ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 original scientific article DOI 10.19233/ASHN.2018.17 received: 2018-07-14 SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL GENOTOXICITY Nina REUTOVA Kabardino-Balkarian Scientific Center, Russian Academy of Sciences, 2, Balkarova, Nalchik, 360002 KBR Russia Petimat DZHAMBETOVA Department of Cell Biology, Morphology and Microbiology, Chechen State University, 32, Sheripova, Grozny 364907, Russia E-mail: dzhambetova.csu@yandex.ru Serikbay ABILEV Vavilov Institute of General Genetics, Russian Academy of Sciences, 3, Gubkina, Moscow 119991, Russia ABSTRACT The present study evaluated the applicability of wild species in investigating environmental genotoxicity. The level of mutation in cells of the root meristem was examined in 8 wild plant species growing in environments affected by different types of pollution. Seeds collected in the disposal areas of a wolfram-molybdenum factory (Kabardino-Balkaria, Russia) were used to study the influence of heavy metal pollution. In the case of pollution with products of oil combustion and refining, the seeds were collected in areas adjacent to mini-factories for domestic oil refining (Chechen Republic, Russia). Pollution induced a 2.5-to-10.2-fold increase in mutation frequency in plant seedlings from contaminated zones compared to plants from "clean" zones. Thus, species of wild flora have proven as convenient and sensitive objects for estimating environmental genotoxicity. Key words: environmental genotoxicity, heavy metals, oil pollution, wild plant species SPECIE DI FLORA SELVATICA QUALI INDICATRICI DI GENOTOSSICITÀ AMBIENTALE SINTESI Il presente studio ha valutato l'applicabilità delle specie selvatiche nello studio della genotossicità ambientale. Il livello di mutazione nelle cellule del meristema radicale è stato esaminato in 8 specie di piante selvatiche che crescono in ambienti affetti da diversi tipi di inquinamento. I semi raccolti nelle aree di smaltimento di una fabbrica di wolfram-molibdeno (Kabardino-Balkaria, Russia) sono stati utilizzati per studiare l'influenza dell'inquinamento da metalli pesanti. Nel caso dell'inquinamento da prodotti di combustione e raffinazione di petrolio, i semi sono stati raccolti in aree adiacenti alle mini-fabbriche per la raffinazione del petrolio (Repubblica Cecena, Russia). L'inquinamento ha indotto un aumento da 2,5 a 10,2 volte della frequenza di mutazione nelle piantine delle zone contaminate rispetto alle piante da zone "pulite". Le specie di flora selvatica si sono pertanto dimostrate utili e sensibili per la stima della genotossicità ambientale. Parole chiave: genotossicità ambientale, metalli pesanti, inquinamento da petrolio, specie di piante selvatiche 137 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et al.: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 INTRODUCTION Understanding the influence of increasing environmental pollution on living organisms has been one of the most important problems to date. The progress of humankind is inseparably linked to the development of industry, which, in turn, inevitably leads to the contamination of the atmosphere, hydrosphere, lithosphere (soil) and biosphere. Water, air and soil pollution adversely affect living organisms, including humans. A contaminated environment has both toxic and genetic effects on living organisms. While the toxic influences, as a rule, can easily be assessed, the impact of pollution on the genetic structures of an organism, including possible remote consequences, is difficult to determine. There are a number of effective methods currently developed to estimate the quality of the environment. The classic methods include sampling of water, air and soil probes, and their laboratory analysis using physical-chemical procedures. This approach, however, does not allow the estimation of the hazardous effect of the detected pollutant on living organisms. Test systems designed to assess the genotoxic effects of pollution, on the other hand, can indicate the possible mutagenic, carcinogenic, teratogenic and other harmful consequences. An impressive number of newly synthesized chemicals and environmental components (soil, air, water and sediments) containing various types of pollutants have already been examined with this method throughout the world (Chen & White, 2004; Ohe et al., 2004; White & Claxton 2004). However, even this approach cannot provide complete information about how a genotoxic effect