RU2314349C2 - Method for evaluating ecological agrobiocenosis condition in anthropogenic influence zone - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves concurrently taking Scotch pine needle samples growing on case and control test areas, determining chlorophyll concentration and catalase activity therein. Metabolism intensity coefficient is calculated next in case and control samples as ratio of chlorophyll concentration and catalase activity. The comparison results are interpreted in terms of anthropogenic influence degree.

EFFECT: high result data reliability; low costs and labor inputs level.

3 cl, 5 tbl

Description

The invention relates to the ecology of sanitation of forest agrobiogeocenoses and is intended for sanitary-hygienic testing of the state of forest ecosystems and predicting the dynamics of their development in areas of technogenic pollution with pollutants of various chemical nature, including waste from poultry farms and pig farms.

The well-known "Method for determining and predicting the sanitary and hygienic condition of the soil in the industrial pig breeding zone" (see RF Patent No. 2129160, C12Q 1/02. Publ. 20.04.99. Bull. No. 11). With good reliability of the results in this way.

This method does not reflect the state of the main components of forest ecosystems - plants growing in a contaminated area.

It is also known that environmental pollution most strongly affects conifers. Characteristic signs of environmental troubles and atmospheric gas composition are the appearance of chlorosis and necrosis, a decrease in the size of needles, shoots of the current year and past years, their thickness, size and number of established buds. There is a thickening of the needles themselves, their life span is reduced. All these signs together give a fairly objective picture. Conifers are convenient in that they can serve as bio-indicators year-round. In addition, the use of conifers makes it possible to carry out bioindication in large areas. In forestry, a system for assessing the state of the environment by a set of coniferous characters has been developed, not only morphological indicators are used, which are very variable, but also a number of biochemical changes. In the system, a special place is occupied by the main indicator of assimilation processes in vegetative organs - the level of chlorophyll, but in the needles of common pine, growing in technogenic zones, this indicator can vary significantly, and it is not always lower in pollution zones compared to that in trees growing in environmentally friendly areas. The latter is due to the compensatory mechanisms of homeostasis, which ensures the survival of plants in adverse conditions. The mechanisms of homeostasis are primarily manifested in the intensity of metabolic processes, and therefore, additional refinement indicators are needed to increase the accuracy of forecasts for assessing the state of anthropogenic impact on forests. The objective of the invention is to develop a method for further assessing the prediction of the state of agrobiogeocenoses, in particular, in areas of contamination by airborne pollutants of industrial and poultry facilities, with which it would be possible to test agroecosystems operating in contaminated natural environments. At the same time, analyzes should be carried out in the conditions of existing chemical laboratories, using specialists of average qualification on conventional laboratory equipment and when obtaining results of high reliability with minimal labor and funds for analysis.

The problem is solved in that, as an express test analysis, the activity of the catalase enzyme is additionally determined in needles samples taken in parallel in areas of anthropogenic impact and a conditionally clean plot located in the same natural conditions. The chlorophyll content is determined in the same samples, after which calculations are carried out, determining the ratio of the chlorophyll content in pine needles to the catalase activity indicators in them, which indirectly indicate the ratio of assimilation and dissimilation in tissues (the intensity of the metabolic process in them).

The novelty of the invention is that it is proposed to determine the degree of adaptation of trees to the anthropogenic load not by changing individual indicators of homeostasis, but by changing the ratio of the most significant indicators of the latter.

It is proposed to evaluate the metabolic rate in the assimilation organs of Scots pine by comparing data on the processes of accumulation of organic matter and decomposition of the latter. Moreover, the level of chlorophyll content in it, which indirectly reflects the rate of accumulation of organic matter, is used as an objective indicator of the process of assimilation in the pine needles. It is proposed to judge the degree of compensation of the oppositely directed process of organic matter dissimilation in the pine needles of various technogenic territories by the activity of oxidoreductase enzymes involved in decomposition processes. In particular, the enzyme catalase involved in the decomposition of peroxide compounds formed during anabolism in the tissues of plant assimilation organs. At the same time, the activity indicators of catalase in the needles of common pine were used as bioindicators on the degree of influence of aerogenic pollutants on the agrobiogeocenoses of technogenic zones.

