SU1486100A1 - Method of determining optimized doses of nitrogen-containing fertilizers - Google Patents

Description

The invention relates to agriculture and can be used to determine the optimal doses of ammonium fertilizers. The goal is to improve the accuracy of determining the dose. Plants are grown at different levels of ammonium fertilizers and during the growing season they determine the activity of the enzyme glutamine synthetase. With an increase in the dose of ammonium fertilizers, the activity of glutamine synthetase first increases, reaching a maximum, and then decreases -. sya The optimal dose of nitrogen fertilizers include the one that corresponds to the highest activity of the enzyme. 4 ill., 2 tab. _but

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The invention relates to forestry and agriculture and can be used to determine the optimal doses of ammonium fertilizers.

The purpose of the invention is to improve the accuracy of determination.

The method is as follows.

Plants are grown in the field in separate areas with the introduction of increasing doses of ammonium fertilizers. During the growing season, the activity of glutamine synthetase (HS) is determined. The highest activity of HS corresponds to the optimal dose of nitrogen fertilizers, at which the highest yield and low yield is achieved. nitrate content in plant material.

Figure 1 and 2 presents graphs of changes in the specific activity of HS (in

needles - 1, in the roots - 2) and the ammonia content (in the needles - 3 and in the roots - 4 pines); on fig.Z and 4 - graphs of changes in the specific activity of HS (in the leaves - A and roots - B barley).

Example 1 (for pine). The experiments were carried out in the training and experimental forestry of the Mari Polytechnic Institute for several seasons. The soil is sod-podzolic on top loam. The plots were under the layer of perennial grasses. Soil analysis: pH _x1 6.0-6.5; the content of P ^ O, according to the Kirsanov method, 8–15 mg / 100 g;

K _g 0 according to the Heive method 8-12 mg / 100 g soil. The ammonia content in the soil was determined by Nessler reagent, taking into account hygroscopic moisture. It ranged from 20-40 mg / 1 kg of dry soil. Sowing of seeds and agrotechnical cultivation complied with the norms

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adopted in forest nurseries. Doses - Kvr and Pby are selected on the basis of literary data. Field experiments were performed in triplicate, with a sequential arrangement of plots, the area of which was 4 m ² . Calculation of fertilizer doses was carried out according to GOST in terms of the active substance. Potassium sulfate, ammonium superphosphate was made according to the following scheme;

Option I Option II

one - without fertilizer 1a - K bo R in (Background) 2 “Ν · η> 2a - background + N46 3 “ _Μ Behind - background + Ν " four - ν ₄₀ 4a - background + Ν „ _σ five - Ν ₆₀ 5a - background + Ν " 6 “ ₄ ω 6a - background + N 450 7 7a - background + N4 (0 ' Ammonium sulfate was added to dissolve renno condition for 40 days grow up pine seedlings for day before pick out ·

ra samples at the rate of 4 liters of solution per 1 m ² . The concentration of ammonium fertilizer in the highest doses did not exceed 2%. In the control variants, instead of ammonium sulphate solution, 4 liters of water per 1 m ^{2 were used} . Other fertilizers were applied in a dissolved state. Moreover, their concentration did not exceed 0.5%.

Plants were collected at 8-9 am. The prepared samples of needles and roots were frozen at (-10) - (- 15) ^ 0 for 1 h and ground in the cold with quartz sand in a 1: 1 ratio. After extraction for 1 hour, 0.05 M Tris-HC1 buffer, pH 7.4 (1: 3 ratio), in the presence of 3 mM EDTA, 8 mM cysteine, 2 mM Mg80 ₄ , centrifugation was carried out for 3000 minutes at 3000 rpm The supernatant was used as an enzyme preparation. The activity of HS in the natural reaction of glutamine synthesis was determined by orthophosphate, formed in equimolar amounts with glutamine in the experimental mixture at pH 7.2 for 10 min at 37 ° C. The composition of the experimental mixture (0.8 ml): 12.5 mm IMIDOSAL - HCl buffer, pH 7>2; 5 mM ATP; 25 mM Mg50 ₄ ; 25 mM ΝΉ ^ ΟΙ; 62.5 W monoglutamate sodium. The reaction was stopped with 2 ml of 1% Fe8O ₄ > 711 ^ 0 in 0.3 n.

H _g 80 ₄ . Then 0 »2 ml of 6.6% is added (ΝΗ ^^ Μο ^ 0 ₄₄ , 41 ^ 0 of 7.5 n.

H ^ ZO ^ and received molybdenum blue, the color intensity of which is

at 690 nm on a photoelectric calorimeter.

The value of optical density was compared with a calibration curve. Control experiments differed in the absence of glutamate. Per unit of enzyme activity took the amount of enzyme catalyzing the formation of one

^ 0 micromolar orthophosphate for 10 min in

appropriate test mix. The specific activity of the enzyme was carried out in quadruplicate (analytical parallel). Determination of protein in extracts was carried out according to Lowry with the formulation of appropriate controls on the content of free amino acids.

