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Beni-Suef University Journal of Basic and Applied Sciences
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Assessment of genotoxic effects of wastewater of Kitchener pool, Nile Delta Region, North Egypt, using Allium test

Beni-Suef University Journal of Basic and Applied Sciences volume 12, Article number: 23 (2023) Cite this article

Abstract

Background

In the present study, Allium test bioassay was utilized to evaluate the effects of mixed wastewater of agricultural and sewage effluents at Kitchener pool, Gharbia governate, Middle Delta region, North Egypt. Germination indices, mitotic index and aberrations, α, β-esterase isozymes and inter-simple sequence repeat (ISSR) fingerprinting were tested by different concentrations of the wastewater (tap water as control, 25%, 50% and 100% wastewater).

Results

Water analysis recorded high levels of electrical conductivity, cations and anions compared to control, but were in the permitted limits according to FAO (Food and Agricultural Organization) except Mg2+ and K1+ were above the limits. P, N and heavy metals as Pb, Mn and Ni were also higher than the control. Germination indices showed reduction for all parameters studied (root and shoot lengths, root and shoot fresh and dry weights, and tolerance index). Mitotic index decreased, and the percentage of mitotic aberrations increased as the concentration of treatments increased and the time prolonged. Different types of aberrations were recorded in all treatments and its percentage is time and dose independent. Goat cells are the most common type recorded after different times in all treatments. The expression of α, β-esterase enzymes showed variation in different treatments compared to control and ISSR profiles showed considerable polymorphism. Concentration of 25% mixed water induced different profiles for expression of both α- and β-esterase from other treatments, and the cluster analysis based on polymorphism in ISSR fingerprinting revealed the distinction of plants treated with this concentration and the control plants from those treated with high concentrations.

Conclusion

It was suggested that concentration of 25% mixed water may be suitable for growth and act as fertilizer. Mixed water from this pool may be genotoxic for Allium cepa plants at early growth if it is used for irrigation in its present form and usage of this wastewater for agricultural purposes may be harmful and must be partially treated and biologically tested before use.

1 Background

The biotic and abiotic elements of the environment are negatively impacted by pollution. Water resource pollution is a major environmental challenge that affects the balance of natural ecosystems because of the increased release of dangerous chemicals and pollutants into the environment [21, 33]. The excessive influx of garbage from many sources into streams, pools and rivers has increased pollution, which negatively impacts both plants and animals as well as people [45]. Heavy metals may be released into the water due to wastes of industries and are slowly wiped off from ecosystems and may accumulate in different organisms causing series diseases. Exposure to chemical agents and pollutants has negative consequences on exposed species and have an impact on subsequent generations because these modifications may be inheritable, these consequences have emerged as being concerning [15]. Several researchers and governmental organizations are interested in this worldwide issue and are investigating its direct and indirect effects on the health of living organisms [30, 45].

There is no doubt that irrigation from rivers and lakes is a major source of support for agriculture in many nations. Unfortunately, water supply is insufficient in semiarid and arid regions. As a result, wastewater used to irrigate gardens, fields and many other agricultural purposes as an easy source of water without any consideration for its consequences. People that consume plants cultivated in contaminated soils may have health issues such as mental retardation, diarrhea, liver and kidney damage [4, 51]. Pollutants not only have an immediate negative impact on health, but they can also be mutagenic or toxic, which can result in human afflictions including cancer, atherosclerosis, cardiovascular disease and early aging [41].

According to recent studies, contaminants stimulate the generation of reactive oxygen species (ROS), which causes oxidative damage. With the purpose of quenching ROS and preventing oxidative cell damage under stressful conditions, organisms activate both non-enzymatic and enzymatic defensive mechanisms [46]. A possible biomarker of heavy metal pollution is esterase enzyme variation [38]. Genotoxicity has been detected using molecular markers [7] which have been effectively used to screen the drainage water's mutagenic effects. ISSR assay was mentioned as a more repeatable procedure and became more beneficial. This tool becomes extremely crucial since screening for genotoxic effects requires comparing patterns made by untreated (control) and treated plants [27, 54]. Furthermore, ISSR markers are highly polymorphic and regarded as effective in assessing the genotoxic potential of heavy metals [10].

Plant bioassays as Allium cepa, Vicia faba and Pisum sativum tests have been utilized for monitoring the potential synergistic effects of pollutants including chemicals [32, 50, 51], sewage wastewater, agricultural drainage wastewater and industrial wastes [33, 34, 44] and nanoparticles [28, 29], respectively. Allium cepa is a great test plant model for detecting of the harmful effects of substances. In this study, A. cepa aberration bioassay was utilized as a short-term test and low cost-effective indicator of toxicity in monitoring of water pollution. Germination indices, mitotic index and aberrations, α, β-esterase isozymes and ISSR fingerprinting were tested by different concentrations of the wastewater.

