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DOI: https://doi.org/10.23670/IRJ.2017.58.027 Берсенева С. А.,1 Белоусова Н.М.2 1,2Кандидат биологических наук, Приморская государственная сельскохозяйственная академия ОЦЕНКА ФИТОТОКСИЧНОСТИ ПОЧВ ПРИМОРСКОГО КРАЯ (ДАЛЬНЕВОСТОЧНЫЙ ФЕДЕРАЛЬНЫЙ ОКРУГ) НА СЕЛЬСКОХОЗЯЙСТВЕННЫЕ ПОЛЛЮТАНТЫ С ПОМОЩЬЮ ВЫСШИХ
РАСТЕНИЙ
Аннотация
Загрязнение почв сельскохозяйственного назначения в Приморском крае происходит за счет увеличения масштабов и территориального распространения поллютантов, которые оказывают влияние на почву и биоту, в целом.
Для получения полной картины загрязнения агроценозов сельскохозяйственными поллютантами проведены исследования с применением ряда биоиндикаторов, относящихся к разным таксономическим группам растений: яровая пшеница (Triticum aestivum L.), кресс-салат (Lepidium sativum L.), овс (Avena L.), соя (Glycine L.), редис (Raphanus sativus L.). Помимо семенных культур нами использован рясковый тест, т.е. в качестве тестирующего объекта выступили водные высшие растения семейства рясковых Lemnaceae, которые характеризуются простотой строения, быстрой скоростью размножения и высокой чувствительностью.
Ключевые слова: фитотоксичность почв, сельскохозяйственные поллютанты, высшие растения, ряска малая.
Berseneva S.A.1, Belousova N.M. 2 1,2PhD in Biology, Primorskaya State Akademy of Agriculture ESTIMATION OF PHYTOTOXICITY OF SOILS OF PRIMORSKY KRAY (FAR EASTERN FEDERAL DISTRICT) ON AGRICULTURAL POLLUTANTS BY HIGHER PLANTS
Abstract
The pollution of agricultural soils in Primorsky Kray takes place due to the increase of the scale and territorial distribution of pollutants affecting the soil and biota in general.
In order to obtain a complete picture of the contamination of agrocenoses with agricultural pollutants, The authors studied a number of bioindicators which belong to different taxonomic groups of plants: spring wheat (Triticum aestivum L.), cockweed (Lepidium sativum L.), oat (Avena L.), soybean (Glycine L.), and radish (Raphanus sativus L.). Besides the seed cultures, we used a rickberry test, i.e. the water plants of the Lemnaceae family were as test objects, they are characterized by simplicity of structure, high reproduction speed and high sensitivity.
Keywords: phytotoxicity of soils, agricultural pollutants, higher plants, duckweed.
Introduction
T he soil - the indicator of long-term natural processes, and its state is a result of long influence of various sources of pollution. Emissions in the atmosphere from the industrial enterprises and motor transport, an irrigation of lands the polluted waters, violations of production requirements at production, processing and use of oil products, numerous accidents on oil pipelines, lead unbalanced use of mineral fertilizers and pesticides to pollution of soils, deterioration of their physical state and as a result loss of fertility and inability to carry out the ecological functions.
From the 70-ties of the past century onwards concern has been raised about levels of contaminants in the environment. Main sources of contamination are industrial processes, waste disposal and intentionally applied chemicals in agriculture, such as fertilizers, pesticides and veterinary drugs.
Regulations dealing with the prevention and remediation of contaminated soils are aimed at the soil as an environmental compartment (soil protection act), industries (environmental regulations), chemicals (REACh, pesticide regulation, Nitrate directive) or waste management (Waste Directive) [4].
The method of vegetative bioindication was applied to definition of anthropogenous impact on an ecosystem in various points of a landscape. The plants growing on the soil are exposed to influence of toxicants and characterize the first stages of pollution of soils, they very accurately react to natural and anthropogenous changes. Bioindicators integrate biologically significant effects of pollution. They allow to define speed of occurring changes, ways and congestion places in ecosystems of various toxicants, to do conclusions about degree of danger to the person and useful biota of concrete substances or their combinations [i].
Their account at carrying out an ecological assessment is important especially. Soil is a complex dynamic system notable for various properties and regimes.
For the characteristic of an ecological condition of an agrocnosis resort to a complex the methods establishing the level of its pollution. Methodical approach of the complex characteristics consists, first of all, of a toxicological and sanitary assessment of each chemical or connection used for increase and preservation of a crop and an ecological assessment of their influence on soil to a component and quality of agricultural production. The individual characteristic of the applied substances doesn't correspond to their cumulative influence in field conditions.
