The paper studied the effects of different nitrogen rates (0, 30, 60, 90 kg N/ha) on the soybean cultivar Proteinka, inoculated with the NS-Nitragin microbial fertilizer. Trials were set up at Backi Petrovac on a soil with no previous history of soybean cultivation. Samples of plant material were taken twice, once at soybean flowering and once at maturity. At flowering, the largest mass and length of the above-ground plant parts were recorded in the treatment with 60 kg N/ha, while the largest nodule number, mass and N content were obtained with 30 kg N/ha. The largest pod, grain numbers and grain mass and N content were produced by the inoculated treatment with no N added, and chat was the treatment that produced the largest yield as well

Aćimović, Radivoje

Marinković, Jelena

Mrkovački, Nastasija

Notulae Botanicae Horti Agrobotanici Cluj-Napoca

English

The paper studied the effects of different nitrogen rates (0, 30, 60, 90 kg N/ha) on the soybean cultivar Proteinka, inoculated with the NS - Nitragin microbial fertilizer. Trials were set up at Backi Petrovac on a soil with no previous history of soybean cultivation. Samples of plant material were taken twice, once at soybean flowering and once at maturity. At flowering, the largest mass and length of the above-ground plant parts were recorded in the treatment with 60 kg N/ha, while the largest nodule number, mass and N content were obtained with 30 kg N/ha. The largest pod, grain numbers and grain mass and N content were produced by the inoculated treatment with no N added, and that was the treatment that produced the largest yield as well

