Glyphosate effects on phenolic metabolism of nodulated soybean (Glycine max L. merr.).

Glyphosate is a herbicide that blocks the shikimic acid pathway. Three Bradyrhizobium japonicum strains with different sensitivities to glyphosate were used to test the effect of this herbicide on the phenolic metabolism of nodulated soybeans and on the bacteroid nitrogenase activity. Glyphosate caused an inhibition in the bacteroid nitrogenase activity that was related with the sensitivity of the nodule-forming strains. Both leaves and nodules accumulated huge amounts of shikimate and phenolic acids (mainly protocatechuic acid), indicating that the herbicide was translocated to the nodule and disturbed phenolic metabolism. However, this accumulation was not clearly related to the sensitivity of the different strains. Bacteroids from control plants were incubated with the same concentration of shikimate, and phenolic acid accumulated in glyphosate-treated plants. Despite the high levels found in nodules, they were not responsible for the decrease of the nitrogenase activity. Glyphosate by itself caused a small inhibition of the bacteroid nitrogenase activity.     

Tolerance and accumulation of shikimic acid in response to glyphosate applications in glyphosate-resistant and nonglyphosate-resistant cotton (Gossypium hirsutum L.).

Measurement of shikimic acid accumulation in response to glyphosate inhibition of 5-enolpyruvylshikimate-3-phosphate synthase is a rapid and accurate assay to quantify glyphosate-induced damage in sensitive plants. Two methods of assaying shikimic acid, a spectrophotometric and a high-performance liquid chromatography (HPLC) method, were compared for their accuracy of recovering known amounts of shikimic acid spiked into plant samples. The HPLC method recovered essentially 100% of shikimic acid as compared with only 73% using the spectrophotometric method. Relative sensitivity to glyphosate was measured in glyphosate-resistant (GR) and non-GR cotton leaves, fruiting branches, and squares (floral buds) by assaying shikimic acid. Accumulation of shikimic acid was not observed in any tissue, either GR or non-GR, at rates of 5 mM glyphosate or less applied to leaves. All tissues of non-GR plants accumulated shikimic acid in response to glyphosate treatment; however, only fruiting branches and squares of GR plants accumulated a slight amount of shikimic acid. In non-GR cotton, fruiting branches and squares accumulated 18 and 11 times, respectively, more shikimic acid per micromolar of translocated glyphosate than leaf tissue, suggesting increased sensitivity to glyphosate of reproductive tissue over vegetative tissue. GR cotton leaves treated with 80 mM of glyphosate accumulated 57 times less shikimic acid per micromolar of translocated glyphosate than non-GR cotton but only 12.4- and 4-fold less in fruiting branches and squares, respectively. The increased sensitivity of reproductive structures to glyphosate inhibition may be due to a higher demand for shikimate pathway products and may provide an explanation for reports of fruit abortion from glyphosate-treated GR cotton.     

Simulated glyphosate drift influences nitrate assimilation and nitrogen fixation in non-glyphosate-resistant soybean

Nontarget injury from glyphosate drift is a concern among growers using non-glyphosate-resistant (non-GR) cultivars. The effects of glyphosate drift on nitrate assimilation and nitrogen fixation potential, nodule mass, and yield of non-GR soybean were assessed in a field trial at Stoneville, MS. A non-GR soybean cultivar 'Delta Pine 4748S' was treated with glyphosate at 12.5% of use rate of 0.84 kg of active ingredient/ha at 3 (V2), 6 (V7), and 8 (R2, full bloom) weeks after planting (WAP) soybean to simulate glyphosate drift. Untreated soybean was used as a control. Soybeans were sampled weekly for 2 weeks after each glyphosate treatment to assess nitrate assimilation and N2 fixation potential. Nitrate assimilation was assessed using in vivo nitrate reductase assay in leaves, stems, roots, and nodules. Nitrogen fixation potential was assessed by measuring nitrogenase activity using the acetylene reduction assay (ARA). Nitrogen content of leaves, shoots, and seed and soybean yield were also determined. In the first sampling date (4 WAP), glyphosate drift caused a significant decrease in NRA in leaves (60%), stems (77%), and nodules (50%), with no decrease in roots. At later growth stages, NRA in leaves was more sensitive to glyphosate drift than stems and roots. Nitrogenase activity was reduced 36-58% by glyphosate treatment at 3 or 6 WAP. However, glyphosate treatment at 8 WAP had no effect on nitrogenase activity. Nitrogen content was affected by glyphosate application only in shoots after the first application. No yield, seed nitrogen, protein, or oil concentration differences were detected. These results suggest that nitrate assimilation and nitrogen fixation potential were significantly reduced by glyphosate drift, with the greatest sensitivity early in vegetative growth. Soybean has the ability to recover from the physiological stress caused by glyphosate drift.     

