Delayed inoculation and starter nitrogen for enhancing early growth and nitrogen status ofLespedeza cuneata

Summary Two growth chamber experiments were conducted to determine the response ofLespedeza cuneata (Dumont) G. Don. (sericea lespedeza) to delayed inoculation and low levels of nitrogen fertilization. Nitrogen was supplied either as NH ₄ ⁺ or as NO ₃ ^– in solution. At 0.5 and 5.0 ppm nitrogen early growth and N₂(C₂H₂) fixation was inhibited by NH ₄ ⁺ and promoted by NO ₃ ^– . Inoculation at seeding did not negatively affect growth prior to the onset of N₂(C₂H₂) fixation. Delayed inoculation until the trifoliate stage thus did not increase growth or N₂ fixation during the first 40 days of growth. After 40 days, specific nitrogenase activity was highest for plants inoculated at the first trifoliate stage of growth. In contrast, growth and total shoot nitrogen accumulation were higher in plants inoculated at planting. The experimental results suggest that delaying inoculation is not a useful technique for improving early growth ofL. cuneata for surface mine reclamation.     

Invasive Lespedeza cuneata and native Lespedeza virginica experience asymmetrical benefits from rhizobial symbionts

Background and Aims

Lespedeza cuneata (Dum. Cours.) G. Don is an invasive legume that displaces populations of native N. American congeners. Our aims are to determine the growth benefits of different rhizobacterial strains for L. cuneata and native Lespedeza virginica (L.) Britton, and to determine if these strains influence competition between these plants.

Methods

Plants were grown under nitrogen-limiting conditions in sterilized soil in pairs consisting of two L. cuneata, two L. virginica, or one of each species, and then plants were inoculated with one of seven rhizobial isolates, or with a no-strain control. After 3 months, plants were harvested for determination of biomass and nodulation rate.

Results

Five of the assayed stains improved L. cuneata biomass over uninoculated controls, but none of the strains benefited L. virginica. L. cuneata plants had more biomass and root nodules when grown in competition with L. virginica than with a conspecific.

Conclusions

Asymmetrical benefits from these symbionts accrued to invasive L. cuneata but not to native L. virginica, and this may provide the invader with a growth advantage in the field. Changes in the availability of effective symbionts in the soils of invaded sites can shape performance of native and invasive plants.     

Carbon mineralization in soil of roots from twenty crop species,as affected by their chemical composition and botanical family

Background and aims

Plant growth-promoting rhizobacteria (PGPR) have been widely studied for agricultural applications. One aim of this study was to isolate cadmium (Cd)-tolerant bacteria from nodules of Glycine max (L.) Merr. grown in heavy metal-contaminated soil in southwest of China. The plant growth-promoting (PGP) traits and the effects of the isolate on plant growth and Cd uptake by legume and non-legume plants in Cd-polluted soil were investigated.

Methods

Cd-tolerant bacteria were isolated by selective media. The isolates were identified by 16S rRNA gene and phylogenetic analysis. The PGR traits of the isolates were evaluated in vitro. Cd in soil and plant samples was determined by ICP-MS.

Results

One of the most Cd-tolerant bacteria simultaneously exhibited several PGP traits. Inoculation with the PGPR strain had positive impacts on contents of photosynthesis pigments and mineral nutrients (Fe or Mg) in plant leaves. The shoot dry weights of Lolium multiflorum Lam. increased significantly compared to uninoculated control. Furthermore, inoculation with the PGPR strain increased the Cd concentrations in root of L. multiflorum Lam. and extractable Cd concentrations in the rhizosphere, while the Cd concentrations in root and shoot of G. max (L.) Merr. significantly decreased.

Conclusions

This study indicates that inoculation with Cd-tolerant PGPR can alleviate Cd toxicity to the plants, increase Cd accumulation in L. multiflorum Lam. by enhancing Cd availability in soils and plant biomass, but decrease Cd accumulation in G. max (L.) Merr. by increasing Fe availability, thus highlighting new insight into the exploration of PGPR on Cd-contaminated soil.     

