Evaluation of the cytotoxic and genotoxic effects of 2 Fluquinconazole using Allium cepa L. as a bioindicator

doi:10.21203/rs.3.rs-2568737/v1

Download PDF

Research Article

Evaluation of the cytotoxic and genotoxic effects of 2 Fluquinconazole using Allium cepa L. as a bioindicator

https://doi.org/10.21203/rs.3.rs-2568737/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

Fluquinconazole is the active substance of a synthetic fungicide which is used extensively in agricultural areas in the world and Turkey. In this study, 30, 60, 90 and 100 mg/l doses of the substance were applied to Allium cepa root tips for 12, 24 and 48 hours. Distilled water was used as a negative control while methyl methane sulfonate (MMS, 10 ppm) was used as a positive control. As a result of the application, it was observed that the substance caused mitotic inhibition by decreasing the mitotic index, leading to changes in mitotic stage ratios. It was also observed that this substance caused chromosomal (anaphase bridges, stickiness, c-mitosis, laggards, and breakages) and nuclear abnormalities (binucleus and micronucleus). At the end of the statistical analysis and examinations, it was detected that the substance caused cytotoxic and genotoxic effects.

Cytotoxicity

genotoxicity

Mitotic index

Allium assay

Micronuclei

Fluquinconazole

Substances such as chemicals, some organic compounds, and disinfectants that are used to destroy the harmful effects of bacteria, viruses, and pests, to increase the quality of life of plants and to help them grow, develop and produce are called "Pesticides". There are many genotoxic, cytotoxic, or mutagenic studies on pesticides, which are categorized into different groups according to their area of use and purpose. As a result of these studies, which focused especially on insecticides and herbicides, it was found that the pesticides used caused many adverse conditions, the most common ones of which were an increase or decrease in the mitotic index and structural and/or numerical abnormalities in chromosomes (de Sousa et al. 2019; de Morais et al. 2019; Ilyushina et al. 2020; Mercado et al. 2020; Gallego and Olivero-Verbel 2021). Another group of pesticides are fungicides. Fungicide, also called antimycotic, is an antimycotic agent, a toxic substance used to kill or inhibit the growth of fungi, fungal spores, and fungal infections. Fungicides, which can be either in natural or in synthetic form, are often used to control parasitic fungi that cause economic damage to crop or ornamental plants. Usually found in synthetic form, commercial fungicides are widely used in agricultural activities to kill fungal pathogens infecting plants.

Fungal pathogens are the number one cause of crop loss worldwide. They can cause serious damage and loss of both quality and profit in agricultural production. Applied in powder or in liquid form, fungicides can be applied at different time periods, ranging from seed to storage. Currently in world agriculture, approximately 140 different fungicidal active substances are sold under different trade names that are several times as the number of substances themselves. One of these, Fluquinconazole, is preferred as a wheat seed treatment. Preferred especially in the applications in seed stage, this substance is used to kill pathogens on the seed or to protect the young plant from pathogens in the soil. Whether fungicides have direct or indirect toxic effects on organisms other than the target organisms is still a widely researched topic (Shishatskaya et al. 2018; Wua et al. 2018; Maurya et al. 2019; Marinho et al. 2020; Pitombeira de Figueirêdo et al. 2020). Since fungicides are used regularly every year, they have a different usage pattern from other herbicides and insecticides, which are used intermittently or as needed. Therefore, the genotoxic effects of these substances need further examination.

A flowable concentrate (FS) formulation for seed dressing, Fluquinconazole is available in the market under different trade names and is regularly used to control the root rot pathogen induced by Gaeumannomyces graminis especially in wheat and to help control seed-borne diseases caused by various fungal species (BKU 2023). The fungicide Fluquinconazole has a molecular formula of C16H8Cl2FN5O, a molecular weight of 376.2 g/mol, a CAS number 136426-54-5, and an open formula 3-(2,4-dichlorophenyl)-6-fluoro-2-(1H-1,2,4-triazol-1-yl)-4(3H)-quinazolinone, belonging to the triazole class (PUBCHEM 2023). In studies conducted with this fungicide, it has been shown to have protective properties against diseases caused by fungi at varying rates and that it helps increase the productivity by significantly reducing disease occurrence (Bateman et al. 2004; Gutteridge et al. 2007; Scherm et al. 2009). In literature review on toxicology studies with Fluquinconazole, studies on residue contamination in water, soil, and products caused by the substance were generally found (EFSA 2011; Szpyrka and Walorczyk 2013; Ramanauskienė et al. 2018).

Allium test is an excellent genetic model and biomarker for the investigation of genotoxic, mutagenic, and cytotoxic effects caused by environmental pollutants. Factors such as lower cost, obtaining similar results to animal studies, the possibility of conducting studies with less use of experimental animals, chromosome size and easy examination under light microscope have led researchers to use Allium cepa and plants with similar characteristics in this field. Introduced to the science world in 1938 by Levan, this test method was developed with some modifications by Fiskesjö in 1985 and Rank in 2003 and has been used for many years. The genotoxic effects of a lot of chemicals (pesticides, food additives, environmental pollutants, etc.) have been investigated with this test, which is also accepted by the United Nations Environment Program (UNEP), the International Program on Plant Testing (IPPB), the U.S. Environmental Protection Agency Genetic Toxicology Program (EPA Gen-Tox Program), and the World Health Organization (WHO) (Stapulionytė et al. 2019; Bellani et al. 2020; Khan et al. 2020; Souza et al. 2020; Lovinskaya et al. 2021; Thesai et al. 2021). Although a great number of articles on the subject can be found in the literature review, articles from recent years were mentioned in this study.

Although there are genotoxic studies with various fungicides in the literature, there are no studies showing the effect of this substance on mitotic index and chromosomes. For this reason, it was aimed to investigate the positive and/or negative effects of the active substance Fluquinconazole on mitotic index, mitotic stages, and chromosomes with the Allium test method, which serves as a marker in genotoxic and cytogenotoxic studies. While changes in mitotic index and mitotic stages indicate the cytogenetic effect of the substance used, chromosomal abnormalities will give us information about the genotoxic effect of the same substance. In addition, results regarding the mutagenic effect of the substance will be obtained by looking at whether it causes micronucleus formation in the cell.

Onion tubers (A. cepa L., 2n = 16) with a diameter of 25–30 mm were purchased from a local market in Sivas. The seed treatment Galmano FS 167, which has been marketed by Bayer Company and whose active substance is Fluquinconazole (167 g/L Fluquinconazole), was used in the study. A growth inhibition test was performed to determine the EC50 concentration (effective concentration = EC50 value) of the substance purchased from an agrochemical company. Determination of the EC50 value has been shown to be a useful parameter for selecting test concentrations in genotoxicity tests (Chauhan et al. 1999; Seth et al. 2007). For this purpose, onions were treated with 10 different concentrations of Fluquinconazole for 4 days. These selected concentrations were prepared considering the amount of active substance 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100 mg/L. For each concentration, 5 different onion tubers were used and the length of 10 roots was measured in each tuber. The control group was treated with distilled water. As a result of the evaluations, it was found that the dose that caused a 50% shortening in root length compared to the control group, i.e., the EC50 value, was 60 mg/L. After identifying this value, 4 concentrations (120, 90, 60 and 30 mg/L) were determined as EC50x2, EC50x3/2, EC50x1 and EC50x1/2 respectively. Kihlman (1971) reported that the cell cycle of root tip meristem cells of Allium cepa is 20–24 hours. Considering this, the application periods of the fungicide for mitotic index and chromosomal aberrations were chosen as 12, 24 and 48 hours. Since the highest mitotic frequency in onion was obtained between 6.00 and 9.00 in the morning (Sharma 1983), sampling of root tip materials was completed between 7.00 and 8.00 in the morning.