detected with a test system can influence plants, animals or their associations. Currently, another approach is being developed to estimate the quality of the environment: the assessment of the genotoxic effect of pollution in situ, directly on plants and animals living in a determinate area (Vardar et al., 2014; Watanabe et al., 2014). In this case, the whole complex of hazardous compounds will be taken into consideration, including their synergism and antagonism. Since the 1970s, higher plants have been widely used to screen mutagens and monitor genotoxicants in the environment (de Serres, 1978; Grant & Salamone, 1994; Sandhu et al., 1994; Ma et al., 2005; Sposito et al., 2015; Amato-Lourenco et al., 2017; Sposito et al., 2017). Most experiments in situ were performed using standard test systems with Tradescantia, Allium cepa and Vicia faba (White & Claxton, 2004). Plant test systems have proven to be highly sensitive to heavy metals (Knasmul-ler et al., 1998; Minissi et al., 1998; Majer et al., 2002; Correia et al., 2014), products of oil-processing industry (Minissi et al., 1998; Morais Leme & Aparecida Marin-Morales, 2008) and other organic substances (Herrero et al., 2012; Mesi & Kopliku, 2013; Goujon et al., 2015; Rodríguez et al., 2015), as well as nanoparticles (Ghosh et al., 2016; Panda et al., 2016). Nevertheless, plant test systems do have their limitations (Lazutka et al., 2003), so it is preferable to use naturally growing plant species for genetic monitoring (Gers'kin et al., 2005; Vardar et al., 2014; Watanabe et al., 2014). The aim of this work was to study the suitability of wild flora species to assess the genotoxicity of an environment. MATERIAL AND METHODS Characterization of the studied area and sampling We used species of wild flora to test both inorganic (heavy metals) and organic (products of oil burning and refining) pollutants. The seeds collected at the tailing dump of a wolfram-molybdenum factory (Kabardino-Balkaria, Russia) were used to study the influence of heavy metal pollution. The factory was engaged in wolfram-molybdenum field exploitation and ore concentration until 2000, when it was closed down. The tailing dump we studied had been put into operation in 1967 and was located at an altitude of 1000-1200 meters above sea level. Surrounding areas similar to the studied areas in geo-climatic conditions (slope exposure and plants) were defined as the "clean zone" (background landscape). In the case of pollution by the products of oil burning and refining, the seeds were collected in the Chechen Republic, in the surrounding areas of villages in which mini-factories for domestic oil refining were located. These mini-factories are usually built in the marginal areas of settlements or in woodland belts; their number can vary. Several villages in which primary oil refining was practised for over 13 years were chosen as investigation sites. These were Dolynsk (Groznensky District), Alkhan-Yurt (Urus-Martanovsky District), Mesker-Yurt and Tsotsan-Yurt (Shalinsky Shalinskyi), and Geldegen (Kurchaloevsky District). They were selected for analysis because they are located in the same natural-climatic region and characterized by similar landscape features and vegetation. These villages were defined as the contaminated area. The village of Goity (Urus-Martanovsky District), which is known to be free of this type of industry and belongs to the same natural-climatic region was chosen as the "relatively clean" zone (termed as "relatively" because the benz(a)pyrene content in the soil was 0.03 mg/kg, which exceeds the legal limit of 0.02 mg/kg). The number of mini-factories in the studied polluted settlements ranged from 0 (Goity) to 62 (Dolynsk). The plant collection proceeded as follows: a square territory of 10,000 ± 200 m2 was marked off virtually and divided along the diagonal into areas of approximately 50 m2. In each area, we selected 10 plants characterised by approximately the same behaviour at the stages of flowering and seed ripening, and then collected their seeds. Altogether, at each investigation site we collected seeds from at least 30 specimens of the same plants spe- 138 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et al.: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 cies, and mixed them. Where only a few specimens of a plant species were available, the seeds were collected from all plants growing in that territory. Test organisms Plants of broadly distributed species