Symbiovars in rhizobia reflect bacterial adaptation to legumes

Legume specificity is encoded in rhizobial genetic elements that may be transferred among species and genera. Dissemination (by lateral transfer) of gene assemblies dictating host range accounts for the existence of the same biological variant (biovar) in distinct microbiological species. Different alternative biovars may exist in a single species expanding their adaptation to different niches (legume nodules). A review of all reported biovars is presented. Instead of the term biovar, symbiotic variant (symbiovar) is proposed as a parallel term to pathovar in pathogenic bacteria. Symbiovars should be determined based on the symbiotic capabilities in plant hosts, distinguished by the differences in host range and supported by symbiotic gene sequence information.     

Effect ofAzospirillum inoculation on nitrogen fixation and growth of several winter legumes

Summary Inoculation of naturally nodulatedPisum sativum L. (garden pea) withAzospirillum in the greenhouse caused a significant increase in nodule numbers above controls. Field inoculation of garden peas in the winter 1981–1982 andCicer arietinum L. (chick pea), in winter 1982–1983, withAzospirillum one week after plant emergence, produced a significant increase in seed yield, but did not affect plant dry matter yield. ForVicia sativa L. (vetch) grown in soil in the greenhouse and in the field for forage, winter 1980–1981, inoculation significantly increased dry matter yield, %N, N-content, and acetylene reduction (nitrogen fixation) activity. InHedysarum coronarium L. (sulla clover), winter 1981–1982, inoculated with both its specificRhizobium (by the slurry method) andAzospirillum, 7 days after emergence, there was an increase in acetylene reduction above controls inoculated withRhizobium alone. These results suggest that it is possible, under conditions tested in this work, to increase nodulation, nitrogen fixation, and crop yields of winter legumes by inoculation withAzospirillum.     

Nitrate effects on the nodulation of legumes inoculated with nitrate-reductase-deficient mutants of Rhizobium

The effect of nitrate on the symbiotic properties of nitrate-reductase-deficient mutants of a strain of cowpea rhizobia (32H1), and of a strain of Rhizobium trifolii (TA1), were examined; the host species were Macroptilium atropurpureum (DC.) Urb. and Trifolium subterraneum L. Nitrate retarded initial nodulation by the mutant strains to an extent similar to that found with the parent strains. It is therefore unlikely that nitrite produced from nitrate by the rhizobia, plays a significant role in the inhibition of nodulation by nitrate. Nitrite is an inhibitor of nitrogenase, and its possible production in the nodule tissue by the action of nitrate reductase could be responsible for the observed inhibition of nitrogen fixation when nodulated plants are exposed to nitrate. However, the results of this investigation show that nitrogen fixation by the plants nodulated by parent or mutant strains was depressed by similar amounts in the presence of nitrate. No nitrite was detected in the nodules. Nodule growth, and to a lesser extent, the nitrogenase specific activity of the nodules (mol C₂H₄g^–1 nodule fr. wt. h^–1), were both affected by the added nitrate.     

Nodulation studies on legumes exotic to Australia: Hedysarum coronarium

Abstract Symbiotic experiments in glasshouse, controlled environment cabinet, and field were conducted with four lines of sulla ( Hedysarum coronarium ) and 15 strains of Rhizobium spp. This plant is highly Rhizobium -specific and appropriate strains are most unlikely to occur naturally in Australia. Under several sets of experimental conditions, H. coronarium nodulated abundantly and effectively with homologous rhizobia introduced from Spain and Italy. The optimum temperature for nitrogen fixation was relatively low (approx. 21°C) but significant interactions between line of host, strain of rhizobia, and growth temperature were frequent. The rhizobia were persistent in soil.     

Salinity tolerance of the tree legumes: Mesquite (Prosopis glandulosa var.torreyana,P. velutina andP. articulata) Algarrobo (P. chilensis), Kiawe (P. pallida) and Tamarugo (P. tamarugo) grown in sand culture on nitrogen-free media

Summary Sand culture pot experiments were carried out with Proposis seedlings in the greenhouse on a nitrogen free nutrient solution with increasing levels of sodium chloride. All species tolerated a 6,000 mg/l salinity with no reduction in growth.P. velutina was the only species that poorly tolerated the 12,000 mg/l salinity level.P. articulata, P. pallida, andP. tamarugo tolerated 18,000 mg/l NaCl with little reduction in growth and grew slightly in a salinity (36,000 mg/l NaCl) greater than seawater. This is the first legume known to grown in salinities equivalent to seawater.     