Germinated for 24 hours in distilled water at room temperature, the onions were treated with 4 different doses of Fluquinconazole for 12, 24, and 48 hours. Onions in the negative control were treated with distilled water for indicated periods while those in the positive control were treated with Methyl methanesulfonate (MMS, Sigma–Aldrich, CAS No 66-27-3) (10 mg/L). 10 onions were used for each concentration in all application groups. At the end of the application periods, the lengths of the root tips were measured and recorded. Then, the roots were cut with sterile scissors, washed in distilled water, and fixated in carnoy solution for 24 hours. After fixation, the roots were stored in 70% alcohol in the refrigerator until use. In order to separate the meristem tissue cells from each other and to ensure better observation of the cells in microscopic examinations, the roots were placed in 1N HCl and hydrolyzed in a water bath at 60ºC for 7 minutes after they were removed from 70% alcohol. After hydrolysis, the roots were placed in distilled water to stop the effect of HCl and kept for 15 minutes, replenishing the water every 5 minutes. The roots were then stained with aceto orcein and well-dispersed preparates were made into permanent for microscopic examination. Microscopic analyses were performed on an Olympus microscope at 100X magnification and mitotic index, chromosomal aberrations in anaphase and telophase, and micronuclei in interphase cells were observed. In identifying the mitotic index of root tip meristem cells, cells divided were counted in a total of approximately 5000–6000 cells, over 1000 cells in each of the 5 preparates prepared for each application, and the formula of General Mitotic Index (GMI) = (number of mitotic cells/total number of cells) X 100 was used. To determine the ratio of mitotic phases, cells in prophase, metaphase, anaphase, and telophase were counted separately and the ratios of Prophase Mitotic Index (PMI), Metaphase Mitotic Index (MMI), Anaphase Mitotic Index (AMI), and Telophase Mitotic Index (TMI) were calculated according to the formula below (Causil Vargas et al. 2017; Salazar and Maldonado 2019; Mercado and Caleño 2020).

PMI = (Number of cells in prophase/number of dividing cells) X 100

MMI = (Number of cells in metaphase/ number of dividing cells) X 100

AMI = (Number of cells in anaphase/number of dividing cells) X 100

TMI = Number of cells in telophase/number of dividing cells) X 100

After mitotic index calculations, in order to identify the ratio of chromosomal aberrations, 5 preparates were selected for each application group and 500 cells, 100 of which were anaphase or telophase cells, were examined in each preparate and the number of chromosomal abnormalities (CA), nuclear abnormalities (NA) and micronucleus ratios were determined. Chromosomal abnormalities such as stickiness, C-mitosis, anaphase bridges, breakages, and laggards were analyzed. In the investigation of nuclear abnormalities, abnormalities in interphase nuclei were taken into consideration. Chromosomal aberrations and nuclear abnormalities were photographed at 10×100 magnification using a Kodak CX-6630 digital camera.

Analysis of variance (ANOVA) was applied to the data obtained and statistical analyses were performed using SPSS v.23 package program. Significant differences between data means were determined by Tukey Test.

In this study where the cytotoxic, genotoxic, and mutagenic effects of Fluquinconazole were investigated, root growth inhibition, which is both a macroscopic observation and an indicator of toxic effect, was first investigated. Table 1 shows the root inhibition ratios of Allium cepa treated with 10 different doses of Fluquinconazole. Among the doses applied to the test material, the 60 mg/L one was determined as the EC50 value and the doses to be used in the study were selected accordingly. In general, root growth retardation has been identified at doses above this dose. Color and structure changes in the roots, growth retardation, and little or no development of lateral roots are macroscopic indicators of the symptoms caused by the toxic effect (Fusconi et al. 2006; W ́ojicik and Tukendorf 1999). It is believed that the elongation inhibition in root growth may be due to a decline in root apical meristematic activity, which may be the result of a cytotoxic effect (Webster and MacLeod 1996; Yıldız et al. 2009). Moreover, it is stated that an inhibition in protein synthesis may also be effective in root growth (Seth et al. 2007). In the light of these opinions and results that we obtained, we think that Fluquinconazole fungicide inhibited protein synthesis in Allium cepa root cells and slowed or prevented cell division by causing a cytotoxic effect, resulting in a decrease in mitotic index, which in turn caused root growth inhibition.

Table 1

Table 2

The proportional changes in mitotic index as a result of the application of different doses of Fluquinconazole to A. cepa roots with 12, 24, and 48 hours of application periods are given in Table 2. Changes in mitotic phase are also shown in the same table. At the end of the evaluations, it was identified that the mitotic index declined at all application doses and times compared to the negative and positive control groups, which was statistically significant. Statistical analysis for mitotic phases showed a decrease in all phases compared to the control group. Increases or decreases in mitotic index and mitotic phases are important indicators that are used to show the cytotoxic effect of the substances tested (Marcano et al. 2004; Fernandes et al. 2007; Leme and Marin-Morales 2009). Although there are many studies showing the changes in mitotic index and mitotic stage ratios caused by pesticides, there is no study on this effect of Fluquinconazole (Bernardes et al. 2015; Verma and Srivastava 2018; Salazar-Mercado and Caleno 2019; Sheikh et al. 2020; Kalefetoğlu Macar 2020; Aragão et al. 2021; Gallego and Olivero-Verbel 2021; Khan et al. 2021). Mitotic index is an acceptable cytotoxic criterion for all living organisms. The cytotoxic level can be determined by a decrease in the mitotic index ratio. Reductions of less than 50% usually indicate a sublethal effect. If the reduction in mitotic index ratio exceeds 50%, it may have a lethal effect on the test organism. Inhibition of the mitotic index may be caused by the suppression of any of the G1, S, or G2 phases of DNA or by the impairment of RNA and/or protein synthesis (Grossmann et al. 2001; Sudhakar et al. 2001; Saxena et al. 2005; Kaymak 2005; Barriuso et al. 2010; Siddiqui et al. 2012; Singh and Roy 2017; Fioresi et al. 2020). In addition, mutations in cyclin and kinase proteins involved in the control of cell division also cause inhibition of mitotic index and mitotic stages (Marc et al. 2002, 2004; Aragão et al. 2021; Altman 2018). We believe that Fluquinconazole used in this study effectively decreased the mitotic index in A. cepa root tip cells compared to the control. The inhibition in root growth rates also supports this conclusion.

Table 3

Table 3 shows the types and ratios of chromosomal abnormalities caused by Fluquinconazole in A. cepa root tip cells and the total abnormality ratio. As can be also seen in the table, all doses we used in the study at 12, 24 and 48 hours of application caused more abnormalities than the control. In the statistical evaluations, it was identified that this situation was significant compared to the negative control group. There was no statistical difference between the 120 mg/L dose in the 12-hours application and the 30 mg/L dose in the 24-hours application. It was also detected that these groups showed similar ratios with the positive control group. In addition, 90 and 120 mg/L doses in the 24-hours application were found to be statistically insignificant. When the types of fungicide-induced chromosomal abnormalities were analyzed, stickiness was observed to occur at the highest rate. This was followed by C-mitosis, anaphase bridges, laggards, and chromosome breakages. At 48 hours of application, the number of these abnormalities increased. Used as a biological indicator of chromosomal damage and genome instability, the chromosomal abnormality test is one of the most widely applied and rapid tests in populations exposed to genotoxic agents (El-Zein et al. 2011; Suspiro and Prista 2011; Yüzbaşıoğlu et al. 2014). Chromosomal abnormalities are changes in normal chromosome structure (structural abnormality) or number (numerical abnormality) that occur spontaneously or as a result of exposure to chemicals/radiation (Russel 2002). The assessment of chromosomal abnormalities takes into account the disturbances in the four phases of the cell cycle (prophase, metaphase, anaphase, and telophase). Chromosomal abnormality analysis not only allows the prediction of genotoxic effects, but also helps in the evaluation of clastogenic and aneugenic formations. Chromosomal abnormalities include changes in chromosomal structure or chromosomal number. Changes in chromosomal structure can result from DNA breakdown, inhibition of DNA synthesis, and modification of DNA replication. The clastogenic effect causes various chromosomal abnormalities, including chromosomal breakages and bridges. Aneuploidy and polyploidy are numerical abnormalities caused by abnormal chromosome segregation (stickiness, C-metaphases, laggards, chromosome losses and multipolarity) either under the influence of aneugenic agents or spontaneously.