Moreover, when researching, it is necessary to take into account the hydrothermal conditions of the growing season, which are different in different years. Therefore, to conduct an objective assessment of the influence of anthropogenic effects on the level of metabolic processes in the needles of pine, an additional coefficient was introduced that shows the ratio of the processes of assimilation and dissimilation in the needles of pine.

The ratio of the indicator of chlorophyll content (mg / 1 g) in pine needles to the activity index of catalase (cm ³ O ₂ ^1g / _2min ) in the same substrate was taken as the coefficient of metabolic intensity (KIM).

Where:

a - the numerical value of the content of chlorophyll in 1 g of absolutely dry needles;

c - the value of catalase activity in 1 g of absolutely dry needles.

The introduction of the metabolic coefficient in the tissues of the assimilation organs of Scots pine makes it possible to reveal in them a regular increase in the intensity of metabolic processes, which characterizes the focus of homeostasis on the restoration of disorders in biological systems caused by the influence of aerotechnogenic pollutants.

The method is as follows.

Trees for sampling are planned at the test site. Branches of coniferous trees of the same breed and of the same age are cut at a height of at least 2 m from a part of the crown adjacent to areas with polluted air. Sampling on the test area (PP) is carried out at five points located diagonally, in triplicate on each tree. The indicated parameters were adopted based on the results of numerous practical experiments as ensuring the required forecast accuracy.

The samples are packaged and delivered to the laboratory for analysis of the content of chlorophyll in the needles and the activity of the catalase enzyme, which is determined by a gasometric method based on the oxygen released as a result of the catalytic effect of the chopped needles tissue on hydrogen peroxide using a catalase in which 1 g is placed test needles, 500 mg of calcium carbonate, a glass with 5 ml of 3% hydrogen peroxide and tightly closed with a rubber stopper connected to the burette (see analysis).

The following reagents are used to determine catalase activity by a gasometric method:

1. 3% hydrogen peroxide solution;

2. CaCO ₃ .

Analysis progress: Add a sample of ground needles of 1 g to a 100 ml flask, add 0.5 g of CaCO ₃ . Carefully put a glass of 5 ml of a 3% hydrogen peroxide solution on the bottom of the flask. Close the flask tightly with a stopper having a tube connected through a tee equipped with a clamp or tap to the burette. The latter is reported with a pear. The burette and pear are filled with water. The water level in the burette and pear is balanced and the pear is fixed at a certain height. Close the tap. It is necessary to ensure that the water level in the burette remains motionless, which indicates the achievement of temperature equilibrium in the device and the room.

The start of the experiment is noted by the stopwatch at the moment when the glass with hydrogen peroxide is knocked over and after that the contents of the flask are shaken. Shaking the mixture should be continued during the experiment, without touching the flask directly with your hands. The evolved oxygen displaces water from the burette, the level of which must be noted. The needles sterilized by dry heat are the control.

The amount of molecular oxygen released is taken into account at a temperature of 18-20 ° C after 30 s for 1-2 minutes.

Catalase activity is expressed in milliliters of oxygen released per 1 g of needles over a certain period of time (2 minutes).

Definition error is allowed up to 5%.

The method of determining chlorophyll is photometrically based on the extraction of chlorophyll from a sheet with solvents (alcohol, acetone) and its determination on a photoelectrocolorimeter or spectrophotometer. Determination of chlorophyll in pine needles is carried out on fresh material.

For work, take a sample of 1 g of needles. Pre-determine the humidity of the needles. A portion of the plant material is carefully ground in a porcelain mortar with broken glass, adding chalk or magnesium carbonate. Extraction from plant material is carried out with 90% alcohol.

A solvent is added to the pulverized plant material and the material continues to be ground together with the solvent.

In a Bunsen flask, a glass filter is strengthened in the plug hole (the diameter of the filter must correspond to the amount of material to be studied). The flask is connected to the pump and the liquid is sucked out. The solvent is poured into the mortar and triturated again for a minute, then the whole plant mass is transferred to the filter, compacted with a stick and sucked off. Rinsing is carried out until the flowing solution becomes colorless. Then the extract is transferred to a 50 ml volumetric flask, rinsing several times with a Bunsen flask and pouring into a volumetric flask. The hood is brought to hell with a solvent.

The colorimetric solution is carried out on a photoelectric colorimeter with a red light filter (FEK-56M). If the liquid is colored intensely green, it must be diluted, since at high concentrations the values on the FEK can go beyond the resolution of the device.

