4–10.3 mg N g⁻¹ and 1.6–1.7 mg P g⁻¹). Substantial differences between species were found in resorption of nutrients, mainly P, at the ends of growing seasons. While P concentrations in Arrhenatherum fresh litter were twice and three times higher (1.6–2.5 mg P g⁻¹) than in Calamagrostis (0.7–0.8 mg P g⁻¹), N concentrations were nearly doubled in Arrhenatherum (13.1–15.6 mg N g⁻¹) in comparison with Calamagrostis (7.4–8.7 mg N g⁻¹). Thus, the nutrients (N and mainly P) were retranslocated from the aboveground biomass of Calamagrostis probably more effectively in comparison with Arrhenatherum at the end of the growing season. On the other hand, Arrhenatherum litter was decomposed faster and consequently nutrient release (mainly N and P) was higher in comparison with Calamagrostis which pointed to different growth and nutrient use strategies of studied grass species.     

Greater nitrogen and/or phosphorus availability increase plant species’ cover and diversity at a High Arctic polar semidesert

Enhanced nitrogen (N) deposition at high latitudes is a circumpolar phenomenon. Low soil phosphorus (P), however, may limit vegetation responses to increased N inputs. From 2000 to 2002, the effects of N at 0, 0.5 (a rate occurring in Greenland and Iceland) and 5 (equivalent to deposition in areas of Europe) g N m⁻² a⁻¹ and P (0.1 g m⁻² a⁻¹) treatments on plant species’ cover and diversity were determined at a polar semidesert site (ambient deposition c 0.1 g N m⁻² a⁻¹) in Svalbard (79°N). The largest response was to combined 5 g N plus 1 g P m⁻² a⁻¹, where cover of Saxifraga oppositifolia increased c fourfold, density of Salix polaris leaves c ninefold, seedlings of several ‘new’ species (Draba oxycarpa, Saxifraga caespitosa, Sagina nivalis) were established and ‘immigration’ of Bryum arcticum and ‘extinction’ of Schistidium apocarpum were observed. There were fewer, less pronounced, effects on the plant community at 0.5 g N m⁻² a⁻¹. Low P availability did indeed appear to restrict vegetation response to N. There was a trend for plant species’ richness and diversity to increase with 1 g P m⁻² a⁻¹ at 0 and 0.5 g N m⁻² a⁻¹, but not at 5 g N m⁻². Plant species showed individualistic responses so that generalisation by functional type was not possible. Such increased colonisation by moss species of bare soil, and greater densities of previously unrecorded angiosperm seedlings, are not usually observed in more closed (subarctic) tundra as a response to N and P additions. These changes are likely to influence significantly nutrient cycles, whole system carbon budgets and surface energy and water balances. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Seasonal variation in the nitrogenase activity of aPanicum maximum var.trichoglume pasture and identification of associated bacteria

Summary The influence of seasonal variation on nitrogenase (N₂-ase) activity of undisturbed soil-plant cores ofPanicum maximum var.trichoglume was measured using the C₂H₂ reduction assay. The largest N₂-ase activity in the field, 14.7 g N ha⁻¹ day⁻¹, occurred in spring when soil moisture was high, soil temperature was low and nitrogenous fertiliser influence was at a minimum. The potential N₂-ase activity of the cores, measured under controlled conditions, reached a maximum of 27.2 g N ha⁻¹ day⁻¹ and averaged 26.3 g N ha⁻¹ day⁻¹ over the 14 month sampling period. N₂-ase activity was positively correlated (P=0.05) with field soil moisture and negatively correlated with field soil temperature (r=0.59 and −0.78 respectively). Multiple regression showed that 69% of the variation of N₂-ase activity in the field was associated with the combined effects of soil moisture and soil temperature. Nitrogen fixing bacteria were isolated from the roots ofP. maximum and based upon morphology, biochemical tests and fluorescent antibody reaction, were found to be closely related toAzospirillum lipoferum.