The growth and nitrogen uptake of Rhodes grass grown on soils with varying histories of legume cropping

Summary Rhodes grass (Chloris gayana) was grown under glasshouse conditions on soils that had previously grown from 1 to 6 soybean (Glycine max) or Siratro (Macroptillium atropurpureum) crops. Soil mineral N contents at sowing were higher in Siratro-cropped than soybean-cropped soils and increased with cropping history.Yields of Rhodes grass at the first harvest (14 weeks) were related to soil mineral N levels at sowing by the relationship Y=138.7 (1–0.917e^–0.012x). Grass grown on all soybean soils was responsive to N at both harvests (14 and 28 weeks). Grass grown on soil that had grown three or more Siratro crops was non-responsive to N at the first harvest but responses to N were recorded on all Siratro soils at the second harvest.The amount of N removed by the grass crops was small in relation to the total amount present in the soil system. This resulted in no change in soil total N levels over the two crop periods.These results highlight the need to study N dynamics in crop systems rather than continue to measure N pool sizes when evaluating the contribution of biologically fixed N to the nutrition of subsequent non-legume crops.     

Soil nutrient status of KwaZulu–Natal savanna and grassland biomes causes variation in cytokinin functional groups and their levels in above-ground and underground parts of three legumes

Cytokinins (CKs) are involved in several developmental stages in the life-cycle of plants. The CK content in plants and their respective organs are susceptible to changes under different environmental conditions. In the current study, we profiled the CK content in the above and underground organs of three legumes (Lessertia frutescens, Mucuna pruriens and Pisum sativum) grown in soils collected from four locations (Ashburton, Bergville, Hluhluwe and Izingolweni) in KwaZulu-Natal province, South Africa. The quantified CK contents in the three legumes were categorized on the basis of their side chains (isoprenoid, aromatic and furfural) and modifications (e.g. free bases and glucosides). Legume and soil types as well as their interaction significantly influenced the concentrations of CKs. Lessertia frutescens, Mucuna pruriens and Pisum sativum had CK content that ranged from 124–653, 170–670 and 69–595 pmol/g DW, respectively. Substantial quantity (> 600 pmol/g DW) of CK were observed in plants grown in Bergville (above-ground part of Lessertia frutescens) and Izingolweni (underground part of Mucuna pruriens) soils. A total of 28 CK derivatives observed in the legumes comprised of isoprenoid (22), aromatic (5) and furfural (1) side-chain CKs. However, the 16 CK derivatives in Mucuna pruriens were isoprenoid-type based on the side-chain. Generally, a higher ratio of cis-zeatin (cZ) relative to the trans-zeatin (tZ) was evident in the above-ground part of Lessertia frutescens and Pisum sativum for the four soil treatments. In terms of functional and physiological importance of the CKs, the free bases (active form) and ribosides (translocation form) were the most abundant CK in Lessertia frutescens and Pisum sativum. However, N-glucoside, a deactivation/detoxicification product was the most dominant CK in Mucuna pruriens from Hluhluwe and Izingolweni soils. The total CKs in the underground parts of the legumes had a positive significant correlation with the total phosphorus and nitrogen content in the plant as well as the soil nitrogen. Overall, the CK profiles of the legumes were strongly influenced by the soil types.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01021-2.     

Rock-phosphate mobilization induced by the alkaline uptake pattern of legumes utilizing symbiotically fixed nitrogen

Summary Soybean and alfalfa were grown on sand and soil to which P was added in the form of finely ground rock phosphates. When the legumes depended on NO₃ as N source, more anionic than cationic nutrients were absorbed. This resulted in a pH increase in the growth medium and in very low availability of P added as rock phosphate. When, however, the legumes made use of symbiotically fixed N, more cationic than anionic nutrients were absorbed leading to an acidification of the growth medium and an ensuing mobilization and higher availability of the rock phosphates.Symbiotic N fixation which initiates the chain of reactions leading to an increased availability of rock phosphate-P is dependent on photosynthate supply and on the availability of phosphate. Therefore, in a separate experiment it was investigated whether a priming effect exerted by a small quantity of added easily soluble phosphate, could enhance the availability of rock phosphate-P to legumes. Results obtained indicated that easily soluble phosphate might indeed be effective in this respect.     

