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Mutagenic impact of picric acid on chloroplast genome and a selection of biological attributes of Brassica napus L. (Brassicaceae)

BMC Genomics volume 26, Article number: 491 (2025) Cite this article

Abstract

The present study was conducted to investigate the possible mutagenic effects of picric acid on the chloroplast genome and certain biological properties of Brassica napus L. under field conditions. The experimental design for this study was a randomized complete block trial, based on 3 factors (dose, duration of priming, and three oilseed rape (cultivars). The seeds of oilseed rape cultivars (Abasin- 95, Dur e Nifa, and Nifa Gold) were exposed to five doses (0 mM, 5 mM, 10 mM, 15 mM, and 20 mM) of picric acid for different soaking times (3 h, 6 h, and 9 h) and the data on agromorphological characteristics were recorded. Control and test samples were collected and compared for genomic chloroplast studies. The results confirmed the inhibitory effect of picric acid on days to emergence, completion of germination, and percent emergence. Seeds treated with a dose of 20 mM delayed seedling emergence (6.15) compared to other treatments. In addition, the differences between seeds primed for 3 h (6.07), and 6 h (5.89) were also highly significant. Similarly, the effects of 0 mM (7.52), 5 mM (8.26) and 10 mM (8.11) doses on germination time were not significant. Comparison of the mean values with respect to germination showed that picric acid led to a reduction in the percentage emergence of the oilseed rape seedlings. A significant increase in leaf size was observed in the Nifa Gold cultivar (10.27). On the other hand, the differences in leaf size between the cultivars Dur e Nifa (9.51) and Abasin-95 (9.20) were not significant. The inhibitory effect of picric acid on the number of leaves could be due to the fact that the meristematic cells are damaged by the mutagen, leading to a reduction in the number of leaves. The Nifa Gold cultivar showed the highest value for plant height (17.53), and the highest number of siliqua (14.69) compared to other genotypes. The number of siliqua confirms that Nifa Gold is best adapted to picric acid in contrast to the other genotypes. However, the differences between the Abasin-95 cultivar (0.068) and the Nifa Gold cultivar (0.062) in terms of seed weight were not significant. The dose of 15 mM (37.72) showed the highest moisture content, followed by 10 mM (37.08), 0 mM (33.87), 5 mM (33.11) and 20 mM (32.73). The chloroplast genome of plants grown from picric acid-treated seeds was mutated. Therefore, annotation of the genome was not possible. Numerous single nucleotide substitutions (187) were detected between the control and test samples. In addition, 43 additions/deletions were observed. The present study suggests that picric acid has a mutagenic effect, which is confirmed by numerous mutations in chloroplast samples. Subsequently, the doses have stimulating effects on plant height and seed weight. Furthermore, priming can improve the emergence and certain biological characteristics of B. napus. In the present study, all parameters were investigated and the significant response of the selected genotypes was confirmed.

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Introduction

B. napus is a bright-yellow flowering member of the Brassicaceae family with genus Brassica and includes 40 species [1,2,3,4,5]. Rapeseed is the third largest source of vegetable oil and biofuel and the third largest oil-producing crop in the world after oil palm (Elaeis guineensis Jacq.) and soybean (Glycine max (L.) Merr.) [6]. It is the second largest source of protein food in the world. Rapeseed is better suited to places with heavy rainfall, and its yield suffers dramatically when drought strikes. In these places, water scarcity is prevalent throughout critical periods of plant growth, resulting in decreased photosynthetic capacity and leaf chlorophyll levels [1, 7]. B. napus reaches a height of about 100 cm. The lower leaves are petiolated, hairless, fleshy and pinnatifid. The upper leaves have no petioles. The leaf color is a darker, bluish-green. A single leaf is attached to the stem at each node, with about 15 to 20 internodes per plant, 5 to 10 mm apart. The flowers of B. napus are bisexual and develop in terminal racemes. The flowers are regular with 4 sepals and 4 petals. The diagonally opposite, yellow petals narrow at the basal end and form a cross, hence the original name of the family Cruciferae. The flowers also contain 6 stamens (2 of which are shorter and lower than the others), a pistil with 2 carpels and superior ovary. The seeds develop in an elongated, two-celled capsule called a siliqua, which has a pronounced midrib [8]. The production and quality of rapeseed oil depends primarily on the growth of the siliques, i.e. the number, size and volume of the fully matured siliques. This is due to the crucial function of siliques within the rapeseed plant. It is not only the carrier of the oilseeds but also contributes to photosynthesis and transmits developmental cues to the seeds during maturation [9,10,11,12].

Mutation breeding, a type of plant breeding, has been used to improve rapeseed (B. napus) varieties by creating genetic mutations through physical or chemical means. This has led to genetic diversity and enabled the development of desired traits [13]. Mutation breeding oilseed rape began in the mid-twentieth century when it was realized that induced mutations could be used to improve crops. Scientists exposed oilseed rape seeds to X-rays or gamma rays to induce random mutations in the plant's DNA [14]. These mutations can affect numerous traits, including oil content, seed yield, disease resistance, and plant shape. Mutant lines exhibiting the desired traits were selected for further analysis and inbreeding. Mutation breeding was used to develop improved agronomic traits in oilseed rape varieties. To combine the desired traits and improve overall performance, better varieties were often crossed with mutation lines created by mutation breeding. Through hybridization and rigorous selection, breeders succeeded in breeding rapeseed varieties with higher yield, better oil quality, and resistance to biotic and abiotic stress factors [15]. Thanks to the commercial release of oilseed rape varieties created through mutation breeding, farmers now have more choices for oilseed cultivation. The improved resistance to pests and diseases, better oil composition, and higher yield potential of these varieties have helped to promote oilseed rape cultivation worldwide [16].

Fluctuations in climatic conditions have a negative effect on the productivity of crops and oil plants [17]. The vulnerability of oilseed rape (B. napus) yields to increased temperatures is a major challenge in the quest for global food security. Yields have been reported to decrease by up to 40% under elevated temperatures [18, 19]. This problem emphasizes the critical impact of climatic variability on the growth stages, seedling emergence, and reproductive performance of B. napus, leading to a significant decline in productivity [20,21,22]. These findings have far-reaching implications as the global agricultural sector is faced with the need to significantly increase food production. More than 2 billion tons of cereals are produced annually worldwide, which are important for both human consumption and livestock feed, accounting for about two-thirds of total direct and indirect protein consumption. Interestingly, only about 10% of this immense production, i.e. 200 million tons, is available for international trade [23]. Given the expected population increase to 9.3 billion people in 2050 [24] and to 12.3 billion people in 2100 [25, 26], there is an urgent need to double current food production, particularly grain production, from 2 billion to over 4 billion tons annually [23]. Although this ambitious target is challenging, it is considered achievable if crop yields are further increased without an equal increase in the exploitation of arable land [27, 28]. In order to meet the demand caused by population growth and changing eating habits, the actual food supply must be increased by 70% [29]. Although the focus is on reducing the distribution gap, the pressure on existing agricultural frameworks to meet this demand indicates that either a sustained increase in crop yields, an expansion of agricultural land, or a synergistic approach combining both is required [30]. The introduction of new crops and oil plants is therefore of immense importance. Genotype changes induced by nature or selection are a slow process. Therefore, techniques to produce new varieties need to be explored. The only solution in this scenario seems to be mutation breeding [31]. Charles Darwin highlighted the importance of genetic variation for evolutionary adaptation and stressed that diversity is crucial not only for the survival of plant species but also for improving the quality of different crops [32]. This genetic diversity plays a central role in breeding programs as it enables plants to adapt effectively to environmental changes [33]. Therefore numerous methods have been introduced in recent years to increase genetic diversity in fruit trees [34]. Mutation, an important source of diversity, refers to unexpected changes in DNA sequences that occur without segregation or recombination. These changes are not limited to nucleotide substitutions but also include deletions/insertions, inversions, and translocations [35]. The concept of plant mutation can be traced back to 300 BC, but it was Hugo de Vries in the late nineteenth century who introduced the term"mutation"in 1903 in his work “The Mutation Theory”, and explained it as the cause of phenotypic changes in Oenothera lamarckiana Ser. [36]. Mutation breeding as a field for the creation of new cultivars was first recognized by Stadler [37] in his work on maize, barley and wheat. In the early 1930 s, the first commercial tobacco variety produced by mutation breeding was released. To date, more than 3088 mutagenic varieties of different fruit trees (e.g. apples, citrus, fruits and peaches), food crops (e.g. rice, barley, wheat, maize, and peas), oil crops (e.g. soybeans, rapeseed, sunflowers) and ornamental plants (e.g. chrysanthemums, dahlias, and poinsettias) have been released [38]. In addition, mutations are considered the basis of modern plant breeding techniques [34]. These mutations occur naturally in organisms to repair genetic material, including DNA and RNA, that has been damaged by physical, chemical, or biological factors [33, 39]. Hugo De Vries coined the term and described mutations as sudden changes in the DNA or RNA of a plant organ without recombination or segregation [36]. The resulting genetic variation, referred to as a"mutant", is a cornerstone of evolutionary progress [38, 40]. These mutations cause heritable changes in DNA and RNA that lead to phenotypic changes [38]. The induction of genetic variability by mutations, either by irradiation or by chemical mutagens, differs from genetic modification in that it does not involve the insertion of genes, which distinguishes it from genetically modified organisms (GMOs) [41]. Since 1925, when it was discovered that mutations in plants can be induced by irradiation or chemical treatments, mutation breeding has become a standard procedure for plant breeders. This approach is favored in regions where transgenic plants are banned and offers a non-transgenic alternative for plant development [42]. While mutations certainly occur in nature, but their impulsive frequency is not sufficient to achieve the rapid progress required in plant breeding. Nonetheless, mutations can be deliberately induced in various plant and animal species using physical and chemical means. These induced mutations range from extensive DNA deletions to minor point mutations. Physical methods of mutation induction, such as irradiation with non- ionising radiation (e.g. UV light) or ionising radiation (including X-rays, gamma rays and fast and slow neutrons), often lead to significant DNA deletions and changes in chromosome structure. In contrast, chemical mutagens usually attack individual nucleotide pairs. Chemical mutagens frequently used in plant breeding include ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), hydrogen fluoride (HF), sodium azide, N-methyl-N-nitrosourea (MNU), and hydroxylamine. The extent of the mutation depends on the exposed tissue and the level of exposure (taking into account dose and duration). Mutations of single nucleotide pairs are usually the focus of interest for breeders, as major chromosomal changes usually have adverse effects. However, the use of mutagens that alter chromosome structure can help to increase the number of recombination events and break unwanted genetic linkages, which is of considerable value for genetic improvement [43]. The present study aimed to investigate the efficacy of a little- studied mutagen called picric acid on an important oilseed crop. Picric acid is an organic compound with the formula (O2N)3 C6H2OH. Its IUPAC name is 2, 4, 6-trinitrophenol (TNP). The name"picric acid"comes from the Greek (pikros) and means"bitter", which refers its bitter taste. It is one of the most acidic phenols [44]. Among the nitroaromatic compounds, picric acid is a very stable molecule [44]. The mutagenic effect of nitroaromatic compounds begins with their entry into cellular systems, the resulting interactions and binding to specific enzymes, the formation of important complexes (e.g. hydroxylamine and electrophilic intermediates), which in turn react with nucleophiles such as proteins or DNA and form an addition product that makes them a potent DNA-altering substance [45]. The mutagenic tendencies of picric acid have already been studied by several authors [44, 45]. Thus, the current scenario of genetic diversity in oilseed rape is influenced by conservation measures, breeding programmes, genomic research, genetic engineering, and the use of wild forms. To ensure the resilience, sustainability, and long-term viability of oilseed rape cultivation in the face of changing agricultural and environmental challenges, it is essential to maintain and improve genetic diversity. The goal of the study is presumably to better understand how picric acid exposure affects the chloroplast genome and associated biological traits of B. napus. For this reason, variability was created by mutation with the following objectives: 1) evaluation of mutagenic effects, 2) investigation of chloroplast function, 3) identification of stress responses, and 4) characterization of genetic diversity.