To convert chlorophyll to standard values, a Getry solution is used, which is prepared as follows:

1) 1% solution of CuSO ₄ · 5H ₂ O (take only blue crystals),

2) 2% solution of K ₂ Cr ₂ O ₇ ,

3) 7% ammonia solution (for 7 ml of 18% ammonia you need to take 11 ml of water).

To make a standard, solutions are accurately measured in a 100 ml volumetric flask (CuSO ₄ · 5H ₂ O - 28.5 ml, K ₂ CrO ₇ - 50 ml, NH ₄ OH - 10 ml), adjusted to the mark with distilled water and mixed.

Getri's solution is colorimetrically equivalent in color to a solution of crystalline chlorophyll, with a concentration of the latter of 85 mg / L.

A calibration curve is constructed by diluting a standard solution, where the chlorophyll content (mg / l) is plotted along the abscissa, and the optical density is plotted along the ordinate. A calibration curve is constructed from 0.085 mg / L (1 ml of the stock solution and 99 ml of water) to 7.65 mg / L (90 ml of the stock solution and 10 ml of water).

The measurements on the FEK are performed several times, calculating the average. According to the data obtained, the concentration of chlorophyll in the test samples is determined by the calibration curve. The percentage of chlorophyll is then calculated.

The proposed determination of metabolic rate (CIM) in the tissues of pine needles is carried out according to the above formula.

Example 1. Assessment of the state of agrobiogeocenoses in the industrial pollution zones of the Polevskoy cryolite (PCZ) and Sredneuralskiy smelter (SUMZ) plants at a distance of 1.5-2.0 km from the pollution sources towards the prevailing wind rose (north-east direction). Control: 26 km from the PKZ and 24 km from the SUMZ.

Needle samples were taken in accordance with the above rules in 3 replicates.

1. The results of a study evaluating the state of agrobiogeocenoses in the zones of influence of SCZ and SUMZA by determining the activity of catalase and the content of chlorophyll in pine needles in trees growing near plants were analyzed by comparing the data obtained in the studied and control sample plots (Table 1) . The degree of degradation of agrobiogeocenoses was determined by the change in these indicators relative to the control, due to the adaptation of woody plants to pollutants. The data obtained (Table 1) indicate an increase in the intensity of adaptation processes with increasing pollution.

Table 1 The results of determining the indicators of homeostasis in pine needles at different levels of pollution with aerotechnogenic pollutants Indicators Service counter Impact zone PKZ Impact zone SUMZ Metabolism ratio * Needles age 1 year of life 2 years of life 1 year of life 2 years of life 1 year of life 2 g of life PKZ
1 g. 2 g. SUMZ
1 g. 2 g. Chlorophyll "a", mg / g 2.01 1.7 1.9 1,58 1.85 1.45 0.94 0.93 0.9 0.85 Catalase, cm ³ About ₂ ^1g / _2min 26,4 31 20,4 22.0 16.5 17.5 0.81 0.71 0.63 0.56 Kim 0,076 0,055 0,093 0,072 0,112 0,083 1.22 1.31 1.47 1.51 CMM increase with increased pollution 22% 31% 47% 51% * The ratio of the indicators of the needles metabolism of 1 and 2 years of life in the impact zones to the indicators on the conditionally control PP **. ** Conditional control trial area (PP) at a distance of 26 km from the PKZ. Impact zone PKZ - at a distance of 1.5-2 km from the plant. Impact zone PRPU - at a distance of 2 km from the SUMZ, Mount Magnitka.

Thus, according to the data presented (in Table 1), the content of chlorophyll in the needles of Scots pine in the impact zones is 85-94% of that in the conditionally control trial area (PP). Catalase activity is also reduced in impact zones compared with the conditionally control trial area and makes up 56-81% of the enzyme activity in the needles of common pine in trees growing in more favorable environmental conditions. However, the ratios of these indicators (CIM) indicate an increase in metabolic processes in the zones of technogenic pollution compared with that in the conditionally control trial area, and the higher the level of pollution, the more significant the differences in the metabolic rate in technogenic zones compared with the control. Therefore, to assess the state of pine agrobiogeocenoses, it is more expedient to use coefficients derived from observations as bioindicators of environmental pollution: ratios of indicators reflecting the vital processes of biological systems (proposed by KIM).