Nodulation and growth ofLeucaena leucocephala (Lam.) de Wit as affected by inoculation and N fertilizer

Leonard jar, pot and field experiments examined the effects of inoculation and the influence of nitrogen fertilizer on nodulation, nitrogen fixation and growth ofLeucaena leucocephala (Lam.) de Wit at IITA, Ibadan, Nigeria. Leucaena responded to both inoculation and/or nitrogen application. Shoot growth and total N and P of inoculated plants were comparable to those of the highest N treatment, and the values were about 55% greater than those of uninoculated ones. Field data indicated that toal N yields of inoculated leucaena were increased by 50% with 40 or 80 kg ha^–1 of N fertilizer. However, N fertilizer depressed N fixation by 56% as was expected from nodule mass data. N-fixation was delayed for about 8 weeks in the plots without N. Application of small amounts of N starter (20 ppm) proved to be beneficial to satisfy the plant need during the early stage of leucaena growth. The rhizobial strains IRc 1045 and IRc 1050 were effective, competitive and survived well in the field one year after their establishment.     

Different interaction among Glomus and Rhizobium species on Phaseolus vulgaris and Zea mays plant growth, physiology and symbiotic development under moderate drought stress conditions

Even though the positive interactions between arbuscular mycorrhizal (AM) fungi and rhizobial bacteria in legume plants are well documented, their interactions under drought conditions could be negative in some species. In the present study, we examined six different strains of Rhizobiun in combination with two AM fungi (Glomus mosseae and Glomus intraradices) on the responses of Phaseolus vulgaris plants to moderate drought conditions. Moreover, to discriminate between direct competition for carbon resources from direct inhibition processes, a non-legume plant (Zea mays) was also used. Although all inoculants (single or double) increased P. vulgaris growth, only one double combination further increased total or pod dry weights. On the other hand, three double combinations decreased pod dry weight compared to plants inoculated with a single AM fungus. In Z. mays plants, one double inoculation treatment further increased shoot dry weight, but another double inoculation treatment decreased root dry weight in plants inoculated with G. mosseae. In addition, in both plant species, a higher percentage of decrease in AM root colonization by some rhizobial strains was observed. This was most likely caused by a direct inhibition of AM fungal growth by the rhizobial strains and also depended on the host plant involved. Further research is needed to elucidate on the mechanisms behind this inhibition.     

Inoculation and production of container-grown red alder seedlings

Summary The inoculation ofAlnus rubra (red alder) withFrankia sp. can lead to a highly efficient symbiosis. Several factors contribute to the successful establishment of nitrogenfixing nodules: (1) quantity and quality ofFrankia inoculant; (2) time and method of inoculation; (3) nutritional status of the host plant.Frankia isolates were screened for their ability to nodulate and promote plant growth of container-grown red alder. Inoculations were performed on seedlings and seeds. Apparent differences in symbiotic performance could be seen when seeds or seedlings were inoculated. Plants inoculated at planting performed significantly better than those inoculated four weeks later in terms of shoot height, nodule number and shoot dry weight. If inoculation was delayed further, reduction in shoot height, nodule number and shoot dry weight resulted. The effect of fertilizer was also investigated with regard to providing optimal plant growth after inoculation. Plants receiving 1/5 Hoagland's solution minus nitrogen showed maximal plant growth with abundant nodulation. Plants receiving 1/5 Hoagland's solution with nitrogen showed excellent plant growth with significantly reduced nodulation.     

