Advances in bread wheat production through CRISPR/Cas9 technology: a comprehensive review of quality and other aspects.

MAIN CONCLUSION: This review provides a comprehensive overview of the CRISPR/Cas9 technique and the research areas of this gene editing tool in improving wheat quality. Wheat (Triticum aestivum L.), the basic nutrition for most of the human population, contributes 20% of the daily energy needed because of its, carbohydrate, essential amino acids, minerals, protein, and vitamin content. Wheat varieties that produce high yields and have enhanced nutritional quality will be required to fulfill future demands. Hexaploid wheat has A, B, and D genomes and includes three like but not identical copies of genes that influence important yield and quality. CRISPR/Cas9, which allows multiplex genome editing provides major opportunities in genome editing studies of plants, especially complicated genomes such as wheat. In this overview, we discuss the CRISPR/Cas9 technique, which is credited with bringing about a paradigm shift in genome editing studies. We also provide a summary of recent research utilizing CRISPR/Cas9 to investigate yield, quality, resistance to biotic/abiotic stress, and hybrid seed production. In addition, we provide a synopsis of the laboratory experience-based solution alternatives as well as the potential obstacles for wheat CRISPR studies. Although wheat's extensive genome and complicated polyploid structure previously slowed wheat genetic engineering and breeding progress, effective CRISPR/Cas9 systems are now successfully used to boost wheat development.

Biosynthesis of capsaicinoids in pungent peppers under salinity stress.

The synthesis of capsaicinoids occurs in the placenta of the fruits of pungent peppers. However, the mechanism of capsaicinoids' biosynthesis in pungent peppers under salinity stress conditions is unknown. The Habanero and Maras genotypes, the hottest peppers in the world, were chosen as plant material for this study, and they were grown under normal and salinity (5 dS m-1 ) conditions. The results showed that salinity stress harmed plant growth but increased the capsaicin content by 35.11% and 37.00%, as well as the dihydrocapsaicin content by 30.82% and 72.89% in the fruits of the Maras and Habanero genotypes, respectively, at 30 days after planting. The expression analysis of key genes in capsaicinoids biosynthesis revealed that the PAL1, pAMT, KAS, and PUN1 genes were overexpressed in the vegetative and reproductive organs of pungent peppers under normal conditions. However, under salinity stress, overexpression of PAL1, pAMT, and PUN1 genes was identified in the roots of both genotypes, which was accompanied by an increase in capsaicin and dihydrocapsaicin content. The findings showed that salinity stress caused an enhancement in the capsaicin and dihydrocapsaicin contents in the roots, leaves, and fruits of pungent peppers. Nonetheless, it was found that the production of capsaicinoids is generally not restricted to the fruits of pungent peppers.

Characterization of Two-Component System gene (TCS) in melatonin-treated common bean under salt and drought stress.

The two-component system (TCS) generally consists of three elements, namely the histidine kinase (HK), response regulator (RR), and histidine phosphotransfer (HP) gene families. This study aimed to assess the expression of TCS genes in P. vulgaris leaf tissue under salt and drought stress and perform a genome-wide analysis of TCS gene family members using bioinformatics methods. This study identified 67 PvTCS genes, including 10 PvHP, 38 PvRR, and 19 PvHK, in the bean genome. PvHK2 had the maximum number of amino acids with 1261, whilst PvHP8 had the lowest number with 87. In addition, their theoretical isoelectric points were between 4.56 (PvHP8) and 9.15 (PvPRR10). The majority of PvTCS genes are unstable. Phylogenetic analysis of TCS genes in A. thaliana, G. max, and bean found that PvTCS genes had close phylogenetic relationships with the genes of other plants. Segmental and tandem duplicate gene pairs were detected among the TCS genes and TCS genes have been subjected to purifying selection pressure in the evolutionary process. Furthermore, the TCS gene family, which has an important role in abiotic stress and hormonal responses in plants, was characterized for the first time in beans, and its expression of TCS genes in bean leaves under salt and drought stress was established using RNAseq and qRT-PCR analyses. The findings of this study will aid future functional and genomic studies by providing essential information about the members of the TCS gene family in beans. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01406-5.

Determination of antimicrobial and antimutagenic properties of some Schiff bases.