Studies have shown that chromosome stickiness is induced by most of the pesticides (Pulate and Tarar 2014a; Kutluer et al. 2019; Verma and Srivastava 2018). Chromosome stickiness can also be observed at high frequency due to the disruption of the nucleic acid metabolism of the cell. The sticky chromosomes caused abnormal unwinding of chromosomes during the transition from anaphase to telophase. Chromosome stickiness may be due to the delay in chromosome movement caused by fungicide application. Thus, the chromosome could not reach the poles and remained dispersed in the cytoplasm, resulting in a dense and sticky appearance. Chromosome stickiness is caused by the misfolding of chromosome fibers into single chromatids and chromosomes. As a result, the fibers become tangled, and the chromosomes are connected to each other via subchromatid bridges. The stickiness may occur due to the effect of the fungicide on the polymerization process or may result in the fragmentation of chromosomes (Chidambaram et al. 2009; Yüzbaşıoglu et al. 2003; Pulate and Tarar 2014b). It is stated that the stickiness of chromosomes is the result of a cytotoxic effect, which is irreversible and ultimately leads to cell death (Dizdari and Kopliku 2013; Goujon et al. 2014, Basu and Tripura 2021; Kundu and Ray 2016).

Another chromosome abnormality caused by Fluquinconazole is C-mitosis (Table 3). The term C-mitosis was coined by Levan (1938) and refers to a type of abnormality in which colchicine inhibits the formation of spindle apparatus, causing chromosomes to disperse within the cell. C-mitosis formation has been found in many studies investigating the genotoxic effects of pesticides (Türkoğlu 2007; Dizdari and Kopliku 2013; Fatma et al. 2018; Datta et al. 2018; Zeyad et al. 2019; Gallego and Olivero-Verbe 2021). Hsu et al. (1986) suggested that c-mitosis induced by mutagens is due to the blockage of tubulin polymerization or aggregation of microtubules and tubulin into crystalline forms. Fluquinconazole might have a colchicine-like effect, causing a change in the protein structure of spindle apparatus.

Another chromosome abnormality that occurred as a result of the study was recorded as anaphase bridge (Table 3). It has been reported that bridges are formed by the breakage and reassembly of chromosomes. The adhesion of chromosomes prevents the chromatids from separating from each other, leading them to remain attached to each other by bridges (Kabarity et al. 1974, Badr et al. 1992). It has been reported that adherent bridges may be the result of incomplete replication of chromosomes due to defective or underactivated replication enzymes (Sinha 1979) or late replication of telomeric heterochromatin DNA sequences (Bennet 1977). If heterochromatin blocks have not completed DNA replication when the nucleus is ready to divide, bridge formation might occur (Kaltsikes et al. 1984). In our study, the bridges observed in ana-telophase were usually single whereas double and triple bridges were also observed. Double and triple bridges are also noted to occur as a result of unequal polarization or dyscentric chromosomes, or due to chromosome breakages or fragmentation and failure of terminalisation (El-Ghamery et al. 2000, Luo et al. 2004).

Another abnormality caused by the chemical under investigation is laggard (Table 3). These chromosomes may have resulted from chromosomes that could not be attached to the spindle apparatus and moved towards each other from the two poles, or from acentric fragments. C-mitosis formations, which are among the abnormalities we obtained as a result of our study, are also due to disruption of the spindle apparatus structure. In this case, it can be thought that the substance we used disrupted the spindle apparatus structure and interfered with chromosome attachment in the metaphase phase of mitosis, resulting in laggards (Patil and Bhat 1992; Pulate and Tarar 2014 a, b; El-Ghamery and Mousa 2017). Laggards can lead to micronuclei formation and sometimes contribute to chromosomal evolution by causing aneuploid gametes.

Chromosome breakages occur when chromosomes are exposed to physical or chemical agents outside of normal conditions (Table 3). Different chemicals have been found to cause breakage by many researchers (İnceer et al. 2000; Kumar and Pannverselvan 2007; Khan et al. 2020; Hobs et al. 2017). Rieger et al. (1973) reported that heterochromatic regions of chromosomes are primarily broken. Fluquinconazole showed a clastogenic effect and caused breakages and fragments in the chromosomes of A. cepa root tip meristem cells. In this case, it can be said that this substance is also effective especially on heterochromatic regions.

Table 4

The effects of the substance used in this study on the nucleus were also investigated and the nuclear abnormalities observed are given in Table 4. The observation of NA in A. cepa root tip cells is an indicator of cytotoxic and genotoxic effect. Nuclear abnormalities in addition to chromosomal abnormalities have been examined in recent studies, paving the way for a better understanding of the effects of the test material used on the DNA of the organism to which it is applied and a more sensitive analysis of the results (Leme et al. 2008; Fernandes et al. 2009; Abdel Migid et al. 2007; Caritá and Marin-Morales 2008; Leme and Marin-Morales 2009; Nefic et al. 2013; Kassa 2021; Bonciu et al. 2018; Adrovic et al. 2021). NAs are categorized into five main groups: binuclei, notched nuclei, blebbed nuclei, MN, and lobed nuclei. In addition, cellular abnormalities with vacuoles, which is a sign of cytotoxic effect, can also be included in this group. These formations are observed in the interphase phase of the cell cycle. In our study, binucleated cells and vacuolated cells were observed. Binuclear cells are the result of a faulty cell division process in which karyokinesis is complete while cytokinesis is incomplete. According to Leme et al. (2008), the presence of these abnormal nucleated cells indicates a process of cell death. In addition, the presence of NAs can also occur as a result of clastogenic and aneugenic effects, which supports the toxic effect of the substance we used. Disruptions in the structure of the spindle apparatus and microtubules involved in the formation of the middle lamellae with the effect of the substance used cause errors in the anaphase phase, resulting in the formation of multinucleated cells as a result of karyokinesis followed by unsuccessful cytokinesis (Verma and Srivastava 2018; Fernandes et al. 2007; Fenech 2000, 2005; Fernandes et al. 2009, Nefic et al. 2013). Another nuclear abnormality we detected in the study was micronucleus (Table 4). Micronuclei (MN) are formations consisting of whole chromosomes or chromosome fragments without centromere, independent of the main nucleus, and located in the cytoplasm during mitosis. They are caused by deficiencies in genes involved in the control of the cell cycle, by defects in spindle apparatus, kinetochore or other parts of the mitotic apparatus, or by defects that cause chromosomal damage. Aneuploidy-inducing agents cause MN formation by preventing the centromeres from dividing properly and the spindle apparatus from functioning properly while clastogens cause chromosome breakages (Fenech et al. 2010, Kisurina-Evgenieva et al. 2016; Fernandes et al. 2007). Micronucleated cells (MNs) represent permanent DNA damage due to the gradual loss of genetic material induced by the first mitotic cycle after chemical treatment (Kopliko and Mesi 2012). This DNA damage can also be observed as chromosomal abnormalities (CAs) when cells divide. Table 4 shows that the fungicide used in the study triggered micronucleus formation in Allium and micronucleus rates increased significantly compared to the control, which was statistically significant. According to these results, Fluquinconazole triggered micronucleus formation in Allium cepa stem cells by causing disruptions in the structure of DNA and/or spindle apparatus or by causing breakages in chromosomes. In addition to micronucleus formation, the presence of binucleated cells and the observation of laggards in chromosome abnormalities also support this theory.

