Skip to main content
Download PDF

Effects of copper sulphate stress on the morphological and biochemical characteristics of Spinacia oleracea and Avena sativa

BMC Plant Biology volume 24, Article number: 889 (2024) Cite this article

Abstract

Plants are subjected to various biotic and abiotic stresses that significantly impact their growth and productivity. To achieve balanced crop growth and yield, including for leafy vegetables, the continuous application of micronutrient is crucial. This study investigates the effects of different concentrations of copper sulphate (0, 75, 125, and 175 ppm) on the morphological and biochemical features of Spinacia oleracea and Avena sativa. Morphological parameters such as plant height, leaf area, root length, and fresh and dry weights were optimized at a concentration of 75 ppm copper sulfate. At this concentration, chlorophyll a & b levels increased significantly in Spinacia oleracea (462.9 and 249.8 𝜇𝑔/𝑔), and Avena sativa (404.7 and 437.63𝜇𝑔/𝑔). However, carotenoid content and sugar levels in Spinacia oleracea were negatively affected, while sugar content in Avena sativa increased at 125 ppm (941.6 µg/ml). Protein content increased in Spinacia oleracea (75 ppm, 180.3 µg/ml) but decreased in Avena sativa. Phenol content peaked in both plants at 75 ppm (362.2 and 244.5 µg/ml). Higher concentrations (175 ppm) of copper sulfate reduced plant productivity and health. Plants exposed to control and optimal concentrations (75 and 125 ppm) of copper sulpate exhibited the best health and growth compared to those subjected to higher concentrations. Maximum plant height, leaf area, root length, fresh and dry weights were observed at lower concentrations (75 and 125 ppm) of copper sulfate, while higher concentrations caused toxicity. Optimal copper sulfate levels enhanced chlorophyll a, chlorophyll b, total chlorophyll, protein, and phenol contents but inhibited sugar and carotenoid contents in both Spinacia oleracea and Avena sativa. Overall, increased copper sulfate treatment adversely affected the growth parameters and biochemical profiles of these plants.

Peer Review reports

Introduction

Plant stress is any undesirable situation, element that disturbs or blocks the metabolism of a plant [1, 2]. Stress-inducing factors can be biotic, like insects and fungi or abiotic, such as temperature, water scarcity, pollution, and extreme saline environments [3]. The production of various crop varieties has decreased worldwide due to salinity, making it a major abiotic stress factor micronutrient application [4]. Approximately 5% of cultivated land is severely affected due to this factor as it reduces plant growth and production [5]. The primary causes of salinity in controlled environments include irregular watering practices, excessive and improper fertilization and inadequate drainage systems [6, 7]. These factors lead to an accumulation of salts in the soil, which can have detrimental effects on plant physiology. Salt stress impacts plant both at the whole organism level and at the cellular level, depending on the osmotic and ionic concentrations of the salts present [8,9,10]. Copper sulphate, also known as blue stone or blue vitriol, forms bright blue crystals containing five molecules of water [CuSO4.5H2O]. The copper sulphate is widely used as a pesticide in agriculture and also in the leather industry [11]. Copper is a critical micronutrient for the growth and development of plants. This is a structural and catalytic component of many enzymes and proteins that are involved in different metabolic activities, including carbohydrates and nitrogen metabolism [12]. At higher concentrations, it accelerates reactive oxygen species and free radical development, which can damage the plasma membrane through the binding of membrane proteins (sulfhydryl groups) [13]. In addition to the fungicidal activity of copper salts, they enhance plant growth [14]. Copper intake in higher plants occurs in the chloroplast mostly as copper oxide [15, 16]. Moreover, it is also a constituent of plastocyanin in the electron transport chain. Thus, copper deficiency affects photosynthetic components [17].

Leafy vegetables are a crucial component of the human diet, and Spinach (Spinacea oleracea L.) member of the goosefoot family (Chenopodiaceae), is key winter leafy vegetable that grow close to the ground [18, 19]. Rich in bioactive compounds and essential minerals including manganese, magnesium, iron, betaine, folate, potassium, calcium, copper, folic acid, zinc, and phosphorous, spinach is also a major source.

Various vitamins (vit), such as A, B6, B2, C, E, and K. In 2007, global production reached 893,494 hectares, yielding 14,044,816 tons. However, food species cultivated using traditional methods are often considered less nutritious and healthy compared to those grown organically [20, 21]. Spinach is a common vegetable in south Asia and is cultivated worldwide. Traditionally, it is used for various medicinal purposes, such as cooling, flatulence laxatives, diuretics, and as a carminative drug [22, 23]. They are mainly pollinated by the wind and have male and female reproductive organs in separate plants (dioecious), although some monoecious species do occur. Their description is centered on their morphology and sex because of their flexible sex expression. Spinach is tolerant to abiotic stresses, including osmotic and salt stresses, and although the genetics behind this feature remains understudied [24]. several ongoing studies on this have shown that two genes are responsible for their tolerance to osmotic, heavy metal, and temperature stresses, respectively [25, 26].

