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EFFECT OF FERTILIZING PSEUDOGLEY SOIL WITH CAN ON NITROGEN CONTENT IN ROOT NODULE OF GLYCINE MAX

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  • Institute of Field and Vegetable Crops

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Due to excellent quality of its grain, the soybean (Glycine max (L.) Merr.) is in the fourth place among grown cultivated plant species, after wheat, corn and rice. A small-plot field trial was conducted in order to investigate the effect of fertilization of pseudogley soil with CAN on the mass and nitrogen content of root nodules in soybean. Pseudogley is an acid, water holding soil of low fertility, and the trial was performed in a humid year. Seed of soybean cv. Bachka was routinely coated with micro-biological preparation NS Nitragin. The CAN (calcium-ammonium nitrate), a widely used nitro-limestone fertilizer, is suitable for acid soils due to its pH-buffering effect. CAN was applied in three doses: 1) 0 kg ha-1 , control; 2) 50 kg ha-1 ; 3) 100 kg ha-1 , and 4) 150 kg ha-1. Maximum mass of root nodules was in control variant from 7.48 mg per nodule while the variant with 100 kg ha-1 N fertilizer was the minimum weight of nodules, 4.89 mg per nodule. The highest nitrogen content in nodules was in the variant with the application of 100 kg ha-1 and amounted to 3.46%, while the lowest content of nitrogen in nodules was in variant 2. Variability in the average nitrogen content (CV) in nodules and mass of nodule ranged 3.47% < Cv < 17.65%. Insignificant negative correlation was achieved between the weight of nodules and nitrogen content in nodules (r = 0.24).
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382
ECOLOGICAL CONDITION OF THE ENVIRONMENT AND THE SCIENTIFIC AND
PRACTICAL ASPECTS OF MODERN RESOURCE – SAVING TECHNOLOGIES IN
AGROINDUSTRIAL COMPLEX, International Conference on the
16-17.2.2017 Ryazan, Russian Federation.
ISBN 978-5-98660-279-0
Original Scientific paper
UDK 631.4
EFFECT OF FERTILIZING PSEUDOGLEY SOIL WITH CAN
ON NITROGEN CONTENT IN ROOT NODULE OF GLYCINE MAX
Popović M.V.1, Stevanović R.P.2, Vučković M.S.3, Radivojević D.M.3, Živanović R.Lj.3, Ikanović
N.J.3, Simic Ž.D.4, Bojović R.R.3
1Institute of Field and Vegetable Crops, Maksima Gorkog 30, 21000 Novi Sad, Serbia;
2Inspection Affairs Administration of Republic Srpska, Square 8, Banja Luka, Bosnia-Herzegovina;
3University in Belgrade, Faculty of Agriculture, Nemanjina 6, 11000 Belgrade, Serbia;
4 Institute of PKB Agroeconomik, Padinska Skela, 11000 Belgrade, Serbia;
*Corresponding authors: e-mail: bravera@eunet.rs; nikola.pavle@teol.net;
Key words: calcium-ammonium-nitrate, legumes, nitrogen-fixing Rhisobia, NS Nitragin, soybean
Summary: Due to excellent quality of its grain, the soybean (Glycine max (L.) Merr.) is in the fourth
place among grown cultivated plant species, after wheat, corn and rice. A small-plot field trial was
conducted in order to investigate the effect of fertilization of pseudogley soil with CAN on the mass
and nitrogen content of root nodules in soybean. Pseudogley is an acid, water holding soil of low
fertility, and the trial was performed in a humid year. Seed of soybean cv. Bachka was routinely coated
with micro-biological preparation NS Nitragin. The CAN (calcium-ammonium nitrate), a widely used
nitro-limestone fertilizer, is suitable for acid soils due to its pH-buffering effect. CAN was applied in
three doses: 1) 0 kg ha-1, control; 2) 50 kg ha-1; 3) 100 kg ha-1, and 4) 150 kg ha-1.
Maximum mass of root nodules was in control variant from 7.48 mg per nodule while the variant with
100 kg ha-1 N fertilizer was the minimum weight of nodules, 4.89 mg per nodule. The highest nitrogen
content in nodules was in the variant with the application of 100 kg ha-1 and amounted to 3.46%, while
the lowest content of nitrogen in nodules was in variant 2. Variability in the average nitrogen content
383
(CV) in nodules and mass of nodule ranged 3.47% < Cv < 17.65%. Insignificant negative correlation
was achieved between the weight of nodules and nitrogen content in nodules (r = 0.24).