2 Methods

2.1 Study area and water samples and analysis

Qotour district is located at Gharbia Governorate in the Nile Delta region in Egypt (Fig. 1). Water samples were collected from Kitchener pool (30° 57′ 44.903″ E, 30° 57′ 59.618″ N) from Qutour district (Fig. 2) during year 2021. Three concentrations of mixed water (25%, 50% and 100%) were prepared and applied for the experiment and tap water used as control. Water analysis was carried out at central laboratory of Kafrelsheikh University and genetic and molecular biology laboratory at faculty of science Kafrelsheikh University, Egypt. PH number was measured using 3510 PH meter (Jenwary model). Electrical conductivity was measured using 4510 conductivity meter (Jenwary model). Total nitrogen was measured using Kjeldahl apparatus (VELP model). Na1+, K1+, Ca2+, Mg2+, Mn, Cu, Zn, Co, Ni, Cd and Pb were measured using atomic absorption spectrophotometry (GBC Avanta Σ model). Cl1−, HCO31− and SO42− were measured by titration method as in Gaagai et al. [17].

Fig. 1
figure 1

Map of the Nile Delta in Egypt including study area (Qotour district)

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Fig. 2
figure 2

A Map of the study area in Gharbia Governorate (Qutour district); B small map of the location of samples at Kitchener pool in Qutour district in the Governorate

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2.2 Germination studies

Onion bulbs of medium sizes were immersed in water to the middle with each concentration (25%, 50% and 100% of mixed water) and tap water as control. Three replicates were used for each treatment as well as control. Germination indices were recorded as length, fresh and dry weights for roots and shoots at the end of 12-day period of treatment. The tolerance index of seedlings was calculated according to [52] by the following formula:

$${\text{Tolerance index }} = \frac{{\text{Mean length of longest root in Sample}}}{{\text{Mean length of longest root in control}}}$$

2.3 Cytological studies

Onion bulbs were immersed in water to the middle with above concentrations and tap water as control for three periods of time 12, 24 and 36 h for cytological studies. 1–2 cm long of each root was cut and fixed in a freshly prepared Carney’s fixative solution for 3 h. Fixed roots were kept in 70% ethyl alcohol in the refrigerator until use. Treated roots were hydrolyzed in 1 N HCl at 60 °C for 10 min and stained in leuco-basic fuchsin stain for 3 h at room temperature. Five well-spread slides of each treatment were examined carefully using binuclear light microscope. Mitotic indexes and mitotic abnormalities were calculated as percentage. Types and percentages of abnormalities were also recorded.

2.4 Isozyme analysis

At the end of 12-day period of germination, 500 mg of leaves for each treatment was grinded in a mortar on ice bath using 1 ml of 0.025 sodium phosphate buffer pH = 7.25 with 20% (W/V) sucrose, left for two hours in the refrigerator and mixed using vortex every 15 min. Samples were centrifuged at 16,000 rpm for 20 min at 4 °C, and supernatants were kept at − 20 °C until used. Native polyacrylamide gel electrophoresis at 10% (w/v) was utilized for separation of isozymes (α- and β-esterase), and the gels were dyed substrates for the desired enzyme according to Soltis et al. [49].

2.5 DNA analysis

At the end of 12-day period of germination, DNA was extracted from young leaves according to manufacturer protocol of GeneJET Genomic DNA Purification Kit (K0721/ Thermo fisher). Total genomic DNA was amplified through Gene Amp Polymerase Chain Reaction (PCR) system cycler through specific ISSR primers (Table 5) in 20 μl reaction volume containing 1 μl from the primer, 2 μl genomic DNA (20 ng), 10 μl Dream Taq PCR Master MIX (Thermo Fisher Scientific, Inc.) and 7 μl dd H2O. Cycling condition was conducted as follows: initial denaturation for 4 min at 94 °C, 45 cycles of denaturation at 94 °C for 1 min, 1 min for annealing at 58 °C, extension for 2 min at 72 °C and finally extension at 72 °C for 8 min. DNA was visualized using 10 μl from PCR products on 1.5% agarose in TBE buffer with ethidium bromide at 100 V for 1 h and photographed by gel documentation system (Geldocit, UVP, England).

2.6 Data analysis

Data were statistically analyzed using one-way ANOVA (analysis of variance). DNA ISSR Bands were scored in binary matrices as 1 for presence and 0 for absence, and the similarity between different treatments was calculated using the Dice coefficient of similarity [13] using the NTSYS-pc software version 2.02 [43]. Building a distance tree to show the distances between the plants under study was done using the unweighted pair group approach using unweighted pair group method with arithmetic mean (UPGMA) [48] as implemented in the NTSYS-pc.