Protective effect of jasmonic acid and potassium against cadmium stress in peas (Pisum sativum L.)

A combination of mineral nutrients and plant growth regulators should be assessed to improve crop performance under various abiotic stresses. There is a need to include plant growth regulators in fertilization regime of various crops along with essential mineral nutrients, especially when they are irrigated with polluted water with higher levels of heavy metals. The performance of pea was evaluated under cadmium (Cd) stress coupled with potassium (K) and jasmonic acid (JA) supplementation. The Cd stress (50 μM) was applied to soil (sandy loam) grown pea plants as basal dose after a month of sowing. The control and stressed plants were then supplemented with K (5 M), JA (0.5 mM) and their collective application along with control as distilled water. Cd stress showed a marked reduction in growth pattern, however, the collective supplementation sufficiently improved the growth pattern of stressed peas plants as evidenced by improvement in shoot length (cm), root length (cm), number of leaves per plant, leaf area (cm²), plant fresh and dry weight (gm). Potassium application under Cd stress significantly enhanced internodal distance (cm) while the number of seeds per pod and relative water contents remained nonsignificant. The applied treatment (JA + K) under Cd stress prominently improved enzymatic activities, which were measured as nitrate reductase activity (NRA), nitrite reductase activity (NiRA), superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Cd stress impacted the biochemical profile by enhancing antioxidant capacity (AC), antioxidant activity (AA), total phenols (TP), while reducing total soluble protein (TSP), chlorophyll ‘a’, chlorophyll ‘b’ and carotenoids. The combined application of JA and K under Cd stress enhanced AC, AA, TP, Chl a and b, TSP and carotenoids. The results indicate that foliar application of JA and K efficiently negated the harmful effects of Cd stress on peas.     

Effect of long-term salinity on cellular antioxidants,compatible solute and fatty acid profile of Sweet Annie (Artemisia annua L.)

Impact of long-term salinity and subsequent oxidative stress was studied on cellular antioxidants, proline accumulation and lipid profile of Artemisia annua L. (Sweet Annie or Qinghao) which yields artemisinin (Qinghaosu), effective against cerebral malaria-causing strains of Plasmodium falciparum. Under salinity (0.0–160 mM NaCl), in A. annua, proline accumulation, contents of ascorbate and glutathione and activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT) increased, but the contents of reduced forms of glutathione (GSH) and ascorbate declined. The fatty-acid profiling revealed a major salinity-induced shift towards long-chain and mono-saturated fatty acids. Myristic acid (14:0), palmitoleic acid (16:1), linoleic acid (18:2) and erucic acid (22:1) increased by 141%, 186%, 34% and 908%, respectively, in comparison with the control. Contents of oleic acid (18:1), linolenic acid (18:3), arachidonic acid (22:0) and lignoceric acid (24:0) decreased by 50%, 17%, 44% and 78%, respectively. Thus, in A. annua, salinity declines ascorbate and GSH contents. However, increased levels of proline and total glutathione (GSH + GSSG), and activities of antioxidant enzymes might provide a certain level of tolerance. Modification in fatty-acid composition might be a membrane adaptation to long-term salinity and oxidative stress.     

A further extension of the method for independently measuring the amount of nitrogen fixed by a legume crop

Summary The combination of using¹⁵N for determining the amount of nitrogen fixed by a legume crop in field experiments and the labelling of only one treatment at a time in each treatment combination is shown to be conceptually and experimentally valid for determining the effect of cultural practices on the amount of nitrogen fixed by a legume crop.     