In this study, we aimed to investigate for the first time antimicrobial and antimutagenic activities new two Schiff bases, obtained from a primary amine (p-toluidine, o-toluidine) and an aldehyde (Helicin). Synthesized compounds characterized with elemental analysis, fourier transform infrared spectroscopy, ultraviolet-visible spectrophotometry. 1H-13C nuclear magnetic resonance spectroscopy. Antimutagenic activity was evaluated by micronuclei assay. Antimicrobial activity of Schiff bases have been demonstrated against pathogenic four Gram-positive bacteria (Staphylococcus aureus, Staphylococcus epidermis, Micrococcus luteus, Bacillus cereus) and four Gram-negative bacteria (Pseudumonas aeroginosa, Salmonella typhi H, Brucella abortus, Escherichia coli) and two yeasts (Candida albicans and Saccharomyces cerevisiae). The results showed that both Schiff bases have antimutagenic activity. Especially, high concentration (20 µM) of (E)-2-(hydroxymethyl)-6-(2-((p-tolylimino)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triol (Compound I) and (E)-2-(hydroxymethyl)-6-(2-((o-tolylimino)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triol (Compound II) have strong antimutagenic activity against aflatoxin B1. On the other hand, both of studied compounds were found effective against pathogenic bacteria and yeasts. Compound I exhibited more activity against P. aeroginosa, S aureus, S.typhi H and C. albicans comparable to Compound II and standard antibiotics. Additionally, Compound II showed better inhibitory activity than Compound I against Candida albicans and Br. Abortus. Therefore, these compounds can be used in phytotherapeutic due to theirs antimutagenic and antimicrobial activities.

Novel polymeric microspheres: Synthesis, enzyme immobilization, antimutagenic activity, and antimicrobial evaluation against pathogenic microorganisms.

New polymeric microspheres containing azomethine (1a-1c and 2a-2c) were synthesized by condensation to compare the enzymatic properties of the enzyme glucose oxidase (GOx) and to investigate antimutagenic and antimicrobial activities. The polymeric microspheres were characterized by elemental analysis, infrared spectra (FT-IR), proton nuclear magnetic resonance spectra, thermal gravimetric analysis, and scanning electron microscopy analysis. The catalytic activity of the glucose oxidase enzyme follows Michaelis-Menten kinetics. Influence of temperature, reusability, and storage capacity of the free and immobilized glucose oxidase enzyme were investigated. It is determined that immobilized enzymes exhibit good storage stability and reusability. After immobilization of GOx in polymeric supports, the thermal stability of the enzyme increased and the maximum reaction rate (Vmax ) decreased. The activity of the immobilized enzymes was preserved even after 5 months. The antibacterial and antifungal activity of the polymeric microspheres were evaluated by well-diffusion method against some selected pathogenic microorganisms. The antimutagenic properties of all compounds were also examined against sodium azide in human lymphocyte cells by micronuclei and sister chromatid exchange tests.

Nitric oxide mitigates salt stress effects of pepper seedlings by altering nutrient uptake, enzyme activity and osmolyte accumulation.

This study was planned to evaluate the role of exogenous application of sodium nitroprusside (SNP), a NO donor, on the deleterious effect of salinity in Capsicum annum L. seedlings. Different NO doses (0, 50, 100 and 150 µM SNP) were foliarly applied to pepper seedlings grown under the non-saline and saline conditions (50, 100 and 150 mM of NaCl). The photosynthetic rate (Pn), stomatal conductance (gs), intercellular CO2 concentration (Ci), transpiration rate (Tr), mineral element (Zn, Fe, B, K, Ca and Mg) uptake, plant growth and leaf relative water content (LRWC) were decreased by NaCl treatment, but NO treatments generally improved the observed parameters. 150 mM NaCl treatment caused overaccumulation of hydrogen peroxide (H2O2) and malondialdehyde (MDA) by 87 and 100% respectively as compared to control. However, NO application (150 µM SNP) at 150 mM of NaCl significantly decreased H2O2 and MDA to 34 and 54%, respectively. The present study clarified that the exogenous NO treatment supported pepper seedlings against salinity stress by regulating the mineral nutrient uptake, antioxidant enzyme activity, osmolyte accumulation, and improving the LRWC and photosynthetic activity.