Materials and Methods

Research site details and experimental layout

The study was conducted in the Department of Botany, Islamia College Peshawar, Khyber Pakhtunkhwa, Pakistan. The trial was designed in a randomized complete block design based on 3 factors (1 st factor: doses, 2nd factor: priming duration, 3rd factor: canola cultivars). Each treatment was replicated three times. The canola genotypes (Abasin- 95, Dur e Nifa, and Nifa Gold) were obtained from Nuclear Institute for Agriculture, Peshawar (NIFA), Khyber Pakhtunkhwa, Pakistan. The seeds were soaked in beakers of distilled water and allowed to stand at room temperature for 6 h. Aeration was encouraged by occasional shaking. The seeds were removed from the water and the excess moisture was removed by pressing in folded filter paper. They were treated with the required concentrations (0 mM, 5 mM, 10 mM, 15, and 20 mM) of picric acid solution for three time durations (3 h, 6 h, and 9 h). After each treatment, the seeds were washed under cold running tap water for about 20 min to remove excess picric acid. Excess moisture was removed by spreading the seeds on blotting paper. Ten seeds were sown in pots filled with soil. Each pot contained about 7 kg of soil. The soil was prepared by mixing clay and loam in almost equal proportions. The soil sample was well-aerated, drained and sterilized. Data on the number of leaves were recorded by counting the leaves of all plants in each pot. Similarly, the length of the 4 th leaf from the base of each plant was recorded. In the same way, the height of the plants (in inches) was determined using a common scale. The siliques were collected at the time of plant maturity and data regarding the number of siliqua/plant, average siliqua weight/replicate, seed weight/siliqua, fresh/dry biomass and moisture content were recorded.

Molecular study

The plants were grown in pots to study the chloroplast genome. Plants grown from untreated seeds served as controls. The plants treated with picric acid were the treated samples. Plant tissue was taken from both the control and test samples and subjected to DNA extraction.

DNA extraction

Leaf samples of each genotype were collected, and DNA was extracted from these samples using a standard protocol. The samples were ground in liquid nitrogen, and the warmed (65 ◦C) CTAB buffer (800 µL) was added to each tube, and the resulting mixture was vortexed thoroughly to homogenize. Tubes were then incubated in a water bath for 1 h at 65 ◦C. Chloroform and phenol-isoamyl-alcohol (800 µL) at a ratio of 24:1 were added to each tube, and the solution was mixed gently by inverting tubes for 2 min. The samples were centrifuged at 14,000 rpm for 15 min. The supernatant was transferred to new 1.5 µL Eppendorf tubes. Cold isopropanol (2/3rd volume of supernatant) was added to each tube and mixed gently. Samples were again centrifuged at 14,000 rpm for 10 min to pellet DNA. The supernatant was discarded, and the sample was washed with the help of ethanol (70%). Ethanol was discarded, and the DNA pellets were allowed to dry at 27 ◦C for 10 min. The DNA samples were dissolved in 100 µL TE (pH 8.0) and stored at − 20 ◦C [46].

Study of the chloroplast genome

The approach of Ahmed et al. 2009 [47], was used to extract the whole genome DNA. Hydrogenated ether is then used to remove polysaccharides and other impurities from the DNA. The extracted whole genome DNA was checked for quality and quantity using a 1% agarose gel and Nanodrop (Thermo Scientific). The high-quality DNA was sent to Novogene, Hong Kong. The control sample was assembled using NovoPlasty [48] and annotated with GeSeq [49]. The reads of treated samples were mapped through Bowtie 2 [50] to the assembled genome, and the number of indels and substitutions were counted by inspecting the alignment manually with care. The number of chloroplast genomes is higher in the leaf than in the nuclear genome, which provides high coverage for the chloroplast genome even from low-depth sequencing [51]. Here, despite the conventional approach in which isolation or enrichment of the chloroplast genome by amplification with long-range PCR is done [46, 52, 53], followed by DNA extraction [54, 55], we used the whole genome sequencing approach and assembled the genome successfully. The same has been reported for several plant lineages [47, 56,57,58,59,60].

Statistical analysis

The experimental data were analysed using MSTAT-C software to establish statistical significance between treatments. An analysis of variance (ANOVA) was used to discover variations in the measured parameters. The Least Significant Difference (LSD) test was used at a 5% probability level (p ≤ 0.05) to compare mean values and assess the significance of detected differences. To facilitate interpretation, results were provided as mean ± standard error [61].

Results and discussion

Statistical interpretation

The effect of doses on seedling emergence, completion of germination, percentage emergence, number of leaves, leaf size, plant height, number of siliqua, weight of siliqua and moisture content of oilseed rape was highly significant. Seed weight and fresh/dry biomass showed no-significant responses to the doses. In contrast, priming resulted in significant changes in seedling emergence, number of leaves, leaf size, plant height, siliqua weight and fresh biomass of oilseed rape. However, the priming techniques were not able to produce significant changes in percentage emergence, number of siliquae, seed weight, dry biomass and seedling moisture content. Significant differences were observed between the selected varieties for The traits such as seedling emergence, germination completion, percentage emergence, fresh/dry biomass and moisture content, number of leaves, leaf size, plant height, number of siliqua and seed weight. The effect of cultivar differences on siliqua weight was not significant. The 1 st order interaction resulted in significant variations in leaf size, number of siliqua and moisture content. However, attributes such as seedling emergence, germination, number of leaves, plant height, siliqua weight, seed weight and fresh/dry biomass did not vary-significantly under the influence of 1 st order interaction. The 2nd order interaction was found to be significant for percentage emergence, number of leaves, leaf size, fresh biomass and moisture content. On the other hand, the 3rd order interaction did not result in significant changes in germination, number of leaves, leaf size, plant height, number of siliqua, fresh seed weight, and dry biomass. Similarly, the studied attributes (except germination, sprouting, number of leaves, seeds, and weight) showed no-significant changes under the 4 th order interaction (Table 1).