The advantage of the introduced bioindicator (CMM) compared to previously used ones (chlorophyll, catalase, polyphenol oxidase and others) is a more objective assessment of the state of assimilation organs of Scots pine in technogenic zones than by individual indicators. So, the values of this coefficient given in Table 1 show a decrease in the metabolic rate in needles for 2 years compared with needles for 1 year (which is natural for all older biological systems), while for some indicators (catalase activity and chlorophyll content) this the pattern is not expressed clearly enough.

In addition, KIM allows one to substantiate a reduction in the life span of pine needles in impact zones (Table 2) by an increase in metabolic processes in it. Therefore, this indicator has prognostic value for the functioning of the assimilation organs of Scots pine in technogenic zones, since a negative correlation rxy = -0.8 between the life span of the needles and CMM was reliably revealed.

table 2 The life span of pine needles on trial plots with different levels of anthropogenic load, metabolic rate coefficient (CIM) Indicators Conditionally control. trial Area Impact zone PKZ Impact zone SUMZ Will continue. of life 3.0-3.5 2.0-2.5 1.5-2.0 KIM needles 1 year of life. 0,076 0,093 0,112

Thus, the studies allow us to draw the following conclusions.

1. Based on the analysis of data on metabolic rates in pine needles, a new original approach to the bioindication of the environment is proposed to determine the coefficients that reflect the main metabolic processes in the tissues of the assimilation organs of trees growing under conditions of industrial pollution.

2. The use of the introduced metabolic intensity coefficient (CIM) for assessing the state of metabolism in the tissues of the assimilation organs of Scots pine reveals a decrease in vital activity in the needles of 2 years of life compared to 25.5% on average in the needles of 1 year of life.

3. The study of metabolic processes in the needles of trees of common pine populations growing in the technogenic territories of the Sverdlovsk Region, by determining the metabolic intensity coefficients (CMM), revealed the highest values of these indicators in the impact zone of the SUMZ with a high level of technogenic pollution.

4. The results of determining the increase in the metabolic rate in the tissues of common pine in areas of technogenic pollution with an increase in the level of the latter are consistent with data on the life expectancy of needles in the impact zones of the SUMZ and SCZ *, which makes it possible to recommend a metabolic rate coefficient (CMM) for biotesting the anthropogenic load on forest ecosystems and predicting their condition (* there is a close negative correlation between the life span of the needles and CMM in the needles of 1 year of life r = -0.8).

Example 2. Determining the state of agrobiogeocenoses in the zone of influence of the Federal State Unitary Enterprise PPZ Sverdlovsky at various distances of 10-1000 meters from it.

The anthropogenic impact of airborne pollutants of poultry enterprises on the state of the environment can be judged by various indicators obtained by bioindication methods, during which the control point is taken outside the zone of significant pollution of the agricultural enterprise.

Processing of taken needle samples according to the proposed method for assessing the state of agrobiogeocenosis and using standard methods was carried out in the laboratory of the Department of Ecology and Pet Hygiene of the Ural State Agricultural Academy of Yekaterinburg. The results of studies after mathematical processing are summarized in tables 3 and 4.

Studies to determine the content of chlorophyll "a" and the activity of the catalase enzyme in the tissues of pine needles in the area of activity of the Federal State Unitary Enterprise PPZ Sverdlovsky were carried out in winter, spring and autumn at different distances as they moved away from the source of pollution, taking into account the prevailing winds in this area (southwest direction).

The results of determining the indicators of catalase activity and the content of chlorophyll in the needles of common pine in trees growing in the Sverdlovsk region, in the forest of the Sysert district, which, according to regionalization according to the severity of the ecological situation, refers to areas with a conditionally satisfactory environmental situation. An exception is the results of determining the chlorophyll content in the needles of trees in autumn: at the end of the growing season, a significant increase to 1.6% was observed, which exceeds the upper limit of fluctuations in the chlorophyll content in the needles of trees, indicated in the literature.

Data on the chlorophyll content (Table 3) in the needles of trees growing on test plots in the zone of influence of Sverdlovsky FSUE PPZ in winter is either reduced compared to the lower limit of the value range (0.3-1.3% according to published data) of the chlorophyll content in pine needles, or slightly exceed this lower limit (0.3%) in all, except for the control, trial plots.