were preferentially used for the investigation of genotoxicity of the environment in the Chechen Republic. These were waybread (Plantago major L.), medicinal dandelion (Taraxacum officinale Wigg. s.l.), Russian dock (Rumex confertus Willd.), and mayweed (Matricaria recutita L.). When studying the genotoxic effect of the waste from the wolfram-molybdenum factory, we used plants that grew in the surrounding regions and had ripe seeds at the moment of analysis (July-August). From the plants growing in dump areas we chose five species, specifically, cheat-grass (Anisantha tectorum); junegrass (Koeleria cristata); black henbane (Hyoscyamus niger); Jurinea ciscaucasica and medicinal dandelion (Taraxacum officinale Wigg. s.l.) to study the level of mutations; these species belong to distinct genera. Tests procedure The anaphase-telophase method was used to assess the level of chromosomal aberrations in plant seedlings. This method is based on the registration of chromosomal aberration at the anaphase and telophase stages. Owing to dormancy, the seeds only germinated in 1-3 months (or more) after harvesting, depending on plant species. Dandelion and grass seeds can germinate earlier, probably due to a shorter dormant period. It was important for the seeds of the same botanical species collected in "clean" and "contaminated" zone to have the same time of storage. The seeds were germinated in glass Petri dishes on filter paper soaked with tap water in a thermostatic chamber at +26 °C. Germination times varied from 4 to 10 days, depending on the plant species. When necessary, the filter paper was additionally moisturized. The 5-10 mm long roots were fixed in a mix of ethanol and glacial acetic acid (3:1) for at least 2-3 hours. The material fixed this way can be stored up to 1-2 months in a refrigerator at 4 °C. The fixed seedlings were stained in acetocarmine (a 2% solution of carmine in 45% acetic acid) during a 10-12 min water bath. Temporary squash slides were prepared from the root meristem according to the standard method (Dubinina, 1978). At least 1000 anaphases were scored. Chromosome fragments, single and double "bridges" were registered. The percentage of abnormal anaphases against the total number of scored anaphases was calculated. Chemical analysis Soil samples were taken during dry summer days on the terraces of the dump area or in territories located ap- proximately 100±20 m away from the oil-refinery factories, when the seeds of wild plants were also collected. Portions of soil were removed by probe from the upper 0-20 cm horizon in five repeats, then 5 kg of soil was mixed to prepare the sample for analysis (1 kg). The soil samples were put in tightly sealed polyethylene containers and marked with a tag indicating the place and time of sampling. For the purposes of chemical analysis, the soil was dried to obtain air-dry mass. The level of heavy metals in the soils examined was determined using atomic-adsorption spectrophotometry (AAS). Analysis of soil samples for the presence of oil products was carried out in collaboration with the Laboratory of the Soil Sciences Department, Lomono-sov Moscow State University by method of infrared spectroscopy (Orlov & Grishina, 1981). The method of low-temperature fluorescence spectroscopy developed by E. V. Shpolsky was used for the assessment of benz(a) pyrene content in soil (Shpolsky et al., 1968). Statistical processing To determine the reliability of the observed differences we used Fisher's conversion for the match against shares, and two-factor dispersion analysis. RESULTS Tables 1 and 2 summarize the obtained data on heavy metal content in the soils of the studied areas. The content of metals in the tailing dump of the tungsten-molybdenum factory (Kabardino-Balkaria, Russia) was significantly higher (Tab. 1). In the Chechen Republic, where the main pollutants are oil products, the content of heavy metals in the soil in the "relatively clean" village of Goity was not significantly different from other studied villages, in some cases it was even higher (Tab. 2). As for benz(a)pyrene and other petroleum products, their contents in the contaminated zone were 60-80 times higher than those in the "relatively clean" zone of Goity (Tab. 3). In Table 3 and all subsequent tables, the Tab. 1: Heavy metal concentrations in the soils of the disposal areas of a wolfram-molybdenum factory (Kabardino-Balkaria, Russia). Tab. 1: Koncentracije težkih kovin v prsti na odlagališčih tovarne