花生主要碳代谢指标与根瘤固氮能力的关系

[目的]比较花生叶片碳代谢指标与根瘤固氮的关系,以寻找与根瘤固氮能力关系最为密切的叶片碳代谢指标,为花生育种提供可靠的判定方法.[方法]盆栽试验在山东省花生研究所莱西试验站进行,试验以19个花生品种(系)为材料,包括山东、河南、河北、四川等省份近年来审定或育成的品种(系),辅助利用15N示踪技术,测定了花生叶片叶绿素含...     

Effect of dinoseb on nitrogen fixation of red clover (Trifolium pratense)

The effect of the herbicide dinoseb (2-sec-butyl-4,6-dinitrophenol) on the symbiotic nitrogen fixation of red clover (Trifolium pratense cv. Venla) was studied in field, greenhouse and laboratory experiments. In the field, the herbicide reduced the levels of nitrogenase (C₂H₂) activity of the plants up to 18 days after treatment, but increased the yields compared to hand-weeded and unweeded controls. Rhizobium strains isolated from root nodules of plants treated with dinoseb showed resistance to the herbicide when grown on laboratory media containing dinoseb. However, in a pot experiment, inoculation with a strain which grew in the presence of 200 μg dinoseb ml^?1 did not improve the performance of red clover treated with dinoseb. In the pot experiment dinoseb inhibited nitrogenase activity when sprayed on the leaves, but not when added to the growth medium. Application of starter nitrogen did not influence the effect of dinoseb. The results suggest that dinoseb did not act directly on the nodules, but affected nitrogen fixation by damaging the photosynthetic parts of the plant.     

Effects of changes in applied nitrogen concentrations on nodulation,nitrogen fixation and nitrogen accumulation during the soybean growth period

ABSTRACT

The specific mechanism by which nitrogen application affects nodulation and nitrogen fixation in legume crops remains uncertain. To further study the effects of nitrogen application on soybean nodulation and nitrogen accumulation, three consecutive tests were performed during the VC-V4, V4-R1 (10 days), and R1-R2 (10 days) growth periods of soybean. In a dual-root soybean system, seedlings on one side were watered with a nutrient solution containing NH₄⁺ or NO₃^? as the N source (N+ side), and those on the other side were watered with a nitrogen-free nutrient solution (N- side). During the VC-R2 period, on the N+ side, high nitrogen treatment inhibited nodule growth and nitrogenase activity (EC 1.18.6.1), and the inhibition was significantly increased with increasing high nitrogen supply time (10 days, 20 days). When the high nitrogen treatment time reached 20 days, the specific nitrogenase activity (C₂H₄ μmol^?1 g^?1 nodule dry mass h^?1) was similar to that in the low nitrogen treatment, indicating that the nitrogen fixation capacity per gram of dry mass nodules was almost the same. Therefore, it is assumed that long-term high nitrogen treatment mainly reduces nitrogen fixation by reducing the nodule number. The effect of nitrogen concentration on the roots on the N+ side was greater than that on the N- side. Taken together, these results indicate that nitrogen application affects a contact-dependent local inhibition of root nodule growth, nitrogenase activity, and nitrogen accumulation. The whole plant systematically regulates specific nitrogenase activity, and high nitrogen inhibition is recoverable.     