3 Results

In the present study, there was no difference in pH value between tap water and mixed water where the values (Table 1) were in the permitted limit according to MWE [39]. The electric conductivity value is high in mixed water recording 1.24 ds/m compared to 0.39 in control, but in the permitted limit according to FAO [6]. In relation to cations, concentrations of Mg2+ and K1+ measured in mixed water (6.33 and 0.28 meq/l) are high compared to control (2.76 and 0.16 meq/l) and FAO limits (4.94 and 0.05 meq/l), but Ca2+, Na1+ and all anions (Cl1−, SO42− and HCO31−) were in the permitted limits according to FAO (Table 1). The analysis of macroelements (P, N) and heavy metals also shows high values compared to control except Cu and Zn and N content is very high recording 12.53 mg/l against 2.8 mg/l in control which is probably due to sewage effluents (Table 2).

Table 1 Water analysis of PH, EC, cations and anions in mixed wastewater and tap water (control)
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Table 2 Water analysis of macroelements and heavy metals in tap water (control) and mixed wastewater
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Root length significantly decreased as the concentration increased after twelve days of germination (Table 3, Fig. 3) recording significant values of 5.00 and 4.7 cm compared to the control (8.67 cm). Shoot length measurement significantly decreased in mixed water as the concentration increased compared to control recording the highest value (7.83 cm) at 25% and the lowest values at the high concentrations (50 and 100%) with values (3.44 and 2.53 cm), respectively, compared to control (13.39 cm).

Table 3 Measurements of root and shoot indices (root and shoot length (cm), fresh and dry weights (gm) of Allium cepa treated with tap water as control, 25%, 50% and 100% of mixed wastewater at 12th day after germination
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Fig. 3
figure 3

Allium cepa treated with 1 tap water as control, 2 25%, 3 50% and 4 100% of mixed wastewater at 12th day after germination

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Root fresh and dry weights decreased as the concentration increased (Table 3), showing significant reduction in root fresh weight for highest concentration 100% with value of 0.91 gm compared to control (1.84 gm). All concentrations showed nonsignificant reduction in root dry weight compared to control. Shoot fresh weight and shoot dry weight also decreased as the concentration of mixed water increased, showing significant reduction in shoot fresh weight for treatments of 50% and 100% of mixed water with values of (4.8 gm and 2.95 gm) compared to control (15.34 gm). Different treatments of mixed water showed highly significant low values of tolerance index (0.57, 0.53 and 0.54) for concentrations 25%, 50% and 100% of mixed wastewater, respectively.

Table 4 presents low values of MI except concentration of 25% at time 12-h treatment (12.40%) and 24 h (11.10) compared to control (11.69%) and enhancement in percentage of total abnormalities with increase in concentration of treatments. Different types of abnormalities such as goat cells, micronuclei, sticky cells, bridges, c-mitosis (metaphase and anaphase), vacuolated and disturbed cells were frequently observed at various stages of mitosis at all treatments with time- and concentration-independent manner (Table 4, Fig. 4a–i). Goat cells (Fig. 4a) are the most common type recorded after different times at all treatments.

Table 4 Number of cells examined, mitotic index, percentage of total abnormalities, types and percentage of abnormalities of Allium cepa roots after different treatments of mixed water and control
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Fig. 4
figure 4

Different types of mitotic aberrations recorded by treated mixed water on Allium cepa root cells. a Goat cell, b vacuolated cells, c sticky cell and micronuclei, d c-metaphase, e c-anaphase, f anaphase, bridge, g multipolar cell, h micronucleus and i disturbed anaphase

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In this study, α-esterase is expressed as three isoforms (Fig. 5); α-EST (esterase) I isoform is present only in treatment of 25% mixed water, and α-EST II is expressed in all treatments and control except treatment of 25%. The isoform α-EST III is expressed in all treatments with different intensity and thickness and treatment of 25% mixed water records the highest intensity and thickness band. The β-esterase was expressed as five isoforms. The isoform β-EST I was absent in treatments of 50% and 100% mixed water with low intensity in treatment of 25% compared to control. The isoform β-EST II disappeared only in treatment of 25% mixed water, and β-EST III was expressed only in treatment of 25% mixed water. The isoforms β-EST IV and β-EST V were expressed with faint bands in all treatments and absent in control.