Synthesis, anti-microbial and anti-mutagenic activities of some Schiff bases derivatives containing thiophene group.

The aim of this study is to investigate for the first time in vitro antimicrobial and antimutagenic activities of Schiff bases included the azomethine group. Antimutagenic activity was evaluated by micronucleus (MN) assay. These group have been examined for antibacterial activity against pathogenic strains, Staphylococcus aureus, Escherichia coli, Salmonella typhi H, Brucella abortus, Micrococcus luteus, Bacillus cereus, Pseudomonas aeroginosa and antifungal activity against Candida albicans and Saccharomyces cerevisiae. The results of MN showed that Schiff bases ((E)-N-(4-chlorophenyl)-1-(5-nitrothiophen-2-yl)methanimine ; (E)-N-(2,4-dichlorophenyl)-1-(5-nitrothiophen-2-yl) methanimine) different concentrations decreased the toxic effects of Aflatoxin B1. Especially, high concentration (20µM) of (E)-N-(4-chlorophenyl)-1-(5-nitrothiophen-2-yl)methanimine (compound 1) has strong antimutagenic activity. In our in vitro test systems, it was observed that Schiff bases had antimutagenic effects on human lymphocytes. On the other hand these compounds were also found to possess antimicrobial activity against some test bacteria and yeast. The antimicrobial test results of these Schiff bases included the azomethine group exhibited better activity than some known antibiotics. In particular, Compound 1 were more potent bactericides than all of the substances synthesized. In conclusion, this Schiff bases included the azomethine group can be use pharmacy industries as recognized with their noncytotoxic, antimicrobial and antimutagenic features.

Static magnetic field induced epigenetic changes in wheat callus.

Deoxyribonucleic acid (DNA) is always damaged by endogenous and exogenous factors. Magnetic field (MF) is one of these exogenous factors. When repair mechanisms are not sufficient, mainly because of imbalance in damage or mistakes in repair mechanisms, methylation of DNA results in polymorphism-related abnormalities. In this study, low intensity static magnetic field-induced DNA damage and methylation in wheat calli were investigated by using Random Amplified Polymorphic DNA and Coupled Restriction Enzyme Digestion-Random Amplification techniques. Calli were derived from mature embryos of wheat. Both 7- and 14-day-old wheat calli were exposed to 7 mT (millitesla) static MF for 24, 48, 72, 96, or 120 h of incubation period. The highest change in polymorphism rate was obtained in calli exposed to 7 mT MF for 120 h in both 7- and 14-day-old calli. Increase in MF duration caused DNA hypermethylation in both 7- and 14-day-old calli. Polymorphism and DNA methylation ratio were higher in 7-day-old calli. The highest methylation level with a value of 25.1% was found in 7-day-old calli exposed to MF for 120 h. Results suggested that low intensity static magnetic field may trigger genomic instability and DNA methylation. Bioelectromagnetics. 37:504-511, 2016. © 2016 Wiley Periodicals, Inc.

Determination of genotoxic effects of fipronil in Vicia faba using random amplified polymorphic DNA analysis.

This study was undertaken to appreciate genotoxic potential of fipronil herbicides in the roots of Vicia faba seedlings. Fipronil was clearly dose dependent on root growth inhibitors, total soluble protein, and random amplified polymorphic DNA (RAPD) assays. Increase in concentration of fipronil resulted in decreased amount of root length and increased level of protein. For the RAPD analyses, 15 RAPD primers were found to produce unique polymorphic band patterns and were subsequently used to produce a total of 110 bands of 45-1250 bp. Each primer generated 4-11 RAPD bands across 15 primers. The changes occurring in RAPD profiles following fipronil treatment included variation becoming evident as disappearance and/or appearance of DNA bands compared with the normal seedlings. These results indicated that genomic template stability was significantly affected at the above fipronil concentration. This study further confirmed that the RAPD assays are useful in determining potential genotoxicity of fipronil.

The antioxidant and antigenotoxic potential of methanol extract of Cladonia foliacea (Huds.) Willd.