Elevated CO2 induces differences in nodulation of soybean depending on bradyrhizobial strain and method of inoculation

High CO₂ has been shown to increase plant growth and to affect symbiotic activity in many legumes species, including soybean (Glycine max [L.] Merr.). In order to assess the interaction between elevated CO₂ and rhizobial symbionts on soybean growth and nodulation, we combined the effects of CO₂ with those of different bradyrhizobial strains and methods of inoculation. Soybean seeds were sown in agricultural soil in pots and inoculated with three strains of Bradyrhizobium japonicum (5Sc2 and 12NS14 indigenous to Quebec soils, and 532c, a reference strain), the inoculum being either applied directly to the seed or incorporated into the soil. Plants were grown in growth chambers (22/17ºC) for 6 weeks, under either near ambient (400 μmol mol^?1) or elevated (800 μmol mol^?1) concentrations of CO_2. Elevated CO₂ increased mass (63%) and number (50%) of soybean nodules, particularly medium and large, allowed a deeper nodule development, and increased shoot dry weight (+30%), shoot C uptake (+33%) and shoot N uptake (+78%), compared to ambient CO₂. The two indigenous strains induced more medium and large nodules under elevated CO₂ than the reference strain and showed the greatest increases in shoot dry weight. Soil inoculation induced higher number of small nodules than seed inoculation, specifically for the two indigenous strains, but did not affect plant growth parameters. We conclude that soybean yield enhancements due to elevated CO₂ are associated with the production of large and medium-size nodules and a deep nodulation, that the two indigenous strains better respond to elevated CO₂ than the reference strain, and that the method of inoculation has little influence on this response.     

Induction of antioxidant enzymes is involved in the greater effectiveness of a PGPR versus AM fungi with respect to increasing the tolerance of lettuce to severe salt stress

This study investigated the influence of inoculation with a plant growth-promoting rhizobacterium, Pseudomonas mendocina Palleroni, alone or in combination with an arbuscular mycorrhizal (AM) fungus, Glomus intraradices (Schenk & Smith) or Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe, on antioxidant enzyme activities (catalase and total peroxidase), phosphatase activity, solute accumulation, growth and mineral nutrient uptake in leaves of Lactuca sativa L. cv. Tafalla affected by three different levels of salt stress. Salinity decreased lettuce growth, regardless of the biological treatment and of the salt stress level. The plants inoculated with P. mendocina had significantly greater shoot biomass than the control plants at both salinity levels, whereas the mycorrhizal inoculation treatments only were effective in increasing shoot biomass at the medium salinity level. At the highest salinity level, the water content was greater in leaves of plants treated with P. mendocina or G. mosseae. At the medium salinity level, G. intraradices- or G. mosseae-colonised plants showed the highest concentrations of foliar P. The P. mendocina- and G. mosseae-colonised plants presented higher concentrations of foliar K and lower concentrations of foliar Na under high salt conditions. Salt stress decreased sugar accumulation and increased foliar proline concentration, particularly in plants inoculated with the PGPR. Increasing salinity stress raised significantly the antioxidant enzyme activities, including those of total peroxidase and catalase, of lettuce leaves compared to their respective non-stressed controls. The PGPR strain induced a higher increase in these antioxidant enzymes in response to severe salinity. Inoculation with selected PGPR could serve as a useful tool for alleviating salinity stress in salt-sensitive plants.     

Symbiotic performance of selectedCyclopia Vent. (honeybush) rhizobia under nursery and field conditions

Three newly selected strains ofCyclopia rhizobia together with an inoculant strain, which has never been tested in the field with adequate experimental design, was assessed under both nursery and field conditions for symbiotic performance. The three new test strains were initially selected for their superior N₂-fbcing abilities under glasshouse conditions, and then evaluated in this study for field performance.Cyclopia subternata Vogel andCyclopia genistoides (L.) R. Br., which have the potential for producing high quality honeybushtea, were used as host plants in both the nursery and field studies. The effect of seedling inoculation at the nursery level was also examined for the four test strains under nursery conditions. The inoculation of cuttings under nursery conditions produced.significant increases in shoot biomass, shoot %N and shoot N content. More specifically, inoculatingC. subternata with strains UCT44b and UCT61a significantly increased shoot biomass and N content relative to strain PPRICI3. Strains UCT44b and UCT61a also showed better nodulation withC. subternata cuttings compared to strains UCT40a and PPRICI3. Field inoculation ofCyclopia increased all growth parameters relative to the uninoculated control, except for leaf %N.Cyclopia subternata inoculated with strains UCT44b, UCT40a and UCT61a produced significantly lower δ¹⁵N values than the uninoculatedC. subtemata reference plant. Using the¹⁵N natural abundance method,C. subternata was estimated to be gaining about half of its N from N₂ fixation, whileC. genistoides obtained less than half of its N from symbiotic nutrition.     