Table 1 Mean-square values and significance tests for emergence (days), completion of germination (days), number of leaves, leaf size (cm), plant height (inches), number of siliqua, siliqua weight (g), seed weight (g), fresh biomass (g), dry biomass (g) and moisture content (%) of three oilseed rape cultivars evaluated with different doses treated with picric acid
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Impact on germination traits

Effect on days to seedling emergence

Seeds treated with a dose of 20 mM delayed seedling emergence (6.15) compared to other treatments. However, the effects of 15 mM (5.96) and 20 mM (6.15) were statistically similar. The doses of 0 mM (5.63), 5 mM (5.56) and 10 mM (5.44) also did not result in significant seedling emergence. The data on days to emergence confirmed two statistical groups at the applied doses. The control, 5 mM and 10 mM belong to the first group and 15 mM and 20 mM to the 2nd group.

Priming the rapeseed for 9 h resulted in significantly higher seedling emergence (5.29) compared to the other priming durations. In addition, the differences between seeds primed for 3 h (6.07), and 6 h (5.89) were also highly significant. Seeds of Dur e Nifa cultivar required the least time for emergence (5.53) compared to Abasin- 95 (5.87) and Nifa Gold (5.84). However, the response of Abasin- 95 cultivar (5.87) and Nifa Gold cultivar (5.84) was statistically similar.

The 3rd order interaction (A × C) delayed the emergence (6.33) of Nifa Gold cultivar seeds treated with 20 mM picric acid. In contrast, seeds of Dur e Nifa cultivar treated with 0 mM and 10 mM took least time to emerge (5.11). The effect of 4 th order interaction (A × B × C) was found to be inhibitory for the seeds of Abasin- 95 cultivar (6.67) treated with 0 mM picric acid for 6 h. In contrast, the cultivar Dur e Nifa showed the fastest emergence (4.33) under the influence of a 0 mM dose and a soaking time of 9 h (Table 2).

Table 2 Effect of various doses of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) on the days to seedling emergence of three oilseed rape cultivars
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The results on germination characteristics showed that higher doses of picric acid delayed the emergence of the oilseed rape varieties. In contrast, pre-soaking of the seed accelerated seedling emergence. Among the oilseed rape genotypes, cultivar Dur e Nifa showed rapid seedling emergence and proved to be more adaptable to picric acid than the other cultivars. The delayed emergence of the oilseed rape seedlings can be attributed to damage to the embryo by picric acid [62]. It is also possible that the mutagen caused some physiological changes (disruption of enzymes) in the oilseed rape seeds that slowed down seedling emergence [63,64,65]. Mutations in genes involved in metabolic pathways responsible for the conversion of fatty acid to glucose could be another reason for delayed seedling emergence [66, 67]. In addition, there is a possibility that treatment with picric acid creates stress conditions for the embryos that lead to dormancy, which in turn results in delayed seedling emergence [47]. Many researchers have reported negative effects of mutagens on the germination of different plants, which are in complete agreement with the results of the present study [65, 68,69,70,71]. Some researchers reported rapid growth of various plants grown from seeds treated with many other mutagens, which is in contradiction with our results [67, 72,73,74,75,76,77,78].

Effect on days taken to germination completion

The untreated rapeseed (7.52) took the fewest days to germinate compared to the other doses. However, the effects of the doses of 0 mM (7.52), 5 mM (8.26) and 10 mM (8.11) on germination time were not significant. The seeds treated with a dose of 20 mM took the longest time to germinate (9.41). However, the doses of 15 mM and 20 mM had a statistically similar effect on germination. In contrast to the other cultivars, rapid germination was observed in the Abasin− 95 cultivar. In contrast, the cultivar Nifa Gold (8.89) took the longest time to complete germination. However, the differences between the Dur e Nifa cultivar (8.31) and the Nifa Gold cultivar (8.89) were highly insignificant. Conversely, the responses of Abasin− 95 cultivar and Nifa Gold cultivar were highly significant in terms of germination rate. The 4 th order interaction increased the germination rate (6.33) of Abasin- 95 cultivar under the influence of 0 mM dose and soaking time. However, the maximum time to complete germination was 12 (days), which could not be specifically assigned to any cultivar or dose (Table 3).

Table 3 Effect of various doses (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) of picric acid on days taken to germination completion of three oilseed rape cultivars
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The results confirmed the inhibitory effect of picric acid on the number of days to full germination. Among the three oilseed rape varieties Abasin- 95 seed germinated the fastest and proved to be the most adaptable to picric acid for the same parameter. According to reports by Roychowdhury & Tah [65], interference with the formation of enzymes involved in the germination process may be one of the physiological effects of mutagens. Secondly, picric acid could physically damage the cells of seed embryos, and disrupt many physiological processes required for rapid germination [62] [62,63,64,65]. There is also the possibility that picric acid affects some genes necessary for fatty acid metabolism, leading to slow completion of germination [66, 79]. In [65, 68,69,70], negative effects of various other mutagens on the germination behavior of different plants were reported, which is fully consistent with our results. In contrast to our results, some researchers have reported rapid germination of various plants grown from seeds treated with different other mutagens [66, 67, 72,73,74,75,76, 80].

Effect on germination percentage (%)

The seeds grown in the control group showed the highest germination (94.44) compared to the other doses. However, the dosages of 0 mM (94.44), 5 mM (92.59) and 10 mM (92.22) had a similar statistical effect. Similarly, the effect of 10 mM (92.22) and 15 mM (86.30) was not significant. The dose of 20 mM (76.30) caused a significant inhibition of percent incidence compared to the other doses. The lethal dose that caused 50% inhibition was 42.597 mM.

The cultivar Dur e Nifa (91.78) recorded the highest percentage of field emergence compared to the other genotypes. In addition, the Dur e Nifa cultivar (91.78) and the Nifa Gold cultivar (84.67) showed significant differences in percentage field emergence. However, the Dur e Nifa cultivar (91.78) and the Abasin-95 cultivar (88.67) did not show any significant variations. Similarly, the differences between the Abasin-95 cultivar (88.67) and the Nifa Gold cultivar (84.67) were not significant.

The 2nd order interaction (B × C) increased the germination of the Dur e Nifa cultivar primed for a period of 3 h duration (95.33). However, the Nifa Gold cultivar, which was primed for 9 h recorded the lowest germination (79.33). The 3rd interaction stimulated the percent emergence (97.78) of the Abasin-95 cultivar and the Dur e Nifa cultivar treated with a dose of 5 mM and 15 mM doses of picric acid, respectively. In contrast, the interaction between dose and cultivar (A × C) resulted in maximum inhibition of percent emergence (68.89) in Nifa Gold cultivar treated with a dose of 20 mM (Table 4).

Table 4 Impact of various treatment levels (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) on germination percentage (%) of three oilseed rape cultivars treated with picric acid
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Comparison of mean values with respect to germination showed that picric acid resulted in a reduction in percentage emergence of rapeseedlings. Among the three rapeseed varieties, Dur e Nifa cultivar showed the highest percentage of germination. The reduction in seed germination induced by mutagenic treatments may be the result of damage to cellular components at the molecular level or altered enzyme activity [63,64,65]. Micco et al. [79] have indicated abnormalities in the mitotic cycles and metabolic pathways of cells due to mutagens as the cause of low germination. Kovacs & Keresztes, [62] reported that the reduction in germination and survival could be due to the incorporation of mutagens into biological materials that act directly on critical targets in the cell. Most researchers reported a decrease in germination rate due to the application of mutagens in various plants [67, 69, 72, 74, 75, 78, 81,82,83,84,85]. Abdul Rahaman et al. [80] studied the effects of sodium azide and nitric acid on the growth parameters of Citrullus and Moringa and concluded that the mutagens have no effect on germination, while Roychowdhury & Tah [65] and El-Nashar & Asrar [70] reported stimulatory effects of various mutagens on germination of different plants in contrast to the present findings.

Response of growth and agronomic traits

Effect on number of leaves

Plants grown from seeds treated with a picric acid dose of 20 mM (11.37) produced most leaves compared to the control (11.06). However, the differences between the doses of 0 mM and 20 mM were not significant. At the same time, a dose of 10 mM (10.48) caused a reduction in the number of leaves compared to the control. Furthermore, the differences between the plants grown from seeds treated with 5 mM (10.67), 10 mM (10.48) and 15 mM (10.81) were not significant. Similarly, no significant differences were observed for the parameter in question in seeds treated with 0 mM (11.06), 5 mM (10.67) and 15 mM (10.81) picric acid.

Soaking the seeds for 6 h resulted in a maximum number of leaves (11.16) compared to other priming durations. At the same time, seeds primed for 3 h (11.06) and 6 h (11.16) resulted in non-significant variations in the number of leaves. In contrast, the differences between plants grown from seed with a soaking time of 9 h (10.49) and the other two soaking times were highly significant.