In this article, the genotoxic and antigenotoxic effects of methanol extract of of Cladonia foliacea (Huds.) Willd. (CME) were studied using WP2, Ames (TA1535 and TA1537), and sister chromatid exchange (SCE) test systems. The results of our studies showed that 5 µM concentration of aflatoxin B1(AFB1) changed the frequencies of SCE and malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GPx) activities. When 5 and 10 µg/mL concentrations of CME was added to AFB1, the frequencies of SCE and MDA level were decreased and SOD, GSH, and GPx levels were increased. The extract CME did not show any mutagenicity on Ames (Salmonella typhimurium TA1535, TA1537) and WP2 (Escherichia coli) test systems. On the other hand, CME has antimutagenicity on the mentioned test systems. The results of this experiment have clearly shown that CME has a significant antioxidative and antigenotoxic effect, which is thought to be due to the antigenotoxic activities of antioxidant enzymes.

Determination of genotoxic effects of boron and zinc on Zea mays using protein and random amplification of polymorphic DNA analyses.

In this research, we aimed to determine genotoxic effects of boron (B) and zinc (Zn) on Zea mays by using total soluble protein content and random amplification of polymorphic DNA (RAPD) analyses. For the RAPD analysis, 16 RAPD primers were found to produce unique polymorphic band profiles on treated maize seedlings. With increased Zn and B concentrations, increased polymorphism rate was observed, while genomic template stability and total soluble protein content decreased. The treatment with Zn was more effective than that of B groups on the levels of total proteins. The obtained results from this study revealed that the total soluble protein levels and RAPD profiles were performed as endpoints of genotoxicity and these analyses can offer useful biomarker assays for the evaluation of genotoxic effects on Zn and B polluted plants.

Antigenotoxic potencies of a lichen species, Evernia prunastri.

In this article, the genotoxic and antigenotoxic effects of methanol extract of Evernia prunastri (Huds.) Willd. (MEP) were studied using WP2, Ames (TA1535 and TA1537) and sister chromatid exchange (SCE) test systems. The results obtained from bacterial test systems demonstrated that MEP has strong antimutagenic potencies on TA1537 and WP2 strains. The highest inhibition rates for MEP on TA1537 and WP2 strains were 37.70% and 69.70%, respectively. According to the SCE test system, MEP reduced the genotoxic effects of aflatoxin. In order to clarify the mechanism underlying the antigenotoxic effects of MEP, the antioxidants were determined. Cotreatments of 5, 10 and 20 µg/mL concentrations of MEP with aflatoxin B1 decreased the frequencies of SCE and the malondialdehyde level and increased amount of superoxide dismutase, glutathione and glutathione peroxidase which were decreased by aflatoxin. The data obtained from this work have clearly shown that MEP has significant antigenotoxic effects which are thought to be partly due to the antioxidant activities and antioxidant inducing capability of MEP. This is the first report indicating the antigenotoxic activities of MEP against several mutagen agents such as N-methyl-N'-nitro-N-nitrosoguanidine, acridin and aflatoxin.

Genotoxic, antigenotoxic and antioxidant properties of methanol extracts obtained from Peltigera horizontalis and Peltigera praetextata.

Now-a-days, there is a big need to reduce genotoxic effects of mutagenic and carcinogenic agents in environment, which are increased by the technological development. Lichens produce a wide variety of unique metabolites due to being in various extreme areas and being symbiotic organisms of fungi and algae. Therefore, this study was planned to search new sources having antimutagenic activity by researching two different lichen species and to determine whether their usage is safe. With this respect, the mutagenic and antimutagenic properties of methanol extracts of the lichens were determined by the bacterial reverse mutation and sister chromatid exchange assays. Furthermore, the malondialdehyde level, superoxide dismutase, glutathione and glutathione peroxidase activities against aflatoxin B1 were determined for understanding the ways in which the lichens showed their genotoxic properties.

Phytotoxical effect of Lepidium draba L. extracts on the germination and growth of monocot (Zea mays L.) and dicot (Amaranthus retroflexus L.) seeds.