Effects of Bradyrhizobium japonicum and Soybean (Glycine max (L.) Merr.) Phosphorus Nutrition on Nodulation and Dinitrogen Fixation

Cells of Bradyrhizobium japonicum were grown in media containing either 1.0 mM or 0.5 μM phosphorus. In growth pouch experiments, infection of the primary root of soybean (Glycine max (L.) Merr.) by B. japonicum USDA 31, 110, and 142 was significantly delayed when P-limited cells were applied to the root. In a greenhouse experiment, B. japonicum USDA 31, 110, 122, and 142 grown with sufficient and limiting P were used to inoculate soybeans which were grown with either 5 μM or 1 mM P nutrient solution. P-limited cells of USDA 31 and 110 formed significantly fewer nodules than did P-sufficient cells, but P-limited cells of USDA 122 and 142 formed more nodules than P-sufficient cells. The increase in nodule number by P-limited cells of USDA 142 resulted in significant increases in both nodule mass and shoot total N. In plants grown with 1 mM P, inoculation with P-limited cells of USDA 110 resulted in lower total and specific nitrogenase activities than did inoculation with P-sufficient cells. Nodule numbers, shoot dry weights, and total N and P were all higher in plants grown with 1 mM P, and plants inoculated with USDA 31 grew poorly relative to plants receiving strains USDA 110, 122, and 142. Although the effects of soybean P nutrition were more obvious than those of B. japonicum P nutrition, we feel that it is important to develop an awareness of the behavior of the bacterial symbiont under conditions of nutrient limitation similar to those found in many soils.     

Promotion of Peanut Growth by Co-inoculation with Selected Strains of <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Azospirillum</Emphasis>

The ability of inoculated rhizobial strains to increase root nodulation of host legumes often depends on their competitiveness with existing native soil strains. Results of studies to date on rhizobial inoculation for improvement of peanut (Arachis hypogaea L.) production in Argentina have been inconsistent and controversial. In many cases, nodulation and yield of peanut crops have been increased by inoculation of specific rhizobial strains. Native peanut-nodulating strains are generally present in soils of agricultural areas, but their growth-promoting effect is often lower than that of inoculated strains. Many species of the genus Bradyrhizobium interact in a host-specific manner with legume species and form nitrogen-fixing root nodules. Other free-living rhizobacteria such as species of the genus Azospirillum are facultatively capable of interacting with legume roots and promoting plant growth. We evaluated and compared the effects of various single inoculation and co-inoculation treatments on peanut growth parameters in greenhouse and field experiments. In the greenhouse studies, co-inoculation with various Bradyrhizobium strains (native 15A and PC34, and recommended peanut inoculant C145), and Azospirillum brasilense strain Az39 generally resulted in increases in the measured parameters. The growth-promoting effect of 15A was similar to or higher than that of C145. In the field studies, 15A-Az39 co-inoculation had a greater promoting effect on measured growth parameters than did C145-Az39 co-inoculation. Our findings indicate that careful selection of native rhizobacterial strains adapted to peanut soils is useful in strategies for growth promotion, and that 15A in particular is a promising candidate for future inoculant formulation.     