Avena sativa L., commonly known as the common covered white oat, is a crucial cereal crop, particularly in the developing world [27]. It belong to the Poaceae family (grass), which includes approximately 11,500 species and 786 genera, making it the fifth largest family of flowering plants [26]. It is frequently used as animal feed and, to some extent, human food [28]. It is widely cultivated for both animal feed and to lesser extent human consumption, Avena sativa is an annual plant that thrives in arid and semi-arid region. The crop is valued for its resilience to salt stress, drought, mineral insufficiency, and cold allowing it to grow in a variety of soil types. Nutritionally, oats are rich in vitamins, minerals, balanced proteins, and soluble fibers, which contribute to their health benefits [29, 30]. The grains also contain unsaturated fatty acids with strong antioxidant properties [31]. Despite a significant decline in global oat production over the past 50 years, there has been a resurgence in cultivation due to the health benefits of their soluble fibers, particularly in lowering cholesterol levels [32, 33]. This study aims to investigate the physiological effects of copper sulfate on the growth of Spinacia oleracea and Avena sativa. By examining how different concentrations of copper sulfate influence various growth parameters, this research seeks to understand the role of this micronutrient in plant development. The findings will contribute to optimizing agricultural practices for these crops, enhancing their growth and nutritional value.

Materials and methods

Experimental design

The present study was conducted in the botanical garden of Bacha Khan University Charsadda using a Randomized Complete Block Design (RCBD) in natural conditions with an average temperature of 23℃. The seeds of Spinacia oleracea and Avena sativa were sown at a depth of 2 cm in pots measuring 20 cm in height and 15 cm in diameter. Each pot was filled with 1 kg of clay loamy soil, which was analyzed to contain 27.15% sand, 19.86% silt, and 52.98% clay, according to the method described by Bouyoucos (1962). Initial soil analysis revealed no traces of copper, and no additional copper supplements were added.

Copper sulfate was applied at concentrations of 75, 125, and 175 ppm/kg of soil. Each treatment was replicated in three pots, and pots without copper sulfate served as controls. Initially, eight seeds were sown per pot. The plants were maintained in the soil until the completion of their life cycle, with climatic conditions controlled in a greenhouse to ensure consistency throughout the study.

Growth parameters

The growth parameters of Spinacia oleracea and Avena sativa under copper sulphate stress were analyzed during the vegetative stage. The parameter measured including plant height (cm), root length (cm), leaf area (mm2), fresh and dry weight (mg), were analyzed during the vegetative stage. Data were obtained from three plant for each parameter across all treatments. This approach ensured consistency in the measurement of dry weight, both the selected varieties were oven-dried at 37℃ for 72 h and then weighed [34].

Biochemical analysis

Protein content

Protein content of leaf was measured by Bradford protein assay (BSA) was used as standard. Fresh leaves 0.1 g was ground with the help of mortar and pestle in 1 ml of potassium phosphate buffer pH 7.5 and, centrifuged for 10 min at 3000 rpm. The supernatant (0.1 ml) of given sample containing unknown amount of protein was poured in the test tubes and total volume of 1 ml was made by distilled water. The extraction buffers reagent A (0.75 g sodium carbonate (Na2CO3) 0.15 g sodium hydroxide (NaOH) (0.1 N) and 0.37 g Na-K tartrate was dissolved in 40 ml of distilled water) and reagent B copper sulfate pentahydrate (CuSO4.5H2O) (0.03 g) was dissolved in 8 ml of distil water) were prepared. 1 ml of reagent C (Solution A (38 ml) and Solution B (0.7 ml) were mixed) was added in extracted supernatant. After shaking for 10 min, 0.1 ml of reagent D (Folin phenol reagent was diluted with distilled water in 1:1 ratio) was added. The absorbance of each sample was recorded at 650 nm after 30 min incubation. The concentration of protein contents was determined with the reference to the standard curve made by using standard BSA. The BSA of different concentrations viz. 20,40,60,80,320, and 640 mg, respectively was prepared. The absorbance of BSA was recorded at 650 nm.

Sugar content

Fresh leaves were used to measure the sugar contents following the method described by [34]. The fresh plant material (0.5 g) was homogenized with 10 ml of distilled water in a clean mortar and then centrifuged at 3000 rpm for 5 min. To 0.1 ml of supernatant, 1 ml of 30% (w/v) phenol was added. After incubation at room temperature, 5 ml concentrated sulphuric acid was added to the mixture. The sample was then incubated for 4 h. Subsequently, the absorbance of each sample was recorded at 420 nm. The concentration of unknown sample was calculated with reference to standard curve made by using glucose.