In the spring, the content of chlorophyll in the needles of Scots pine at the test plots under study in the influence zone of the poultry plant at different distances from it was significantly higher than in winter and amounted to 0.52-0.76%. The highest levels of chlorophyll in this period, as well as in winter, were found in the needles of trees growing on the control trial area (5 km in the south-west direction) and exceed the data obtained on trial areas in the zone of influence of the pollution source 1.1-1.6 time.

In the autumn period, the content of chlorophyll in the needles of trees was the highest in all test areas, and in the test areas located on the leeward side (south-west direction) with respect to the source of pollution, its level exceeded the upper limit of fluctuations indicated in the literature.

Thus, the studies showed that the process of photosynthesis is carried out by the assimilation organs of Scots pine especially actively at the end of the growing season, its intensity decreases sharply in winter and increases by spring.

Table 3 The results of photocolorimetric determination of chlorophyll in needles 2 years of life Place of sampling in relation to the source of pollution Chlorophyll content based on abs. dry mass of needles (%) Direction Distance December 2003 March 2004 September 2004 NW 10 m 0.2 ± 0.018 0.52 ± 0.050 0.78 ± 0.076 50 m 0.15 ± 0.013 0.54 ± 0.052 0.92 ± 0.091 200 m 0.13 ± 0.011 0.52 ± 0.051 0.81 ± 0.82 500 m 0.33 ± 0.032 0.6 ± 0.058 0.57 ± 0.058 S-z 10 m 0.18 ± 0.015 0.55 ± 0.053 1.44 ± 0.120 50 m 0.11 ± 0.010 0.60 ± 0.059 1.5 ± 0.140 200 m 0.37 ± 0.035 0.52 ± 0.052 1.14 ± 0.110 1000 m 0.36 ± 0.035 0.63 ± 0.062 1.6 ± 0.160 The control 5000 m 0.48 ± 0.046 0.86 ± 0.087 1.6 ± 0.159

The most favorable conditions for the functioning of the assimilation organs of Scots pine were identified according to the data obtained in the control trial plot, where during all periods of observation the readings of chlorophyll in the needles of Scots pine are highest. Adverse conditions for the assimilation process were found already at a distance of 200 m from the source of pollution in both the north-west and south-west directions.

Another objective method for assessing the physiological state of the assimilation organs of trees growing under anthropogenic conditions can be an enzymatic analysis of pine needles. Under favorable growth conditions, any plants, as a rule, have stable indicators of enzymatic activity, and in varying - on the contrary, the range of fluctuations of the values of the indicators is higher, the stronger is any effect. Thus, an increase in the activity of oxidoreductases in the needles' tissues indicates deep violations of the most important physiological processes in them, the formation of peroxides, as well as significant changes in redox reactions (N.M. Shebalova, L.G. Babushkina, 1999; O.P .Neverova, 2003).

Studies to determine the activity of catalase in pine needles showed that the highest values of enzyme activity were in the winter season (Table 4). The revealed regularity is due to the attenuation of assimilation processes with a predominance of anabolism (destruction of organic substances of dissimilation, accumulated during the vegetative period by assimilation processes of organic substances: sugar, ascorbic acid, etc.

Table 4 The results of the determination of the catalase enzyme in the needles of common pine at different distances from the federal state unitary enterprise PPZ Sverdlovsky Sampling location Distance from PPZ \ on the right. The activity of the enzyme catalase in terms of abs. dry needles weight (cm ³ O ₂ ^1g / _{2 min} ) December 2003 March 2004 September 2004 Needles 1 g. Needles 2 g. Needles 1 g. Needles 2 g. Needles 1 g. Needles 2 g. NW 10 m 17.2 ± 1.56 15.0 ± 1.50 8.7 ± 0.85 10.7 ± 1.10 8.5 ± 5.37 11.2 ± 1.1 50 m 17.0 ± 1.65 13.4 ± 1.26 7.5 ± 0.75 6.5 ± 5.56 7.4 ± 5.79 7.3 ± 0.69 200 m 32.4 ± 3.20 27.0 ± 2.78 19.3 ± 1.95 21.1 ± 2.10 18.7 ± 15.5 18.3 ± 1.8 500 m 47.4 ± 4.50 48.0 ± 4.59 13.4 ± 1.23 10.7 ± 1.56 14.1 ± 14.1 13.3 ± 1.10 S-z 10 m 29.7 ± 2.89 40.5 ± 4.10 21.2 ± 2.1 1.78 ± 1.50 17.6 ± 1.70 16.4 ± 1.50 50 m 54.0 ± 5.0 60.7 ± 6.50 20.0 ± 2.0 20.6 ± 2.10 23.3 ± 2.23 23.2 ± 2.23 200 m 31.7 ± 3.10 34.6 ± 3.56 21.0 ± 2.0 20.0 ± 2.0 22.5 ± 2.20 22.6 ± 2.10 1000 m 50.0 ± 4.97 40.0 ± 3.71 20.6 ± 2.0 20.7 ± 1.98 20.0 ± 2.0 22.2 ± 2.20 Control plot 32.5 ± 3.10 32.8 ± 3.26 24.6 ± 2.20 33.8 ± 3.30 20.6 ± 2.10 25.3 ± 2.50