Fig. 5
figure 5

Expression of α-EST and β-EST zymogrames in roots of Allium cepa treated with (1) tap water as control, (2) 25%, (3) 50% and (4) 100% of mixed wastewater at 12th day after germination

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A great variation in banding pattern profile of ISSR fingerprinting in onion plants was recorded using 10 primers (Table 5). The primers produced a total of 108 bands including 54 polymorphic and 54 monomorphic markers in the control and treated plants with different concentrations of mixed water. Primer UBC 886 recorded the highest percentage of polymorphism (63.64%) with bands in range of size (94–977 bp). Differences in band intensity and thickness were also observed (Figs. 4 and 5). New markers were appeared in the used ISSR primers in the treated plants that were absent in the control such as band formed by primer UBC 808 with size of 280 bp, 3 bands formed by UBC 809 with sizes 210, 344 and 450 bp, and 5 bands formed by UBC 810 with sizes 407, 433, 469, 489 and 994 (Fig. 6). On the other hand, bands appeared in control and were absent in the other treatments such as 2 bands formed by UBC 857 with sizes 286 and 395, a band formed by UBC 884 with size 346, and 4 bands formed by UBC 886 with sizes 230, 337, 836 and 977 bp (Fig. 7).

Table 5 ISSR profile analysis using 10 primers in Allium cepa treated with tap water as control, 25%, 50% and 100% of mixed wastewater at 12th day after germination
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Fig. 6
figure 6

ISSR profile of Allium cepa treated with (1) tap water as control, (2) 25%, (3) 50% and (4) 100% of mixed wastewater at 12th day after germination, using 5 different primers. M = Standard DNA molecular size marker

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Fig. 7
figure 7

ISSR profile of Allium cepa treated with (1) tap water as control, (2) 25%, (3) 50% and (4) 100% of mixed wastewater at 12th day after germination, using 5 different primers M = Standard DNA molecular size marker

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A cluster analysis (Fig. 8) that was constructed using the arithmetic average (UPGMA) clearly revealed the distinction of plants treated with 100% mixed water and to some extent the plants treated with 50% mixed water from the control plants and plants treated with 25% mixed water.

Fig. 8
figure 8

Dendrogram illustrating differentiation of Allium cepa treated with tap water as control, 25%, 50% and 100% of mixed wastewater at 12th day after germination based on polymorphism in ISSR fingerprinting

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4 Discussion

Cations and anions measured in mixed water are high compared to control, but were in the permitted limits according to FAO [6] except Mg2+ and K1+ were above the limits. N, P and heavy metals recorded high values compared to control which probably due to sewage effluents and this to some extent affects water content and causes a severe problem to crop plants when used for irrigation.

Root length significantly decreased in mixed water as the concentration increased after twelve days of germination that was consistent with Kassa et al. [25] and Admas and Kerisew [1]. This reduction might be caused by accumulation of metals in roots than other plant parts, as previously indicated by [31]. Also, shoot length measurement significantly decreased as the concentration increased and these results were consistent with Goli and Sahu [16] and Kapil and Mathur [24]. Hajihashemi et al. [18] reported that reduction in growth may be due to reduction of the photosynthetic characteristics, nutrient concentrations and carbohydrates in treated plants. With increasing effluent concentrations of industry wastewater, a gradual decline in the germination of seed and seedling growth of Vigna radiata was observed by Kothari et al. [26]. Shoot dry weight recorded nonsignificant reduction in all treatments compared to control as previously stated by Goli and Sahu [16] and Kamlesh [23]. The root length can serve as a crucial tolerance index due to suppression of root elongation is the first noticeable effect of metal toxicity [19]. Different treatments of mixed water showed highly significant low values of tolerance index that agreed with Mehta and Bhardwaj [36] and Kamlesh [23].

For all living organisms, the mitotic index is regarded as an acceptable indicator of cytotoxicity [44]. Abnormalities that were recorded in control (1.3%) may be due to hard handling of slides during squash technique rather any other effects [20, 37]. MI of 25% concentration was, approximately, the same as in control that it was suggested this concentration might be suitable for growth and act as fertilizer and agreed with Maxim and Andrey [35], Dourado et al. [14] and Cresenico et al. [12]. The lowest value recorded for MI by concentration of 100% at 36-h treatment is time and dose dependent, and percentage of mitotic aberrations increased as the concentration increased and the time prolonged as previously recorded by Labeeb [27], Mohamad [37], Cresenico et al. [12] and Admas and Kerisew [1]. This result proves the harmful and toxic effect of mixed water in this pool, especially when used in its present form for long time.