volframa in molibdena (Kabardino-Balkaria, Rusija). Sampling Heavy metal content (mg/kg) area Mo Pb Cu Zn Bi Sn Tailing dump >40 33 37 30-60 6.5 4.5 Clean zone <2 8-13.5 9.5-13 10-15 0 0 139 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et al.: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 Tab. 2: Heavy metal concentration (mg/kg) in the soils of the studied villages of the Chechen Republic. Tab. 2: Koncentracije težkih kovin (mg/kg) v prsti preiskanih naselij v Cečenski republiki. No Site of soil sampling Pb Cd Cr Co Mn 1 Goity 38.21 0.68 56.23 15.41 40.66 2 Tsotsan-Yurt 32.64 1.89 20.14 17.54 18.26 3 Geldegen 59.85 1.35 85.60 18.64 114.68 4 Mesker-Yurt 34.33 1.62 20.28 19.06 15.96 5 Alkhan-Yurt 54.70 1.22 72.44 17.80 136.78 6 Dolynsk 61.22 1.35 95.32 19.48 170.02 villages of the Chechen Republic are listed in order of increasing benz(a)pyrene content. The anaphase-telophase method was used to assess the level of chromosomal aberrations in root meristem cells of plant seedlings. Figures 1a and 1b show a normal anaphase and telophase, respectively. Abnormal anaphases display acentric fragments and "bridges." Fragments of variable sizes emerge as a result of deletions (Fig. 1c) and chromosome lagging (Fig. 1d) during their movement to the poles. The joining of two centromere-containing fragments leads to the formation of a dicentric chromosome, which is affected by two mitotic centres and, being stretched between two daughter groups of anaphase or telophase chromosomes, forms a "bridge." Depending on the type of chromosome damage, different types of "bridges" may occur. The re-joining of two broken sister chromatids leads to the formation of a chromatid (usually single) "bridge" (Fig. Fig. 1: Meristematic cells of Taraxacum officinale Wigg. s.l.: a - normal anaphase; b - normal telophase; c -anaphase with fragments; d - anaphase with lagging chromosomes; e - telophase with single "bridge"; f -anaphase with double "bridge". Fig. 1: Meristemske celice regrata Taraxacum officinale Wigg. s.l.: a - normalna anafaza; b - normalna telo-faza; c - anafaza s fragmenti; d - anafaza z zamikom kromosomov; e - telofaza s posameznim "mostom"; f - anafaza z dvojnim "mostom". 1e), whereas lateral re-joining of two broken chromosomes leads to the formation of a chromosomal (usually double) "bridge" (Fig. 1f). Table 4 shows the results of the estimation of the levels of chromosomal aberrations (CA) in plants collected in the disposal areas of the wolfram-molybdenum factory (Kabardino-Balkaria, Russia) contaminated with heavy metals. The frequency of CA in the root meristem of plants collected in disposal areas was statistically significantly higher than the frequency of CA in plants from the "clean zone." Fragments proved to be the prevailing type of chromosomal aberration in all plant species we studied, with "bridges" found in minor quantities. Wild flora species were also used for genetic monitoring of oil products pollution. Table 5 shows the results of the estimation of mutation level in seedlings obtained from seeds collected in the vicinity of the Chechen villages with mini-factories for domestic oil refining. The study also revealed significantly increased levels of mutation in plant species collected in contaminated zones compared to the "relatively clean" zone of Goity. Of all types of induced alterations, the studied species displayed the highest increase in the frequency of fragments. The highest level of chromosomal rearrangement was recorded in plants from the village of Dolynsk, which is situated in the most contaminated zone. Dispersion analysis has shown that in the case of contamination with oil products, mutation frequency was more dependent on the level of soil pollution than on the plant species used for testing. The influence of soil pollution was 67.1 %, the influence of species 22.5 %, their combined effect only 8.56 %. DISCUSSION In our studies we used 8 plant species, including common (T. officinale, P. major, R. confertus) and rare plants. All of them were quite suitable for genetic monitoring and fairly sensitive to various types of pollutants. All species from contaminated zones showed a significant increase in the level of anaphase/telophase abnormalities compared to the clean zone. In the case of the environmental pollution with heavy metals (dumps 140 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et al.: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 Tab. 3: Oil products and benz(a)pyrene