Effect of salinity on nodule formation by soybean

A split-root growth system was employed to evaluate the effect of NaCl on nodule formation by soybean (Glycine max L. Merr. cv Davis). By applying the salinity stress and rhizobial inoculum to only one-half the root system, the effects of salinity on shoot growth were eliminated in the nodulation process. Rhizobium colonization of inoculated root surfaces was not affected by the salt treatments (0.0, 26.6, 53.2, and 79.9 millimolar NaCl). While shoot dry weight remained unaffected by the treatments, total shoot N declined from 1.26 grams N per pot at 0.0 millimolar NaCl to 0.44 grams N per pot at 79.9 millimolar NaCl. The concentration of N in the shoot decreased from 3.75% N (0.0 millimolar NaCl) to 1.26% N at 79.9 millimolar NaCl. The decrease in shoot N was attributed to a sharp reduction in nodule number and dry weight. Nodule number and weight were reduced by approximately 50% at 26.6 millimolar NaCl, and by more than 90% at 53.2 and 79.9 millimolar NaCl. Nodule development, as evidenced by the average weight of a nodule, was not as greatly affected by salt as was nodule number. Total nitrogenase activity (C₂H₂ reduction) decreased proportionally in relation to nodule number and dry weight. Specific nitrogenase activity, however, was less affected by salinity and was not depressed significantly until 79.9 millimolar NaCl. In a second experiment, isolates of Rhizobium japonicum from nodules formed at 79.9 millimolar NaCl did not increase nodulation of roots under salt stress compared to nodule isolates from normal media (0.0 millimolar NaCl). Salt was applied (53.2 millimolar NaCl) to half root systems at 0, 4, 12, and 96 hours from inoculation in a third experiment. By delaying the application of salt for 12 hours, an increase in nodule number, nodule weight, and shoot N was observed. Nodule formation in the 12- and 96-hour treatments was, however, lower than the control. The early steps in nodule initiation are, therefore, extremely sensitive to even low concentrations of NaCl. The sensitivity is not related to rhizobial survival and is probably due to the salt sensitivity of root infection sites.     

Plant growth-promoting rhizobacteria affect the growth and nutrient uptake of Fraxinus americana container seedlings

Plant growth-promoting rhizobacteria (PGPR) are important catalysts that regulate the functional properties of agricultural systems. However, there is little information on the effect of PGPR inoculation on the growth and nutrient accumulation of forest container seedlings. This study determined the effects of a growth medium inoculated with PGPR on the nutrient uptake, nutrient accumulation, and growth of Fraxinus americana container seedlings. PGPR inoculation with fertilizer increased the dry matter accumulation of the F. americana aerial parts with delayed seedling emergence time. Under fertilized conditions, the accumulation time of phosphorous (P) and potassium (K) in the F. americana aerial parts was 13 days longer due to PGPR inoculation. PGPR increased the maximum daily P and K accumulations in fertilized seedlings by 9.31 and 10.44 %, respectively, but had little impact on unfertilized ones. Regardless of fertilizer application, the root exudates, namely sugars, amino acids, and organic acids significantly increased because of PGPR inoculation. PGPR inoculation with fertilizer increased the root, shoot, and leaf yields by 19.65, 22.94, and 19.44 %, respectively, as well as the P and K contents by 8.33 and 10.60 %, respectively. Consequently, the N, P, and K uptakes increased by 19.85, 31.97, and 33.95 %, respectively. Hence, PGPR inoculation with fertilizer can be used as a bioenhancer for plant growth and nutrient uptake in forest container seedling nurseries.     

Influence of arbuscular-mycorrhizal fungi, Rhizobium meliloti strains and PGPR inoculation on the growth of Medicago arborea used as model legume for re-vegetation and biological reactivation in a semi-arid mediterranean area