It is earlier established that their joint receipt is followed by processes of migration, a translocation, mobilization, an immobilization, etc. It leads to change of a chemical condition of substances, change of their quantitative structure and formation of metabolites in the soil and plants often exceeding toxic action of individual substances [3].
In this regard, except concrete definition of action and an after-effect of the used chemical means, it is expedient to use biotesting methods, and in particular, phytotesting for an ecological assessment of an agrocnosis.
The agriculture including crop branch is one of the priority directions in development of Primorsky region where agricultural production is conducted across all the territory, and its main volumes are concentrated in the southern and southwest zone.
Due to the annual increase of the agricultural areas which are subjected to pollution by various chemicals such as pesticides, herbicides, salts of heavy metals, mycotoxins and other agents there is the necessity to conduct the researches on determination of soil pollution by chemicals harmful to plants.
The assessment of soils phytotoxicity is one of the methods of vegetable systems genetic monitoring since it allows to reveal dependences between soil pollution by the pollutants and their influence on reproductive system of the higher plants.
The most perspective researches of soils phytotoxicity and also water pollution testing by various pollutants is biotesting.
Pollution of agricultural soils in Primorsky region happens due to the increase in scales and territorial distribution of pollutants which have impact on the soil and a biota, in general.
Special biotests to determine the efficiency of agrochemicals systemic effect in genetic monitoring come to an assessment of change degree in biochemical and morphometric indexes, i.e. germination and seeds viability energy, roots growth, plant infestation by diseases under the influence of the pollutant.
The aim of researches - to carry out an assessment of phytotoxicity of soils of Primorsky region on agricultural pollyutant by means of the higher plants.
For realization of the purpose the following tasks were set:
1. To select soil samples, to prepare them for the analysis.
2. To carry out an assessment of level of pollution of soils of Primorsky region by means of bioindicators of various systematic groups.
The research object is the soil of agricultural areas in Primorsky region.
Material and methods
The assessment of phytotoxicity of soils was carried out in the territory of Primorsky region in Hankaysk, Horolsk, Mikhaylovsk, Ussuriisk and Kirovsk areas which specialize on cultivation grain (wheat, oats), the leguminouses (soy), additional potatoes, corn.
For an assessment of spatial pollution of soils tests of soils from the superficial horizons on depth of 10 cm were selected. Each sample on a point of selection was made of 5 pointed samples taken by an envelope method from a platform at least 100 sq.m.
For averaging of results, and also an exception of errors, tests were selected from a platform with the party of 5-10 m. The volume of tests was defined by specific needs for carrying out analytical works, and made in most cases 400-800 g.
Selection of soil material from cuts was made on the genetic horizons, in that case when it was difficult to define borders of the horizons - with a step of 15-30 cm.
When sampling from soil cuts the description of landscape and ecological conditions of the district and physicomechanical properties of soils was made.
All selected tests after preliminary processing (drying, complaint through a sieve of 1 mm, quartation) were directed to laboratories for carrying out analytical researches - a sprouting of seeds of test cultures in water extracts of the studied soils.
The option with the distilled water - control 1 acted as absolute control.
In researches used a rolled method of a sprouting of seeds. For this purpose on a strip to the ruled filter paper previously moistened with a water extract sowed on 20 seeds of each culture in three multiple frequencies, then strips twisted, and the turned-out rolls stacked in rastiln and placed in the thermostat for 7 days.
For the 7th days registered quantity of the sprouted seeds and took measurements of growth characteristics (length of a root and escape) on 10 plants of each sample. Statistical processing was carried with use of the Microsoft Excel 2007 program.
For an objective assessment about action of a pollyutant on properties of the soil according to requirements of a technique of biotesting, test objects have to treats different taxonomical groups.
For a complete picture of agrocoenosis pollution by agricultural pollutants there were research works conducted using a number of biological indicators belonged to different taxonomic groups of plants: spring wheat (Triticum aestivum L.), garden cress (Lepidium sativum L.), oat (Avena L.), soybean ( Glycine L.), radish (Raphanus sativus L.). In addition to seed crops we also used Lemnaceae test, i.e. the test objects were higher aquatic plants of the Lemnaceae family, which are characterized by the simplicity of the structure, the rapid rate of reproduction and high sensitivity.
Results
Depending on results of experience one of 4 levels of pollution is appropriated to substrata:
1. Pollution is absent: viability of seeds of 90-100%, shoots amicable, strong, equal.
2. Weak pollution: viability of seeds of 60-90%, sprouts of almost normal length, strong, equal.
3. Average pollution: viability of 20-60%, sprouts shorter and is thinner, some sprouts have uglinesses.
4. Strong pollution: viability of seeds very weak, less than 20%, sprouts small and ugly [2].
The established dynamic changes on total toxicity testify that use of phytotesting allows to conduct express control which has to be used for an expert assessment and restoration of an ecological condition of an agrocnosis during vegetation. It is necessary and to develop further (to unify and automate) methodical part. Its practical application will provide cultivation of quality agricultural production.