Оригинальный Научный доклад
УДК 631.4
ЭФФЕКТ ВНЕСЕНИЯ УДОБРЕНИЙ PSEUDOGLEY ГРУНТ С CAN
ПО АЗОТНЫМ СОДЕРЖАНИЯ В КЛУБЕНЬКОВЫХ ДЛЯ GLYCINE MAX
Popović M.V.1, Stevanović R.P.2, Vučković M.S.3, Radivojević D.M.3, Živanović R.Lj.3, Ikanović
N.J.3, Simic Ž.D.4, Bojović R.R.3
1 Институт полевых и овощных культур, Maксима Горког 30, 21000 Нови Сад, Сербия;
2 Управление инспекции по делам Республики Сербской, Площади 8, Баня-Луке, Б. и Г.;
3 Университет в Белграде, Факультет сельского хозяйства, Nemanjina 6, 11000 Белград, Сербия;
4 Институт ПКБ Agroekonomik, Padinska Skela, 11000 Белградe, Сербия;
* Корреспондент авторы: Электронная почта: bravera@eunet.rs; nikola.pavle@teol.net;
Ключевые слова: кальций-аммиачная-нитраты, бобовых культур, азотфиксирующих Rhisobia,
Н.С. нитрагина, сои
Резюме: Благодаря отличным качеством зерна, сои (Glycine max (L.) Merr.) находится на
четвертом месте среди выращиваемых культурных растений, вслед пшеницы, кукурузы и риса.
Пробное поле небольшого участка было проведено с целью изучения влияния оплодотворения
pseudogley почвы с CAN на содержание массы и азота в корневых клубеньков сои. Pseudogley
является кислотой, удерживания воды почвы низкого уровня рождаемости, и исследование
было проведено во влажном году. Семян сои сорта Bachka был обычно покрыт
микробиологического препарата NS Нитрагина. CAN (нитрат кальция-аммония), широко
используемый нитро-известняк удобрение, пригоден для кислых почв из-за его рН-буферным
эффектом. CAN был применен в трех дозах: 1) 0 кг га-1, контроль; 2) 50 кг га-1; 3) 100 кг га-1 и 4),
150 кг га-1.
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Максимальная масса корневых узелков был в контрольном варианте, от 7,48 мг узелка-1 в то
время как вариант с 100 кг га-1 азотных удобрений был минимальный вес узелков, 4,89 мг
узелка-1. Самое высокое содержание азота в узелков в варианте с применением 100 кг га-1 и
составила 3,46%, в то время как низкое содержание азота в узелков был во втором варианте.
Незначительная отрицательная корреляция была достигнута между весом узелков и содержания
азота в узелков (г = 0.24).
Introduction
Soybean (Glycine max (L.) Merr.) production is increasing for decades, reaching over 100
millions of hectares worldwide (Faostat, 2015). Due to excellent quality of grain (about protein 40%
and oil 20%), soybean is in fourth place among the grown cultivated plant species, after wheat, corn
and rice. Soybean protein accounted for about 2/3 of the world production of plant proteins while 1/3 of
vegetable oil comes from soybean. The world's largest producers of soybean are the United States, with
a 29% share of total world production, followed by Brazil with 24% and Argentina with the
participation of 17% of the world's surface. Soybean was grown in Europe at around 450.000 ha.
Average perennial soybean yields in Serbia are at the level of average world yield and they are 2.5tha-1.
High and stable yields of soybeans are achieved by selecting varieties of high genetic potential's and
other agronomic properties and correct application of growing technology. The Institute of Field and
Vegetable crops from Novi Sad has developed 136 high yield varieties of soybean. An important
condition for achieving of high and stable soybean yield is correct fertilization and inoculation of
soybean seed by microbiological fertilizer Nitragin before sowing [1, 2, 3, 4, 5, 6].
Bradyrhizobium japonicum, Bradyrhizobium Elkani and Sinorhizobium fredi live in symbiosis
with soybean and form nodules [7]. Nodule bacteria in symbiosis with plant create active nodule-
bumps (biological nitrogen factory) at its root, and by that fixate up to 180 kg N ha-1 from the air per
year [8]. The strains of Rhizobia are mesophilic microorganisms. The optimal temperature for growth
and multiplication of Rhizobium is about 25 °C. It does not grow below 10°C or above 37°C [9].