Medicago arborea can be used for re-vegetationpurposes under semiarid conditions. These woody legumes have the ability toforman association with arbuscular mycorrhizal (AM) fungi and rhizobial bacteria,which can be maximised by microorganisms producing certain stimulatingmetabolites acting as plant growth promoting rhizobacteria (PGPR). The effectsof single and combined inoculations using microorganisms with different andinteractive metabolic capacities, namely three Glomusspecies, two Rhizobium meliloti strains (a wild type, WTand its genetically modified derivative GM) and a plant growth promotingrhizobacterium, (PGPR), were evaluated. All three inoculated AM fungi affectedMedicago growth in different ways. Differences weremaintained when soil was co-inoculated with each of the rhizobial strains (WTorGM) and the PGPR. Mycorrhizal fungi were effective in all cases, but the PGPRonly affected plant growth specific microbial situations. PGPR increased growthof G. mosseae-colonised plants associated withRhizobium WT strain by 36% and those infected byG. deserticola when associated with the rhizobial GMstrainby 40%. The most efficient microbial treatments involved mycorrhizalinoculation, which was an indication of the AM dependency of this plantspecies.Moreover, PGPR inoculation was only effective when associated with specificmycorrhizal endophytes (G. mosseae plus WT andG.deserticola plus GM rhizobial strain). The reduced root/shoot (R/S)ratio resulting from PGPR inoculation, was an indication of more effective rootfunction in treated plants. AM colonisation and nodule formation wereunaffectedby the type of AM fungus or bacteria (rhizobial strain and/or PGPR). AM fromnatural soil were less infective and effective than those from the collection.The results supported the existence of selective microbial interactionsaffecting plant performance. The indigenous AM fungi appeared to be ineffectiveand M. arborea behaved as though it was highly dependentonAM colonisation, which implied that it must have a mycorrhizal association toreach maximum growth in the stressed conditions tested. Optimum growth ofmycorrhizal M. arborea plants was associated with specificmicrobial groups, accounting for a 355% increase in growth overnodulatedcontrol plants. The beneficial effect of PGPR in increasing the growth of awoody legume, such as M. arborea under stress, was onlyobserved with co-inoculation of specific AM endophytes. As a result of theinteraction, only shoot biomass was enhanced, but not as a consequence ofenhancing of the colonising abilities of the endophytes. The growthstimulation,occurring as a consequence of selected microbial groups, may be critical anddecisive for the successful establishment of plants under Mediterraneanclimaticand soil conditions.     

Tomato Growth in Soil Amended with Sugar Mill By-Products Compost

Sugar mill by-products compost may be a good soil amendment to promote tomato (Lycopersicon esculentum L.) growth. In addition, the compost may further promote plant growth by inoculation with N₂-fixing bacteria. Compost from sugar-mill waste was prepared with and without the N₂-fixing bacteria, Azotobacter vinelandii, Beijerinckia derxii and Azospirillum sp. and incubated for 50 days. Each compost type was added to 10 kg of soil in pots at rates of 0, 15, and 45 g with and without fertilizer N at rates of 0, 0.75, and 1.54 g. A blanket application of P and K was applied to all pots. Shoot and root dry weights and N content of the whole plant was measured at 55 days. Dry weight of tomato shoots was increased by 40% by addition of fertilizer N and root weight was increased by 66%. Without fertilizer N the high rate of inoculated compost increased shoot growth 180% and uninoculated compost increased shoot growth 112%. For most treatments with and without fertilizer N, inoculated compost enhanced shoot growth and nitrogen content more than uninoculated compost. Root weights were nearly doubled by addition of either compost in comparison to the 0 N treatment. At the low rate of compost addition without fertilizer N, root weight was the same for uninoculated and inoculated compost but at the high rate of compost addition root weight was 32% higher for inoculated compost. The N₂-fixing bacteria colonized roots when inoculated compost was used. Sugar mill by-products compost proved to be an effective soil amendment for promoting the growth of tomato plants.     

Genetic variability in <Emphasis Type="Italic">Bradyrhizobium japonicum</Emphasis> strains nodulating soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill]

Brazil has succeeded in sustaining production of soybean [Glycine max (L.) Merrill] by relying mainly on symbiotic N₂ fixation, thanks to the selection and use in inoculants of very effective strains of Bradyrhizobium japonicum and Bradyrhizobium elkanii. It is desirable that rhizobial strains used in inoculants have stable genetic and physiological traits, but experience confirms that rhizobial strains nodulating soybean often lose competitiveness in the field. In this study, soybean cultivar BR 16 was single-inoculated with four B. japonicum strains (CIAT 88, CIAT 89, CIAT 104 and CIAT 105) under aseptic conditions. Forty colonies were isolated from nodules produced by each strain. The progenitor strains, the isolates and four other commercially recommended strains were applied separately to the same cultivar under controlled greenhouse conditions. We observed significant variability in nodulation, shoot dry weight, shoot total N, nodule efficiency (total N mass over nodule mass) and BOX-PCR fingerprinting profiles between variant and progenitor strains. Some variant strains resulted in significantly larger responses in terms of shoot total N, dry weight and nodule efficiency, when compared to their progenitor strain. These results highlight the need for intermittent evaluation of stock bacterial cultures to guarantee effective symbiosis after inoculation. Most importantly, it indicates that it is possible to improve symbiotic effectiveness by screening rhizobial strains for higher N₂ fixation capacity within the natural variability that can be found within each progenitor strain.     