Nitrogenase divarication with histological attributes distinctive for glycosylated aerial stem nodules and nitrosylated root nodules of sesbania

Exceptional symbiotic nitrogen fixation with Sesbania has provided high soil fertility for many past centuries of paddy rice production. Unique stem nodulation results in high nitrogenase activity levels of S. rostrata, Brem, during rapid growth in continuously flooded rice fields that greatly disfavor legume root nodulation and this functional development. The objective of this study was to determine plant nutrient interactions that influence contrasting root and aerial stem nodule histology governing effective nitrogenase activity levels and nitrogen fixation. Top growth, nodulation, and nitrogenase activity levels were significantly increased with increased available soil P. Response to K levels and Ca additions resulted only when soil P was adequate in all treatment combinations. However, there was no significant correlation between fresh nodule weight, nitrogenase activity, and nodules plant^‐1 for both root nodules and aerial stem nodules. Nodule histology was highly contrastive with nodule type and Rhizobium morphology, cytosol composition, and governing enzyme activity levels. Distinctive nonpleomorphic cocci bacteroids of functional aerial stem nodules have tentative designation as Azorhizobium caulinodans gen. nov. sp. nov.     

A novel approach to determine the glyphosate tolerant trait in soybeans

The ability of soybean breeders to accurately, economically, and rapidly determine the transfer of the CP4 gene, the gene which confers soybean tolerance to the herbicide glyphosate, to elite soybean lines is essential to development of new glyphosate tolerant soybean (GTS) cultivars. This research focused on a simple greenhouse screening procedure to replace large, costly, and laborious field screening. Non-GTS seed was determined to be susceptible to soaking in a 1% glyphosate solution for 4 h. This process is quicker, more efficient, and as reliable as field screening for determination of glyphosate susceptibility in soybean seed. Furthermore, this research clearly demonstrates that the metabolic pathway of glyphosate activity, the shikimate acid pathway, is active, and the target enzyme of glyphosate, 5-enol-pyruvyl-shikimate-3-phosphate synthase, is present during seed germination.     

Isoflavone,glyphosate, and aminomethylphosphonic acid levels in seeds of glyphosate-treated,glyphosate-resistant soybean

The estrogenic isoflavones of soybeans and their glycosides are products of the shikimate pathway, the target pathway of glyphosate. This study tested the hypothesis that nonphytotoxic levels of glyphosate and other herbicides known to affect phenolic compound biosynthesis might influence levels of these nutraceutical compounds in glyphosate-resistant soybeans. The effects of glyphosate and other herbicides were determined on estrogenic isoflavones and shikimate in glyphosate-resistant soybeans from identical experiments conducted on different cultivars in Mississippi and Missouri. Four commonly used herbicide treatments were compared to a hand-weeded control. The herbicide treatments were (1) glyphosate at 1260 g/ha at 3 weeks after planting (WAP), followed by glyphosate at 840 g/ha at 6 WAP; (2) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied preemergence (PRE), followed by glyphosate at 1260 g/ha at 6 WAP; (3) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by glyphosate at 1260 g/ha at full bloom; and (4) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by acifluorfen at 280 g/ha plus bentazon at 560 g/ha plus clethodim at 140 g/ha at 6 WAP. Soybeans were harvested at maturity, and seeds were analyzed for daidzein, daidzin, genistein, genistin, glycitin, glycitein, shikimate, glyphosate, and the glyphosate degradation product, aminomethylphosphonic acid (AMPA). There were no remarkable effects of any treatment on the contents of any of the biosynthetic compounds in soybean seed from either test site, indicating that early and later season applications of glyphosate have no effects on phytoestrogen levels in glyphosate-resistant soybeans. Glyphosate and AMPA residues were higher in seeds from treatment 3 than from the other two treatments in which glyphosate was used earlier. Intermediate levels were found in treatments 1 and 2. Low levels of glyphosate and AMPA were found in treatment 4 and a hand-weeded control, apparently due to herbicide drift.     