The high activity of catalase, which is superior to that in the needles of trees growing in the control trial plot, was detected in trial plots remote 200-1000 m in the north-west and 50-1000 m in the south-west from the source of pollution. Moreover, the highest activity of catalase was detected in pine needles of the first year of life on a test area of 500 m in the north-west direction and in needles of the second year of life on the test area of 50 m in the south-west direction. This phenomenon is due to the increase in degradation processes in these trial plots associated with increased litter in adjacent areas. At the same time, the data on the activity of catalase in the test areas of 10–50 m in the northwest direction are reduced compared with those in the control area by 2.2–1.8 times, which characterizes the overall level of decrease in biological processes in the needles of trees and is consistent with data on the accumulation of chlorophyll assimilation process.

Compared to the winter period, the results of determination of catalase in spring were significantly reduced and do not confirm the trend detected in the winter period, namely: the highest level of catalase activity was in pine needles in the control area, while the enzyme activity in the needles of the second year of life was slightly higher, than in the month of December. So, in the needles of the first year of life, the activity of catalase is reduced by 30% compared with the winter period. At the same time, on test plots in the zone of influence of FSUE PPZ Sverdlovskiy, a decrease in catalase activity compared to the winter period was observed from 1.5 to 5 times (1000 m in the south-west direction). With the beginning of the growing season, assimilation processes sharply increase, which is the reason for the decrease in catalase activity in growing assimilation organs. Obviously, the biological processes in the needles of the second year of life are most stable throughout the year compared with the needles of the first year of life.

Data on the activity of catalase in pine needles, obtained in September 2004, are consistent with those in the spring. Significant differences were observed in the results of determination of catalase at the test site in the needles of the second year of life compared to the spring period (25.4 versus 33.8), while on the trial area in the waste influence zone of the Federal State Unitary Enterprise “Sverdlovskiy” indicators of catalase activity in pine needles ordinary autumn slightly differed from the results obtained in the spring. At the same time, the trends in activity depending on the distance from the pollution source persisted in the fall.

Thus, studies to determine the catalase in pine needles in the zone of influence of the Federal State Unitary Enterprise PPZ Sverdlovsky show that the enzyme activity in the control trial plot is the most stable. Deviations were detected in the pollution zones, both in the direction of strengthening (500 m, 1000 m), and in the direction of decreasing (10 m, 50 m) of catalase activity depending on the degree of adaptation of trees to contaminants (waste from the poultry farm). The dynamics of the activity of the catalase enzyme in pine needles during the growing and post-growing periods indicates an increase in the activity of the enzyme in the post-growing season at all trial plots, which indicates the seasonality of metabolic processes in the assimilation organs of common pine.

To conduct an objective assessment of the influence of anthropogenic effects on the level of metabolic processes in pine needles, additional coefficients are used that show the ratio of the processes of assimilation and dissimilation in pine needles.

The ratio of the indicator of chlorophyll content (mg / 1 g) in the needles of ordinary pine to the activity indicator of catalase (cm ³ O ₂ ^1g / _{2 min} ) in the same substrate was taken as a coefficient of metabolic intensity (KIM).

The dynamics of the intensity of the metabolic process in pine needles revealed an increase in photosynthesis from winter to autumn and is expressed by the corresponding coefficients (Table 5). The metabolic rate was studied during the year - in winter, spring (beginning of the growing season), in autumn at the end of the growing season.