The results of this study have shown that the fungicide with Fluquinconazole active substance, which is widely used in Turkish agriculture especially in wheat production and sold in the market under different trade names, causes cytotoxic and genotoxic effects in Allium cepa root tip meristem cells. Reduction in root growth, inhibition of mitotic index, and chromosomal and nuclear abnormalities were identified, indicating that this substance should be considered an environmental hazard. The EC50 value is a useful parameter for selecting test concentrations in genotoxicity assays. The root inhibition observed in this process is detected as a decrease in meristematic activity, which is associated with inhibition of DNA and/or protein synthesis. In this case, it can be said that the fungicide used negatively affected root elongation by inhibiting protein synthesis and apical meristematic activity in A. cepa root tip meristem cells. The cytotoxicity levels of different chemicals can be identified by the variation in MI. A mitotic index lower than the negative control may indicate that the growth and development of the test organisms were affected by the test chemicals. The fungicide used was found to effectively reduce the mitotic index. This effect of the fungicide may occur at different stages of the cell cycle as well as at later stages. Based on the data we obtained, this substance is thought to be cytotoxic due to its mitodepressive effect. In addition, chromosomal (anaphase bridges, stickiness, c-mitosis, breakages, and laggards) and nuclear abnormalities (binucleus, micronucleus) observed at the end of the study indicate that the fungicide used is aneugenic and clastogenic. Furthermore, the A. cepa test proved once again to be a rapid and sensitive test in detecting environmental genotoxics and mutagens.

Fungi have evolved over one billion years and can be found in many habitats worldwide thanks to their adaptability and flexibility. Among a lot of species of fungi, some are pathogenic. People use fungicides to reduce production losses. However, while fungicides have contributed to a significant increase in agricultural productivity, their overuse has led to both health and environmental impacts. The first cytotoxic and genotoxic data related with Fluquinconazole have been obtained in this study. This fungicide and others containing Fluquinconazole will continue to be a cornerstone of the agricultural industry. However, in order to advocate for sustainable and environmentally friendly agriculture, it is recommended to develop safer formulations and to conduct research on lower concentrations and shorter exposure times. Since the data obtained in this study indicate that the fungicide has a cytogenotoxic effect, caution should also be exercised against unintended damage to both the applicators and the crops applied.

Table 1

*A. cepa* root growth subjected to different concentrations of fluquinconazole.
Concentrations (mg/L)	Root length of A. cepa (cm)	Growth (%)
Control	3.46	100
10	3.05	88
20	2.65	77
30	2.49	72
40	2.06	60
50	1.97	57
60	1.72	50
70	1.58	46
80	1.63	47
90	1.24	36
100	1.08	31

The means ± SD values with different letter (a, b, c, d) indicate statistically significant differences, according to Tukey (P ≤ 0.05). SD: Standard deviation.

Table 2

Mitotic indexes of the cells from *A. cepa* roots, submitted to different concentrations of fluquinconazole.
Time of treatment (h)	Doses (mg/L)	Total cells observed	Dividing cells	GMI ± SD *	MI % to control	PMI (%) ± S.D	MMI (%) ± S.D	AMI (%) ± S.D	TMI (%) ± S.D
	Control	5054	3296	65.21 ± 0.62 a	100	33.54 ± 0.19 a	12.85 ± 0.84 a	9.63 ± 1.64 a	9.19 ± 0.55 a
	MMS	5048	2046	40.54 ± 0.24 b	62	17.19 ± 0.11 b	11.93 ± 0.73 b	5.49 ± 0.87 b	5.93 ± 0.45 bg
	30	5107	2476	48.48 ± 0.15 c	74	31.27 ± 0.23 c	10.24 ± 0.89 c	3.51 ± 0.12 c	3.46 ± 0.58 c
12 h	60	5066	2285	45.10 ± 0.20 d	69	32.78 ± 0.16 a	9.12 ± 1.51 d	1.09 ± 0.17 d	2.11 ± 1.02 df
	90	5041	2193	43.50 ± 0.19 e	67	31.28 ± 0.14 c	8.17 ± 0.70 ej	3.07 ± 0.49 c	0.98 ± 0.56 ef
	120	5123	1925	37.57 ± 1.64 f	58	29.62 ± 1.43 d	6.11 ± 1.39 f	1.30 ± 0.84 d	0.54 ± 0.91 e
	Control	5049	3461	68.54 ± 1.25 g	100	36.91 ± 1.49 e	17.67 ± 1.67 g	7.51 ± 1.73 e	6.45 ± 0.97 b
	MMS	5061	2084	41.17 ± 1.35 b	60	18.35 ± 0.75 f	10.89 ± 1.92 h	6.64 ± 1.54 f	5.29 ± 1.25 g
	30	5072	1772	34.93 ± 0.76 h	51	21.18 ± 0.28 g	8.21 ± 1.34 ej	3.0 ± 0.65 c	2.54 ± 0.46 d
24 h	60	5165	1619	31.34 ± 1.11 i	46	18.57 ± 0.47 f	9.45 ± 0.45 d	1.87 ± 1.14 g	1.45 ± 0.95 dfk
	90	5082	1411	27.76 ± 0.94 j	40	11.83 ± 0.59 h	9.36 ± 0.89 d	4.12 ± 0.37 hk	2.45 ± 1.76 dhj
	120	5097	1245	24.42 ± 1.79 k	36	9.35 ± 1.94 i	8.29 ± 1.25 ej	4.31 ± 0.96 hjk	2.47 ± 1.14 d
	Control	5034	3247	64.50 ± 1.54 a	100	35.84 ± 0.98 j	13.47 ± 1.32 i	10.51 ± 1.24 i	4.68 ± 0.76 g
	MMS	5097	1859	36.47 ± 0.39 l	57	21.18 ± 0.73 g	8.51 ± 0.28 e	4.81 ± 0.78 j	1.97 ± 1.81 df
	30	5059	1172	23.16 ± 0.38 m	36	9.15 ± 0.33 ikm	7.82 ± 0.18 j	4.16 ± 0.41 k	2.03 ± 0.97 dhi
48 h	60	5124	955	18.63 ± 0.72 n	29	9.76 ± 1.10 im	5.17 ± 0.23 k	2.11 ± 1.28 g	1.59 ± 0.25 hfj
	90	5098	845	16.57 ± 0.47 o	26	8.42 ± 0.93 ik	4.92 ± 0.35 k	2.14 ± 0.64 g	1.09 ± 0.96 ef
	120	5024	528	10.50 ± 1.36 p	16	5.13 ± 1.29 n	3.68 ± 1.56 l	1.05 ± 0.87 d	0.64 ± 1.25 ek

The means ± SD values with different letter indicate statistically significant differences, according to Tukey (P ≤ 0.05). SD: Standard deviation. GMI: General mitotic index. PMI: Prophase mitotic index. MMI: Metaphase mitotic index. AMI: Anaphase mitotic index. TMI: Telephase mitotic index.