The Dur e Nifa cultivar (12.02) produced the highest number of leaves. It was also found that the Abasin-95 cultivar (9.25), Dur e Nifa cultivar (12.02), and Nifa Gold cultivar (11.43) showed highly significant variations for the parameter in question. The 2nd order interaction study (B × C) showed that priming for 6 h is stimulated the number of leaves of Dur e Nifa (13.03). Subsequently, Abasin-95 cultivar primed for 3 h recorded the highest inhibition (8.90) for the attribute in question (Table 5).

Table 5 The number of leaves of oilseed rape cultivars under the influence of different doses (0, 5 mM, 10 mM, and 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) of picric acid
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The inhibitory effect of picric acid on the number of leaves could be due to the fact that the meristematic cells are damaged by the mutagen, leading to a reduction in the number of leaves. Since mutagens are known to induce physical, physiological and genetic changes in plants, the reduction in the number of leaves of oilseed rape plants could be related to the inhibition of polymerization of tubulin into microtubules or RNA synthesis, which affects the mitotic divisions of cells, and leads to a reduction in leaf production [67]. Soaking the seeds for 6 h was found to be favorable for improving leaf number. The Dur e Nifa cultivar showed sufficient adaptability to picric acid in terms of leaf number. Our findings on the decrease in leaf number of oilseed rape plants with picric acid are confirmed by [17, 74, 83] and Olawuyi & Okoli [69] when they studied the effects of different mutagens on horse gram, cowpea, tomato, and maize respectively. In contrast, Kumar et al. [84], Kumar et al. [86], Barman et al. [87] and El-Nashar & Asrar [70] reported positive effects of chemical mutagens on mulberry, brassica, jamun and calendula respectively.

Effect on leaf size (cm)

The effect of picric acid on the seeds to increase leaf size proved to be as effective as with untreated seeds. The plants with the highest leaf size (10.53) emerged from seed soaked for 3 h, leaf size followed by 6 h (9.60) and 9 h (8.85). A significant increase in leaf size was observed in the Nifa Gold cultivar (10.27). On the other hand, the differences in leaf size between the cultivars Dur e Nifa (9.51) and Abasin-95 (9.20) were not significant.

The 2nd order interaction (B × C) recorded the highest value for leaf size (11.93) in Dur e Nifa cultivar primed for 3 h. On the other hand, Dur e Nifa primed for 9 h recorded the lowest value for leaf size (7.82) (Table 6). The improvement in leaf size with priming could be due to the fact that the oilseed rape made the necessary adaptation to the mutagen before sowing [88]. The pre-exposure of the seeds stimulated the defense mechanisms of the oilseed rape plants against the mutagen, which led to an increase in leaf size. The increase in leaf size would maximize the plants'biomolecule synthesis apparatus of the plants needed to cope with the effects of the mutagen [47, 88]. Researchers such as [72], Mshembula et al. [83] and El-Nashar & Asrar [70] reported an increase in leaf growth of sesame, cowpea and calendula respectively, when chemical mutagens were used. On the other hand, Akhtar [17] and Olawuyi & Okoli [69] reported deleterious effects of chemical mutagens on leaf growth of tomato and maize plants.

Table 6 Leaf size (cm) of oilseed rape cultivars as a function of different doses (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) of picric acid
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Effect on plant height (inches)

The increase in seedling height was observed and the maximum increase over the control (12.85) was recorded at 15 mM (15.10) and it was not significantly different from the 5 mM (14.38), 10 mM (14.74) and 20 mM (14.50) doses. Soaking the seeds for 6 h (15.19) resulted in a significant increase in plant height compared to 3 h (13.24). Similarly, soaking the seeds for 3 h and 9 h resulted in significant variations in plant height. On the other hand, the effects of 6 h (15.19) and 9 h (14.51) duration on plant height were not significant.

The Nifa Gold cultivar showed the highest value for plant height (17.53), followed by the Dur e Nifa cultivar (16.36) and the Abasin- 95 cultivar (9.05) (Table 7). The stimulatory effect of picric acid confirmed that it affects the plant growth hormone genes that enable plants to reach maximum height [63, 64, 69]. Since many researchers have reported the effects of mutagens on cytological aspects of plants, a link between the growth of picric acid and the meristematic stem cells of oilseed rape plants could also be hypothesised in the present case [62]. It is likely that, picric acid-induced rapid mitosis in the meristematic tissue, which helped the plants to reach maximum height [69, 89]. It is also possible that picric acid increases the stomata density of plants [76]. As the number of stomata increases, so does the number of guard cells. We know that guard cells contain chloroplasts, which are the assimilation factories of plants. The more assimilation machines there are, the more carbon is fixed and the more raw materials are made available for the plants, which gain height as a result. Researchers such as Mshembula et al. [83], Birara et al. [76], Barman et al. [87], El-Nashar & Asrar [70], and Abdul Rahaman et al. [80] reported an increase in the height of plants grown from seeds treated with different chemical mutagens. In contrast, Obadoni et al. [72], Bolbhat et al. [74], Jagajanantham et al. [75], Arisha et al. [90], Akhtar [17], Olawuyi & Okoli [69] reported a decrease in plant height when different chemical mutagens were used, which contradicts the results of the present study.

Table 7 Plant height of oilseed rape cultivars as a function of different doses (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) of picric acid
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Effect on the number of siliqua/plant

The analysis of variance regarding the number of siliqua/plant confirmed the non-significant reaction of the plants to picric acid. Priming the seeds for 6 h did not significantly increase the number of siliqua compared to other durations. The Nifa Gold cultivar (14.69) produced the highest number of siliqua compared to the other genotypes. At the same time, the number of siliquas did not show-significant variations between the Dur e Nifa cultivar (14.36) and the Nifa Gold (14.69) cultivar. In contrast, the Abasin- 95 cultivar (11.58) recorded a significant decrease in the number of siliqua observed in the Abasin- 95 cultivar (Table 8).

Table 8 Number of siliqua in oilseed rape cultivars influenced by various doses of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h)
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The number of siliqua confirms that Nifa Gold is best adapted to picric acid in contrast to the other genotypes. It appears that picric acid has the least effect on the genes responsible for the synthesis of siliqua in oilseed rape plants. It can also be concluded that oilseed rape plants are mature enough at the fruiting stage and less susceptible to the effect of mutagens, which makes them a good crop for cultivation under stress conditions [36]. A reduction in the number of fruits/plants was observed by Bolbhat et al. [74], Mshembula et al. [83] and Jagajanantham et al. [75] using different chemical mutagens. In contrast, Birara et al. [76] reported an increase in the number of fruits/plants in sesame with chemical mutagens.

Effect on siliqua weight of the husk (g)

The dose of 20 mM (0.120) significantly increased the weight of rapeseed compared to the control (0.105). However, the changes induced by the doses of 5 mM (0.117), 10 mM (0.110), 15 mM (0.113) and 20 mM (0.120) were highly non-significant. Priming the priming of seeds for 9 h resulted in a significant increase in siliqua weight compared to 3 h duration (0.108). However, the differences between 6 h (0.114) and 9 h duration were not significant (Table 9).

Table 9 Siliqua weight (g) in oilseed rape cultivars influenced by various doses of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h)
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It should be clear that both the weight of the seeds seed and that of the pods contribute to the total weight of the siliqua. And it was the weight of the husk of the siliqua, not that of the seeds, that was higher than that of the untreated seeds. The increase in husk mass of oilseed rape plants treated with picric acid could be due to the production of large protein molecules to cope with the altered conditions caused by the mutagen. Similar to the present results, Birara et al. [24] reported an increase in capsule weight in sesame using hydroxylamine. In contrast to the present study, Bolbhat et al. [74], Mshembula et al. [83], Jagajanantham et al. [75], Arisha et al. [90], and Olawuyi & Okoli [69] confirmed a decrease in pod weight in many plants due to chemical mutagens.

Effect on seed weight (g)

The Abasin- 95 cultivar (0.068) had the highest seed weight compared to the other genotypes. However, the differences between the Abasin-95 cultivar (0.068) and the Nifa Gold cultivar (0.062) in terms of seed weight were highly non-significant. In contrast, the Dur e Nifa cultivar (0.059) recorded a significant decrease in seed weight. The Nifa Gold cultivar (0.100) showed the highest seed weight at a dose of 10 mM for 9 h. On the other hand, priming of the Nifa Gold cultivar (0.043) at a dose of 10 mM for 6 h had an inhibitory effect on seed weight (Table 10).

Table 10 Seed weight (g) of oilseed rape cultivars under the effect of different doses (0, 5 mM, 10 mM, 15 mM, and 20 mM) and various priming durations (3 h, 6 h, and 9 h) of picric acid
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The results suggest that picric acid has the least effect on the genes responsible for seed production in oilseed rape plants [91]. It can also be deduced from this that genes responsible for seed production in oilseed rape plants counteract the mutagenic effects of picric acid [47]. A similar result, namely a non-significant increase in seed weight in chickpea genotypes, was reported by Wani & Anis [92]. In contrast to our results, Bolbhat et al. [74], Mshembula et al. [83], Jagajanantham et al. [75], and Arisha et al. [90] reported a reduction in seed weight of horse gram, cowpea, lady finger and bell pepper by the application of chemical mutagens. In contrast, some researchers reported positive effects of chemical mutagens on the weight of sesame and hibiscus seeds [75].