Salinity effects on osmoregulation and growth of the euryhaline flounder Paralichthys orbignyanus

The flounder, Paralichthys orbignyanus, is found in coastal and estuarine waters of the Western South Atlantic Ocean. It is being considered for aquaculture due to its high market price and wide tolerance to environmental factors such as salinity, pH, and nitrogenous compounds. The objective of this study was to characterize the ionic and osmotic regulation of P. orbignyanus over the range of its tolerated ambient salinities (0-40‰) and to evaluate the survival and growth in freshwater (0‰) and seawater (30‰) over 90 days. After 15 days of exposure to different salinities (0‰, 10‰, 20‰, 30‰ and 40‰), plasma osmolality and ionic (Na⁺, Cl⁻, K⁺ and Ca²⁺) concentrations slightly increased with salinity. The isosmotic point was estimated as 328.6 mOsm kg⁻¹ H₂O and corresponded to 10.9‰ salinity. After 90 days, survival was similar in freshwater and seawater, but osmo- and ionoregulation was significantly affected in freshwater and flounders reared in this medium showed a lower growth rate than those reared in seawater. Based on the results from this study, P. orbignyanus can be characterized as a marine/estuarine euryhaline teleost capable of hyper/hypo iono- and osmoregulation over the fluctuating salinity regime faced by this species in the environment. Furthermore, results suggest that the lower growth rate exhibited by P. orbignyanus in freshwater could be due, at least partially, to a higher energy expenditure associated to a higher branchial Na⁺, K⁺-ATPase activity in this environment.     

盐度对裸项栉鰕虎鱼繁殖和生长的影响

目的探索裸项栉鰕虎鱼繁殖和育苗的适宜盐度。方法比较不同盐度梯度条件下裸项栉鰕虎鱼的产卵率、孵化率和生长存活情况。结果裸项栉鰕虎鱼性腺成熟、产卵和孵化的适宜盐度为10‰-20‰,过低或过高盐度该鱼产卵量少,孵化率极低;适宜的盐度有利于裸项栉鰕虎鱼的生长。结论裸项栉鰕虎鱼适盐范围广,适宜的繁殖、生长盐度较低。     

Species variation in the fixation and transfer of nitrogen from legumes to associated grasses

Summary Three legume species (alfalfa, red clover, and birdsfoot trefoil) in combination with five grass species (timothy, bromegrass, red fescue, tall fescue, and orchardgrass) were used to study N transfer in mixtures, using the 15_N dilution technique. The advantage of grass-legume mixtures was apparent. Total herbage and protein yields of grasses in mixtures were higher than those alone, especially at the later cuts. This benefit of mixed cropping is mainly due to N transfer from legumes to associated grasses. N₂-fixation and N transfer by alfalfa rated highest, red clover intermediate, and birdsfoot trefoil lowest. The importance of each pathway of N transfer from legumes appeared to differ between species. Alfalfa and red clover excreted more N than trefoil, while the latter contributed more N from decomposition of dead nodule and root tissue. The greatest advantage from a grass-legume mixture, with respect to the utilization of N released from the legume, varied with early maturing tall fescue (Kentucky 31), orchardgrass (Juno), and bromegrass (Tempo), to intermediate timothy (Climax), and least with late maturing red fescue (Carlawn). Contribution no. 817 of the Ottawa Research Station.     

Salt effects on urea hydrolysis and nitrification during leaching through laboratory soil columns

Summary We studied the transport and transformation of urea under steady-state conditions in two soils and at three water salinities (1.0, 5.0, and 10.0 dS/m) using laboratory soil columns. A mathematical model that considers diffusion, convection, adsorption and first-order kinetic transformation of nitrogen was used to describe measured effluent concentration of the two nitrogen species. Increasing salt levels in the applied water decreased the hydrolysis of urea in the two soils studied with first-order rate coefficients decreasing from 0.015 to 0.009 h^–1 in the fine sandy loam, and from 0.075 to 0.015 h^–1 in a silty loam. Similarly, the nitrification rate decreased by 50% and 70% in the two soils as salinity increased. The rate coefficients measured in the leaching studies were much smaller than measured in incubation-type experiments. Calculated half-lives for urea and NH ₄ ⁺ provided a method interpreting the kinetic rate coefficients as a function of the experimental conditions.     