Table 3

Frequency of Chromosomal Anomalies (CA) at *A. cepa* tip of the root treated with different concentrations of fluquinconazole.
Time of treatment (h)	Concentrations (mg/L)	Total studied cells	Anaphase bridges (%)	Stickiness (%)	C-mitosis (%)	Laggards (%)	Breaks (%)	Chromosomal aberration (%)
	Control	500	0.74	1.01	0.38	0.28	-	2.41 ± 1.27 a
	MMS	500	4.56	6.14	3.54	0.31	0.95	15.50 ± 2.14 b
	30	500	3.21	5.24	3.16	0.70	1.57	13.88 ± 2.49 b
12 h	60	500	4.87	7.61	6.25	0.98	1.78	21.49 ± 1.04 c
	90	500	4.96	8.23	5.38	2.45	2.76	23.78 ± 0.57 cd
	120	500	5.19	7.86	6.31	3.95	2.17	25.48 ± 2.04 d
	Control	500	0.12	0.89	0.51	0.60	0.23	2.35 ± 0.23 a
	MMS	500	7.21	8.17	6.87	1.72	1.09	25.06 ± 0.18 b
	30	500	5.23	10.45	4.39	2.43	2.64	25.14 ± 0.15 b
24 h	60	500	8.47	15.28	9.51	5.71	3.28	42.25 ± 0.21 c
	90	500	9.34	19.79	13.47	7.36	2.69	52.65 ± 1.14 d
	120	500	11.63	20.87	15.41	3.00	2.36	53.27 ± 0.22 d
	Control	500	0.24	1.05	0.87	0.39	0.16	2.71 ± 0.28 a
	MMS	500	12.02	11.35	11.16	3.05	1.51	39.09 ± 0.11 b
	30	500	13.21	15.69	15.48	10.27	1.75	56.40 ± 1.25 c
48 h	60	500	17.24	24.03	19.94	10.78	3.05	59.04 ± 1.97 c
	90	500	16.17	19.36	14.26	10.65	2.09	62.53 ± 2.48 d
	120	500	12.64	21.67	15.97	11.28	3,41	64.97 ± 1.76 d

The means ± SD values with different letter indicate statistically significant differences, according to Tukey (P ≤ 0.05). SD: Standard deviation.

Table 4

Frequency of Nucleer Anomalies (NA) at *A. cepa* tip of the root treated with different concentrations of fluquinconazole.
Time of treatment (h)	Concentrations (mg/L)	Total studied cells	Micronuclei (%)	Binucleate cell (%)	Total Nuclear aberration (%)
	Control	500	0.00	0.00	0.00 a
	MMS	500	2.18	1.61	3.79 b
	30	500	5.34	4.29	9.63 c
12 h	60	500	8.21	8.35	16.56 d
	90	500	7.36	9.21	16.57 d
	120	500	9.17	10.87	20.04 e
	Control	500	0.00	0.00	0.00 a
	MMS	500	4.35	1.42	5.77 b
	30	500	1.24	4.54	5.78 b
24 h	60	500	6.38	7.63	14.01 c
	90	500	9.46	8.97	18.43 d
	120	500	13.41	11.13	24.54 e
	Control	500	0.00	0.00	0.00 a
	MMS	500	7.56	1.87	9.43 b
	30	500	13.43	6.51	19.94 c
48 h	60	500	17.51	9.28	26.79 d
	90	500	24.02	13.49	37.51 e
	120	500	25.98	12.98	38.96 e

The means ± SD values with different letter indicate statistically significant differences, according to Tukey (P ≤ 0.05). SD: Standard deviation.

Authors and Affiliations

Sivas Cumhuriyet University, Faculty of Science, Department of Biology, 58070, Sivas, Turkey.

Şifa Türkoğlu

Data availability

The data that supports the findings of this study is available for sharing from the corresponding author upon reasonable request.

Contributions

All of the scientific studies in this article were carried out by the author, Şifa Türkoğlu, and he critically revised the manuscript, commented on the drafts of the manuscript, and approved the final manuscript.

Corresponding author

Correspondence to Şifa Türkoğlu.

Ethics approval and consent to participate.

Not applicable.

Consent for publication

Not applicable.

Competing interests

The author declares no competing interests.