Effect on fresh and dry weight (gm)

In contrast to the other soaking times, the seeds soaked for 6 h yielded the plants with the highest fresh weight (37.58) in contrast to other priming durations. At the same time, soaking times of 3 h (35.77) and 6 h (37.58) were found to have no significant effect on fresh biomass of oilseed rape. Likewise, pre-soaking the seeds for 3 h and 9 h did not result in significant changes in the fresh biomass of oilseed rape. Conversely, plants grown from seeds pre-soaked for 6 h (37.58 gm) and 9 h durations (34.03 g) showed significant changes in fresh weight.

The Dur e Nifa cultivar recorded the highest fresh/dry weight (36.91/24.31), followed by the Nifa Gold cultivar (36.22/23.71) and the Abasin-95 cultivar (34.25/20.93). At the same time, the Dur e Nifa cultivar and the Nifa Gold cultivar did not show significant differences in fresh/dry biomass. In contrast, the differences between the Dur e Nifa cultivar and the Abasin-95 cultivar were highly significant in fresh biomass. However, the Abasin- 95 cultivar showed significant differences in dry biomass compared to the other genotypes. The 2nd order interaction (B × C) increased the fresh biomass (40.2) of the Nifa Gold cultivar, primed for 6 h. On the other hand, Abasin- 95 cultivar, primed for 9 h recorded the lowest fresh biomass (31.73) (Table 11 & 12).

Table 11 Effect of various treatment levels of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) on the fresh weight (g) of three oilseed rape cultivars
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Table 12 Effect of various treatment levels of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) on the dry weight (g) of three oilseed rape cultivars
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The compiled results on biomass attributes showed no-significant effects of picric acid on fresh/dry weight of oilseed rape plants. At the same time, priming the seeds for 6 h resulted in significant changes in the fresh weight of in the plants. There were also significant differences between the rapeseed varieties in the fresh and dry weights of the plants. An increase in fresh weight due to the application of various mutagens has already been observed in many plants such as mulberry [84], sesame [76], calendula [70] and kenaf [93]. In contrast, studies by Jagajanantham et al. [75] confirmed the inhibitory effect of mutagens on the fresh weight of plants and Mshembula et al. [83] and Jagajanantham et al. [75] also reported a decrease in dry weight in plants such as cowpea and Hibiscus sp. from seeds treated with some mutagens. On the other hand, [76, 84] reported an increase in dry weight of mulberry, sesame and hibiscus by mutagens.

Effect on moisture contents (%)

Moisture contents showed significant variations for the applied doses, the selected cultivars and the order 1 st (A × B), 2nd (B × C) and 3rd (A × C) orders interactions. At the same time, priming and the 4 th order interaction had no significant effects on the moisture content of rapeseed.

The dose of 15 mM (37.72) showed the highest moisture content, followed by 10 mM (37.08), 0 mM (33.87), 5 mM (33.11) and 20 mM (32.73). At the same time, the dosages of 10 mM and 15 mM did not result in significant changes in the moisture content compared to the other dosages. Likewise, the dosages of 0 mM and 10 mM did not show significant differences in moisture content. The same trend of non-significance in moisture content was observed at the 0 mM, 5 mM and 20 mM doses. In contrast, a dose of 15 mM picric acid resulted in significant changes in moisture content in contrast to the 0 mM, 5 mM and 20 mM treatment levels. Similarly, a dose of 10 mM resulted in significant effects on moisture content compared to 5 mM and 20 mM.

The Abasin-95 cultivar (37.92) had the highest moisture content compared to the other genotypes. At the same time, the Dur e Nifa cultivars (33.24) and the Nifa Gold cultivar (33.55) showed highly non-significant variations in moisture content. The 1 st order interaction (A × B) resulted in a significant increase in the moisture content of rapeseed (40.28) compared to the other treatments under the effect of the 15 mM dose and the 9 h presoaking. In contrast, the moisture content decreased to 28.04% when the seeds were soaked for 9 h with a 5 mM dose. The 2nd order interaction (B × C) increased the elevated moisture content of the Abasin- 95 cultivar (42.27) primed for 3 h in contrast to the other treatments. Priming the Dur e Nifa cultivar for 3 h decreased the moisture content (30.39) compared to the other treatments. The 3rd order interaction (A × C), stimulated the moisture content (41.13) of the Abasin- 95 cultivar exposed to a dose of 10 mM. In contrast, the Nifa Gold cultivar treated with a dose of 5 mM showed the lowest moisture content (26.66) (Table 13).

Table 13 Effect of various treatment levels of picric acid (0, 5 mM, 10 mM, 15 mM, and 20 mM) and different priming durations (3 h, 6 h, and 9 h) on the moisture content (%) of three oilseed rape cultivars
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In contrast to traits such as fresh/dry weight, picric acid can increase or decrease the moisture content of oilseed rape plants. Moreover, oilseed rape cultivars showed significant variations between varieties in terms of moisture content. The increase in moisture content at some picric acid doses (10 mM and 15 Mm), indicates that some anatomical variations (increase in trichomes) are induced by the mutagens in oilseed rape plants. Trichomes and stomata play a very important role in the conservation of water in the plant [77]. Trichomes are the epidermal outgrowths of plants. Trichomes increase the reflection of sunlight which helps to lower the internal temperature and thus reduces transpiration in plants growing under water deficit conditions. This helped the plants to intercept the dew drops, reduce transpiration and conserve water [80]. Studies by Kumar et al. [84], Akhtar [17] and Birara et al. [76] confirm the possibility of increasing the moisture content of plants by using mutagens [94] also reported five genotypes of B. napus and their physiological attributes and growth characters under drought induced osmotic stress [95] examined the transgenic rape seed lines and their phenophysiological traits under glyphosate treatment.

Effect on chloroplast genome

Control sample

The chloroplast genome of the control sample (contig 1) consists of 153,452 base pairs including a total of 10 rRNA and 38 t -RNA genes. It has two inverted repeat regions (a,b), which can be clearly seen in Fig. 1.

Fig. 1
figure 1

Diagrammatical representation of the chloroplast genome (153, 452 bp), illustrating gene annotations i.e. green represented as protein-coding genes, purple represented as tRNA genes, red represented as rRNA genes, gray represented as inverted repeat regions. Arrows demonstrated the direction of gene transcription

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Treated sample

The chloroplast genome of plants grown from picric acid-treated seeds was mutated. Therefore, annotation of the genome was not possible. Numerous single nucleotide substitutions (187) were detected between the control and test samples (Table 14). In addition, 43 additions/deletions were observed (Table 15).

Table 14 Comparison of control and treated samples based on single nucleotide polymorphism
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Table 15 Comparison of control and treated samples based on number of indels (insertions/deletions)
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Future research directions

Future research on the carcinogenic effects of picric acid on Brassica napus L. (Brassicaceae)’s chloroplast genome and biological characteristics may go in the following directions:

  • 1. Examine whether these mutations enhance or decrease the plant's resistance to environmental stressors such as heat, salinity, or drought.

  • 2. Examine the possibility of creating new Brassica cultivars with desired features by using the produced mutations.

  • 3. Examine alterations in secondary metabolites that may affect therapeutic usage or pest resistance.

  • 4. Research possible hazards related to picric acid emission into the environment and how it affects organisms that are not the intended targets.

  • 5. Integrate research with CRISPR/Cas9 or other gene-editing tools to implement targeted modifications based on mutagenesis outcomes.

Conclusion

This study shows that picric acid has a strong mutagenesis effect on the chloroplast genome and biological properties of Brassica napus cultivars. Picric acid's mutagenic influence was obvious in delayed seedling emergence, lower percentage emergence, and significant genomic alterations, including 187 single nucleotide changes and 43 insertions/deletions. Despite its inhibitory effects on early growth metrics, specific doses and priming durations had stimulatory effects, particularly on plant height and seed weight, with the Nifa Gold cultivar surpassing others in terms of adaptation and productivity under mutagenic conditions. These findings highlight picric acid dual role as an inhibitor and potential promoter of agronomic traits, depending on the dosage and exposure period. The findings also point to its possible utility in mutation breeding programs targeted at boosting specific agricultural attributes. More research into optimising its use and understanding its broader effects on genomic stability and plant physiology is required. Moreover, priming can improve germination and certain morpho-agronomic attributes of B. napus. The studied parameters confirmed the variations among the selected cultivars and can be used for further studies. The chloroplast genome studies categorically confirmed the mutagenic effect of picric acid. Taken together, these results provide new information to understand the mutagenic interactions on the chloroplast genome, while the application of picric acid, which can affect the growth and development of B. napus, has a positive morpho-agronomic effect.

Data availability

Availability of chloroplast genome data and materials has been deposited in NCBI GenBank Database and under the accession number PV294938 (https://www.ncbi.nlm.nih.gov/nuccore/2930385756?log$ = activity). All data generated or analyzed during this study are included in this published article.