Nodulation of Soybeans as Affected by Half-root Infection with Heterodera glycines

A split-root technique was applied to soybean, Glycine max (L.) Merr. cv. Lee 68, to characterize the nature of the nodulation suppression by race 1 of the soybean cyst nematode (SCN), Heterodera glycines. Root-halves of each split-root plant were inoculated with Rhizobium japonicum, and one root-half only was inoculated with various numbers of SCN eggs. Nodulation (indicated by nodule number, nodule weights, and ratio of nodule weight to root weight) and nitrogen-fixing capacity (indicated by rate of acetylene reduction) were systemically and variously suppressed on both root-halves of the split-root plant 5 weeks after half-root inoculation with 12,500 SCN eggs. Inoculation with 500 eggs caused this suppression only on the SCN-infected (+NE) root-half; nodulation on the companion uninfected (-NE) root-half was stimulated slightly. The +NE root-halves inoculated with 5,000 eggs were excised at 2-week intervals; nodulation on the remaining -NE root-halves was not different from that of the noninoculated control when measured 6 weeks after the SCN inoculation. Thus, the systemic suppression of nodulation was reversible upon the removal of the SCN. Similarly, application of various levels of KNO₃ to the -NE root-halves of the split-root plant did not alleviate the suppressed nodulation on the companion +NE root-halves, even though plant growth was much improved at certain levels of nitrogen (125 μg N/g soil). This indicated that the localized suppression of nodulation by SCN was caused by factors in addition to poor plant growth.     

Modeling Symbiotic Performance of Introduced Rhizobia in the Field by Use of Indices of Indigenous Population Size and Nitrogen Status of the Soil

The ability to predict the symbiotic performance of rhizobia introduced into different environments would allow for a more judicious use of rhizobial inoculants. Data from eight standardized field inoculation trials were used to develop models that could be used to predict the success of rhizobial inoculation in diverse environments based on indices of the size of indigenous rhizobial populations and the availability of mineral N. Inoculation trials were conducted at five diverse sites on the island of Maui, Hawaii, with two to four legumes from among nine species, yielding 29 legume-site observations. The sizes of indigenous rhizobial populations were determined at planting. Soil N mineralization potential, total soil N, N accumulation and seed yield of nonnodulating soybean, and N derived from N₂ fixation in inoculated soybean served as indices of available soil N. Uninoculated, inoculated, and fertilizer N treatments evaluated the impact of indigenous rhizobial populations and soil N availability on inoculation response and crop yield potential. The ability of several mathematical models to describe the inverse relationship between numbers of indigenous rhizobia and legume inoculation responses was evaluated. Power, exponential, and hyperbolic functions yielded similar results; however, the hyperbolic equation provided the best fit of observed to estimated inoculation responses (r² = 0.59). The fact that 59% of the observed variation in inoculation responses could be accounted for by the relationship of inoculation responses to numbers of indigenous rhizobia illustrates the profound influence that the size of soil rhizobial populations has on the successful use of rhizobial inoculants. In the absence of indigenous rhizobia, the inoculation response was directly proportional to the availability of mineral N. Therefore, the hyperbolic response function was subsequently combined with several indices of soil N availability to generate models for predicting legume inoculation response. Among the models developed, those using either soil N mineralization potential or N derived from N₂ fixation in soybean to express the availability of mineral N were most useful in predicting the success of legume inoculation. Correlation coefficients between observed and estimated inoculation responses were r = 0.83 for the model incorporating soil N mineralization potential and r = 0.96 for the model incorporating N derived from N₂ fixation. Several equations collectively termed “soil N deficit factors” were also found to be useful in estimating inoculation responses. In general, models using postharvest indices of soil N were better estimators of observed inoculation responses than were those using laboratory measures of soil N availability. However, the latter, while providing less precise estimates, are more versatile because all input variables can be obtained through soil analysis prior to planting. These models should provide researchers, as well as regional planners, with a more precise predictive capability to determine the inoculation requirements of legumes grown in diverse environments.     