Laboratory experiments were performed to determine phytotoxic potentials of white top (Lepidium draba) methanol extracts (root, stem and leaf) on germination and early growth of corn (Zea mays) and redroot pigweed (Amaranthus retroflexus). Furthermore, the effects of different methanol extracts of L. draba on the phytohormone (indole-3-acetic acid (IAA), gibberellic acid (GA), abscisic acid (ABA) and zeatin) levels of corn and redroot pigweed were investigated. It was observed that all concentrations of methanol extracts of root, stem and leaf of L. draba inhibited germination, radicle and plumule elongation when compared with the respective controls. Besides this, the degree of inhibition was increased in concert with increasing concentrations of extracts used. On the other hand, phytohormone levels changed with the application of different extract concentrations. Comparing with the control, the GA levels significantly decreased while the ABA levels increased in all the application groups. Zeatin and IAA levels showed changes depending upon the applied extracts and concentrations.

Genotoxic and antigenotoxic potentials of two Usnea species.

For ages, lichens have long been investigated popularly for biological roles, mainly antitumor, antimicrobial and antioxidant activities. Many positive results were obtained in these previous research. Thus, in this study, we aimed to determine whether extracts of Usnea articulata (UAE) and Usnea filipendula (UFE) possessing a protection against aflatoxin B1 (AFB1)-induced genotoxic and oxidative damage. The results of our studies showed that 5 µM concentrations of AFB1 increased the frequencies of sister chromatid exchange (SCE) and the level of malondialdehyde (MDA) and decreased the activities of superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GPx). However, when 5, 10 and 20 µg/mL concentrations of UAE and UFE was added to AFB1, the frequencies of SCE and MDA level were decreased and SOD, GSH and GPx level were increased. The Ames (Salmonella typhimurium TA1535, TA1537) and WP2 (Escherichia coli) test systems carried out evinced that UAE and UFE possess any mutagenicity, but have antimutagenic effects. Consequently, the results of this experiment have clearly shown that UAE and UFE have strong antioxidative and antigenotoxic effects that are associated with its antioxidant nature. A detailed study can be performed to determine the antioxidant properties of each compound that will extend the use of lichen extracts in food and pharmacy industries.

Chemical Composition and Antibacterial Activity of Essential Oils of Two Species of Lamiaceae against Phytopathogenic Bacteria.

In this study, we aimed to determine chemical composition and antibacterial activities of Satureja hortensis and Calamintha nepeta against to 20 phytopathogenic bacteria causing serious crop loss. The essential oils of S. hortensis and C. nepeta were isolated by the hydrodistillation method and the chemical composition of the essential oils were analyzed by GC-MS. The antibacterial properties of the essential oils were evaluated against 20 phytopathogenic bacteria through Disc diffusion assay and micro dilution assay. The results revealed that the essential oils of S. hortensis and C. nepeta have significant antibacterial activity. Furthermore, the findings of the study are valuable for future investigations focusing on the alternative natural compounds to control plant diseases.

5-Aminolevulinic acid treatment mitigates pesticide stress in bean seedlings by regulating stress-related gene expression and retrotransposon movements.

Overdoses of pesticides lead to a decrease in the yield and quality of plants, such as beans. The unconscious use of deltamethrin, one of the synthetic insecticides, increases the amount of reactive oxygen species (ROS) by causing oxidative stress in plants. In this case, plants tolerate stress by activating the antioxidant defense mechanism and many genes. 5-Aminolevulinic acid (ALA) improves tolerance to stress by acting exogenously in low doses. There are many gene families that are effective in the regulation of this mechanism. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. In this study, the expression levels of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), and stress-associated protein (SAP) genes were determined by Q-PCR in deltamethrin (0.5 ppm) and various doses (20, 40, and 80 mg/l) of ALA-treated bean seedlings. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. It was determined that deltamethrin increased the expression of SOD (1.8-fold), GPX (1.4-fold), CAT (2.7-fold), and SAP (2.5-fold) genes, while 20 and 40 mg/l ALA gradually increased the expression of these genes at levels close to control, but 80 mg/l ALA increased the expression of these genes almost to the same level as deltamethrin (2.1-fold, 1.4-fold, 2.6-fold, and 2.6-fold in SOD, GPX, CAT, and SAP genes, respectively). In addition, retrotransposon-microsatellite amplified polymorphism (REMAP) was performed to determine the polymorphism caused by retrotransposon movements. While deltamethrin treatment has caused a decrease in genomic template stability (GTS) (27%), ALA treatments have prevented this decline. At doses of 20, 40, and 80 mg/L of ALA treatments, the GTS ratios were determined to be 96.8%, 74.6%, and 58.7%, respectively. Collectively, these findings demonstrated that ALA has the utility of alleviating pesticide stress effects on beans.