References

  1. Aboodeh H, Bakhshandeh A, Moradi Telavat MR, Siadat SA, Moosavi SA, Alami Saeid K. The Physiological Response of Rapeseed (Brassica napus L.) Genotypes to Drought Stress. Agrotechniques in Industrial Crops. 2024;4(1):38–47.

  2. Bakhshi B. Heat and drought stress response and related management strategies in oilseed rape. Agrotechniques Industrial Crops. 2021;1(4):170–81.

    Google Scholar 

  3. Dolatparast B, Ahmadvand G, Mehrshad B, Hamzei J, Yazdandoost Hamedani M. The Effect of Delay in Planting Date on the Traits of the Rosette and Phenological Stages of Four Winter Oilseed Rape Cultivars in Hamadan. Iran Agrotechniques in Industrial Crops. 2021;1(3):129–38.

    Google Scholar 

  4. Amin F, Al-Huqail AA, Ullah S, Khan MN, Kaplan A, Ali B, Iqbal M, Elsaid FG, Ercisli S, Malik T. Mitigation effect of alpha-tocopherol and thermo-priming in Brassica napus L. under induced mercuric chloride stress. BMC Plant Biol. 2024;24(1):108.

  5. Khan NA, Khan MN, Razzaq A, Khalid S, Kaplan A, Iqbal M, Bibi M, Razak SA. Allelopathic Potential of Acacia modesta Wall., on the Growth of three Brassica Species. RADS J Biol Res Appl Sci. 2023;14(1):18–28.

  6. Wang H, Li X, Meng B, Fan Y, Khan SU, Qian M, Zhang M, Yang H, Lu K. Exploring silique number in Brassica napus L.: Genetic and molecular advances for improving yield. Plant Biotechnol J. 2024;22(7):1897-912.

  7. Kakaei M, Chaghakaboodi Z, Zebarjadi A, Kahrizi D. Exploring the Physiology and Genetic Stability of Rapeseed Plants for Assessing Oil Content in Western Iran. Agrotechniques in Industrial Crops. 2024;5(1):34–45.

    Google Scholar 

  8. Bailey LH, Bailey EZ. Hortus third: a concise dictionary of plants cultivated in the United States and Canada. 1976.

  9. Oleksy A, Zając T, Klimek-Kopyra A, Pustkowiak H, Jankowski K. Relative siliques position in a crop layer as an indicator of yield and quality in winter rape. Pak J Agric Sci. 2018;55(4):727-38.

  10. Wang C, Hai J, Yang J, Tian J, Chen W, Chen T, Luo H, Wang H. Influence of leaf and silique photosynthesis on seeds yield and seeds oil quality of oilseed rape (Brassica napus L.). Eur J Agronomy. 2016;74:112–118.

  11. Wolko J, Dobrzycka A, Bocianowski J, Bartkowiak-Broda I. Estimation of heterosis for yield-related traits for single cross and three-way cross hybrids of oilseed rape (Brassica napus L.). Euphytica. 2019;215(10):156.

  12. Ma Z, Du R, Xie J, Sun D, Fang H, Jiang L, Cen H. Phenotyping of Silique Morphology in Oilseed Rape Using Skeletonization with Hierarchical Segmentation. Plant Phenomics. 2023;5:0027.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Channaoui S, Labhilili M, Mouhib M, Mazouz H, El Fechtali M, Nabloussi A. Development and evaluation of diverse promising rapeseed (Brassica napus L.) mutants using physical and chemical mutagens. OCL. 2019;26:35.

  14. Shu QY, Forster BP, Nakagawa H, Nakagawa H. Plant mutation breeding and biotechnology, CABI. Plant Breeding and Genetics Section. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture. International Atomic Energy Agency, Vienna, Austria. 2012. ISBN, 978-2.

  15. Snowdon RJ, Iniguez Luy FL. Potential to improve oilseed rape and canola breeding in the genomics era. Plant Breeding. 2012;131(3):351–60.

    Article  CAS  Google Scholar 

  16. Bailey-Serres J, Parker JE, Ainsworth EA, Oldroyd GE, Schroeder JI. Genetic strategies for improving crop yields. Nature. 2019;575(7781):109–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Akhtar N. Effect of physical and chemical mutagens on morphological behavior of tomato (Solanum Lycopersicum) CV.“Rio Grande” under heat stress conditions. Plant Breed Seed Sci. 2014;70:69–79.

  18. Brown JK, Beeby R, Penfield S. Yield instability of winter oilseed rape modulated by early winter temperature. Sci Rep. 2019;9(1):6953.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Wu W, Shah F, Duncan RW, Ma BL. Grain yield, root growth habit and lodging of eight oilseed rape genotypes in response to a short period of heat stress during flowering. Agric For Meteorol. 2020;287: 107954.

    Article  Google Scholar 

  20. Weymann W, Böttcher U, Sieling K, Kage H. Effects of weather conditions during different growth phases on yield formation of winter oilseed rape. Field Crop Res. 2015;173:41–8.

    Article  Google Scholar 

  21. Marjanović-Jeromela A, Terzić S, Jankulovska M, Zorić M, Kondić-Špika A, Jocković M, Hristov N, Crnobarac J, Nagl N. Dissection of year related climatic variables and their effect on winter rapeseed (Brassica napus L.) development and yield. Agronomy. 2019;9(9):517.

  22. Boter M, Pozas J, Jarillo JA, Piñeiro M, Pernas M. Brassica napus roots use different strategies to respond to warm temperatures. Int J Mol Sci. 2023;24(2):1143.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Tubiello FN, Soussana J-F, Howden SM. Crop and pasture response to climate change. Proc Natl Acad Sci. 2007;104(50):19686–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Alkema L, Gerland P, Raftery A, Wilmoth J. The United Nations probabilistic population projections: An introduction to demographic forecasting with uncertainty. Foresight (Colch). 2015;2015(37):19.

  25. Gerland P, Raftery AE, Ševčíková H, Li N, Gu D, Spoorenberg T, Alkema L, Fosdick BK, Chunn J, Lalic N. World population stabilization unlikely this century. Science. 2014;346(6206):234–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kim C, Guo H, Kong W, Chandnani R, Shuang L-S, Paterson AH. Application of genotyping by sequencing technology to a variety of crop breeding programs. Plant Sci. 2016;242:14–22.

    Article  CAS  PubMed  Google Scholar 

  27. Fischer G, Shah MM, Van Velthuizen H. Climate Change and Agricultural Vulnerability. Laxenburg: IIASA; 2002.

  28. Fischer G, Shah M, N. Tubiello F, Van Velhuizen H. Socio-economic and climate change impacts on agriculture: an integrated assessment, 1990–2080. Philosophical Transactions of the Royal Society B: Biol Sci. 2005;360(1463):2067–2083.

  29. Bruinsma J. Agriculture: Towards 2015/2030. A FAO Perspective. London: Earthscan; 2003; p. 432.

  30. Porfirio LL, Newth D, Harman IN, Finnigan JJ, Cai Y. Patterns of crop cover under future climates. Ambio. 2017;46:265–76.

    Article  PubMed  Google Scholar 

  31. SAGEL Z. Determination of effect of chemical mutagen EMS on TAEK A-3 and TAEK C-10 mutant soybean varieties in M1 generation. Ekin J Crop Breed Genet 2017;3(1):19–24.

  32. Swarup S, Cargill EJ, Crosby K, Flagel L, Kniskern J, Glenn KC. Genetic diversity is indispensable for plant breeding to improve crops. Crop Sci. 2021;61(2):839–52.

    Article  Google Scholar 

  33. Ulukapi K, Nasircilar AG. Induced mutation: creating genetic diversity in plants. In: Genetic Diversity in Plant Species-Characterization and Conservation. IntechOpen; 2018. p. 41-55.

  34. Sattar MN, Iqbal Z, Al-Khayri JM, Jain SM. Induced genetic variations in fruit trees using new breeding tools: Food security and climate resilience. Plants. 2021;10(7):1347.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Lamo K, Bhat DJ, Kour K, Solanki SPS. Mutation studies in fruit crops: a review. Int J Curr Microbiol Appl Sci. 2017;6(12):3620–33.

    Article  Google Scholar 

  36. de Vries H. Mutations theorie. English; The Mutation Theory: Experiments and Observations on the Origin of Species in the Vegetable Kingdom. In: Kegan P, Trench T, editors. Open Court; 1909.

  37. Stadler LJ. Some genetic effects of X-rays in plants. J heredity. 1930;21:3-20.

  38. Shu Q-Y, Forster BP, Nakagawa H. Plant mutation breeding and biotechnology: Cabi; 2012. Plant Breeding and Genetics Section. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture. International Atomic Energy Agency, Vienna, Austria. ISBN, 978-2.

  39. Wagner A. The role of robustness in phenotypic adaptation and innovation. Proceed Royal Society B: Biological Sciences. 2012;279(1732):1249–58.