Different types of abnormalities such as goat cells, micronuclei, sticky cells, bridges, c-mitosis (metaphase and anaphase), vacuolated and disturbed cells were frequently observed at various stages of mitosis at all treatments with time- and concentration-independent manner. Goat cells are the most common type recorded after different times at all treatments; to our knowledge, this aberration was not previously recorded by any treatment with polluted water. To some extent, this may be due to the double action of mixed water rich in different types of pollutants and heavy metals from either drainage water or sewage effluents. The high percentage of goat cells may also be due to interaction of some abiotic factors [3, 4]. The other types of aberrations were previously recorded by many authors using different plants irrigated with polluted water [1, 21, 25, 27, 37].

The results of α- and β-esterase isozymes illustrate that concentration of 25% mixed water induces different profiles for expression of both α- and β-esterase from other treatments that may be agree with our previous suggestion that this concentration is suitable for growth and may act as fertilizer. The variation in expression of esterase isozymes in our study may be a sign that plant defense systems have been activated to withstand the stress caused by heavy metals and other contaminants present in these effluents as previously stated by Mukherjeea et al. [38], Mattar et al. [34] and Labeeb et al. [29].

A great variation in banding pattern profile of ISSR fingerprinting in onion plants was recorded, and differences in band intensity and thickness were also observed. Following exposure to environmental pollutants, polymorphism of molecular markers created by DNA fingerprinting using the randomly amplified polymorphic DNA (RAPD) and ISSR linked genotoxicity to genetic variation [47]. Also, Bajpai et al. [8], Bakry et al. [9] and Badr et al. [7] used ISSR-PCR technique to investigate genotoxicity. Changes in DNA banding profiles indicate DNA modifications from single base alterations to chromosomal rearrangements [5], possibly because of the mutagenicity and toxicity of the heavy metals in the effluent [2]. Also, the stress causes a quick and temporary overproduction of reactive oxygen species (ROS), which damages DNA in plants [7]. New markers were appeared in the used ISSR primers in the treated plants that were absent in the control and bands appeared in control and were absent in the other treatments. The alternation of DNA profiles including loss or appearance of some bands occurs as a result of changes in oligonucleotide priming sites that impact the activity and interaction of DNA polymerase with changed DNA [11, 40, 42].

A cluster analysis clearly revealed the distinction of plants treated with 100% mixed water and to some extent the plants treated with 50% mixed water from the control plants and plants treated with 25% mixed water. These results were consistent with Kamal et al. [22]. Concentration of 25% mixed water is the most relative to control (tap water) that may be agree with our previous suggestion that this concentration is suitable for growth and may act as fertilizer.

As this pool is frequently considered an easy source of water for irrigation, as well as the ignorance of many farmers, this study illustrates how much toxicity of this water on crops and vegetables when used directly or indirectly or incorporated in food chain of the ecosystem as previous studies of plants were affected by industrial waste [53, 55].

5 Conclusion

Different treatments of mixed water showed significant decrease in root and shoot lengths as the concentration increased and highly significant low values of tolerance index. Different types of abnormalities were frequently observed at various stages of mitosis at all treatments with time- and concentration-independent manner and goat cells were the most common type. MI of 25% concentration of mixed water was, approximately, the same as in control, this concentration induced different profiles for expression of both α- and β-esterase, and a cluster analysis based on polymorphism in ISSR fingerprinting revealed the distinction of plants treated with it and the control plants from those treated with high concentrations. It was suggested that concentration of 25% mixed water may be suitable for growth and act as fertilizer. Water from this pool may be genotoxic for Allium cepa plants at early growth if it is used for irrigation in its present form and usage of this wastewater for agricultural purposes may be harmful and must be partially treated and biologically tested before use.

Availability of data and materials

Not applicable for this section.

Abbreviations

ISSR:

Inter-simple sequence repeat

EC:

Electric conductivity

MI:

Mitotic index

ROS:

Reactive oxygen species

PCR:

Polymerase chain reaction

ANOVA:

Analysis of variance

UPGMA:

Unweighted pair group method with arithmetic mean

Micro:

Micronuclei

Stick.:

Stickiness

Vac. cell:

Vacuolated cell

C-Mito:

C-Metaphase

Dist.:

Disturbed chromosome

EST:

Esterase

RAPD:

Randomly amplified polymorphic DNA

FAO:

Food and Agricultural Organization

References

  1. Admas T, Kerisew B (2022) Assessment of cytotoxicity and genotoxicity potential of effluents from Bahir Dar Tannery using Allium cepa. Adv Public Health 7:1–10

    Article  Google Scholar 

  2. Ahmed HH, Abonama OM (2017) Molecular identification effect of wastewater on plant and bacterial DNA. Int J Adv Res 5:1113–1121

    Article  CAS  Google Scholar 

  3. Amal MEA (2016) Physiological and biochemical responses to heat stress on barley seedlings and their impact on growth and yield. Egypt J Bot 56(1):319–334