content in the soils of some villages of the Chechen Republic. Tab. 3: Derivati nafte in vsebnost benz(a)pirena v prsti nekaterih naselij v Cečenski republiki. No Soil sample site PH benz(a)pyrene content (mg/kg) Hydrocarbons (%) Oil products (%) 1 Goity 8.30 0.03 0.03 0.02 2 Tsotsan-Yurt 8.42 0.06 0.86 0.73 3 Geldegen 7.79 0.14 0.63 0.52 4 Mesker-Yurt 8.63 0.15 1.17 1.10 5 Alkhan-Yurt 8.42 0.26 1.48 1.48 6 Dolynsk 7.86 1.83 1.72 1.56 of the tungsten-molybdenum factory), the frequency of CA was 1.5-5 times higher compared to the clean zone. In the case of oil and benz(a)pyrene pollution (Chechen Republic), the frequency of mutations increased 5-10 times. A comparison of results related to the two types of pollutants (heavy metals and oil products) revealed both similarities and differences in their actions. In the case of oil product influence, we observed an emergence of double "bridges" in seedlings, which was not detected in the case of contamination with heavy metals. In both cases, fragments were the predominant type of disturbance. The results obtained using species of wild flora are in agreement with data obtained through standard plant test systems (Reutova, 2005, 2017). In these works the genotoxicity of the soil from the tailings of the same tungsten-molybdenum factory was studied using standard plant test systems, and the results indicated a 2-fold increase in the frequency of mutations. The same studies were performed on soils of the villages in the Chechen Republic (Dzambetova & Reutova 2006). Soils contaminated with petroleum products caused a 5-fold increase in the frequency of mutations in plant test systems. Virtually any plant species growing in a particular territory can be used for estimating genotoxic effects on the environment. It should be noted, however, that all species are not equally suitable objects for analysis. It is easier to work with species from the Poaceae and Asteraceae genera, as their seeds germinate easily and the roots are not very thick, which allows for good squash preparations. It is more difficult to work with seeds which have a dense, thick peel, as they germinate poorly and very slowly (e.g. the seeds of black henbane). Mutagenic effects of HM, hydrocarbons, oil products and benz(a)pyrene have been studied for several dec- Tab. 4: Level of chromosomal aberration in a wild plant from areas around a wolfram-molybdenum factory (Kabardino-Balkaria, Russia). Tab. 4: Stopnja kromosomskih aberacij na divje rastočih rastlinah iz predelov okoli tovarne volframa in molibdena (Kabardino-Balkaria, Rusija). Species No of cells (ana/telo) cells with aberrations Aberrations (%) P Yurinea ciscaucasica Clean zone dumps 976 1264 17 46 1.76 3.64 <0,01 Anisantha tectorum Clean zone dumps 1024 1004 15 51 1.46 3.09 <0,05 Koeleria cristata Clean zone dumps 1043 1187 8 41 0.77 3.45 <0,001 Hyoscyanus niger Clean zone dumps 270 322 3 9 1.10 2.79 <0,05 Taraxacum officinale Clean zone dumps 1033 1007 41 194 3.97 19.72 <0,001 141 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et al.: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 ades, using various test systems and providing detailed reports of the results in the reviews (Chen & White, 2004; Ohe et al., 2004; White & Claxton, 2004). But there are very few works devoted to the study of environmental genotoxicity employing wild flora species growing in contaminated areas. In the past years, there have been some studies in which species of wild flora (mostly wood plants) were used for genetic monitoring and determining genotoxic effect on the environment. To this end, a micronuclei (MN) test was applied to the root meristem of pine (Geras'kin et al., 2005) and silver birch (Kalaev et al., 2006) seedlings. In all cases an increase of micronuclei frequencies was observed, even when the types of pollutants were different: ionizing radiation and heavy metals. Herbaceous plants - Arabidopsis thaliana and Crepis tectorum - have previously been used to study the genetic processes in permanently irradiated plant populations. These studies revealed that plants with rearranged karyotypes started to appear two years after the Chernobyl accident, their frequency correlated with the frequency of CA in the cells of the root meristem. Thirty-eight years after the Kyshtym nuclear accident no increase in the frequency of plants with rearranged karyotype has been detected, because such plants did not have any selective advantages and were subjected Tab. 