    Article  Google Scholar 

  40. Mba C, Afza R, Bado S, Jain SM. Induced mutagenesis in plants using physical and chemical agents. Plant cell culture: essential methods. 2010;20:111–30.

    Article  Google Scholar 

  41. Prudencio AS, Devin SR, Mahdavi SME, Martínez-García PJ, Salazar JA, Martínez-Gómez P. Spontaneous, Artificial, and Genome Editing-Mediated Mutations in Prunus. Int J Mol Sci. 2022;23(21):13273.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Joung JK, Sander JD. TALENs: a widely applicable technology for targeted genome editing. Nat Rev Mol Cell Biol. 2013;14(1):49–55.

    Article  CAS  PubMed  Google Scholar 

  43. Parry MA, Madgwick PJ, Bayon C, Tearall K, Hernandez-Lopez A, Baudo M, Rakszegi M, Hamada W, Al-Yassin A, Ouabbou H. Mutation discovery for crop improvement. J Exp Bot. 2009;60(10):2817–25.

    Article  CAS  PubMed  Google Scholar 

  44. Ding Y-L, Lyu Y-C, Leong MK. In silico prediction of the mutagenicity of nitroaromatic compounds using a novel two-QSAR approach. Toxicol In Vitro. 2017;40:102–14.

    Article  CAS  PubMed  Google Scholar 

  45. Ismail M, Khan M, Khan SB, Akhtar K, Khan MA, Asiri AM. Catalytic reduction of picric acid, nitrophenols and organic azo dyes via green synthesized plant supported Ag nanoparticles. J Mol Liq. 2018;268:87–101.

    Article  CAS  Google Scholar 

  46. Din IU, Khan S, Khan FU, Khan M, Khan MN, Hafeez A, Wahab S, Wahid N, Ali B, Qasim UB. Genetic Characterization of Advance Bread Wheat Lines for Yield and Stripe Rust Resistance. ACS Omega. 2023;8(29):25988–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Ahmed I, Islam M, Arshad W, Mannan A, Ahmad W, Mirza B. High-quality plant DNA extraction for PCR: an easy approach. J Appl Genet. 2009;50:105–7.

    Article  CAS  PubMed  Google Scholar 

  48. Dierckxsens N, Mardulyn P, Smits G. NOVOPlasty: de novo assembly of organelle genomes from whole genome data. Nucleic Acids Res. 2017;45(4):e18–e18.

    PubMed  Google Scholar 

  49. Tillich M, Lehwark P, Pellizzer T, Ulbricht-Jones ES, Fischer A, Bock R, Greiner S. GeSeq–versatile and accurate annotation of organelle genomes. Nucleic Acids Res. 2017;45(W1):W6–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods. 2012;9(4):357–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Nock CJ, Waters DL, Edwards MA, Bowen SG, Rice N, Cordeiro GM, Henry RJ. Chloroplast genome sequences from total DNA for plant identification. Plant Biotechnol J. 2011;9(3):328–33.

    Article  CAS  PubMed  Google Scholar 

  52. Tahir O, Bangash SAK, Ibrahim M, Shahab S, Khattak SH, Ud Din I, Khan MN, Hafeez A, Wahab S, Ali B. Evaluation of agronomic performance and genetic diversity analysis using simple sequence repeats markers in selected wheat lines. Sustainability. 2022;15(1):293.

    Article  Google Scholar 

  53. Khattak SH, Bangash SAK, Kaleem I, Begum S, Noor S, Ali GM, Hafeez A, Ali B, Khan MN, Kaplan A. Diversity Analysis for Wheat Genotypes Against Rust Resistant Genes. Polish J Environ Stud. 2024;33(3):2129-40.

  54. Amiryousefi A, Hyvönen J, Poczai P. The chloroplast genome sequence of bittersweet (Solanum dulcamara): Plastid genome structure evolution in Solanaceae. PLoS ONE. 2018;13(4):e0196069.

    Article  PubMed  PubMed Central  Google Scholar 

  55. Cai J, Ma P-F, Li H-T, Li D-Z. Complete plastid genome sequencing of four Tilia species (Malvaceae): a comparative analysis and phylogenetic implications. PLoS ONE. 2015;10(11):e0142705.

    Article  PubMed  PubMed Central  Google Scholar 

  56. Henriquez CL. Abdullah, Ahmed I, Carlsen MM, Zuluaga A, Croat TB, McKain MR: Molecular evolution of chloroplast genomes in Monsteroideae (Araceae). Planta. 2020;251:1–16.

    Article  Google Scholar 

  57. Henriquez CL, Ahmed I, Carlsen MM, Zuluaga A, Croat TB, McKain MR. Evolutionary dynamics of chloroplast genomes in subfamily Aroideae (Araceae). Genomics. 2020;112(3):2349–60.

    Article  CAS  PubMed  Google Scholar 

  58. Abdullah, Mehmood F, Rahim A, Heidari P, Ahmed I, Poczai P. Comparative plastome analysis of Blumea, with implications for genome evolution and phylogeny of Asteroideae. Ecol Evol. 2021;11(12):7810–7826.

  59. Shahzadi I, Mehmood F, Ali Z, Ahmed I, Mirza B. Chloroplast genome sequences of Artemisia maritima and Artemisia absinthium: Comparative analyses, mutational hotspots in genus Artemisia and phylogeny in family Asteraceae. Genomics. 2020;112(2):1454–63.

    Article  CAS  PubMed  Google Scholar 

  60. Mehmood F, Shahzadi I, Ahmed I, Waheed MT, Mirza B. Characterization of Withania somnifera chloroplast genome and its comparison with other selected species of Solanaceae. Genomics. 2020;112(2):1522–30.

    Article  CAS  PubMed  Google Scholar 

  61. Steel RGD, Torrie JH. Principles and procedures of statistics. Principles and procedures of statistics. 1960. xvi + 481 pp.

  62. Kovacs E, Keresztes A. Effect of gamma and UV-B/C radiation on plant cells. Micron. 2002;33(2):199–210.

    Article  CAS  PubMed  Google Scholar 

  63. Khan S, Goyal S. Improvement of mungbean varieties through induced mutations. African Journal of Plant Science. 2009;3(8):174–80.

    Google Scholar 

  64. Verma A, Reddy K, Dhansekar P, Singh B. Effect of acute gamma radiation exposure on seed germination, survivability and seedling growth in cumin cv. Gujarat Cumin-4. Int J Seed Spices. 2017;7(1):23–28.

  65. Roychowdhury R, Tah J. Chemical mutagenic action on seed germination and related agro-metrical traits in M1 Dianthus generation. Current Botany. 2011;2(8):19-25.

  66. Damude HG, Kinney AJ. Enhancing plant seed oils for human nutrition. Plant Physiol. 2008;147(3):962–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Maamoun M, El-Mahrouk M, Dewir Y, Omran S. Effect of radiation and chemical mutagens on seeds germination of black cumin (Nigella sativa L). Journal of Agricultural Technology. 2014;10(5):1183–99.

    CAS  Google Scholar 

  68. Rajib Roychowdhury RR, Alam M, Sunanda Bishnu SB, Tinkari Dalal TD, Jagatpati Tah JT. Comparative study for effects of chemical mutagenesis on seed germination, survivability and pollen sterility in M1 and M2 generations of Dianthus. 2012;65:29-38.

  69. Olawuyi O, Okoli S. Genetic variability on tolerance of maize (Zea mays L.) genotypes induced with sodium azide mutagen. Molecular Plant Breeding. 2017;8(3):27-37.

  70. El-Nashar Y, Asrar A. Phenotypic and biochemical profile changes in calendula (Calendula officinalis L.) plants treated with two chemical mutagenesis. Genet Mol Res. 2016;15(2):27173326.

    Article  Google Scholar 

  71. Sable AD, Sable AD, Shegokar SP, Gore NS, Harke S. Effect of sodium azide induction on germination percentage and morphological growth in two varieties of okra. Int J Curr Microbiol App Sci. 2018;7(6):3586–93.

    Article  Google Scholar 

  72. Mensah J, Obadoni B, Akomeah P, Ikhajiagbe B, Ajibolu J. The effects of sodium azide and colchicine treatments on morphological and yield traits of sesame seed (Sesame indicum L.). Afr J Biotechnol. 2007;6(5):534-8.

  73. Emrani S, Arzani A, Saeidi G. Seed viability, germination and seedling growth of canola (Brassica napus L.) as influenced by chemical mutagens. Afr J Biotechnol. 2011;10(59):12602–13.

    Article  CAS  Google Scholar 

  74. Bolbhat S, Bhoge V, Dhumal K. Effect of mutagens on seed germination, plant survival and quantitative characters of horsegram (Macrotyloma uniflorum (Lam.) Verdc). Int J Life Sci Pharm Res. 2012;2(4):129-36.

  75. Jagajanantham N, Dhanavel D, Gnanamurthy S, Pavadai P. Induced on chemical mutagens in Bhendi Abelmoschus esculentus L Moench. Int J Curr Sci. 2013;5:133–7.