Nastasija MRKOVACKI

Jelena MARINKOVI

R. ACIMOVIC

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Available online at www.notulaebotanicae.roNot. Bot. Hort. Agrobot. Cluj 36 (1) 2008, 48-51Print ISSN 0255-965X; Electronic ISSN 1842-4309Notulae Botanicae Horti AgrobotaniciCluj-NapocaEffect of N Fertilizer Application on Growth and Yield of Inoculated SoybeanNastasija MRKOVAČKI1) , Jelena MARINKOVIĆ1) , R. AĆIMOVIĆ2)1)Institute of Field and Vegetable Crops, 30 Maksima Gorkog  St., 21000 Novi Sad; Serbia, e-mail: nastasija@ifvcns.ns.ac.yu2)Concern Farmakom MB, bb Hajduk Veljkova St., 15000 Šabac; SerbiaAbstractThe paper studied the effects of different nitrogen rates (0, 30, 60, 90 kg N/ha) on the soybean cultivar Proteinka, inoculated with the NS–Nitragin microbial fertilizer. Trials were set up at Bački Petrovac on a soil with no previous history of soybean cultivation. Samples of plant material were taken twice, once at soybean flowering and once at maturity. At flowering, the largest mass and length of the above-ground plant parts were recorded in the treatment with 60 kg N/ha, while the largest nodule number, mass and N content were obtained with 30 kg N/ha. The largest pod, grain numbers and grain mass and N content were produced by the inoculated treatment with no N added, and that was the treatment that produced the largest yield as well.  Keywords: B. japonicum, cultivar, NS–Nitragin, total microbial abundance IntroductionSoybean (Glycine max (L) Mer) is an economically im-portant leguminous crop on a worldwide scale and also the most important legume in China (Gan et al., 2003). Soy-bean nitrogen (N) requirements are met in a complex  man-ner, as this crop is capable of utilizing both soil N (mostly in the form of nitrate) and atmospheric N (through symbiotic nitrogen fixation) (Harper 1974; Milić et al., 2002). Stud-ies of the effects combined N (soil N + mineral fertilizer N) has on the physiology of the Rhizobium-legume symbiosis have shown that a large amount of N in the fertilizer re-duces root infections (Dazzo and Brill, 1978), nodule mass and number (Streeter, 1988), levels of N-fixing activity in the nodules (Gibson, 1974), and the total amount of fixed N.  The degree of N-fixation inhibition varies depending on the form of N applied (Dart and wildou, 1970), season (Pate and Dart, 1961), light intensity, temperature (Gip-son, 1974), and other environmental factors (Pankhurst, 1981). Symbiotic nitrogen fixation begins after the for-mation of nodules, reaches maximum activity in the early stages of pod filling, and subsides in the late reproductive stage  (Imsande, 1988; George and Singleton, 1992).  for increased legume yields and nitrogen fixation levels, it is es-sential to understand the interactions taking place between inorganic N supply and plant growth throughout the en-tire period of crop development.In order for us to obtain information needed for the formulation of optimal N addition levels for soybean culti-vation, experiments were carried out to determine:- the effects adding different N amounts has on nodula-tion and nitrogen fixation at flowering;- the effects of incorporation of different N amounts on yield parameters and grain yield.Materials and methodsUsed in this paper were Bradyrhizobium japonicum mi-crobial strains that are used for the production of the NS–Nitragin microbial fertilizer. The Bradyrhizobium japoni-cum strains were grown on a Demolon liquid medium in a thermostat on a shaker at 300 C for four days and injected onto a sterile peat carrier. The trials were carried out on an experimental plot at Bački Petrovac in 2005 and 2006. A randomized block design with four replications was used in each of the treatments, which were as follows: No N added30 kg N/ha 60 kg N/ha 90 kg N/ha Seeds from all the treatments were inoculated with NS–Nitragin just before sowing.The seeds were of the soybean cultivar Proteinka, devel-oped at the Institute of field and Vegetable Crops in Novi Sad.At flowering, plant samples were taken from five plants per replicate in each treatment. Nodule mass and number per plant, length and mass of the above-ground plant parts and root, and the N content of nodules and above-ground plant parts were determined. At the end of the growing season, pod and grain number, grain N content, and grain mass and yield were analyzed. N content was determined using the standard kjeltec equipment.Mrkovački, N. et al. /  Not. Bot. Hort. Agrobot. Cluj 36 (1) 2008, 48-5149Results obtained at maturityTable 3 shows pod number, grain number, grain mass and N content, and yield at 14% moisture. The greatest number of pods and grains and the largest grain mass were obtained in treatment 1 (no N), while the smallest values of these traits were recorded in treatment 3 (60 kg N/ha). Table 4 shows abundance at soybean maturity of the microbial groups studied. The largest total number of mi-croorganisms and Azotobacter abundance were found in the treatment with no N added, which correlates with the indicators shown in Table  3. According to these findings concerning pod and grain number and grain mass and N content as indicators of yield and total microbial abundance and Azotobacter numbers as indicators of soil biogenicity, the inoculated treatment with no N added was the most effective.The largest average yield in the 2005/2006 period was produced by the inoculated treatment without N, which