Total chlorophyll content

Total chlorophyll content of leaf was analyzed and crude preparation 1 ml was mixed with 4 ml of 80% (w/v) acetone and allowed to stand in dark at room temperature. It was centrifuged at 2000 rpm for 5 min to clear the suspension. Supernatant was used for chlorophyll determination. Absorbance of solution was read at 645 nm (chlorophyll a) and at 663 nm (chlorophyll b) on spectrophotometer against 80% (v/v) acetone blank. Total chlorophyll content was determined for the equation given by [35].

$$\begin{gathered}\:Total\:chlorophyll\left( {\frac{{mg}}{1}} \right) \hfill \\\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\,\, = \left( {20.2 \times \:A645} \right) + (8.02 \times \:B663\:V*W*/1000 \hfill \\ \end{gathered}$$
(1)

Carotenoid content

Carotenoid content were analyzed as described by [36]. The determination of Carotenoid contents, 50 mg of fresh leaf sample was homogenized with excess of acetone in a mortar with pestle. The supernatant was decanted and filtered a Buchner funnel using Whitman No.42 filter paper. The content was transferred from mortar to the Buchner funnel and washed with 80% acetone until colorless. Filtrate were pooled and the volume was made up to 25 mL. The absorbance of this diluted extract was measured at 480 and 510 nm, respectively.

Phenol content

Total phenol contents were estimated the Folin-ciocalteu method [36]. 50 mg freeze dried plant material was homogenized with 70% methanol, and the homogenate was then centrifuged at 3000 rpm for 10 min. The supernatant was separated and brought to a final volume 25 ml with 70% methanol. 1 ml of this extract was mixed with 1 ml of Folin-ciocalteu reagent and then 2 ml of 20% Na2CO3 solution was added. The tube was shaken and heated in a boiling water bath for 1 min followed by rapid cooling under running tap water. The transmittance was measured at 650 nm, and the total phenolic contents was quantified using a standard curve.

Data Analysis

The complete data were collected in three biological repeats and analyzed by using randomize complete block design (RCBD). Data were analyzed and expressed in the form of mean ± standard deviation (SD) to find the variations in a single treatment and by using SPSS.16.0 as a statistical tool to find the differences among the groups.

Results

Effect of copper sulphate on growth parameters of Spinacia oleracea and Avena sativa

The study on the effect of copper sulfate on growth parameters reveals that both Spinacia oleracea and Avena sativa exhibited optimal growth responses at a 75 ppm concentration. At, this level, Spinacia oleracea achieved a maximum plant height of 41 cm and Avena sativa reached 19.1 cm, while root length was longest in the control condition (Table 1). Increasing copper sulfate concentrations beyond 75 ppm led to a reduction in plant height and a decrease in root length, although the leaf area peaked at 75 ppm with measurements of 4.60 mm² for Spinacia oleracea and 4.49 mm² for Avena sativa (Fig. 1, A). Minimum leaf area was observed at concentration of 175 ppm in term of Spinacia oleracea keeping Avena sativa as control. The observed results display that application of copper sulphate at optimum level will enhance the growth parameters however the higher concentration had shown inhibition.

Table 1 (a). Effect of Copper Sulphate on growth parameters of S. oleracea
Full size table
Table 2 (b). Effect of Copper Sulphate on growth parameters of Avena sativa
Full size table
Fig. 1
figure 1

Effects of copper sulphate on the growth parameters of (A) Spinacia oleracea (B) Avena sativa

Full size image

The statistical analysis revealed that Spinacia oleracea had the highest fresh weight (5.119 g) under controlled condition, while the lowest fresh weight (3.850 g) was recorded at 175 ppm copper sulfate, likely due to the plant’s higher concentration. In contrast, Avena Sativa showed increased fresh weight (1.095 g) at 75 ppm, but a significant reduction (0.506 g) at 175 ppm. For dry weight, Spinacia oleracea exhibited the maximum value (0.7866 g) under control conditions and the minimum (0.1373 g) at 175 ppm. Avena sativa showed increase in dry weight at 75 ppm, but the dry weight was inhibited at higher concentration higher concentration of 125 and 175 ppm (Fig. 1A&B).

Effect of copper sulphate on biochemical parameters of Spinacia oleracea and Avena sativa

The complete biochemical assessment of Spinacia oleracea and Avena sativa were carried out, which showed large variations under copper sulphate stress. The application of copper sulphate on various biochemical parameters i.e., protein, sugar, chlorophyll a and b, total chlorophyll and phenol contents were observed showing copper sulphate concentration of (75 and 125 ppm) can enhance majority of biochemical related parameters except carotenoid contents whereas, excess of copper sulphate treatment of 175 ppm cause toxicity.