Funding

Not applicable

Abdel Migid HM, Azab YA, & Ibrahim WM (2007) Use of plant genotoxicity bioassay for the evaluation of efficiency of algal biofilters in bioremediation of toxic industrial effluent. Ecotoxicology and Environmental Safety 66(1): 57–64. DOI: 10.1016/j.ecoenv.2005.10.011
Adrovic J, Eminovic I, Stojko Vidovic S, and Feriz Adrovic F (2021) Chromosomal aberrations and nuclear anomalies in root tip cells of Allium cepa L. caused by radon in water. IJMBR (Int. J. Mod. Biol. Res.) 9: 25-36. doi.org/10.33500/ijmbr.2020.9.003
Altman J (2017) Pesticide interactions in crop production: Beneficial and deleterious effects. CRC Press. ISBN 9781315896359, 591 Pages.
Aragão FB, Duarte ID, Fantinato DE, Galter IN, Silveira GL, dos Reis GB, Andrade-Vieira LF, & Matsumoto ST (2021) Toxicogenetic of tebuconazole based fungicide through Lactuca sativa bioassays. Ecotoxicology and Environmental Safety 213, 111985. https://doi.org/10.1016/j.ecoenv.2021.111985
Badr A, Ghareeb A and El-Din HM (1992). Cytotoxicity of some pesticides in mitotic cells of V. faba roots. Egyptian Journal of Applied Sciences 7: 457–468.
Barriuso J, Marín S, & Mellado RP (2010) Effect of the herbicide glyphosate on glyphosate-tolerant maize rhizobacterial communities: A comparison with pre-emergency applied herbicide consisting of a combination of acetochlor and terbuthylazine. Environmental Microbiology 12(4): 1021–1030. DOI: 10.1111/j.1462-2920.2009.02146.x
Basu S and Tripura K (2021). Differential sensitivity of Allium cepa L. and Vicia faba L. to aqueous extracts of Cascabela thevetia (L.) lippold. South African Journal of Botany 139: 67–78. https://doi.org/10.1016/j.sajb.2021.01.033
Bateman GL, Gutterıdge RJ and Jenkyn JF (2004). Take-all and grain yields in sequences of winter wheat crops testing fluquinconazole seed treatment applied in different combinations of years. Ann. Appl. Biol. 145: 317-330. https://doi.org/10.1111/j.1744-7348.2004.tb00389.x
Bellani L, Muccifora S, Barbieri F, Tassi E, Ruffini Castiglione M, & Giorgetti L (2020). Genotoxicity of the food additive E171, titanium dioxide, in the plants Lens culinaris L. and Allium cepa L. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 849, 503142. https://doi.org/10.1016/j.mrgentox.2020.503142
Bennet MD (1977) Heterochromatin, aberrant endosperm nuclei and grain shriveling in wheat-rye genotypes. Heredity 39: 411-419.
Bernardes PM, Andrade-Vieira LF, Aragão FB, Ferreira A, & da Silva Ferreira MF (2015) Toxicity of Difenoconazole and tebuconazole in Allium cepa. Water, Air, & Soil Pollution 226(7). DOI: 10.1007/s11270-015-2462-y
Bonciu E, Roșculete E, Olaru AL, & Roșculete CA (2018) Evaluation of the mitodepressive effect, chromosomal aberrations and nuclear abnormalities induced by urea fertilization in the meristematic tissues of Allium cepa L. Caryologia 71(4): 350–356. https://doi.org/10.1080/00087114.2018.1473918
BKU (2023) https://bku.tarimorman.gov.tr/AktifMaddeGrup/Details/159?csrt=10387680295507611051&undefined=undefined (Last Access Date: 01/24/2023)
Caritá R and Marin-Morales MA (2008) Induction of chromosome aberrations in the Allium cepa test system caused by the exposure of seeds to industrial effluents contaminated with azo dyes. Chemosphere 72(5): 722–725. DOI: 10.1016/j.chemosphere.2008.03.056
Causil Vargas LA, Coronado JL, Verbel LF, Vega J, MF, Donado E, KA, & Pacheco GC (2017). Efecto Citotóxico del Hipoclorito de Sodio (naclo), en células ápicales de raíces de cebolla (Allium cepa L.). Revista Colombiana De Ciencias Hortícolas 11(1): 97–104. https://doi.org/10.17584/rcch.2017v11i1.5662.
Chauhan LKS, Saxena PN & Gupta SK (1999) Cytogenetic effects of cypermethrin and fenvalerate on the root meristem cells of Allium cepa. Environmental and Experimental Botany 42(3): 181–189. https://doi.org/10.1016/S0098-8472(99)00033-7
Chidambaram A, Sundaramoorthy P, Murugan A, Ganesh SK, Baskaran L (2009) Chromium induced cytotoxicity in Black gram (Vigna mungo L.). Ind. J. Environ. Health. Sci. Eng. 6 (1): 17-22.
Datta S, Singh J, Singh J, Singh S & Singh S (2018) Assessment of genotoxic effects of pesticide and vermicompost treated soil with Allium cepa test. Sustainable Environment Research 28(4): 171–178. https://doi.org/10.1016/j.serj.2018.01.005
de Morais CR, Pereira BB, Almeida Sousa PC, Vieira Santos VS, Campos CF, Carvalho SM, Spanó MA, de Rezende AAA, Bonetti AM (2019) Evaluation of the genotoxicity of neurotoxic insecticides using the micronucleus test in Tradescantia pallida. Chemosphere 227: 371-380. DOI: 10.1016/j.chemosphere.2019.04.073
de Sousa FA, de Morais CR, Vieira JS, Maranho LS, Machado FL, Pereira S, Barbosa LC, Coelho HE, Campos CF, Bonetti AM (2019) Genotoxicity and carcinogenicity of ivermectin and amoxicillin in vivo systems. Environ Toxicol Pharmacol. 70, 103196. DOI: 10.1016/j.etap.2019.103196
Dizdari AM and Kopliku D (2013) Cytotoxic and genotoxic potency screening of two pesticides on Allium cepa L. Procedia Technology 8: 19–26. https://doi.org/10.1016/j.protcy.2013.11.005
EFSA (2011) Conclusion on the peer review of the pesticide risk assessment of the active substance fluquinconazole, EFSA Journal 9(5): 2096
El-Ghamery AA and Mousa MA (2017) Investigation on the effect of benzyladenine on the germination, radicle growth and meristematic cells of Nigella sativa L. and Allium cepa L. Annals of Agricultural Sciences 62(1): 11–21. https://doi.org/10.1016/j.aoas.2016.11.002
El-Ghamery AA, El Nahas AI and Mansour MM (2000) The action of atrazine herbicide as an inhibitor of cell division on chromosomes and nucleic acids content in root meristems of Allium cepa and Vicia faba. Cytologia 55: 209–215. https://doi.org/10.1508/cytologia.65.277
El-Zein R, Vral A, & Etzel CJ (2010) Cytokinesis-blocked micronucleus assay and cancer risk assessment. Mutagenesis 26(1): 101–106. DOI: 10.1093/mutage/geq071
Fatma F, Verma S, Kamal A & Srivastava A (2018). Monitoring of morphotoxic, cytotoxic and genotoxic potential of Mancozeb using Allium assay. Chemosphere 195: 864–870. DOI: 10.1016/j.chemosphere.2017.12.052
Fenech M (2005) In vitro micronucleus technique to predict chemosensitivity. Methods Mol Med. 111:3-32. DOI: 10.1385/1-59259-889-7:003
Fenech M (2000). The in vitro micronucleus technique. Mutat Res. 455(1-2):81-95. https://doi.org/10.1016/S0027-5107(00)00065-8
Fenech M, Kirsch-Volders M, Natarajan AT, Surralles J, Crott JW, Parry J, Norppa H, Eastmond DA, Tucker JD & Thomas P (2010) Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells. Mutagenesis 26(1): 125–132. DOI: 10.1093/mutage/geq052
Fernandes TC, Mazzeo DE & Marin-Morales MA (2009) Origin of nuclear and chromosomal alterations derived from the action of an aneugenic agent—Trifluralin herbicide. Ecotoxicology and Environmental Safety 72(6): 1680–1686. DOI: 10.1016/j.ecoenv.2009.03.014
Fernandes, T.C.C., Mazzeo, D.E., & Marin-Morales, M.A., 2007. Mechanism of micronuclei formation in polyploidizated cells of Allium cepa exposed to trifluralin herbicide. Pesticide Biochemistry and Physiology, 88(3), 252–259. https://doi.org/10.1016/j.pestbp.2006.12.003
Fioresi VS, de Cássia Ribeiro Vieira B, de Campos JM & da Silva Souza T (2020) Cytogenotoxic activity of the pesticides imidacloprid and iprodione on Allium cepa root meristem. Environmental Science and Pollution Research 27(22): 28066–28076. DOI: 10.1007/s11356-020-09201-5
Fiskesjö G (1985) The Allium cepa test as a standard in environmental monitoring. Hereditas 102: 99-112.