Nitrogen metabolism and seed composition as influenced by glyphosate application in glyphosate-resistant soybean

Previous research has demonstrated that glyphosate can affect nitrogen fixation or nitrogen assimilation in soybean. This 2-year field study investigated the effects of glyphosate application of 1.12 and 3.36 kg of ae ha(-1) on nitrogen metabolism and seed composition in glyphosate-resistant (GR) soybean. There was no effect of glyphosate application on nitrogen fixation as measured by acetylene reduction assay, soybean yield, or seed nitrogen content. However, there were significant effects of glyphosate application on nitrogen assimilation, as measured by in vivo nitrate reductase activity (NRA) in leaves, roots, and nodules, especially at high rate. Transiently lower leaf nitrogen or (15)N natural abundance in high glyphosate application soybean supports the inhibition of NRA. With the higher glyphosate application level protein was significantly higher (10.3%) in treated soybean compared to untreated soybean. Inversely, total oil and linolenic acid were lowest at the high glyphosate application rate, but oleic acid was greatest (22%) in treated soybean. These results suggest that nitrate assimilation in GR soybean was more affected than nitrogen fixation by glyphosate application and that glyphosate application may alter nitrogen and carbon metabolism.     

Effects of salinity and nitrogen source on growth and nitrogen fixation in alfalfa

The Interaction between the effects of nitrate (NO₃) and sodium chloride (NaCl) concentration on growth) water relations, nitrogen (N) contents and N fixation were investigated in alfalfa (Medicago sativa L. cv. Magali). The plants were grown hydroponically in a growth chamber, in the presence or absence of 3 mM potassium nitrate (KNO₃) and exposed to various concentrations of NaCl. Increased salinity resulted in a significant decrease in shoot and root biomass, relative water content and water potential. Shoot growth was more inhibited by NaCl than root biomass. The plants grown in the presence of NO₃ were slightly less affected by NaCl than the plants dependent on N fixation for their N nutrition. Nitrogenase activity measured by acetylene reduction activity was substantially inhibited by NaCl, and this inhibition was significantly correlated to the inhibition of shoot growth and total N contents. The comparison of the curves of ARA response to oxygen (O₂) partial pressure showed that the salt‐induced inhibition of nitrogenase activity was associated with a significant increase in the critical O₂ pressure of the nodules exposed to NaCl. This result shows that NaCl decreases the nodule permeability to O₂ diffusion in undeterminate nodule of alfalfa, like previously shown with determinate nodules of soybean.     

Phosphorus‐deficiency effects on response of symbiotic N2 fixation and carbohydrate status in soybean to atmospheric CO2 enrichment

The impact of phosphorus (P) deficiency on response of symbiotic N₂ fixation and carbohydrate accumulation in soybean (Glycine max [L.] Merr.) to atmospheric CO₂ enrichment was examined. Plants inoculated with Bradyrhizobium japonicum MN 110 were grown in growth chambers with controlled atmospheres of 400 and 800 μL CO₂ L^‐1 and supplied either 1.0 mM‐P (P‐sufficient) or 0.05 mM‐P (P‐deficient) nitrogen (N)‐free nutrient solution. When plants were supplied with sufficient P, CO₂ enrichment significantly increased whole plant dry mass (83%), nodule mass (67%), total nitrogenase activity (58%), and N (35%) and P (47%) accumulation at 35 days after transplanting (DAT). Under sufficient P supply, CO₂ enrichment significantly increased starch concentrations in nodules compared to the normal atmospheric CO₂ treatment. Under normal CO₂ levels (400 μL L^‐1) nonstructural carbohydrate concentration (starch plus soluble sugar) was significantly higher in leaves of P‐deficient plants than in leaves of P‐sufficient plants in which nonstructural carbohydrate concentration exhibited a strong diurnal pattern. Under deficient P supply whole plant dry mass, symbiotic N₂‐fixation parameters, and N and P accumulation were not enhanced by atmospheric CO₂ enrichment. Phosphorus deficiency decreased nonstructural carbohydrate accumulation in nodules at the end of a 10‐day period in which functional activity was developing by 86% relative to P‐sufficient controls. While P deficiency elicited significant increases in the nonstructural carbohydrate concentration in leaves, it caused significant decreases in the nonstructural carbohydrate concentration in nodules over the diurnal cycle from 30 to 31 DAT. Collectively, these results indicate that the lack of a symbiotic N₂‐fixation response to atmospheric CO₂ enrichment by P‐deficient plants may be related to the decreased carbohydrate status of nodules.     