Chlorophyll a (𝝁𝒈/𝒈

The results indicated that chlorophyll a level increased to 462.9 in Spinacia oleracea and 249.8 𝜇𝑔/𝑔 Avena sativa at the optimal copper concentration of 75 ppm. Conversely Spinacia oleracea exhibited a low chlorophyll a level of 198.8𝜇𝑔/𝑔 under control condition without copper sulphate treatment. In Avena sativa, an excessive copper concentration of 175 ppm to a decrease in chlorophyll a reducing the level to 136.3 𝜇𝑔/𝑔 (Fig. 2A&B).

Fig. 2
figure 2

Effects of copper sulphate on the chlorophyll-a, b, total sugar, protein and phenolic contents of Spinacia oleracea and Avena sativa

Full size image

Chlorophyll b (𝝁𝒈/𝒈)

The maximum value (404.7 and 437.6 3𝜇𝑔/𝑔) of chlorophyll b in Spinacia oleracea and Avena sativa was recorded at copper sulphate concentration of 75 ppm. In Spinacia oleracea minimum level of chlorophyll b (152.8 𝜇𝑔/𝑔) was assessed under control condition whereas, in Avena sativa the higher concentration of 175 ppm shown low chlorophyll b contents of 333.9 8𝜇𝑔/𝑔, respectively. Therefore, both copper sulphate toxic and copper sulphate deficient treatments reduced the chlorophyll a and b contents both in Spinacia oleracea and Avena sativa (Table 2; Figs. 2A and 3B).

Total Chlorophyll (𝝁𝒈/𝒈)

Similar responses were showed by for total chlorophyll contents both Spinacia oleracea and Avena sativa showed similar results as observed for chlorophyll a and b. The higher level of total chlorophyll content (692.1 and 713.6 𝜇𝑔/𝑔) were recorded at optimum level of (75 ppm) copper sulphate. Whereas, the reduction in total chlorophyll contents (472.5 and 479.4 𝜇𝑔/𝑔) were observed at highest concentration of copper sulphate treatment (Table 2a).

Table 3 (a) Effect of Copper Sulphate on chlorophyll a, b, total chlorophyll and biochemical parameters of Spinacia oleracea
Full size table
Table 4 (b) Effect of Copper Sulphate on chlorophyll a, b, total chlorophyll and biochemical parameters of Avena sativa
Full size table

Carotenoid contents (𝝁𝒈/𝒈)

The result shown carotenoid contents gradually reduced under copper sulphate deficient to copper toxic treatment both in Spinacia oleracea L. and Avena sativa L (Fig. 2&B). The maximum carotenoid contents (44.54 and 92.66 𝜇𝑔/𝑔) were recorded under control condition of copper sulphate. The low (26.28 and 54.24 𝜇𝑔/𝑔) carotenoid contents were found under higher (175 ppm) copper sulphate treatment (175 ppm), respectively. From the above discussion it is clarify that carotenoid contents decrease as the copper sulphate treatment increase.

Soluble Sugar contents (𝝁𝒈/ml)

The sugar contents of Spinacia oleracea were significantly reduced under elevated copper sulphate concentration (Fig. 2, D). The maximum (505.0 𝜇𝑔/ml) sugar contents were recorded under copper sulphate deficient condition whereas, lower (274.5 𝜇𝑔/ml) sugar contents were calculated under high (175 ppm) copper sulphate treatment These results were divergent in Avena sativa which showed 125 ppm concentration of copper sulphate can enhance (941.6 𝜇𝑔/ml) the sugar contents. Whereas, the low sugar (319.1 𝜇𝑔/ml) was assessed under control condition. Therefore, the response of sugar of Spinacia oleracea and Avena sativa was recorded under varying copper sulphate stress.

The highest protein content, 159.3 µg/ml, was noted under control conditions, whereas the lowest protein content, 105.2 µg/ml, was observed at the highest copper sulfate concentration (175 ppm) (Table 2).

Soluble leaf protein contents (𝝁𝒈/ml)

The maximum protein contents in Spinacia oleracea was observed at 75 ppm of copper sulfate, reaching (180.3𝜇𝑔/ml), while the minimum protein content was recorded at 175 ppm (Table 2). In Avena sativa, protein contents decrease as the concentration of copper sulphate increased. The higher proteins contents (159.3 𝜇𝑔/ml) were observed under control conditions, whereas, the lower protein contents (105.2 𝜇𝑔/ml) were observed at the highest copper sulphate treatment (175 ppm) (Table 2).