    Article  Google Scholar 

  4. Alvarenga IFS, Dos Santos FE, Silveira GL, Andrade-Vieira LF, Martins GC, Guilherme LRG (2020) Investigating arsenic toxicity in tropical soils: a cell cycle and DNA fragmentation approach. Sci Total Environ 1:698. https://doi.org/10.1016/j.scitotenv.2019.134272

    Article  CAS  Google Scholar 

  5. Atienzar FA, Venier P, Jha AN, Depledge MH (2002) Evaluation of the random amplified polymorphic DNA (RAPD) assay for the detection of DNA damage and mutations. Mutat Res 521(1–2):151–163

    Article  CAS  PubMed  Google Scholar 

  6. Ayers R, Westcott D (1994) Water Quality for Agriculture. FAO Irrigation and Drainage, Paper 29 Revision 1 Food and Agricultural Organization of the United Nations, Rome, Italy

  7. Badr A, El-Shazly HH, Mohamed HI (2021) Plant responses to induced genotoxicity and oxidative stress by chemicals. In: Khan Z, Ansari MYK, Shahwar D (eds) Induced genotoxicity and oxidative stress in plants. Springer, Singapore, pp 103–131

    Chapter  Google Scholar 

  8. Bajpai R, Shukla V, Singh N, Rana TS, Upreti DK (2015) Physiological and genetic effects of chromium (+VI) on toxitolerant lichen species. Pyxine Cocoe Environ Sci Pollut Res 22(5):3727–3738

    Article  CAS  Google Scholar 

  9. Bakry FA, Ismail SM, El-Atti MSA (2015) Glyphosate herbicide induces genotoxic effect and physiological disturbances in Bulinus truncatus snails. Pestic Biochem Physiol 123:24–30

    Article  CAS  PubMed  Google Scholar 

  10. Bernardes PM, Andrade-Vieira LF, Aragão FB, da Silva FA, Ferreira MF (2015) Toxicity of difenoconazole and tebuconazole in Allium cepa. Water Air Soil Pollut 226(7):207

    Article  Google Scholar 

  11. Correia S, Matos M, Ferreira V, Martins N, Conclaves S, Romano A, Carnide OP (2014) Molecular instability induced by aluminum stress in Plantago species. Mutat Res Genet Toxicol Environ Mutagen 770:105–111

    Article  CAS  PubMed  Google Scholar 

  12. Cresenico C, Cubuga J, Apostado R, Hernando H, Lador C, Havana H (2017) Allium cepa test: an evaluation of genotoxicity. Proc Int Acad Ecol Environ Sci 7(1):12–19

    Google Scholar 

  13. Dice LR (1945) Measure of the amount of ecologic association between species. Ecology 26(3):297–302

    Article  Google Scholar 

  14. Dourado P, Rocha L, Roveda J, Juinor L, Candido C, Cardoso M, Morales M, Olivera P, Grisolia A (2017) Genotoxic and mutagenic polluted surface water in the Midwestern rejoin of Brazilusing animal and plant bioassays. Gen Mol Boil 40(10):123–133

    CAS  Google Scholar 

  15. Elawady M, Marzouk N, Bakry B, Abderhman M, Kenawy A (2017) Treated wastewater impact on rural green farm life cycle, Egypt. Egypt J Chem 60:1–6

    Google Scholar 

  16. Goli J, Sahu O (2014) Effect of distillery industry effluent on fertility of soil and crops. Int J Soil Crop Sci 2:39–45

    Google Scholar 

  17. Gaagai A, Aouissi HA, Bencedira S, Hinge G, Athamena A, Haddam S, Gad M, Elsherbiny O, Elsayed S, Eid MH, Ibrahim H (2023) Application of water quality indices, machine learning approaches, and GIS to identify groundwater quality for irrigation purposes: a case study of Sahara Aquifer, Doucen Plain, Algeria. Water 15:289. https://doi.org/10.3390/w15020289

    Article  CAS  Google Scholar 

  18. Haji Hashemi S, Mbarki S, Skalicky M, Noedoost F, Raeisi M, Brestic M (2020) Effect of wastewater irrigation on photosynthesis, growth, and anatomical features of two wheat cultivars (Triticum aestivum L.). Water 12:607

    Article  Google Scholar 

  19. Han YL, Yuan HY, Huang SZ, Guo Z, Xia B, Gu J (2007) Cadmium tolerance and accumulation by two species of Iris. Ecotoxicology 16:557–563

    Article  CAS  PubMed  Google Scholar 

  20. Haroun SA, Aboulgaith A (2015) Allelopathey effects of Zygophyllum simplex extract on Vicia faba L. Cytologia 80(3):1–9