Relationship between Ureide N and N(2) Fixation, Aboveground N Accumulation, Acetylene Reduction, and Nodule Mass in Greenhouse and Field Studies with Glycine max L. (Merr)

The relationship between ureide N and N₂ fixation was evaluated in greenhouse-grown soybean (Glycine max L. Merr.) and lima bean (Phaseolus lunatus L.) and in field studies with soybean. In the greenhouse, plant N accumulation from N₂ fixation in soybean and lima bean correlated with ureide N. In soybean, N₂ fixation, ureide N, acetylene reduction, and nodule mass were correlated when N₂ fixation was inhibited by applying KNO₃ solutions to the plants. The ureide-N concentrations of different plant tissues and of total plant ureide N varied according to the effectiveness of the strain of Bradyrhizobium japonicum used to inoculate plants. The ureide-N concentrations in the different plant tissues correlated with N₂ fixation. Ureide N determinations in field studies with soybean correlated with N₂ fixation, aboveground N accumulation, nodule weight, and acetylene reduction. N₂ fixation was estimated by ¹⁵N isotope dilution with nine and ten soybean genotypes in 1979 and 1980, respectively, at the V9, R2, and R5 growth stages. In 1981, we investigated the relationship between ureide N, aboveground N accumulation, acetylene reduction, and nodule mass using four soybean genotypes harvested at the V4, V6, R2, R4, R5, and R6 growth stages. Ureide N concentrations of young stem tissues or plants or aboveground ureide N content of the four soybean genotypes varied throughout growth correlating with acetylene reduction, nodule mass, and aboveground N accumulation. The ureide-N concentrations of young stem tissues or plants or aboveground ureide-N content in three soybean genotypes varied across inoculation treatments of 14 and 13 strains of Bradyrhizobium japonicum in 1981 and 1982, respectively, and correlated with nodule mass and acetylene reduction. In the greenhouse, results correlating nodule mass with N₂ fixation and ureide N across strains were variable. Acetylene reduction in soybean across host-strain combinations did not correlate with N₂ fixation and ureide N. N₂ fixation, ureide N, acetylene reduction, and nodule mass correlated across inoculation treatments with strains of Bradyrhizobium spp. varying in effectiveness on lima beans. Our data indicate that ureide-N determinations may be used as an additional method to acetylene reduction in studies of the physiology of N₂ fixation in soybean. Ureide-N measurements also may be useful to rank strains of B. japonicum for effectiveness of N₂ fixation.     

Effects of salt stress on nitrogenase activity and growth of four legumes

The effects of salt stress on nitrogenase (N2-ase) activity, growth and nitrogen content ofVicia faba (L.),Medicago sativa (L.) Merrill,Glycine max andVigna sinensis (L.) were investigated. Four levels of salinity were applied and salt treatments were imposed on inoculated and N-fertilized plants.M. sativa tolerated mild levels of salinity but higher salt concentrations depressed N₂-ase activity of this species. The other three legumes were considerably affected by salt treatments, and N₂-ase activity was significantly reduced by salinity. Vicia faba, carrying elongate nodules, could restore a partial N₂-ase activity upon recovery from salt stress whereasG. max andV. sinesis, both with spherical nodules, could not regain significant activity when salinity was removed. Salt stress retarded growth of both inoculated and N-fertilized plants. The nitrogen content of both treatments was also affected by salinity and the effect was more severe for inoculated than N-fertilized plants.