Table 5 Dynamics of the coefficient of the intensity of metabolic processes in the needles of common pine (2 years of life) in the trial plots, different from the source of pollution. PP removal KIM metabolism coefficient KIM increase from winter to autumn December 2003 March 2004 September 2004 March-December September-march September-december sz 10 m 0.13 0.5 0.7 3.8 1.4 5.3 50 m 0.11 0.8 1.2 7.2 1.5 11.0 200 m 0.05 0.2 0.4 4.0 2.0 8.0 500 m 0.07 0.6 0.4 8.5 2.0 6.0 y-s 10 m 0.04 0.3 0.8 7.5 2.6 20.0 50 m 0.02 0.3 0.6 15.0 2.0 30.0 200 m 0.11 0.3 0.5 2.7 1.7 4.5 1000 m 0.09 0.3 0.7 3.3 2.3 7.7 Control 5 km, southwest 0.14 0.25 0.6 1.8 2.4 4.3

The data of mathematical processing of the results of determining metabolic indicators confirm the above analysis of the results of determining chlorophyll and catalase in the needles of common pine (Tables 1, 2) and allow us to quantify the change in the process of assimilation intensity depending on the distance of trees from the source of pollution.

An analysis of the results of determining the dynamics of the metabolism coefficient (CIM) shows that the latter is the same in all test areas in the waste zone of the Sverdlovsk PPZ waste, namely: a sharp increase in the intensity of metabolic processes from winter to spring (from 2.7 to 15 times), which continues until the fall. However, in the growing season (from spring to autumn), the change in the intensity of metabolic processes is not so pronounced as in the previous period (from winter to spring), only 1.7-2.3 times.

In general, the increase in metabolism during the study period was most significant at the PP in the southwest direction near the pollution source (10-50 m) and at a distance of 500-1000 m.

A different pattern of change in the intensity of metabolic processes (CIM) during the observation period was revealed at the checkpoint. Here, the intensification of this process does not occur as sharply as on the PP of the pollution zone and has a gradual character from spring to autumn with a general increase in CMM by 4.3 times.

Thus, the data obtained allow us to conclude that the influence of anthropogenic stress on natural ecosystems can be detected by determining the intensity of metabolic processes in the needles of common pine, growing in the area of anthropogenic impact.

- According to the results obtained at different times of the year, the relative indicators of the metabolic rate at the checkpoint vary from 0.14 to 0.6. Moreover, in winter in the pollution zone they are much lower than at the checkpoint; on the contrary, in spring they sharply increase in comparison with the checkpoint and remain at a high level in the fall.

- Indicators of the intensity of metabolic processes in the needles of common pine at the checkpoint should be taken as the norm, with which similar indicators are compared at other BCPs and the level of negative impact of agricultural waste is evaluated.

- An assessment of the intensity of metabolic processes in different periods of the annual cycle showed that the most informative results, allowing to isolate the zones of the greatest impact of pollutants on biological systems, were obtained in the spring, which is due to a regular increase in metabolism in plant tissues with the beginning of the growing season.

- Considering the changes in the metabolic processes in the assimilation organs of plants as a reaction of adaptation of living organisms to a negative impact on the environment, we can evaluate the sharp increase in CMM in the pollution zone, 5-6 times higher than that at the checkpoint, as a manifestation of adaptation.

Claims (3)

1. A method for assessing the ecological state of agrobiocenosis in zones of anthropogenic influence, characterized in that it includes the simultaneous sampling of pine needles common on the test area and the control trial area, determining the content of chlorophyll and catalase activity in them, calculating the metabolic rate coefficient (CMM) for test and control samples according to the formula

, where a is the value of the chlorophyll content in 1 g of absolutely dry needles; in - the value of the activity of catalase in 1 g of absolutely dry needles, comparing the CMM of the studied samples with the CMM of the control samples, which judge the degree of anthropogenic influence. 2. The method of assessing the ecological state according to claim 1, characterized in that the needles are sampled on the test area under study at five points located diagonally, cutting branches of uniform trees at a height of at least 2 m from a part of the crown adjacent to contaminated areas air, and the control test area is chosen at a distance of at least 5-6 km from the source of aerotechnogenic pollution or at a distance of at least 1-3 km from the livestock breeding enterprise. 3. The method of assessing the ecological state according to claim 1, characterized in that the chlorophyll content is determined by the instrumental method on a photoelectric calorimeter in parallel with the determination of catalase activity in the samples, and studies are carried out at the beginning of the growing season.