Total phenol contents (𝝁𝒈/ml)

It was observed an increase in phenolic contents under all copper sulphate treatment except 175 ppm concentration. In both Spinacia oleracea and Avena sativa maximum (362.2 and 244.5 𝜇𝑔/ml) phenol contents were recorded at 75 ppm concentration of copper sulphate. However, the elevation in copper sulphate concentration resulted in reduction of phenolic contents, hence the low phenol contents (256.9 and 202.5 𝜇𝑔/ml) were recorded at175 ppm in both Spinacia oleracea and Avena sativa (Table 2; Fig. 2 A & B).

Discussion

In our studies, the different concentrations of copper sulfate have been used in order to assess their effects on plant physiology. In particular, concentrations of 75 to 125 ppm have been identified as the most effective for increasing certain parameters of interest based on an exhaustive analysis. In order to increase the levels of essential components such as chlorophyll a, chlorophyll b, phenols and carotenoids, these concentrations have been shown to be most effective. Furthermore, the use of copper sulphates 75 and 125 ppm lead to very high plant height gains as well as a rise in both new and dried weight for plants [2, 37]. These results show that, in addition to their potential for optimizing plant performance, the use of these particular concentrations of copper sulfate has a significant influence on chosen physiology parameters as well as an overall growth and development of the trials plants [2]. However, the high concentration of 175 ppm was slightly toxic, reduced root length, and caused chlorosis. We also showed that for Spinacia oleracea and Avena sativa the optimum concentration of 75 ppm increased plant height, whereas copper deficiency and toxic concentrations reduced plant height. This result corroborates that of a study by [38], who reported that treatment with different copper concentrations increased plant height in Brassica juncea an.

Brassica napus [39]) found that zinc and copper are effective for increasing the height of Lactuca sativa and Avena sativa plants. In the present study, root length in both Spinacia oleracea and Avena sativa decreased as the copper sulphate concentration increased. These findings are in accordance with those of [40], which indicated that copper stress decreases root length in Trigonella foenum-graecum plants. In another study, the uptake of cadmium was found to reduce root length in Spinacia oleracea [41]. Furthermore, we observed that leaf area showed a positive response to copper sulphate stress. Plants treated with 75 ppm copper sulphate showed the maximum leaf area in both Spinacia oleracea and Avena sativa [42] examined the toxic effect of copper on leaf area and other morphological parameters of Glycine max Contrastingly, treatment of Spinacia oleracea with different metals was shown to have positive effects on various morphological parameters, including leaf area [43].

In this study, both fresh and dry weights decreased significantly as the copper concentration increased. In selected varieties, the maximum fresh and dry weights were assessed in the control and 75 ppm groups. These results similar to those of [44, 45], in which copper treatment decreased dry weight and other morphological parameters of Vitis labrusca plants. The comparative study to determine the effect of copper on Avena sativa and other varieties [46]. Their results indicated that excess copper reduced both the root, shoot, and dry and fresh weights [47]. Heavy metals, such as cadmium, zinc, and lead cause a decrease in the root, shoot, and fresh and dry weights of Spinacia oleracea. Another study reported that application of optimum levels of copper and cadmium increased the dry and fresh weights in Amaranthus caudatus and Spinacia oleracea [48, 49]. Our results showed that applying a sufficient copper concentration (75 ppm) increased chlorophyll a level. Copper is an essential micronutrient that plays an important role in the synthesis of both chlorophyll a and b [41]. Copper plays a critical role in chlorophyll function, as it is integral to the pigments structure responsible for light absorbing in leaves [50]. Our study closely aligns with the finding of Ambrosini et al. (2018), who reported that high copper concentration reduces chlorophyll a content in Vitis labrusca plants. Similarly, wheat has been observed to exhibit a negative response to copper stress, characterized by decrease chlorophyll a levels [51]. Copper and GA3 reduced chlorophyll a level in Spinacia oleracea in other studies [52]. Similar to chlorophyll a level, total chlorophyll and chlorophyll b levels were also enhanced with sufficient copper treatment (75 and 125 ppm). The present work was supported by ancient work that chlorophyll b and total chlorophyll levels were enhanced by subjecting Avena sativa plants to optimum levels of salt stress [53]. Our findings are also compared with previous research that chlorophyl b and total chlorophyll content decreased as nickel concentration increased. Moreover, we reported that as copper concentration increased, carotenoid content decreased [54] showed that different concentrations of copper decreased carotenoid content in Brassica juncea and Brassica napus plants. Similar results were also observed by [55], in which carotenoid contents of Spinacia oleracea decreased as the copper concentration increased. In the present study, the responses of Spinacia oleracea and Avena sativa. to copper stress were different in terms of sugar levels. In Spinacia oleracea soluble sugar level decreased as copper concentration increased, whereas in Avena sativa sufficient levels (75 and 125 ppm) of copper sulphate enhanced sugar content. This result is in agreement with the work of [56], who reported that copper toxicity decreases sugar content in Avena strigosa. In Avena sativa sufficient potassium levels have been found to increase sugar content [57]. The combined effect of copper, cadmium, and aluminum has been shown to increase soluble sugar content in Hordeum vulgare [24, 58].