    Article  Google Scholar 

  21. Juliana S, Luiza F, Bruno D, Alexia B (2019) Influence of land use and cover on toxogenetic potential of surface water from central west Brazilian rivers. Arch Environ Contam Toxicol 76(3):483–495

    Article  Google Scholar 

  22. Kamal MI, El-Hady A, Zaied KA, El-Mohsen A (2020) Assessment of water quality via genetic diversity induced in onion using Inter Simple Sequence Repeats (ISSR) markers. J Agric Chem Biotechnol 11(4):107–115

    Google Scholar 

  23. Kamlesh (2019) Phytotoxic effect of paper mill effluent treatment on seed germination and seedling growth of wheat (Triticum aestivum) and radish (Raphanus sativus). Int J Adv Sci Res Manag 4(2):153–160

    Google Scholar 

  24. Kapil J, Mathur N (2020) The impact of dairy effluent on germination parameters of seeds of mung bean (Vigna radiata) and mustard (Brassica nigra). Int J Econ Plants 7(4):170–175

    Article  Google Scholar 

  25. Kassa BA (2021) Cytotoxicity and genotoxicity evaluation of municipal wastewater discharged into the head of Blue Nile River using the Allium Cepa test. Sci Afr 13:e00911

    CAS  Google Scholar 

  26. Kothari R, Pathak AK, Sharma V, Ahmad S, Singh H, Singh RP, Tyagi VV (2022) Impact of pollutant load from textile dyeing industry wastewater on biometric growth profile of Vigna radiata. Bull Environ Contam Toxicol. https://doi.org/10.1007/s00128-022-03491-w

    Article  PubMed  Google Scholar 

  27. Labeeb M (2014) Impact of polluted irrigation water in Kafrelsheikh governorate on agronomic, cytological and molecular traits of Vicia faba L. Dissertation, University of Menoufia, Egypt.

  28. Labeeb M, Badr A, Haroun SA, Mattar MZ, El-Kholy AS, El-Mehasseb IM (2020) Ecofriendly synthesis of silver nanoparticles and their effects on early growth and cell division in roots of green pea (Pisum sativum L.). Gesunde Pflanz 72:113–121

    Article  CAS  Google Scholar 

  29. Labeeb M, Badr A, Haroun SA, Mattar MZ, El-Kholy AS (2022) Ultrastructural and molecular implications of ecofriendly made silver nanoparticles treatments in pea (Pisum sativum L.). J Genet Eng Biotechnol 20(5):1–14

    Google Scholar 

  30. Leme D, Marin-Morales M (2009) Allium cepa test in environmental mentoring: a review on its application. Mutat Res 682:71–81

    Article  CAS  PubMed  Google Scholar 

  31. Liao L, Xu J, Peng S, Qiao Z, Gao X (2013) Uptake and bioaccumulation of heavy metals in rice plants as affect by water saving irrigation. Adv J Food Sci Technol 5(9):1244–1248

    Article  Google Scholar 

  32. Macar O, Kalefetoğlu Macar T, Yalçın E, Çavuşoğlu K (2022) Acute multiple toxic effects of Trifloxystrobin fungicide on Allium cepa L. Sci Rep 12:15216

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Mattar MZ, Sammour RH, Soliman HA, El-kholy AS, Seada ML (2014) Cytogenetic impact of different types of polluted water on Vicia faba L. at Kafr El-Sheikh governorate, Egypt. J Am Sci 10(8):100–107

    Google Scholar 

  34. Mattar MZ, Sammour RH, El-kholy AS, Seada ML (2015) Detection of genotoxic effect of wastewater on Vicia faba L. by using biochemical assay and RAPD markers. Egypt J Bot 55(2):281–296

    Article  Google Scholar 

  35. Maxim V, Andry R, Irina A, Anna R (2013) Lterations revealed in Allium cepa test system under the action of some xenobiotics. World Appl Sci J 22(3):342–344

    Google Scholar 

  36. Mehta A, Bhardwaj N (2012) Phytotoxic effect of industrial effluents on seed germination and seedling growth of Vigna radiata and Cicer arietinum. GJBB 1(1):1–5

    Google Scholar 

  37. Mohamad N (2019) Effluent quality assessment of selected wastewater treatment plant in Jordan for irrigation purposes: water quality index approach. J Ecol Eng 20(10):206–216

    Article  Google Scholar 

  38. Mukherjeea S, Mukherjeea S, Bhattacharyyab P, Duttaguptaa AK (2004) Heavy metal levels and esterase variations between metal-exposed and unexposed duckweed Lemna minor: field and laboratory studies. Environ Int 30(6):811–814

    Article  Google Scholar 

  39. MWE (2006) Ministry of water and electricity, technical guidelines for the use of treated sanitary wastewater in irrigation for landscape and agriculture irrigation. Saudi Arabia