The agrotechnology of cultivation of crops assumes use of pesticides, herbicides in certain doses and structures. We investigated arable soils on which grew:
1. Control
2. Wheat
3. Corn
4. Potatoes
5. Rice
Each option was analyzed on a condition of phytotests (tab. 1).
Correlation analysis. As a result of the made experiments it was revealed that average of the sprouted seeds of a garden cress and a garden radish on control options I made 19,3 and 19 pieces respectively, average germinating seeds of test objects made - 96,67% and 95% respectively that confirms lack of pollutants in arable soils. Weak pollution of arable soils pollyutant characterizes fields on which cultivate corn and potatoes. Strong pollution of arable soils of Primorsky region, including the Hankaysky area is observed at rice cultivation (the Chinese tenants who don't respect the rules of introduction of herbicides and fertilizers).
Table 1 - Viability and biomass of seeds of a garden cress, garden radish
№ tests Number of tests Number of the sprouted seeds Average of the sprouted seeds Valiability, % Average viability, % Biomass, m (г) Average biomass, mcv (г) Level pollution
Garden cress seeds
1 1.1 19,00 19,33 95,00 96,67 0,5618 0,6256 Pollution no
1.2 19,00 95,00 0,6345
1.3 20,00 100,00 0,6804
2 2.1 19,00 19,67 95,00 98,33 0,5893 0,6330 Pollution no
2.2 20,00 100,00 0,7608
2.3 20,00 100,00 0,5490
3 3.1 15,00 18,00 75,00 90,00 0,4467 0,5285
3.2 19,00 95,00 0,5228 The weak pollution
3.3 20,00 100,00 0,6160
4 4.1 18,00 17,67 90,00 88,33 0,5687 0,5604 The weak pollution
4.2 20,00 100,00 0,6116
4.3 15,00 75,00 0,5009
5 5.1 12,0 10,0 15,7 21,63 0,223 0,2976 Acute pollution
5.2 10,0 22,8 0,34221
5.3 8,0 26,4 0,3277
Garden radish seeds
№ tests Number of tests Number of the sprouted seeds Average of the sprouted seeds Valiability, % Average viability, % Biomass, m (г) Average biomass, mcp (г) Level pollution
1 1.1 17,00 19,00 85,00 95,00 0,6573 0,6674 Pollution no
1.2 20,00 100,00 0,6969
1.3 20,00 100,00 0,6480
2 2.1 15,00 17,33 75,00 86,67 0,6450 0,6631 The weak pollution
2.2 18,00 90,00 0,6498
2.3 19,00 95,00 0,6946
3 3.1 18,00 17,33 90,00 86,67 0,5605 0,5086 The weak pollution
3.2 20,00 100,00 0,5743
3.3 14,00 70,00 0,3909
4 4.1 19,0 18,0 78 85,6 0,450 0,4723 The weak pollution
4.2 15,0 90 0,3788
4.3 20,0 89 0,589
5 5.1 9,0 9,0 13,0 12,3 0,267 0,263 Acute pollution
5.2 10,0 9,0, 0,325
5.3 8,0 15,0 0,197
7 days later after disembarkation of seeds of test cultures to solutions took measurements according to the following characteristics: lengths of a root and escape. The analysis of data showed that on key parameters of test cultures, soils on culture of rice accumulate in a bigger measure of pollyutant that proves decrease in indicators of viability and activity of test plants.
The fulfilled technique allows to hold testing in 7 days as test cultures it is recommended to use seeds of a garden cress, a garden radish, the estimated parameters - length of roots and escapes, and also viability of seeds.
Discussion
In 2016 researches of arable lands of the agricultural enterprises by means of test object - a duckweed small will be continued.
The used biotests allowed to fix the negative phenomena in soils even at rather weak anthropogenous loadings.
The conducted researches showed that the used set of test systems for extent of influence of anthropogenous load of soils allows to estimate determination of toxicity objectively.
The analysis of data showed that the most polluted agricultural pollyutant are the soils used at cultivation of rice on the Chinese technology.
Conclusion
The test cultures chosen for monitoring allow to define authentically in one-factorial experience biological effect of growth inhibition and development of plants at their cultivation in the soils polluted and not polluted by agricultural pollyutant.
For an assessment of presence of pollyutant in the soil it is necessary to consider growth and development of axial bodies of sprouts of test plants, viability and force of growth of seeds of a garden radish and kres-salad in as the indicators.