The protein levels decreased in both varieties as the copper concentration increased. This is similar to the results of [59], in which the protein content in a citrus plant decreased due to copper treatment. Treatment with cadmium decreases protein content in Spinacia oleracea [60], whereas that with copper chloride significantly reduces protein content in Helianthus annuus [61]. The phenolic content of both Spinacia oleracea and Avena sativa was enhanced by 75 and 125 ppm of copper, but 175 ppm showed higher toxicity (Khatun et al.., 2008). Moreover, Withania somnifera. showed a positive response to copper toxicity, as copper treatment increased their phenolic content to some degree [62]. The phenolic content and antioxidant activity of Olea europaea showed a positive response to copper sulphate and copper hydroxide stress. In another study, the phenolic content in wheat cell wall decreased as the copper content increased [63, 64].

Conclusion

Copper sulphate is essential for healthy plants growth and is commonly used as a fungicide. This study found that treatment with an optimum concentration 75 ppm enhances both morphological and biochemical parameters in both crop varieties, while higher concentrations reduce biomass. Therefore, copper sulphate can be used as a fertilizer to increase agricultural yields. The optimal dosage may vary depending on the specific crop species and soil conditions. Excessive application can cause toxicity, leading to adverse effects on plant growth and development. To maximize the benefits of copper sulfate while minimizing risks, careful calibration and monitoring are necessary to determine the appropriate concentration for different crops and environmental conditions.

Data availability

All the data is available within the manuscript.

References

  1. Gaspar T, Franck T, Bisbis B, Kevers C, Jouve L, Hausman JF, et al. concepts in plant stress physiology. Application to plant tissue cultures. Plant Growth Regul. 2002;37:263–285.

  2. Saleem A, Zulfiqar A, Ali B, Naseeb MA, Almasaudi AS, Harakeh SJ. Iron sulfate (FeSO4) improved physiological attributes and antioxidant capacity by reducing oxidative stress of Oryza sativa L. cultivars in alkaline soil. Sustainability. 2022;14(24):16845.

  3. Rizwan N, Khan QU, Khan AAJPJB. Potassium dynamic in the rhizosphere of maize (ZEA MAYS L.) grown under induced saline sodic condition. Pak J Bot. 2022;54(1):25–31.

  4. Ashraf M, Foolad MRJE. Botany e: roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot. 2007;59(2):206–16.

  5. Isik GJPJB. Ecophysiological responses of solanum lycopersicum L. to different levels of salt stress. Pak J Bot. 2022;54(1):1–5.

  6. Wang Z, Huang B, Xu QJJASHS. Effects of abscisic acid on drought responses of Kentucky bluegrass. J Am Soc Hortic Sci. 2003;128(1):36–41.

  7. Yasmeen S, Wahab A, Saleem MH, Ali B, Qureshi KA, Jaremko MJS. Melatonin as a foliar application and adaptation in lentil (Lens culinaris Medik.) Crops under drought stress. 2022, 14(24):16345.

  8. Bahadur A, Ahmad R, Afzal A, Feng H, Suthar V, Batool A, Khan A, Mahmood-ul-Hassan MJC. The influences of Cr-tolerant rhizobacteria in phytoremediation and attenuation of cr (VI) stress in agronomic sunflower (Helianthus annuus L.). 2017, 179:112–9.

  9. Hasegawa PM, Bressan RA, Zhu J-K. Bohnert HJJAropb: Plant cellular and molecular responses to high salinity. 2000, 51(1):463–99.

  10. Muranaka S, Shimizu K, Kato MJP. A salt-tolerant cultivar of wheat maintains photosynthetic activity by suppressing sodium uptake. Photosynthetica. 2002;40:505–15.

  11. Saravu K, Jose J, Bhat MN, Jimmy B, Shastry BJ. Acute ingestion of copper sulphate: a review on its clinical manifestations and management. 2007;11(2):74.

  12. Iqbal S, Khan AM, Dilshad I, Moatter K, Ahmed T, Gilani SAJP, Biology A. 84. Influence of seed priming with CuSO4 and ZnSO4 on germination and seedling growth of oat under NaCl stress. Pure and Appl Biol. 2020;9(1):897–912.