    Google Scholar 

  76. Birara A, Manikandan M, Andargie M. The mutagenic effect of hydroxylamine hydrochloride on the agronomic traits of Sesame (Sesamum indicum L.). Plant Sci Today. 2014;1(1):33–8.

    Article  Google Scholar 

  77. Abdulrahaman A, Oladele F. Response of trichomes to water stress in two species of Jatropha. Insight Botany. 2011;1(2):12.

    Google Scholar 

  78. Ariraman M, Gnanamurthy S, Dhanavel D, Bharathi T, Murugan S. Mutagenic effect on seed germination, seedling growth and seedling survival of Pigeon pea (Cajanus cajan (L.) Millsp). Int Letters Nat Sci. 2014;16:41-9.

  79. De Micco V, Arena C, Pignalosa D, Durante M. Effects of sparsely and densely ionizing radiation on plants. Radiat Environ Biophys. 2011;50:1–19.

    Article  PubMed  Google Scholar 

  80. AbdulRahaman A, Afolabi A, Zhigila D, Oladele F, Al Sahli A. Morpho-anatomical effects of sodium azide and nitrous acid on Citrullus lanatus (Thunb.) Matsum. & Nakai (Cucurbitaceae) and Moringa oleifera Lam.(Moringaceae). Hoehnea. 2018;45:225–237.

  81. Shrivastava P, Tiwari H. Deleterious effects of chemical mutagens on economically Important Plant. J Environ Res Dev. 2008;2(4):633–8.

    CAS  Google Scholar 

  82. Sharifi-Sirchi G, Baghizadeh A, Golestani M, Saffari V. The effects of chemical and physical mutagens on morphological and cytological characters of barley (Iranian cv. Nosrat). Afr J Biotechnol. 2012;11(16):3842–8.

    CAS  Google Scholar 

  83. Mshembula B, Mensah J, Ikhajiagbe B. Comparative assessment of the mutagenic effects of sodium azide on some selected growth and yield parameters of five accessions of cowpea–Tvu-3615, Tvu-2521, Tvu-3541, Tvu-3485 and Tvu-3574. Arch Appl Sci Res. 2012;4(4):1682–91.

    CAS  Google Scholar 

  84. Kumar H, Muralidhar T, Acharya S, Das MJ. EMS induced morphometric biomass and phytochemical variations in Morus species (Genotype RFS135). Am J Exp Agriculture. 2012;3(1):43–55.

    Article  Google Scholar 

  85. Ariraman M. GS, Dhanavel D, Bharathi T, Murugan S. Mutagenic effect on seed germination, seedling growth and seedling survival of Pigeon pea (Cajanus cajan (L.) Millsp). Int Lett Nat Sci. 2014;16.

  86. Kumar A, Singh BK, Meena HS, Ram B, Singh VV, Singh D. Cytogenetical investigations in colchicine-induced tetraploids of Brassica fruticulosa: an important wild relative of cultivated Brassicas. Cytologia. 2015;80(2):223–30.

    Article  CAS  Google Scholar 

  87. Barman P, Rekha A, Pandey AK. Effect of pre-sowing treatments with chemical mutagens on seed germination and growth performance of jamun (Syzygium cumini L. Skeels) under different potting substrates. Fruits. 2015;70(4):239–48.

    Article  Google Scholar 

  88. Ahmed MA, Chakraborty N, Tabana Y, Dahham SS, Shazrina Ismail I, Mohamed R, Yunus MA, Sandai D. The effect of physical and chemical mutagen on tomato plant. Adv Biol Res. 2017;11(2):64–9.

    CAS  Google Scholar 

  89. Joshi N, Ravindran A, Mahajan V. Investigations on chemical mutagen sensitivity in onion (Allium cepa L.). Int J Botany. 2011;7(3):243–8.

    Article  CAS  Google Scholar 

  90. Arisha M, Liang B, Shah SM, Gong Z, Li D. Kill curve analysis and response of first generation Capsicum annuum L. B12 cultivar to ethyl methane sulfonate. Genet Mol Res. 2014;13(4):10049–61.

    Article  CAS  PubMed  Google Scholar 

  91. Kamble GC, Petkar H, Patil A. Biotechnological Study of Mutagenic Effect on Flowering and Fruiting Pattern in Wild Chickpea.

  92. Wani AA, Anis M. Gamma ray-and EMS-induced bold-seeded high-yielding mutants in chickpea (Cicer arietinum L.). Turk J Biol. 2008;32(3):161–6.

    CAS  Google Scholar 

  93. Ibukun EO, Oluwaseyi AE. Effect of sodium azide on agro-morphological traits of four varieties of Kenaf (Hibiscus cannabinus). GSC Biological and Pharmaceutical Sciences. 2019;8(3).

  94. Zebarjadi A, Borjian L, Ghasempour H, Noori A, Kahrizi D, Chaghakaboodi Z. Growth and Physiological Characterization in Five Genotypes of Brassica napus Callus Culture under Drought-Induced Osmotic Stress. Agrotechniques Industrial Crops. 2024;4(3):171–9.

    Google Scholar 

  95. Roeintan A, Safavi SM, Kahrizi D. Investigating the Phenophysiological Traits of Transgenic Rapeseed Lines (Brassica napus) with aroA Gene Harboring a Point Mutation of Proline 101 to Serine (P101S) under Glyphosate Herbicide Treatment. Agrotechniques in Industrial Crops. 2024;5(2):127-36.

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Acknowledgements

This research was funded by Princess Nourah bint Abdulrahman University Researchers Supporting Project number (PNURSP2025R365), Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia. The authors extend their appreciation to the Deanship of Research and Graduate Studies at King Khalid University for funding this work through Large Research Project under grant number RGP2/323/45".

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Authors and Affiliations

  1. Department of Botany, Islamia College Peshawar, Peshawar, 25120, Pakistan

    Ishtiaq Ahmad, Barkat Ullah & Muhammad Nauman Khan

  2. Department of Botany, University of Peshawar, Peshawar, 25120, Pakistan

    Zahir Muhammad

  3. Department of Biology, Faculty of Science, University of Tabuk, Tabuk, 71491, Saudi Arabia

    Dikhnah Alshehri

  4. Department of Crop and Animal Production, Sason Vocational School, Batman University, Batman, 72060, Turkey

    Alevcan Kaplan

  5. Key Laboratory of Ecosystem Network Observation and Modeling, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, 11 A, Datun Road, Chaoyang District, Beijing, 100101, China

    Majid Iqbal

  6. University of Chinese Academy of Sciences, (UCAS), Beijing, 100049, China

    Majid Iqbal

  7. Department of Biology, Faculty of Science, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia

    Laila A. Al-Shuraym

  8. Department of Biology, College of Science, King Khalid University, Abha, 61413, Saudi Arabia

    Uthman Balgith Algopishi

  9. Department of Biology, College of Science and Arts, King Khalid University, Muhayl Asser, 61913, Saudi Arabia

    Sadeq K. Alhag

  10. Department of Basic Sciences, College of Nursing, Taif University, P.O. Box 11099, Taif, 21944, Saudi Arabia

    Meshari A. Alsuwat

  11. Department of Biomedical Sciences, Institute of Health, Jimma University, Jimma, Ethiopia

    Diriba Dereje Olana

  12. School of Agriculture and Environment, UWA Institute of Agriculture, The University of Western Australia, Perth, WA, 6009, Australia

    Muhammad Ammar Javed

  13. Institute of Biological Sciences, Faculty of Science, Universiti Malaya, Kuala Lumpur, 50603, Malaysia

    Sarah Abdul Razak

  14. Biology Laboratory, University Public School, University of Peshawar, Peshawar, 25120, Pakistan

    Muhammad Nauman Khan

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  1. Ishtiaq Ahmad
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Contributions

I.A.: Conceptualization, writing – original draft, data curation, methodology, software; B.U., Z.M.: Visualization, validation, project administration, software, supervision; M.N.K, A.K.: Methodology, formal analysis, investigation, validation; M.A.J.: Writing – review & editing, resources, software, formal analysis, visualization; D.D.O., M.I., U.B.A., S.A.R., M.A.A: Writing – review & editing, resources, data curation, funding acquisition; S.K.A., L.A.A.S & D.A. All authors contributed significantly, and have read and agreed to the published version of the manuscript.”

Corresponding authors

Correspondence to Diriba Dereje Olana or Muhammad Ammar Javed.

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Ethics approval and consent to participate

Oilseed rape genotypes (Abasin- 95, Dur e Nifa and Nifa Gold) were obtained from Nuclear Institute for Agriculture, Peshawar (NIFA), Pakistan. All the experiments were performed in accordance with relevant guidelines and regulations".

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The authors declare no competing interests.

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Ahmad, I., Ullah, B., Muhammad, Z. et al. Mutagenic impact of picric acid on chloroplast genome and a selection of biological attributes of Brassica napus L. (Brassicaceae). BMC Genomics 26, 491 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12864-025-11565-2

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Keywords

BMC Genomics

ISSN: 1471-2164

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