Optimal humidity for nodules to form is 60-70% of full water capacity of the land. In the upper layers
of soil, where it is good aeration and humidity, a larger number of nodules are forming while in the
layer below 30 cm nodule formation is negligible [10]. Soil pH is among the most important
environmental factors that influence the presence and effectiveness of Rhizobium strains. The
representation of these microorganisms is very low in acid soils (pH <5.5) which results in lack of
nodulation and significantly reduce dry matter yield [11, 12]. Due to the high sensitivity of nodule-
385
bacteria on the pH value, the nitrogen fixation does not commes to the fore [13]. By reducing the
acidity microbial activity increases in pseudogley soil [12, 14, 15]. Nodulation is estimated using a
nodulation index scale based on number of nodules per plant: for 0-5 nodules - poor nodulation; 6-10
nodules, medium nodulation; 11-15 nodules, good nodulation; 16-20 nodules, very good nodulation,
and, with over 20 nodules-excellent nodulation (http://vasatwiki.icrisat.org). Root nodulation is a good
indicator of successful symbiosis between legume host-plants and their Rhizobium/Bradyrhizobium
bacteria. The potential for symbiotic nitro-fixation is estimated by symbiotic nitro-fixation indicators,
namely: the number of nodules, nodule weight, mass of above-ground part of the plant, the nitrogen
content in above-ground plant parts and [12, 15, 16].
The aim of this study was to investigate the effect of applying CAN fertilizer on nodules mass and
nitrogen content in soybean nodules, and estimate effectiveness of symbiosis between plants i
Rhizobium/Bradyrhizobium. The results are expected to improve modern technology of soybean
production.
Material and methods
R A small-plot field trial was carried out in order to investigate the influence of fertilizer CAN
on soybean nodules and contents of nitrogen in the nodules of soybean plants at the site of Brezovo
Polje in Brcko, on pseudogley type of soil. The main plot area was 10.8 m2 (6.0m x 1.8m), while
billing plot area was 5.4 m2. The variety Bachka (Bačka in Serbian; maturity group 0), created at the
Institute of Field and Vegetable crops in Novi Sad, was tested. Density of crops was the same for all
variants and amounted to 500,000 plants per hectare. Experiment was set as one-factorial, split plot
method (split-plot) with four replications. During experiment a standard technology for growing
soybean was applied, excluding studied factors. The preceding crop was corn. Soybean preceding crop
was corn. Deep plowing (25 cm) was carried out in the autumn directly after corn harvesting. Seedbed
preparation was carried out in the spring by cultivator. Sowing was done at beginning of April. Just
before sowing, according to the anticipated fertilization plan, the CAN fertilizer was applied and the
seed was inoculated by microbiological agent - NS Nitragin. Researches were conducted with the aim
to investigate the effect of nitrogen fertilization and NS Nitragin on the mass of nodules and nitrogen
content in soybean nodules, on pseudogley type of soil in a humid year. There were four variants of the
applied doses of CAN fertilizer: 1) 0 kg ha-1 in the control; 2) 50 kg ha-1; 3) 100 kg ha-1, and 4) 150 kg
ha-1. Starting from the moment of nodules forming, in 15 days intervals, 10 plants samples were taken
386
per every repetition, from which the nodules number, weight and nitrogen content were determined.
Laboratory analyzes of nitrogen content in soybean nodules were made by the method of Kjeldahl [13,
16, 18]. The results are processed by mathematic-statistical method of analysis of variance and
evaluation of the difference by LSD test. An evaluation of the correlations between the studied
parameters is provided. The results are shown graphically and in tables.
Soil conditions
The experiment was conducted on pseudogley type of soil. The soil had very acid reaction, low
humus, poor calcareous, medium provided with P2O5 and well provided with K2O (table 1).
Dissolubility and accessibility of microelements in the soil is mostly influenced by pH reaction
of soil, content of organic matter, mechanical composition of the soil (quantity of clay fraction),
content of calcium carbonate and accessibility of phosphorus in the soil. Production value of
pseudogley is small and soybean can be successfully grown only with application of melioration
measures along with adequate regime of fertilization which is compatible to the need of soybean on
pseudogley [18, 19].