  13. Elzaawely AA, Xuan TD, Tawata SJE, Botany E. Changes in essential oil, kava pyrones and total phenolics of Alpinia zerumbet (pers.) BL Burtt. & RM Sm. Leaves exposed to copper sulphate. Environ Exp Bot. 2007;59(3):347–53.

  14. Ponmurugan P, Manjukarunambika K, Elango V, Gnanamangai BMJJEN. Antifungal activity of biosynthesised copper nanoparticles evaluated against red root-rot disease in tea plants. 2016;11(13):1019–31.

  15. Kranner I, Minibayeva FV, Beckett RP, Seal CEJNP. What is stress? Concepts, definitions and applications in seed science. 2010, 188(3):655–673.

  16. Salah M, Mansour M, Zogona D, Xu XJFRI. Nanoencapsulation of anthocyanins-loaded β-lactoglobulin nanoparticles: characterization, stability, and bioavailability in vitro. 2020, 137:109635.

  17. Shahbaz M, Ravet K, Peers G, Pilon MJFPS. Prioritization of copper for the use in photosynthetic electron transport in developing leaves of hybrid poplar. 2015, 6:407.

  18. Bergquist S. Bioactive compounds in baby spinach (Spinacia oleracea L.), vol. 2006; 2006.

  19. Ahmad P, Ahanger MA, Alyemeni MN, Wijaya L, Egamberdieva D, Bhardwaj R, Ashraf MJJPI. Zinc application mitigates the adverse effects of NaCl stress on mustard [Brassica juncea (L.) Czern & Coss] through modulating compatible organic solutes, antioxidant enzymes, and flavonoid content. 2017, 12(1):429–37.

  20. Citak S, Sonmez SJS. Effects of conventional and organic fertilization on spinach (Spinacea Oleracea L.) growth, yield, vitamin C and nitrate concentration during two successive seasons. 2010, 126(4):415–20.

  21. Arain SM, Sial MA, Jamali KDJPJB. Identification of wheat mutants with improved drought tolerance and grain yield potential using biplot analysis. 2022, 54(1):45–55.

  22. Rao K, Tabassum B, Babu SR, Raja A. Banji DJWjop, sciences p: preliminary phytochemical screening of Spinacia oleracea L. 2015, 4(6):532–51.

  23. Zeb U, Batool A, Khan H, Ullah H, Gul B. Ethnobotanical assessment of Hazar Nao Hills, District Malakand, Khyber Pakhtunkhwa, Pakistan. Int J App Exp Biol. 2022;1(2):59–66.

  24. Upadhyaya C, Upadhyaya T, Patel. Attributes of non-ionizing radiation of 1800 MHz frequency on plant health and antioxidant content of Tomato (Solanum Lycopersicum) plants. J Radiat Res App Sci. 2022;15(1):54–68.

  25. Ribera A, Bai Y, Wolters A-MA, van Treuren R, Kik CJE. A review on the genetic resources, domestication and breeding history of spinach (Spinacia oleracea L.). 2020;216:1–21.

  26. Zeb U, Wang X, AzizUllah A, Fiaz S, Khan H, Ullah S, et al. Comparative genome sequence and phylogenetic analysis of chloroplast for evolutionary relationship among Pinus species. Saudi J Biol Sci. 2022;29(3):1618–27.

  27. Rasane P, Jha A, Sabikhi L, Kumar A. Unnikrishnan VJJofs, technology: nutritional advantages of oats and opportunities for its processing as value added foods-a review. 2015, 52:662–75.

  28. Syed S, Gadhe K. Katke SJJoP, Phytochemistry: studies on physical, chemical and mineral evaluation of oats (Avena sativa). 2020, 9(5):79–82.

  29. Fan M, Zhang Z, Wang F, Li Z, Hu YJJPN, Science S. Effect of nitrogen forms and levels on β-glucan accumulation in grains of oat (Avena sativa L.) plants. 2009, 172(6):861–6.

  30. Yongguang M, Jixiang L, Chunsheng M, Zhanwu GJNBHAC-N. Effects of NaCl stress on the growth and physiological changes in oat (Avena sativa) seedlings. 2015, 43(2):468–72.

  31. Djuric M, Mladenovic J, Pavlovic R, Murtic N, Murtic S, Milic V, Šekularac GJAJB. Aluminium content in leaf and root of oat (Avena sativa L.) grown on pseudogley soil. 2011, 10(77):17837–40.

  32. Boczkowska M, Tarczyk EJGR, Evolution C. Genetic diversity among Polish landraces of common oat (Avena sativa L.). 2013, 60:2157–69.