Evaluation of spectrophotometric and HPLC methods for shikimic acid determination in plants: models in glyphosate-resistant and -susceptible crops

Endogenous shikimic acid determinations are routinely used to assess the efficacy of glyphosate in plants. Numerous analytical methods exist in the public domain for the detection of shikimic acid, yet the most commonly cited comprise spectrophotometric and high-pressure liquid chromatography (HPLC) methods. This paper compares an HPLC and two spectrophotometric methods (Spec 1 and Spec 2) and assesses the effectiveness in the detection of shikimic acid in the tissues of glyphosate-treated plants. Furthermore, the study evaluates the versatility of two acid-based shikimic acid extraction methods and assesses the longevity of plant extract samples under different storage conditions. Finally, Spec 1 and Spec 2 are further characterized with respect to (1) the capacity to discern between shikimic acid and chemically related alicyclic hydroxy acids, (2) the stability of the chromophore (t1/2), (3) the detection limits, and (4) the cost and simplicity of undertaking the analytical procedure. Overall, spectrophotometric methods were more cost-effective and simpler to execute yet provided a narrower detection limit compared to HPLC. All three methods were specific to shikimic acid and detected the compound in the tissues of glyphosate-susceptible crops, increasing exponentially in concentration within 24 h of glyphosate application and plateauing at approximately 72 h. Spec 1 estimated more shikimic acid in identical plant extract samples compared to Spec 2 and, likewise, HPLC detection was more effective than spectrophotometric determinations. Given the unprecedented global adoption of glyphosate-resistant crops and concomitant use of glyphosate, an effective and accurate assessment of glyphosate efficacy is important. Endogenous shikimic acid determinations are instrumental in corroborating the efficacy of glyphosate and therefore have numerous applications in herbicide research and related areas of science as well as resolving many commercial issues as a consequence of glyphosate utilization.     

Improving salt stress responses of the symbiosis in alfalfa using salt-tolerant cultivar and rhizobial strain

Salt stress can affect alfalfa growth directly by adversely affecting metabolism, or indirectly by its effect on Rhizobium capacity for symbiotic N₂ fixation. Growth and carbohydrate metabolism in leaves, roots and nodules of two alfalfa cultivars (Medicago sativa cv Apica and salt-tolerant cv Halo) in association with two rhizobial strains (A2 and salt-tolerant Rm1521) exposed to different levels of NaCl (0, 20, 40, 80 or 160 mM NaCl) were assessed under controlled conditions. For both cultivars, shoot and root biomasses and shoot to root ratio significantly declined with increasing NaCl concentrations. Under 80 mM NaCl, Halo plants yielded 20% more fresh shoot biomass than Apica while plants inoculated with Rm1521 allocated more biomass to the roots than to the shoots compared to A2. Halo plants maintained a steady shoot water content (about 80%) under the entire range of NaCl concentrations. Shoot water content was more variable in Apica. Apica in association with salt-tolerant strain Rm1521 maintained a better water status than with strain A2, as indicated by the higher shoot water content at 80 mM NaCl. Under salt stress, two major compatible sugars involved in plant osmoregulation, sucrose and pinitol, increased in leaves while a large accumulation of starch was observed in roots. In nodules, pinitol, sucrose and starch increased under salt stress and were much more abundant with strain Rm1521 than with A2. This suggests that there could be an active transport from the shoot to the nodules to help maintain nodule activity under NaCl stress and that strain Rm1521 increases the sink strength toward nodules. Our results show that combining cultivars and rhizobial strains with superior salt tolerance is an effective strategy to improve alfalfa productivity in salinity affected areas.     