Field experiments have shown that the enzyme activity increases with increasing doses of applied ammonium fertilizer, reaching the highest values in the wet season at a dose of Ν _βί . (figure 1). In the dry season, the high activity of the HS is shown at a dose of Ν ₄₀ - Ν ^ ο '(figure 2). At a dose of Ν _β0 and above, regardless of weather conditions, a decrease in the activity of the HS is shown, and at a dose of Ν ₄₆₀ · in the dry season, the absence of activity of the HS in both x and th and cor. yah, and in the wet - only in the roots

(Fig. 1, 2).

PRI me R 2 (for barley). The experiments were carried out on the basis of the Mari sovkhozatkhnikuma breeding area - small-plot (table 1) with an area of 6 m ² and in semi-production conditions (table 2) with an area of 150 m ² in triplicate. Fertilizers used were the same as in example 1, for

with the exception of potash, instead of potassium sulfate used potassium chloride. Data of agrochemical analysis of soils: the reaction of the environment is neutral, ΝΗ ₉ 4045 40 mg / 1 kg of dry soil; P _g Oy 1525 mg, K _g 0 17-25 mg per 100 g of soil. Collection of plants and determination of activity was carried out according to example 1.

The activity of the HS was determined in the roots and in the top sheet of barley varieties "Ray" in the process of vegetation in phases of development. The scheme of the experiments, the results for yield and grain quality are presented in Table 1.2. On fig.Z presents the results of determining the optimal dose of ammonium fertilizer on the activity of HS in the leaves (A) and roots (B) of barley grown on a small plot plot area,

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favorable weather conditions year. The highest activity of HS is shown when introduced before sowing (7th option I diagrams) + Ν # in the form of basal top dressing on shoots (5.7 ® option II diagrams) + Ν ^ ο top dressing extra root during the booting phase (7 variant III diagrams). Other options instead of the last top dressing spray- ιθ whether water. The high activity of HS in the leaves (A) and in the roots (B) of barley correlates with the high yield of barley obtained in the 7th variant in the favorable 15 year weather conditions (Table 1) in the calculation of rm ^{2 of the} reference area. Field experiments in the dry season in semi-production conditions showed that a high level of HS activity (Fig. 4) also correlates to 20 with the highest yield (Table 2) obtained by fractional application of ammonium fertilizers (options 4 and -6) and the largest percentage of total nitrogen in the grain (Table 1, 2). 25

Data on the content of nitrates (Table 1, 2) show that their number increases with an increase in the dose of ammonium fertilizers, but their content does not exceed 33% of the allowable rate.

Claims (2)

The method of determining the optimal doses of nitrogen fertilizers, including growing plants with different levels of nutrient availability and subsequent registration of the test substance produced in plants, according to the results of which judge the optimal doses, characterized in that, in order to increase the accuracy of determination, as a test substances use the enzyme glutamine synthetase, registration is carried out by measuring its activity, with the optimal dose of nitrogen fertilizers It corresponds to the high activity of glutamine synthetase. „Table 1 * Experimental options Average yield Total nitrogen in grain,% Content of nitrates in grain, mg / c'g g / m2% to control Without fertilizer (control 427 100.0 1.46 62.25 role) Background (Р40 К6 (^ 474 11.1 1.55 61.40 Background + Ν,) 00 (before sowing) 550 129.3 1.73 78.80 Background + Ν3 (, (before sowing) 634 148.2 1, 84 Background + Ν ^ „(before sowing + N30 (top-dressing root for shoots) + Ν2ο (extra root dressing in the stage of booting) 595 139.3 1.76 80.60 455 642 106.5 153.5 1.79 2.04 88.20 94 , 20 Blagop best weather conditions year 1486100 8 Table2 Experimental options Average yield c / ha% to control Total nitrogen in grain, 7. Content in grain, mg / kg 1. 2. 3. 4. No fertilizer 20.0 100.0 1, 44 68.7 (control) Background (Pe0K60) 20.4 102.0 1.65 66.9 Background + Ν // 0 (up to 19.3 96.5 2.00 70.0 VA) Background + Νδ0 (up to 21.6 108.0 2.08 73 ^ 5 va) (root dressing for seedlings) + Νίσ (foliar dressing in the pipe phase5. 6. Formations) background + Ν., (Up to '20.8 104.0 2.25 94.8 VA) Background + Ντο (up to 22.9 114.5 2.27 99.2 VA) + Ν4ο · (foliar dressing on shoots) + Ν10 (foliar dressing in the bobbing phase) -K Droughty weather conditions for the year 1486100 ', active activity Sch 1486100 I! “A I ^ o, sho § OM„ 12Ч6} 5, Fig. Do .1-1111, _ 1 I 46 35 _/ option experience (tm / ι. 2)