It should be noted that definition of pollution of soils products of an anthropogenous origin (pesticides and other agrochemicals) and the analysis of phytotoxicity of soils not so obviously, as degradation of soils (salinization, an erosion, reconsolidation, exhaustion, etc.) therefore researches are necessary for a reliable assessment by the developed technique, analytical base which is capable to diagnose negative influence of pollutants at the initial stages.
Список литературы / References
1. Голополосова T.V Биоиндикация как метод определения степени загрязнения окружающей окружающей среды / Т.В. Голополосова, Л.Н. Савинов, В. К. Глушанков, Ситникова Н. С.//Реагенты, 2000. Тезисы XIIII Международной научно-технической конференции. - 2000. - С. 64-66.
2. Другов Ю. С. Экологическая аналитическая химия / Ю. С. Другов. - M.: 2000. - С. 260-266.
3. Контроль Мелехова О. П. Биологический контроль окружающей среды. Биоиндикация и биотестирование / О. П. Мелехова. - M.: Академия, 2007. - 288 с.
4. Опекунова М. Г. Биоиндикационные загрязнения / М. Г. Опекунова. -: Санкт-петербургский государственный университет, 2004. - 266 с.
Список литературы на английском языке / References in English
1. Golopolosova T.V. Bioindikatsiya kak metod opredeleniya stepeni zagriazneniya okruzhayushchey sredy [Bioindication as Method for Determination of Environmental Pollution Degree] / T.V. Golopolosova, L.N. Savinov, VK Glushankov, Sitnikova N. S. // Reagenty, 2000. Tezisy XIIII Mezhdunarodnoy nauchno-tekhnicheskoy konferentsii [Reagents, 2000. Abstracts of the 14th International Scientific and Technical Conference] - 2000. - P. 64-66. [In Russian]
2. Drugov Yu. S. Ekologicheskaya analiticheskaya khimiya [Ecological Analytical Chemistry] / Yu. S. Drugov. - M.: 2000. - P. 260-266. [In Russian]
3. Melekhova O. P. Biologicheskiy kontrol okruzhayushchey sredy. Bioindikatsiya i biotestirovaniye [Biological Control of Environment. Bioindication and Biotesting] / O.P. Melekhova. - M.: Academiya, 2007. - 288 p. [In Russian]
4. Opekunova MG Bioindikatsionnye zagriazneniya [Bioindication Contamination] / M. G. Opekunova.: St. Petersburg State University, 2004. - 266 p. [In Russian]
DOI: https://doi.org/10.23670/IRJ.2017.58.062 Вятчина О.Ф.1, Жданова Г.О.2, Стом Д.И.3, Толстой М.Ю.4
1ORCID: 0000-0002-2205-1971, Кандидат биологических наук, доцент, Иркутский государственный университет,
2ORCID: 0000-0002-8355-9517, младший научный сотрудник, НИИ биологии, Иркутский государственный университет,
3ORCID: 0000-0001-9496-2961, доктор биологических наук, профессор,
Иркутский государственный университет, Иркутский национальный исследовательский технический университет,
Байкальский музей ИНЦ СО РАН,
4ORCID: 0000-0002-1573-060X, кандидат технических наук, профессор, Иркутский национальный исследовательский технический университет Работа выполнена при финансовой поддержке грантов РФФИ р_а № 16-48-030887, № 16-48-030881.
ПЕНООБРАЗУЮЩАЯ АКТИВНОСТЬ SACCHAROMYCES CEREVISIAE КАК НОВАЯ ТЕСТ-РЕАКЦИЯ ДЛЯ БИОТЕСТИРОВАНИЯ СОЛЕЙ ТЯЖЕЛЫХ МЕТАЛЛОВ
Аннотация
Для оценки токсичности солей тяжелых металлов предложена новая тест-реакция, основанная на определении пенообразующей активности Saccharomyces cerevisiae. Показано, что разработанный биотест позволяет обнаруживать хлорид ртути в концентрации 0,0001, сульфат меди, хлорид кадмия, хлорид кобальта - 0,001, сульфат свинца - 0,01, сульфат железа - 0,1, сульфат цинка - 1 г/л. Результаты тестирования сопоставлены с данными, полученными при использовании методов по выживаемости Paramecium caudatum, подавлению люминесценции Photobacterium phosphoreum, прорастанию семян Raphanus sativus, входящих в государственный реестр токсикологических методов контроля природных сред. Составлены ряды токсичности исследуемых солей тяжелых металлов по отношению к используемым тест-объектам. Отличительной особенностью нового биотеста с использованием S. cerevisiae является экспрессность (время тест-отклика 15 мин), отсутствие необходимости в поддержании и выращивании тест-объекта (используется препарат сухих дрожжей), техническая простота исполнения, экономичность, хорошая воспроизводимость.
Ключевые слова: биотестирование, тяжелые металлы, Saccharomyces cerevisiae, пенообразование.