Fusconi A, Repetto O, Bona E, Massa N, Gallo C, Dumas-Gaudot E & Berta G (2006) Effects of cadmium on meristem activity and nucleus ploidy in roots of Pisum sativum L. cv. frisson seedlings. Environmental and Experimental Botany 58(1-3): 253–260. https://doi.org/10.1016/j.envexpbot.2005.09.008
Gallego JL and Olivero-Verbel J (2021) Cytogenetic toxicity from pesticide and trace element mixtures in soils used for conventional and organic crops of Allium cepa L. Environmental Pollution 276, 116558. DOI: 10.1016/j.envpol.2021.116558
Goujon E, Sta C, Trivella A, Goupil P, Richard C & Ledoigt G (2014) Genotoxicity of Sulcotrione pesticide and photoproducts on Allium cepa root meristem. Pesticide Biochemistry and Physiology 113: 47–54. https://doi.org/10.1016/j.pestbp.2014.06.002
Grossmann K, Tresch S and Plath P (2001) Triaziflam and diaminotriazine derivatives affect enantioselectively multiple herbicide target sites. Zeitschrift Für Naturforschung C 56(7-8): 559–569. DOI: 10.1515/znc-2001-7-814
Gutteridge RJ, Jenkyn JF, Bateman GL (2007) The potential of non‐pathogenic Gaeumannomyces spp., occurring naturally or introduced into wheat crops or preceding crops, for controlling take‐all in wheat. Ann Appl Biol., 150: 53–64. https://doi.org/10.1111/j.1744-7348.2006.00107.x
Hobbs CA, Saigo K, Koyanagi M & Hayashi S (2017) Magnesium stearate, a widely-used food additive, exhibits a lack of in vitro and in vivo genotoxic potential. Toxicology Reports, 4: 554–559. DOI: 10.1016/j.toxrep.2017.10.003
Hsu TC, Liang JC and Sataya-Prakash KL (1986) Cytogeneticassays for mitotic poisons using somatic animal cells. In: Serres FJ [ed.], Chemical mutagens, principles and methods for their detection, 10: 155–182. ISBN: 978-1-4613-3694-5, Plenum Press.
Ilyushina NA, Egorova OV, Masaltsev GV, Averianova NS, Revazova YA, Rakitskii VN, Goumenou M, Vardavas A, Stivaktakis P, Tsatsakis A (2020) Genotoxicity of mixture of imidacloprid, imazalil and tebuconazole. Toxicology Reports 7: 1090-1094. DOI: 10.1016/j.toxrep.2020.08.021
Inceer H, Beyazoglu O and Ergul HA (2000) Cytogenetic Effects of Wastes of Copper Mine on Root Tip Cells of Allium cepa L. Pakistan Journal of Biological Sciences 3: 376-377. DOI: 10.3923/pjbs.2000.376.377
Kabarity A, El-Bayoumi AS and Habib AA (1974) Effect of morphine sulphate on mitosis of Allium cepa root tips. Biologia Plantarum 16: 275–282.
Kalefetoğlu Macar T (2020) Investigation of cytotoxicity and genotoxicity of abamectin pesticide in Allium cepa L. Environmental Science and Pollution Research 28(2): 2391–2399. DOI:10.1007/s11356-020-10708-0
Kaltsikes PJ (1984) Breeding vegetable varieties resistant to diseases. Proc. 3rd Meeting on Protected Vegetables and Flowers, May 911, Heraklion, Crete, p. 60. Abstract.
Kassa BA (2021) Cytotoxicity and genotoxicity evaluation of municipal wastewater discharged into the head of Blue Nile River using the Allium cepa test. Scientific African 13, e00911. https://doi.org/10.1016/j.sciaf.2021.e00911
Kaymak F (2004) Cytogenetic effects of maleic hydrazide on Helianthus annuus L. Pakistan Journal of Biological Sciences 8(1): 104–108. DOI: 10.3923/pjbs.2005.104.108
Khan A, Kumar V, Srivastava A, Saxena G & Verma PC (2021) Biomarker-based evaluation of cytogenotoxic potential of glyphosate in Vigna Mungo (L.) Hepper genotypes. Environmental Monitoring and Assessment 193(2): 73. DOI: 10.1007/s10661-021-08865-x
Khan IS, Ali MN, Hamid R & Ganie SA (2020) Genotoxic effect of two commonly used food dyes metanil yellow and Carmoisine using Allium cepa L. as indicator. Toxicology Reports 7: 370–375. DOI: 10.1016/j.toxrep.2020.02.009
Kihlman BA (1971) Root tips for studying the effects of chemicals on chromosomes. In: Hollaender, A., (Ed.), Chemical Mutagens, Plenum Press, New York, pp.489-514.
Kisurina-Evgenieva OP, Sutiagina, OI & Onishchenko GE (2016) Biogenesis of micronuclei. Biochemistry (Moscow) 81(5): 453–464. DOI: 10.1134/S0006297916050035
Kopliku D and Mesi AD (2012) Toxicity screening of water sources in flooded agricultural areas of NënShkodra lowland using Allium cepa L. assay. J. Environ. Sci. & Engin. A, 10: 1197-1202.
Kumar LP and Pannverselvan N (2007) Cytogenetic studies of food preservative in Allium cepa root meristematic cells. Facta Universitatis 14: 60-63.
Kundu LM & Ray S (2016) Mitotic abnormalities and micronuclei inducing potentials of colchicine and leaf aqueous extracts of Clerodendrum viscosum vent. in Allium cepa root apical meristem cells. Caryologia 70(1): 7–14. https://doi.org/10.1080/00087114.2016.1254452
Kutluer F, Çavuşoğlu K, Yalçın E (2019) The investigation of the physiological, anatomical and genotoxic effects in Allium cepa L. of Deltamethrin. Duzce University Journal of Science and Technology 7: 961-972. https://doi.org/10.29130/dubited.457074
Leme DM, Angelis D de & Marin-Morales MA (2008) Action mechanisms of petroleum hydrocarbons present in waters impacted by an oil spill on the genetic material of Allium cepa root cells. Aquatic Toxicology 88(4): 214–219. DOI: 10.1016/j.aquatox.2008.04.012
Leme DM and Marin-Morales MA (2009) Allium cepa test in environmental monitoring: A review on its Application. Mutation Research/Reviews in Mutation Research 682(1): 71–81. https://doi.org/10.1016/j.mrrev.2009.06.002
Levan A (1938) The effect of colchicine on root mitoses in Allium. Hereditas 24: 471-486.
Lovinskaya A, Kolumbayeva S, Begimbetova D, Suvorova M, Bekmagambetova N & Abilev S (2021). Toxic and genotoxic activity of river waters of the Kazakhstan. Acta Ecologica Sinica 41(6): 499–511. https://doi.org/10.1016/j.chnaes.2021.01.011
Luo LZ, Werner KM, Gollin SM & Saunders WS (2004) Cigarette smoke induces anaphase bridges and genomic imbalances in normal cells. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 554(1-2): 375–385. https://doi.org/10.1016/j.mrfmmm.2004.06.031
Marc J, Mulner-Lorillon O, Boulben S, Hureau D, Durand G & Bellé R (2002) Pesticide roundup provokes cell division dysfunction at the level of Cdk1/cyclin B activation. Chemical Research in Toxicology 15(3): 326–331. https://doi.org/10.1021/tx015543g
Marc J, Mulner-Lorillon O & Bellé R (2004) Glyphosate-based pesticides affect cell cycle regulation. Biology of the Cell 96(3): 245–249. DOI: 10.1016/j.biolcel.2003.11.010
Marcano L, Carruyo I, Del Campo A & Montiel X (2004) Cytotoxicity and mode of action of maleic hydrazide in root tips of Allium cepa L. Environmental Research 94(2): 221–226. DOI: 10.1016/s0013-9351(03)00121-x
Marinho MDC, Diogo BS, Lage OM, Antunes SC (2020) Ecotoxicological evaluation of fungicides used in viticulture in non-target organisms. Environ Sci Pollut Res Int. 27(35): 43958-43969. DOI: 10.1007/s11356-020-10245-w
Maurya R, Dubey K, Singh D, Jain AK, Pandey AK (2019) Effect of difenoconazole fungicide on physiological responses and ultrastructural modifications in model organism Tetrahymena pyriformis. Ecotoxicol Environ Saf. 182, 109375. DOI: 10.1016/j.ecoenv.2019.109375
Mercado SAA, Caleño JDQ, Jhan Piero Rojas Suárez JPR (2020) Cytogenotoxic effect of propanil using the Lens culinaris Med and Allium cepa L test. Chemosphere 249, 126193. DOI: 10.1016/j.chemosphere.2020.126193
Mercado SA and Caleño JD (2020) Cytotoxic evaluation of glyphosate, using Allium cepa L. as bioindicator. Science of The Total Environment 700, 134452. DOI: 10.1016/j.scitotenv.2019.134452
Nefic H, Musanovic J, Metovic A & Kurteshi K (2013) Chromosomal and nuclear alterations in root tip cells of Allium cepa L. induced by Alprazolam. Medical Archives 67(6), 388. doi: 10.5455/medarh.2013.67.388-392