Role of exogenously supplied ferulic and p-coumaric acids in mimicking the mode of action of acetolactate synthase inhibiting herbicides

Chlorsulfuron and imazethapyr (herbicides that inhibit acetolactate synthase; ALS, EC 4.1.3.18) produced a strong accumulation of hydroxycinnamic acids that was related to the induction of the first enzyme of the shikimate pathway, 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (EC 2.5.2.54). The exogenous application of two hydroxycinnamic acids, ferulic and p-coumaric acids, to pea plants resulted in their internal accumulation, arrested growth, carbohydrate and quinate accumulation in the leaves, and the induction of ethanolic fermentation. These effects resemble some of the physiological effects detected after acetolactate synthase inhibition and suggest important roles for ferulic and p-coumaric acids in the mode of action of herbicides inhibiting the biosynthesis of branched chain amino acids.     

Biotoxic Effects of Herbicides on Growth,Nodulation, Nitrogenase Activity,and Seed Production in Chickpeas

Abstract

The effects of pre‐emergent (PRE) application of methabenzthiazuron (MBT), terbutryn, and linuron on plant vigor, number of nodules formed, nitrogenase activity (NA), chlorophyll contents, nitrogen (N) uptake, and seed yield were assessed in inoculated chickpeas, grown in sandy clay loam soil, in a controlled environment. The PRE application of MBT, terbutryn, and linuron at 2.5, 1.25 and 2 g a.i. kg^?1, respectively, had no significant adverse effect on plant vitality, yield, and NA of excised nodules in inoculated chickpea plants, but reduced these parameters at 7.5, 3.75, and 6 g a.i. kg^?1 of soil. Among the herbicides tested, linuron at each dose rates significantly (P≤0.05) decreased the grain yield, whereas terbutryn at 1.25 and 2.5 g a.i. kg^?1 improved the grain yield by 19.4% and 11%, respectively, relative to the nonherbicide control. While comparing the sum of mean values of toxicity of all dose rates of each herbicide on grain yield, the toxicity increased in the following order: control=linuron>methabenzthiazuron>terbutryn. Nodulation (nodule number per plant and their dry mass) and chlorophyll contents per plant decreased consistently with increased herbicide rates, except linuron, which at 2 g a.i. kg^?1 improved nodulation at both 45 and 90 days after seeding and corresponded with Mesorhizobium activity. The N content in grain and straw significantly decreased at the highest tested rates of each herbicide. Linuron at 2 and 4 g a.i. kg^?1 improved N content in straw, whereas at 4 g a.i. kg^?1 it significantly reduced N content in grain.     

Manganese in RR Cotton as Affected by Glyphosate

There is evidence that glyphosate application in soybean tolerant to herbicides could interfere in the manganese (Mn) nutrition of the crop, but there is no information on this effect in cotton plants. This study aimed at assessing manganese accumulation and distribution in cotton as affected by glyphosate application. The experiment was conducted in nutrient solution with four Mn concentrations and two cotton cultivars: conventional NuOpal and NuOpal tolerant to glyphosate (RR). Glyphosate was applied or not to the tolerant cultivar. The inclusion of the glyphosate resistance gene in cotton and herbicide, application increased shikimic acid (ShA) concentration in the plants. Glyphosate application decreased cotton leaf area and the dry matter production of the plant structures. The adverse effects of glyphosate were not overcome with higher Mn rates in the solution.     

干旱及复水条件下草甘膦对抗草甘膦大豆幼苗渗透调节物质和莽草酸含量的影响

为探明干旱胁迫及复水条件下不同剂量草甘膦对抗草甘膦大豆(RR1)幼苗叶片渗透调节物质、莽草酸(shikimic acid, SA)含量及根系活力的影响,采用盆栽试验,在大豆的第3复叶期进行水分胁迫5d和除草剂草甘膦处理,研究RR1幼苗叶片可溶性蛋白(soluble protein, SP)、可溶性糖(soluble sugar, SS)、游离脯氨酸(free praline, FP)、莽草酸(shikimic acid, SA)含量和根系活力(RA)的变化。结果表明,干旱胁迫前期RR1叶片的SP含量随草甘膦剂量的增加呈先升高后降低趋势,0.46kg/hm2叶片SP的含量最高,胁迫后期SP含量随草甘膦剂量的增加而降低;SS、FP和SA含量随草甘膦剂量的增加和胁迫时间的延长而增加,RA随草甘膦剂量的增加和胁迫时间的延长而降低。复水12d后,不同剂量草甘膦处理的各指标均有所恢复。干旱条件下,经草甘膦处理的RR1叶片的SP含量和RA低于草甘膦在正常水分条件下的处理,而SS、FP和SA含量相反。相关性分析表明,FP和SA含量与草甘膦剂量的相关关系最明显;而SS和SA含量与干旱胁迫时间的相关关系最明显。说明正常水分条件下,草甘膦对RR1幼苗造成的伤害经过一段时间后有所缓解;干旱胁迫加剧了草甘膦对RR1幼苗叶片渗透调节物质、莽草酸含量和根系活力的影响。抗草甘膦大豆主要通过积累FP、SS和SA对草甘膦和干旱胁迫做出响应。     