Table 1. Agrochemical properties of soil, Brčko, Bosnia and Herzegovina
Parameters
soil type pH CaCO3, % Humus, % P2O5, mg/100g K2O, mg/100g
Pseudogley 4.30 0.60 2.88 14.30 24.50
Meteorological conditions. Meteorological conditions greatly influence the success of crop
production [19]. Meteorological data were obtained from the meteorological station, Brčko, Bosnia-
Hercegovina (graph. 1). In the year of the study, recorded temperatures were in average of 18.2oC,
while precipitation during the vegetation period was 930 mm, and was higher than the long-term
average of 502 mm (Graph. 1).
Quantity and distribution of rainfall during the growing season are of great importance for the
success of soybean production. Total water needs of soybean plants are 450-480 mm, daily needs are 1-
5mm, and in some months are: April 10-40 mm; May 30-60 mm; June 90-110 mm; July 100-125 mm;
August 100-125 mm; September 50-80 mm [1, 5, 6].
387
za Brčko u 2001. godini
0
20
40
60
80
100
120
140
160
180
IV V V I VII VIII IX
M onths
0
40
80
120
160
200
240
280
320
360
Temperature
Precipitation
Average
long year
438,3 mm
Average
long year
17,6 0C
Temperature,
0
C
Precipitation (mm)
Arid period
Humid period
Graph. 1. Air diagrams according to Walter, humid year in Brčko, Bosnia & Herzegovina
RESULTS AND DISCUSSION
Mass of nodules per plant is an indicator of nodulation ability of nitrogen fixation. Differences
of the values of nodules mass between fertilization variants were statistically significant (table 2 and 3).
The weight of nodules on soybean on pseudogley decreased monotonically with increasing
amounts of nitrogen quantity. Largest nodule was in control variant – 7.48 mg, while in the variant with
100 kg ha-1 of applied fertilizer was the minimum weight of nodule – 4.89 mg. Statistically significant
big mass of nodules had control variant in relation to all tested variants (table 2 and 3).
Table 2. Effect of fertilization on the mass of nodules and nitrogen content in soybeans nodules
Fertilization
Parameter Control 50 100 150
X
Change
of rate %
Cv
%
Std.
Dev.
Mass of soybean
nodules, mg/nodul 7.48 6.48 4.89 5.80 6.13 -9.95 17.65 1.01
Nitrogen content of
soybean nodules, % 3.25 3.08 3.46 3.26 3.23 1.03 3.47 0.24
Parameter Mass of soybean nodules Nitrogen content of soybean nodules
0.5
0.568 0.424
LSD 0.1
0.825 0.617
388
Table 3. Anova for tested parameter
Univariate Results for Each DV
Sigma-restricted parameterization
Effective hypothesis decomposition
Degr. of
Freedom
Mass of nodule
SS
Mass of nodule
MS
Mass of nodule
F
Mass of nodule
p
Intercept
Nutrition
Error
Total
1
456,3333 456,3333
5047,005
0,000000
3
10,7183
3,5728
39,515
0,000038
8
0,7233 0,0904
11
11,4417
Univariate Results for Each DV (Rossia Soya stat)
Sigma-restricted parameterization
Effective hypothesis decomposition
Effect
Degr. of
Freedom
Nitrogen
content in
nodule
SS
Nitrogen
content in
nodule
MS
Nitrogen
content in
nodule
F
Nitrogen
content in
nodule
p
Intercept
Nutrition
Error
Total
1
127,7269 127,7269 2523,000
0,000000
3
0,2406 0,0802
1,584
0,267636
8
0,4050 0,0506
11
0,6456
The weight of nodules was decreased when the amount of nitrogen increased to the variants
with 100 kg ha-1- 4.89 mg and started to grow again in the variant with 150 kg ha-1 - 5.80 mg (table 2).
Content of nitrogen in soybean nodules, on average, amounted to 3.23%. The highest content of
nitrogen in nodules was in the variant with the application of 100 kg ha-1 and amounted to 3.46%, while
the lowest nitrogen content in nodules was with the application of 50 kg ha-1, in the second variant
3.08% (table 2 and 3, Graph. 2b).
Nutrit ion; Weighted Means
50 100 Control 150
Nutrit ion
4,0
4,5
5,0
5,5
6,0
6,5
7,0
7,5
8,0
8,5
9,0
9,5
Mass of nodule
Nutriti on; LS Means
50 100 Control 150
Nutriti on
2,6
2,7
2,8
2,9
3,0
3,1
3,2
3,3
3,4
3,5
3,6
3,7
3,8
3,9
Nitrogen content in nodule
a. b.
Graph 2. Effect of nutrition of mass of soybean nodules a, and content of soybean nodules, b.