Pitombeira de Figueirêdo, L, Athayde DB, Daam MA, van Gestel CAM, Guerra GDS, Duarte-Neto PJ, Espíndola ELG (2020) Impact of temperature on the toxicity of Kraft 36 EC® (a.s. abamectin) and Score 250 EC® (a.s. difenoconazole) to soil organisms under realistic environmental exposure scenarios. Ecotoxicol Environ Saf., 194, 110446. DOI: 10.1016/j.ecoenv.2020.110446
PUBCHEM (2023) https://pubchem.ncbi.nlm.nih.gov/compound/Fluquinconazole (Last Access Date: 01/24/2023)
Pulate PV and Tarar JL (2014a) Cytogenetic effects of tilt on root tip meristem of onion Allium cepa L. International Journal of Plant, Animal and Environmental Sciences (IJPAES) 4(2): 53-57.
Pulate PV and Tarar JL (2014b) Cytogenetic effect of systemic fungicide calixin on root meristem cells of Allium cepa L. International Journal of Life-Sciences Scientific Research 2: 341-345
Patil BC and Bhat GI (1992) A comparative study of MH and EMS in the induction of chromosomal aberrations on lateral root meristem in Clitoria ternatea L.. Cytologia 57(2): 259–264.
Ramanauskienė J, Semaškienė R, Jonavičienė A & Ronis A (2018) The effect of crop rotation and fungicide seed treatment on take-all in winter cereals in Lithuania. Crop Protection 110: 14–20. DOI: 10.1016/j.cropro.2018.03.011
Rank J (2003) The method of allium anaphase-telophase chromosome aberration assay. Ekologija Vilnius 1: 38- 42.
Rieger R, Nicolof H, Michaelis A (1973) Intro chromosomal clustering of chromatic aberrations induced by N-Methyl-N-NitrosoUrethan in Vicia faba and Barley. Biol. Zent. 92: 681-189.
Russel PJ (2002) Chromosomal mutation B. Cummings (Ed.), Genetics, Pearson Education Inc., San Francisco, pp. 595-621
Salazar Mercado SA and Maldonado Bayona HA (2019) Evaluation of cytotoxic potential of chlorpyrifos using Lens culinaris med as efficient bioindicator. Ecotoxicology and Environmental Safety 183, 109528. https://doi.org/10.1016/j.ecoenv.2019.109528
Saxena PN, Chauhan LKS & Gupta SK (2005) Cytogenetic effects of commercial formulation of cypermethrin in root meristem cells of Allium sativum: Spectroscopic basis of chromosome damage. Toxicology 216(2-3): 244–252. DOI: 10.1016/j.tox.2005.08.008
Scherm H, Christiano RSC, Esker PD, Del Ponte EM, Godoy CV (2009) Quantitative review of fungicide efficacy trials for managing soybean rust in Brazil. Crop Protection 28, (9): 774-782. DOI: 10.1016/j.cropro.2009.05.006
Seth CS, Chaturvedi PK & Misra V (2007). Toxic effect of arsenate and cadmium alone and in combination on giant duckweed (Spirodela Polyrrhiza L.) in response to its accumulation. Environmental Toxicology 22(6): 539–549. DOI: 10.1002/tox.20292
Sharma CBSR (1983) Plant meristems as Monitors of genetic toxicity of environmental chemicals. Current Science 52(21): 1000–1002.
Sheikh N, Patowary H & Laskar RA (2020) Screening of cytotoxic and genotoxic potency of two pesticides (malathion and cypermethrin) on Allium cepa L. Molecular & Cellular Toxicology 16(3): 291–299. DOI:10.1007/s13273-020-00077-7
Shishatskaya E, Menzyanova N, Zhila N, Prudnikova S, Volova T, Thomas S (2018) Toxic effects of the fungicide tebuconazole on the root system of fusarium-infected wheat plants. Plant Physiology and Biochemistry 132: 400-407. DOI: 10.1016/j.plaphy.2018.09.025
Siddiqui S, Meghvansi MK & Khan SS (2012) Glyphosate, alachor and maleic hydrazide have genotoxic effect on Trigonella foenum-Graecum L. Bulletin of Environmental Contamination and Toxicology 88(5): 659–665. DOI: 10.1007/s00128-012-0570-6
Singh D and Roy BK (2017) Evaluation of malathion-induced cytogenetical effects and oxidative stress in plants using Allium test. Acta Physiologiae Plantarum 39(4), 92. DOI : 10.1007/s11738-017-2391-z
Sinha U (1979) Cytomorphological and macromolecular changes induced by p-flurophenylalanine in Allium cepa and Triticale. Journal of Cytologia and Genetics 14: 198
Souza IR, Silva LR, Fernandes LSP, Salgado LD, Silva de Assis HC, Firak DS, Bach L, Santos-Filho R, Voigt CL, Barros AC, Peralta-Zamora P, Mattoso N, Franco CR, Soares Medeiros LC, Marcon BH, Cestari MM, Sant’Anna-Santos BF & Leme DM (2020) Visible-light reduced silver nanoparticles’ toxicity in Allium cepa test system. Environmental Pollution 257, 113551. https://doi.org/10.1016/j.envpol.2019.113551
Stapulionytė A, Kleizaitė V, Šiukšta R, Žvingila D, Taraškevičius R & Čėsnienė T (2019) Cyto/genotoxicological evaluation of hot spots of soil pollution using allium bioassays in relation to geochemistry. Mutation Research - Genetic Toxicology and Environmental Mutagenesis 842: 102-110. DOI: 10.1016/j.mrgentox.2019.01.001
Sudhakar R, Gowda KN & Venu G (2001) Mitotic abnormalities induced by silk dyeing industry effluents in the cells of Allium cepa. Cytologia 66(3): 235–239.
Suspiro A and Prista J (2011) Biomarkers of occupational exposure do anticancer agents: A minireview. Toxicology Letters 207(1): 42–52. DOI: 10.1016/j.toxlet.2011.08.022
Szpyrka E and Walorczyk S (2013). Dissipation kinetics of fluquinconazole and pyrimethanil residues in apples intended for Baby Food Production. Food Chemistry 141(4): 3525–3530. DOI: 10.1016/j.foodchem.2013.06.055
Thesai AS, Nagarajan G, Rajakumar S, Pugazhendhi A & Ayyasamy PM (2021) Bioaccumulation of fluoride from aqueous system and genotoxicity study on Allium cepa using Bacillus licheniformis. Journal of Hazardous Materials 407, 124367. https://doi.org/10.1016/j.jhazmat.2020.124367
Türkoğlu Ş (2007) Genotoxicity of five food preservatives tested on root tips of Allium cepa L. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 626(1-2): 4–14. DOI: 10.1016/j.mrgentox.2006.07.006
Verma S and Srivastava A (2018) Morphotoxicity and cytogenotoxicity of Pendimethalin in the test plant Allium cepa L. - a biomarker based study. Chemosphere 206: 248–254. DOI: 10.1016/j.chemosphere.2018.04.177
Webster PL and MacLeod RD (1996) The root apical meristem and its margin. Plant Roots. The Hidden Half (second ed.), Y. Waishel, A. Eshel, U. Kafkafi (Eds.), Marcel Dekker, New York, ISBN 0-8247-9685-3 pp. 51-76
Wójicik M and Tukendorf A (1999) CD - tolerance of maize, rye and wheat seedlings. Acta Physiologiae Plantarum 21(2): 99–107. DOI: 10.1007/s11738-999-0063-3
Wu S, Lei L, Liu M, Song Y, Lu S, Li D, Shi H, Raley-Susman KM, He D (2018) Single and mixture toxicity of strobilurin and SDHI fungicides to Xenopus tropicalis embryos. Ecotoxicol Environ Saf.,153: 8-15. DOI: 10.1016/j.ecoenv.2018.01.045
Yıldız M, Ciğerci İH, Konuk M, Fatih Fidan A & Terzi H (2009) Determination of genotoxic effects of copper sulphate and cobalt chloride in Allium cepa root cells by chromosome aberration and comet assays. Chemosphere 75(7): 934–938. DOI: 10.1016/j.chemosphere.2009.01.023
Yüzbaşıoğlu D, Zengin N, Ünal F (2014) Gıda koruyucuları ve genotoksisite testleri. Gıda, 39 (3), 179-186. doi: 10.5505/gida.24861
Yüzbaşioğlu D, Ünal F, Sancak C & Kasap R (2003) Cytological effects of the herbicide racer “flurochloridone” on Allium cepa. Caryologia 56(1): 97–105.
Zeyad MT, Kumar M & Malik A (2019) Mutagenicity, genotoxicity and oxidative stress induced by pesticide industry wastewater using bacterial and plant bioassays. Biotechnology Reports, 24, e00389. Doi: 10.1016/j.btre.2019.e00389
Zuno A, Marcon F, Leopardi P, Salvatore G, Carere A & Crebelli R (1994) An assessment of the in vivo clastogenicity of Erythrosine. Food and Chemical Toxicology 32(2): 159–163. Doi: 10.1016/0278-6915(94)90178-3

Download PDF

Version 1

posted

You are reading this latest preprint version

Evaluation of the cytotoxic and genotoxic effects of 2 Fluquinconazole using Allium cepa L. as a bioindicator

Status:

Version 1

Abstract

Introduction

Material And Methods

Results And Discussion

Conclusion

Declarations

References

Status:

Version 1