  40. Poyraz IE, Poyraz I, Kiyan HT, Öztürk N, Erken S, Gülbağ F (2018) Detection of the genotoxicity of gentiana L. extracts by using RAPD-PCR and ISSR-PCR techniques. Indian J Pharm Educ Res 52(4):133S-139S

    Article  Google Scholar 

  41. Radić S, Stipanicev D, Vujcić V, Rajcić MM, Sirac S, Pevalek-Kozlina B (2010) The evaluation of surface and wastewater genotoxicity using the Allium cepa test. Sci Total Environ 408(5):1228–1233

    Article  PubMed  Google Scholar 

  42. Rai KK, Rai N, Rai SP (2018) Salicylic acid and nitric oxide alleviate high temperature induced oxidative damage in Lablab purpureus L. plants by regulating bio-physical processes and DNA methylation. Plant Physiol Biochem 128:72–88

    Article  CAS  PubMed  Google Scholar 

  43. Rohlf FJ (2002) NTSYS-pc-Numerical taxonomy and multivariate analysis system. Applied Biostatistics Inc, New York

    Google Scholar 

  44. Smaka K, Stegnar P, Lovoka M, Toman M (1996) The evaluation of wastewater, surface and ground water quality using the Allium cepa procedure. Mutat Res 368:171–179

    Article  Google Scholar 

  45. Sana K, Mohammad A, Malik A (2019) Mutagenicity and toxicity evaluation of textile industry wastewater using bacteria and plant bioassay. Toxicol Rep 6:193–201

    Article  Google Scholar 

  46. Sairam RK, Tyagi A (2004) Physiological and molecular biology of salinity stress tolerance in plants. Curr Sci 86:407–420

    CAS  Google Scholar 

  47. Savva D (1998) Use of DNA fingerprinting to detect genotoxic effects. Ecotoxicol Environ Saf 41:103–106

    Article  CAS  PubMed  Google Scholar 

  48. Sokal RR, Michener CD (1958) A statistical method for evaluating systematic relationships. Univ Kansas Sci Bull 28:1409–1438

    Google Scholar 

  49. Soltis D, Haufler C, Darrow D, Gastony G (1983) Starch gel electrophoresis of ferns: a compilation of grinding buffers, gel and electrode buffer, and staining schedules. Am Fern J 73:9–27

    Article  Google Scholar 

  50. Simata K, Sudhir B, Pratibha A, Paliwal J (2019) The evaluation of various organic solvent using A. cepa test. Eur J Biomed Pharm Res 4(12):820–823

    Google Scholar 

  51. Tugce K, Oksal M, Emine Y, Kultigin C (2020) Resveratrol ameliorates the physiological, biochemical, cytogenetic and anatomical toxicities induced by copper11 chloride exposure in Allium cepa. Environ Sci Pollut Res 27(1):657–667

    Article  Google Scholar 

  52. Turner RG, Marshal C (1972) Accumulation of zinc by subcellular root of Agrostis tannis sibth in relation of zinc tolerance. New Phytol 71:671–676

    Article  CAS  Google Scholar 

  53. Uzair M, Ahmad M, Nazim K (2009) Effects of industrial waste on seed bank and growth of wild plants in Dhabeji area, Karachi, Pakistan. Pak J Bot 41(4):1659–1665

    Google Scholar 

  54. Vanijajiva O (2011) Genetic variability among durian (Durio zibethinus Murr.) cultivars in the Nonthaburi province, Thailand detected by RAPD analysis. J Agric Technol 7:1105–1114

    Google Scholar 

  55. Wahid A, Ahmad S, Rehman S, Ahmad S (2004) Growth and biochemical status of wheat seedling treated with industrial effluents. Biologia 50(1):91–98

    CAS  Google Scholar 

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  1. Botany and Microbiology Department, Faculty of Science, Kafrelsheikh University, Kafr Elsheikh, Egypt

    Aziza S. El-Kholy, Soliman A. Haroun & May Labeeb

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  1. Aziza S. El-Kholy
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ASE was involved in experimental investigations, data interpretation, and reviewing and editing the manuscript. SAH was responsible for conceptualization, and reviewing and editing the manuscript. ML contributed to experimental investigations, writing of the original draft, data interpretation, and reviewing and editing the manuscript. All authors read and approved the final version.

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El-Kholy, A.S., Haroun, S.A. & Labeeb, M. Assessment of genotoxic effects of wastewater of Kitchener pool, Nile Delta Region, North Egypt, using Allium test. Beni-Suef Univ J Basic Appl Sci 12, 23 (2023). https://doi.org/10.1186/s43088-023-00364-x

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