correlates with pod and grain numbers, grain mass,  and total microbial and Azotobacter abundance at maturity (Table 4). A decline in yield with increasing N rate was observed as well. This leads to the conclusion that inocu-lated soybean can be successfully grown without the addi-tion of N fertilizer, especially on soils such as the one from At flowering and at the end of the vegetation period, soil samples for microbiological and chemical analyses were taken as well. They were used to determine total mi-crobial abundance on soil agar, the number of ammoni-fiers on MPA, the number of free N-fixing bacteria, and Azotobacter abundance on fjodor’s medium.Results and discussionResults obtained at floweringThe results shown in the tables are the average for the years 2005 and 2006 godina. Table 1 shows the length, mass and N content of the above-ground plant parts and nodule number, mass and N content at flowering. The greatest length, mass and N content of the above-ground plant parts were found in Treatment 3 (60 kg N/ha), while the largest nodule number, mass and  N content were re-corded in Treatment 2 (30 kg N/ha). Table 2 shows total microbial abundance, the abun-dance of ammonifiers and oligonitrophiles, and Azoto-bacter abundance at flowering. The largest total number of microorganisms and Azotobacter abundance were ob-tained with 60 kg N/ha, which  correlates with the length and mass of the above-ground parts of the plant. Table 1 Effect of N levels on the length, mass and N content of the above-ground plant parts and nodule number, mass and N content at soybean floweringN level kg/ha Above-ground length (cm)Above-ground mass (g)Above-ground NContent (mg/plant) Nodule numberNodule mass(mg)Nodele N content (mg/plant)0 99.16 17.87 543.69 60.29 342.00 1.73930 100.25 15.99 483.35 80.87 398.50 2.21460 101.29 19.51 619.65 40.08 313.00 1.76790 91.04 19.46 574.04 30.75 171.50 0.519Table 2 Effect of N levels on total microbial abundance and the number of ammonifiers, oligonitrophiles and Azotobacters at soybean floweringN level kg/ha Total number of microorganisms x 107Number of ammonifiers x 107Number of oligonitrophiles x106Number of Azotobacters x 1020 248.13 150.75 242.32 145.2030 203.88 158.69 257.36 156.6960 285.53 184.48 313.13 181.9590 284.23 147.99 358.81 173.46Table 3 Effect of N levels on pod and grain numbers, grain mass and N content and yieldN level kg/ha Pod number Grain number Grain Ncontent (mg/plant) Grain mass Yield(kg/ha)0 60.67 137.37 197.28 25.41 435030 59.83 134.66 162.38 24.74 425160 53.87 123.88 185.93 22.84 407390 59.96 136.04 193.28 25.01 4014Mrkovački, N. et al. /  Not. Bot. Hort. Agrobot. Cluj 36 (1) 2008, 48-5150arid tropics, as BNf does not operate at the beginning of plant growth and not all plant N requirements can be met by nitrogen fixation, especially in low soil N conditions (wani et al., 1995).In Chinese conditions, according to Gan et al., (2002), the addition of  50 kg N/ha-1 at the reproductive stages R1 through R5 instead of 75 N/ha before sowing or at the V4 stage is recommended for the soybean genotypes stud-ied. The field trial results showed that the addition of N at the reproductive stage, especially at the R5 stage (pod filling), was the most effective in increasing total fixed N amounts, whereas adding high N amounts prior to sow-ing or at the V4 stage had a negative effect on the yield of soybean grains. ConclusionsResults obtained at maturity showed that the largest total number of microorganisms and Azotobacter abun-dance were found in the treatment with no N added.According to these findings concerning pod and grain number and grain mass and N content as indicators of yield and total microbial abundance and Azotobacter numbers as indicators of soil biogenicity, the inoculated treatment with no N added was the most effective.The largest average yield in the 2005/2006 period was produced by the inoculated treatment without N.This leads to the conclusion that inoculated soybean can be successfully grown without the addition of N fertil-izer.ReferencesBhangoo, M. S., D. J. Albritton, 1976, Nodulating and non – nodulating Lee soybean isolines response to applied nitrogen. Agronomy Journal 68, 642 – 645.Cherney, J. H., J. M. Duxbury, 1994, Inorganic nitrogen supply our study. The soil in question is a moderately calcareous, humus-rich chernozem with a medium to good supply of N (leaning more towards the latter) and a pH of  7.25-8.2 (depending on analyses) (Table 5).Smith et al., (1986) have pointed out that «legumes seeded fields do not need nitrogen», while other authors have recomended that small quantities of N should be applied to reach good nodulation and maximum yields (Streeter, 1988; Cherney and Duxbury, 1994).It is known that mineral N and fixed N are neces-sary for maximum N accumulation and yield in legumes (Harper, 1974; Bhangoo and Albritton, 1976; franco et al., 1979) due to the high energy requirements in the de-velopment, maintenance and functioning of the symbiotic system compared with mineral N assimilation (Ryle et al., 1978; Pate et al., 1979; finke et al., 1982).A review by Imsande, (1989) indicates that soybean plants growing with an abundance of mineral N in the medium may accumulate less total N than soybeans that are well nodulated. The results of George and Singleton, (1992), however, have shown the opposite,  as in that study the dry matter mass and total N of soybean