N-acetyl-cysteine mitigates arsenic stress in lettuce: Molecular, biochemical, and physiological perspective.

Agricultural land contaminated with heavy metals such as non-biodegradable arsenic (As) has become a serious global problem as it adversely affects agricultural productivity, food security and human health. Therefore, in this study, we investigated how the administration of N-acetyl-cysteine (NAC), regulates the physio-biochemical and gene expression level to reduce As toxicity in lettuce. According to our results, different NAC levels (125, 250 and 500 µM) significantly alleviated the growth inhibition and toxicity induced by As stress (20 mg/L). Shoot fresh weight, root fresh weight, shoot dry weight and root dry weight (33.05%, 55.34%, 17.97% and 46.20%, respectively) were decreased in plants grown in As-contaminated soils compared to lettuce plants grown in soils without the addition of As. However, NAC applications together with As stress increased these growth parameters. While the highest increase in shoot fresh and dry weight (58.31% and 37.85%, respectively) was observed in 250 µM NAC application, the highest increase in root fresh and dry weight (75.97% and 63.07%, respectively) was observed in 125 µM NAC application in plants grown in As-polluted soils. NAC application decreased the amount of ROS, MDA and H2O2 that increased with As stress, and decreased oxidative damage by regulating hormone levels, antioxidant and enzymes involved in nitrogen metabolism. According to gene expression profiles, LsHIPP28 and LsABC3 genes have shown important roles in reducing As toxicity in leaves. This study will provide insight for future studies on how NAC applications develop resistance to As stress in lettuce.

Molecular determination of genotoxic effects of cobalt and nickel on maize (Zea mays L.) by RAPD and protein analyses.

Assessment of DNA damages stemming from toxic chemicals is an important issue in terms of genotoxicology. In this study, maize (Zea mays L.) seedlings were used for screening the genotoxic effects of cobalt (Co) and nickel (Ni) treatments at various concentrations (5 mM, 10 mM, 20 mM and 40 mM). For this purpose, randomly amplified polymorphic DNA (RAPD) technique was applied to genomic DNA extracted from metal-exposed and unexposed plant materials. Besides, changes in total protein contents were screened by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. For RAPD analysis, 16 RAPD primers were found to produce unique polymorphic band profiles on different concentrations of Co-/Ni-treated maize seedlings. Increased polymorphism resulting from the appearance of new bands or disappearance of normal bands was observed with increasing concentration of Co and Ni treatments. Genomic template stability, a qualitative measurement of changes in RAPD patterns of genomic DNA, decreased with increasing metal concentration. In SDS-PAGE analysis, it was observed that the total soluble protein content decreased by Co treatment, while it increased by Ni treatment. The results obtained from this study revealed that RAPD profiles and total soluble protein levels can be applied to detect genotoxicity, and these analyses can offer useful biomarker assays for the evaluation of genotoxic effects on Co- and Ni-polluted plants.

Determination of the genotoxic effects of Convolvulus arvensis extracts on corn (Zea mays L.) seeds.

In this research, the methanolic extracts of Convolvulus arvensis were tested for genotoxic and inhibitor activity on the total soluble protein content and the genomic template stability against corn Zea mays L. seed. The methanol extracts of leaf, stem and root of C. arvensis were diluted to 50, 75 and 100 µl concentrations and applied to corn seed. The total soluble protein and genomic template stability results were compared with the control. The results showed that especially 100 µl extracts of diluted leaf, stem and root had a strong inhibitory activity on the genomic template stability. The changes occurred in random amplification of polymorphic DNA (RAPD) profiles of C. arvensis extract treatment included variation in band intensity, loss of bands and appearance of new bands compared with control. Also, the results obtained from this study revealed that the increase in the concentrations of C. arvensis extract increased the total soluble protein content in maize. The results suggested that RAPD analysis and total protein analysis could be applied as a suitable biomarker assay for the detection of genotoxic effects of plant allelochemicals.