5: Level of chromosomal aberration in a wild plant from areas contaminated with oil refinery products (Chechen Republic, Russia). Tab. 5: Stopnja kromosomskih aberacij na divje rastočih rastlinah iz predelov pod vplivom onesnaženja iz rafinerije (Cečenska republika, Rusija). Plant species Collection site Number of cells (ana/telo) Cells with aberrations Aberrations (%) P Taraxacum officinale Wigg. s.l. Goity 1056 24 2.27 Tsotsan-Yurt 1007 96 9.53 <0.001 Geldegen 1050 114 10.85 <0.001 Mesker-Yurt 1012 104 10.28 <0.001 Alkhan-Yurt 1026 121 11.7 <0.001 Dolynsk 1044 126 12.07 <0.001 Matricaria recutita L. Goity 1164 18 1.55 Tsotsan-Yurt 1062 106 9.98 <0.001 Geldegen 1018 98 9.63 <0.001 Mesker-Yurt 1013 118 11.65 <0.001 Alkhan-Yurt 1071 89 8.31 <0.001 Dolynsk 1070 121 11.31 <0.001 Rumex confertus Willd. Goity 1002 17 1.69 Tsotsan-Yurt 1027 89 8.67 <0.001 Geldegen 1020 106 10.39 <0.001 Mesker-Yurt 1006 96 9.54 <0.001 Alkhan-Yurt 1082 87 8.04 <0.001 Dolynsk 1051 113 10.75 <0.001 Plantago major L. Goity 1018 9 0.89 Tsotsan-Yurt 1101 53 4.81 <0.001 Geldegen 1059 61 5.76 <0.001 Mesker-Yurt 1006 47 4.67 <0.001 Alkhan-Yurt 1023 36 3.52 <0.001 Dolynsk 1004 91 9.06 <0.001 142 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et aL: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 to negative selection along with a decreasing irradiation dose rate (Abramov et al., 2006). The authors also noticed that the plant species they used in their studies could be recommended as test objects for the registration of genetic effects of low-dose ionizing radiation. Our study, too, has proved the suitability of wild flora species for identifying the genotoxicity of an environment when contaminated with inorganic (HM) and organic (hydrocarbons, oil products and benz(a)pyrene) substances. CONCLUSIONS Species of wild flora are very convenient objects for genetic monitoring, because they allow in situ analysis and an evaluation of the effects of all complex of environmental factors, they are highly sensitive to different types of pollutions, and their testing does not require expensive facilities or highly qualified personnel. To perform a comparative analysis of the results it is necessary to have an indicator species with a fairly broad distribution. From our point of view, medicinal dandelion (Taraxacum officinale Wigg. s.l.) is the best candidate. Dandelion proves to be a universal species, as its flowering or fruiting plants can be found throughout the summer period, with massive flowering typically occurring in May and early June. Dandelion seeds have a short dormant period; their germination rate is high and they germinate fast. The seedling roots are of an optimal size for making good squash preparations and the species is sensitive to various types of pollution (heavy metals, organic pollutants). Given all these factors we suggest that dandelion be used as a standard species for the examination of genotoxic effects on the environment. 143 ANNALES ■ Ser. hist. nat. ■ 28 ■ 2018 ■ 1 Nina REUTOVA et aL: SPECIES OF WILD FLORA AS INDICATORS OF ENVIRONMENTAL, 137-146 RASTLINSKE VRSTE KOT INDIKATORJI OKOLJSKE GENOTOKSIČNOSTI Nina REUTOVA Kabardino-Balkarian Scientific Center, Russian Academy of Sciences, 2, Balkarova, Nalchik, 360002 KBR Russia Petimat DZHAMBETOVA Department of Cell Biology, Morphology and Microbiology, Chechen State University, 32, Sheripova, Grozny 364907, Russia E-mail: dzhambetova.csu@yandex.ru Serikbay ABILEV Vavilov Institute of General Genetics, Russian Academy of Sciences, 3, Gubkina, Moscow 119991, Russia POVZETEK Pričujoča raziskava opredeljuje uporabnost divje rastočih rastlinskih vrst za ugotavljanje okoljske genotoksičnosti. Nivo mutacij v celicah v koreninskem meristemu je bil preiskan na 8 rastlinskih vrstah, ki uspevajo v okolju pod vplivov različnih okoljskih dejavnikov. Za ugotavljanje vpliva onesnaženja s težkimi kovinami so uporabili semena, nabrana na odlagališčih tovarne wolframa in molibdena (Kabardino-Balkaria, Rusija). V primeru onesnaževanja z produkti izgorevanja in rafiniranja nafte so bila semena pobrana v predelih z mini-tovarnami za predelavo nafte (Čečenska republika, Rusija). Onesnaženje je povzročilo 2,5 do 10,2 - kratno povečanje frekvence mutacij na sadikah iz onesnaženega okolja v primerjavi s sadikami v "čistem" okolju. S tega vidika so divje rastoče rastlinske vrste uporabne kot občutljivi objekti za ugotavljanje okoljske genotoksičnosti. 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