and string bean grown in two locations increased significantly when increased amounts of mineral N were incorporated (9 – 120 – 900 kg N/ha).The findings of Seneviratne et al., (2000) have shown that inoculation and fertilizer use promote plant growth and increase grain yields in soybean. Incorporating 23 kg N/ha as the primary fertilizer application and adding 23 kg N/ha at the end of flowering does not inhibit soybean nodulation. This shows the importance of nitrogen fixa-tion in tropical conditions even with fertilizer incorpora-tion included. In all the locations in said study, grain yields increased as a result of inoculation and fertilizer use when compared to uninoculated check treatments. It has been shown that legumes have a highly signifi-cant response to small N rates (20 – 30 kg N/ha) in semi–Table 4 Effect of N levels on total microbial abundance and the number of ammonifiers, oligonitrophiles and  Azotobacters at soybean maturityN level kg/haTotal number of microorganisms x 107Number of ammonifiers x 107Number of oligonitrophiles x106Number of Azotobacters x 1020 364.35 189.98 337.89 167.5030 343.08 239.70 330.81 158.6760 316.86 226.81 348.65 156.8990 340.34 209.92 321.05 149.30Table 5 Chemical soil analysisN level kg/hapH CaCO3% HumusTotal N% P2O5 K2OKCl H2O0 7.27 8.23 4.49 2.90 0.190 46.00 31.6830 7.25 8.20 4.24 2.90 0.195 50.69 33.6160 7.26 8.19 4.13 2.95 0.195 45.29 32.4790 7.23 8.20 4.33 2.90 0.194 46.56 32.27Average 7.25 8.20 4.30 2.91 0.193 47.13 32.51Mrkovački, N. et al. /  Not. Bot. Hort. Agrobot. Cluj 36 (1) 2008, 48-5151new perspective. Agronomy Journal 81, 549 – 556.Milić Vera, Nastasija Mrkovački, Milica Hrustić, 2002, Interrelationship of nitrogen fixation potential and soybean yield. A Periodical of Scientific Research on Field and Vegetable Crops 36, 133 – 139.Pankhurst, C. E., 1981, Effects of plant nutrient supply on nodule effectiveness and Rhizobium strain competition for nodulation of Lotus pedunculatus. Plant and Soil 60, 325 – 339.Pate, J. S., P. J. Dart, 1961, Nodulation studies in legumes. IV, The influence of inoculum strain and time of application of ammonium nitrate on symbiotic response. Plant and Soil 15, 329 – 346.Pate, J. S., D. B. Layzell, C. A. Atkins, 1979, Economy of carbon and nitrogen in a nodulated and nonnodulated (NO3 – grown) legume. Plant Physiology 64, 1083 – 1088.Ryle, G. J. A., C. E. Powell, A. J. Gordon, 1978, Effect of source of nitrogen on the growth of Fiskeby soya bean: the carbon economy of whole plants. Annals of Botany (London) 42, 637 – 648.Seneviratne, G., L. H. J. Van Holm, E. M. H. G. S. Ekanayake, 2000, Agronomic benefits of rhizobial inoculant use over nitrogen fertilizer application in tropical soybean. Field Crops Research 68, 199 – 203.Smith, D., R. J. Bula, R. P. Walgenbach, 1986, Forage Management, 5th ed. Kendall Hunt Publishing Company, Dubuque, IA.Streeter, J., 1988, Inhibition of legume nodule formation and N2 fixation by nitrate. CRC Critical Reviews in Plant Science 7, 1 – 23.Wani, S. P., O. P. Rupela, K. K. Lee, 1995, Sustainable agriculture in the semi – arid tropics though biological nitrogen – fixation in grain legumes. Plant and Soil 174, 29 – 49.and symbiotic dinitrogen fixation in alfalfa. Journal of Plant Nutrition 17, 2053 – 2067.Dart, P. J., D. C. Wildou, 1970, Nodulation and nitrogen fixation by Vigna sinensis and Vicia atropurpurea, The influences of concentration form, and site of application of combined nitrogen. Australian Journal of Agricultural Research 21, 45- 65.Dazzo, F. B., W. J. Brill, 1978, Regulation by fixed nitrogen of host – symbiont recognation in the Rhizobium – clover symbiosis,.Plant Physiology 62, 18 – 21.Finke, R. L., J. E. Harper, R. H. Hageman, 1982, Eficiency of nitrogen assimilation by N – fixing and nitrate grown soybean plants (Glycine max). Plant Physiology 70, 1178 – 1184.Franco, A. A, J. C. Pereira, C. A. Neyra, 1979, Seasonal patterns of nitrate reductase and nitrogenase activities in Phaseolus vulgaris L. Plant Physiology 63, 421 – 424.Gan, Y., I. Stulen, F. Posthuimus, H. Keulen, J. C. Pieter, 2002, Effects of N menagement on growth, N2 fixation and yield of soybean. Nutrient Cycling Agrosyst 62, 163 – 174.Gan, Y., I. Stulen, H. Keulen, J. C. Pieter, 2003, Effect of N fertilizer top – dressing at various reproductive stages on growth, N2 fixation and yield of three soybean (Glycine max L. Merr.) genotypes. Field Crops Research 80, 147 – 155.George, T., P. W. Singleton, 1992, Nitrogen assimilation traits and dinitrogen fixation in soybean and common bean. Agronomy Journal 84, 1020 – 1028.Gibson, A. H., 1974, Consideration of the legumes as a symbiotic association, Indian National Science Academic Production 40B, 741 – 767.Harper, J. E., 1974, Soil and symbiotic requirements for optimum soybean production. Crop Science 14, 255 – 260.Imsande, J., 1988, Interrelationship between plant developmental stage, plant growth rate, nitrate utilization and nitrogen fixation in hydroponically grown soybean. Journal of Experimental Botany 39.Imsande,  J., 1989, Rapid dinitrogen fixation during soybean podfill enhances net photosynthetic output and seed yield. A 

Effect of N Fertilizer Application on Growth and Yield of Inoculated Soybean

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Effect of N Fertilizer Application on Growth and Yield of Inoculated Soybean

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