Molecular and functional characterization of the Azorhizobium caulinodans ORS571 hydrogenase gene cluster

In this work, we report the cloning and sequencing of the Azorhizobium caulinodans ORS571 hydrogenase gene cluster. Sequence analysis revealed the presence of 20 open reading frames hupTUVhypFhupSLCDFGHJK hypABhupRhypCDEhupE. The physical and genetic organization of A. caulinodans ORS571 hydrogenase system suggests a close relatedness to that of Rhodobacter capsulatus. In contrast to the latter species, a gene homologous to Rhizobium leguminosarum hupE was identified downstream of the hyp operon. A hupSL mutation drastically reduced the high levels of hydrogenase activity induced by the A. caulinodans ORS571 wild-type strain in symbiosis with Sesbania rostrata plants. However, no significant effects on dry weight and nitrogen content of S. rostrata plants inoculated with the hupSL mutant were observed in plant growth experiments.     

Numerical taxonomic analysis of cross-inoculation patterns of legumes and Rhizobium

Summary A survey was made of published results of tests of the capacity of Rhizobium derived from one legume genus to nodulate plants from other genera. The data were derived from more than 14,000 separate cross-inoculation trials involving species from 165 genera of legumes. Numerical taxonomic techniques were applied to 113 of the genera for which results of substantial cross-infection tests were available. The data were examined using mean character difference coefficients re-expressed as total and positive-only similarity coefficients. The resulting similarity matrices were clustered by the unweighted pair-group method using arithmetic averages. Eighteen affinity groups were defined at the 70% similarity level. With few exceptions, the physiological and cultural behavior of the rhizobia was consistent within the defined groups. Two broad categories were suggested in the numerical taxonomic analysis, and their validity is discussed in regard to the geographic, physiological and cultural characteristics of the legumes and their Rhizobium microsymbionts. The taxonomic and agronomic value of this approach and the new groupings are discussed.     

Traffic and industrial activities around Riyadh cause the accumulation of heavy metals in legumes: A case study

The objective of this study was to analyse the effect of the continuously increasing anthropogenic activities around Riyadh, Saudi Arabia on the accumulation of heavy metals in leguminous crops. This study determined whether four legume crops, Pisum sativum L., Vicia faba L., Glycine max and Vigna sinensis, could accumulate the heavy metals Cu, Mn, Pb and Zn in their leaves, pods and grains during the summer when grown under conditions with ambient air pollution from heavy traffic and industrial activities in Riyadh, Saudi Arabia. The effect of the air pollution was examined by quantifying the protein and trace element Cu, Mn, Pb and Zn concentrations in the leaves, pods and grains of the four plant species. Analysis of the results indicated that air pollution significantly increased the heavy metal concentrations in the leaves, pods and grains. Toxic concentrations of the heavy metals were found in the plants grown at L3, L4 and L5. In conclusion, the air pollution increases as the traffic, industrial activities and population density increase.     

盐度对企鹅珍珠贝足丝分泌的影响

企鹅珍珠贝（Pteria penguin）是生产附壳珍珠的大型海水经济贝类,其依靠强壮的足丝将自身固定在硬质基底上,抵抗水流的冲击和抵御被捕食等。足丝分泌和足丝的形状很容易受到环境的影响,本实验采用盐度30为低盐度组、盐度35为中盐度组和盐度40为高盐度组,研究这3种盐度对企鹅珍珠贝足丝分泌、足丝直径和足丝拉力的影响,通过单因素方差分析法（LSD法）分析这三个足丝相关指标在3种盐度组间是否存在显著性差异。结果显示,3种盐度下企鹅珍珠贝足丝附着率无显著差异,但在整个实验周期72 h内,中盐度组的足丝分泌总数为（48.7 ± 15.1）根,显著高于低盐度组的（24.7 ± 5.0）根和高盐度组的（13.3 ± 1.5）根。在实验的前6 h内,中盐度组的足丝首次附着率显著高于低盐度组和高盐度组（P < 0.05）,但在后续的12 h、18 h、30 h、42 h、54 h和66 h这6个时间点,3个盐度组的足丝首次附着率均无显著性差异。足丝直径未受盐度变化的影响,但盐度对足丝拉力具有显著影响,中盐度组的足丝拉力显著高于低盐度组和高盐度组（P < 0.05）。上述结果表明,企鹅珍珠贝为适应一定范围内盐度的改变,会在短时间内通过抑制足丝分泌来减少能量消耗,随着对环境的适应足丝分泌会恢复。盐度影响足丝分泌且对足丝拉力影响显著,但对足丝直径无明显影响。本研究可以为企鹅珍珠贝养殖及珍珠插核培育提供理论基础。