389
Variability in the average of nitrogen content in nodules and mass of nodules ranged from 3.47%
<CV <17.65% (table 2).
Correlations of studied properties. Correlations express the degree of dependence between
the properties. It is very important to understand better a functional, morphological or physiological
connection of morphological soybean productive characteristics. Information of morphological
variability of productive properties of soybean and their method of use are very important for breeding
program development [3, 5, 6]. Negative insignificant correlation connection was achieved between the
weight of nodules and nitrogen content in nodules, r = 0.24 (table 4).
Table 4. Correlations of tested traits
Variable Mass of soybean nodules Nitrogen content of nodules
Mass of soybean nodules 1.00 -0.24
Nitrogen content of soybean nodules -0.24 1.00
The weight of nodules and nodules number are representing indicators of efficiency of nitrogen
fixation [20, 12, 15]. The average number of nodules per soybeans plant move up to several hundred
[21] and depends on the water content in the soil, soil pH, temperature, mineral nutrition with nitrogen,
salinity, but also the variety and strain B. japonicum [22, 23, 24]. By application of sowing inoculation
of soybean seed with Nitragin, maximum benefit of natural process of symbiotic nitrogen fixation in
the cultivation of soybeans is achieved, and thus provides environmentally acceptable and economical
reasonable production [12].
Conclusion
According to the established results of research, the following conclusions were made:
Maximum mass of nodules was in control variant, from 7.48 mg nodule-1 while the variant with
100 kg ha-1 N fertilizer was the minimum weight of nodules, 4.89 mg nodule-1.
The highest nitrogen content in nodules was in variant with the application of 100 kg ha-1 and
amounted to 3.46%, while the lowest content of nitrogen in nodules was in 2nd variant.
Variability in the average nitrogen content in nodules and mass of nodule ranged from 3.47%
<Cv <17.65%.
Insignificant negative correlation was achieved between the weight of nodules and nitrogen
content in nodules (r = 0.24).
390
Production value of pseudogley is small and on such soil of low production value, soybean can
be successfully grown only with application of melioration measures along with adequate regime
of fertilization which is compatible to the need of soybean on pseudogley.
Acknowledgements. This study was results of research conducted within the projects: ТР 31022;
31025 and 31078; financed by the Ministry of Education, Science and Technology Development of
Republic of Serbia.
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Competing interest. The authors declare that they have no financial or personal relationship which
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The symbiotic relationship between the soybean plant and rhizobium results in fixation of atmospheric nitrogen (N(2)) in the root nodules, with the result that nitrogenous fertilization of the soybean is unnecessary. The effectiveness of nodule formation and N(2) fixation with rhizobial strains is under genetic control with two general categories identified: (1) promiscuous, which produces functional nodules with cowpea-type rhizobial strains; and (2) nonpromiscuous, which forms no or nonfunctional nodules with these strains. The segregation pattern of this promiscuity trait was studied using nodule dry weight (NDW) and leaf color score (LCS) as indicators of N(2) fixation effectiveness. Individual plants in each of six populations [P(1) = nonpromiscuous, P(2) = promiscuous, F(1) = P(1) x P(2) (and the reciprocal cross), BC(1)(P(1)) = F(1) (female) x P(1), BC(1)(P(2)) = F(1) (female) x P(2), F(2)] were scored for these characters after inoculation with a rhizobial strain that would distinguish between both types. For NDW, nonpromiscuity was found to be partially dominant (h/d = 0.37), controlled by four loci. For LCS, nonpromiscuity was shown to be almost completely dominant (h/d = 0.74), controlled by two loci. LCS was a more meaningful estimate of N(2) fixation because it represented the total effectiveness of nodulation to provide nitrogen for the plant.
Influence of agro-technical and agro-ecological practices on seed production of wheat, maize and soybean
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Popović, V. Influence of agro-technical and agro-ecological practices on seed production of wheat, maize and soybean. Doctoral thesis, University of Belgrade, Faculty of Agriculture, Zemun, 2010. 55-66.
Proizvodnja soje (Glycine max) u svetu i kod nas. XXI Savetovanje o biotehnologiji sa međunarodnim učešćem
  • Živanović Lj
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Živanović Lj. & Popović V. Proizvodnja soje (Glycine max) u svetu i kod nas. XXI Savetovanje o biotehnologiji sa međunarodnim učešćem, Čačak, 2016. vol. 21 (23), 129-135. In Serbian.