Soil fertility effects on growth,yield, nodulation and nitrogenase activity of Austrian winter pea

Austrian winter pea (Pisum sativum subspecies arvense (L.) Poir) is grown as a cool season annual to produce high protein seed and forage as well as for soil fertility improvement. This legume is grown on a wide range of soil types with many different cropping systems. The objective of these studies was to determine the influence of K levels, with and without P and Ca fertilization, for increased growth, yield, nodulation and nitrogenase activity. Results were from 3 years’ field and greenhouse experiments with a Psammentic Paleustalf (Eufaula series) utilizing Rhizobium leguminosarum (Frank), ATCC 10314 as inoculum. Soil fertility effects on composition and histology of field‐grown nodules are presented.

Available soil P was a limiting plant nutrient in field studies with significant response to K resulting with PK combinations for top growth, tillers, pods, seed yield, nodule mass, and nitrogenase activity levels (C₂H₂, red.). Multiple regression for nitrogenase (umol C₂H₄ h^‐1) = 1.09 tiller number + 3.37 nodule weight + 2.29 pod number, R² = 0.837, C.V. = 29.9%. Results from the greenhouse experiments indicated significant responses with increased K application levels when combined with P and Ca fertilization for top growth, nodule weight, number of nodules and nitro‐genase activity. Highly significant correlations resulted with nitrogenase x nodule weight (r=0.538) and nitrogenase x top growth (r=0.359) with multiple regression of treatment effects for nitrogenase (μmol C₂H₄ h^‐1) = 2.73 P + 1.04 K + 4.92 Ca, R² = 0.797 and C.V. = 48.8%. Soil addition of plant nutrients resulted in significantly increased concentrations of those elements within nodules. Magnesium content was not consistently influenced by P, Ca, and K amendments. Sodium decreased with increased K fertilization. Multiple regression of elemental composition (mg g^‐1 nodule) for nitrogenase (pmol C₂H₄ h^‐1) = 0.21 P + 0.86 K + 2.35 Ca ‐ 2.01 Na, R² = 0.772, C.V. = 55.6%. The proportion of plant nutrients in nodules contained within the nodule cytosol was highest for K (56.2%) and lowest for Ca (21.4%) with intermediate levels of Mg (50.2%), P (45.4%), and Na (37.2%).

Practical application from these data include the requirement of adequate available soil K for increased yield and nitrogen fixation with favorable P and Ca soil levels in Austrian winter pea production.     

氮锌复合作用对白三叶草根瘤生长发育和营养元素吸收的影响

采用砂培试验、中子活化和电子显微技术,研究了不同水平氮锌配施对白三叶草根瘤生长发育、营养元素吸收的影响。结果表明,随施氮水平增加,根瘤的数量、固氮酶活性、单瘤重量、细胞内类菌体数量、营养元素吸收量显著下降。与低锌条件(0.1μmol Zn/L)相比,足量锌(2.0μmol Zn/L)显著增加了根瘤的数量、单瘤重量、固氮酶活性、营养元素吸收量和细胞内类菌体数量。与其他处理相比较,低氮和足量锌处理(5mmol N/L、2.0μmol Zn/L)显著增加了根瘤的数量、单瘤重量、固氮酶活性、营养元素吸收量和细胞内类菌体数量。