  33. Achleitner A, Tinker NA, Zechner E, Buerstmayr HJT, Genetics A. Genetic diversity among oat varieties of worldwide origin and associations of AFLP markers with quantitative traits. 2008, 117:1041–53.

  34. Deming J, Carpenter S. Determination of extracellular polymeric substances (EPS) using a modified phenol-sulfuric acid (PSA) assay for sugars. 2023.

  35. Pérez-Patricio M, Camas-Anzueto JL, Sanchez-Alegría A, Aguilar-González A, Gutiérrez-Miceli F, Escobar-Gómez E, Voisin Y, Rios-Rojas C. Grajales-Coutiño RJS: optical method for estimating the chlorophyll contents in plant leaves. 2018, 18(2):650.

  36. Kumar A, Aery NCJJPN. Biochemical changes, biomass production, and productivity of Triticum aestivum as a function of increasing molybdenum application. 2023, 46(10):2351–62.

  37. Khan H, et al. Phytoremediation of selected heavy metals using Bryophyllum daigremontianum (Raym-Hamet & H. Perrier) A. Berger. Bangladesh J Bot. 2020;49(1):21–27.

    Google Scholar 

  38. Alhammad BA, Seleiman MF, Harrison MTJA. Hydrogen peroxide mitigates cu stress in wheat. 2023, 13(4):862.

  39. Stuckey JW, Neaman A, Verdejo J, Navarro-Villarroel C, Peñaloza P, Dovletyarova EAJJSS, Nutrition P. Zinc alleviates copper toxicity to lettuce and oat in copper-contaminated soils. 2021, 21:1229–35.

  40. Parwez R, Aqeel U, Aftab T, Khan MMA, Naeem MJPP, Biochemistry. Melatonin supplementation combats nickel-induced phytotoxicity in Trigonella foenum-graecum L. plants through metal accumulation reduction, upregulation of NO generation, antioxidant defence machinery and secondary metabolites. 2023:107981.

  41. Sheikh L, Younis U, Shahzad AS, Hareem M, Elahi NN, Danish SJPJB. Evaluating the effects of cadmium under saline conditions on leafy vegetables by using acidified biochar. 2023, 55:1537.

  42. Pathak A, Haq S, Meena N, Dwivedi P, Kothari SL, Kachhwaha SJP. Multifaceted Role of Nanomaterials in Modulating In Vitro Seed Germination, Plant Morphogenesis, Metabolism and Genetic Engineering. 2023, 12(17):3126.

  43. Russo C, Nugnes R, Orlo E, di Matteo A, De Felice B, Montanino C, Lavorgna M, Isidori MJEP. Diclofenac eco-geno-toxicity in freshwater algae, rotifers and crustaceans. 2023, 335:122251.

  44. Reshi ZA, Ahmad W, Lukatkin AS, Javed SBJM. From nature to lab: a review of secondary Metabolite Biosynthetic pathways, Environmental influences, and in Vitro approaches. 2023, 13(8):895.

  45. Salah M, Xu XJJMS. Anthocyanin-β-lactoglobulin nanoparticles in acidic media: synthesis, characterization and interaction study. 2021, 1232:129995.

  46. Khan A, Khan AA, Hasan SA, Irfan M, Ahmad MJJPN. Changes in morphology and yield attributes of Nigella sativa L. by copper (cu) induced toxicity. 2023:1–13.

  47. Dotaniya M, Sharma A, Nagar M, Dotaniya C, Doutaniya R. Saha JJBoEC, Toxicology: can application of Pressmud Mediated Plant Nutrient Dynamics under Lead Contaminated Soils of Indian Vertisol? 2023, 110(2):44.

  48. Paschal A. WASTEWATER REUSE FOR IRRIGATION: investigation of heavy metals and pharmaceuticals impact on crop irrigation. In.; 2023.

  49. Zeb U, Aziz T, Azizullah A, Zan XY, Khan AA, Bacha SAS, et al. Complete mitochondrial genomes of edible mushrooms: features, evolution, and phylogeny. Physiol Plant. 2024;176(3):e14363.

  50. Arnon DIJPp. Copper enzymes in isolated chloroplasts. Polyphenoloxidase Beta vulgaris. 1949;24(1):1.

    Google Scholar 

  51. Moustakas M, Ouzounidou G, Symeonidis L, Karataglis SJSS, Nutrition P. Field study of the effects of excess copper on wheat photosynthesis and productivity. 1997, 43(3):531–9.

  52. Gong Q, Li Z-h, Wang L, Zhou J-y, Kang Q, Niu D-dJES, Research P. Gibberellic acid application on biomass, oxidative stress response, and photosynthesis in spinach (Spinacia oleracea L.) seedlings under copper stress. 2021, 28(38):53594–604.

  53. Ahmad F, Jabeen K, Iqbal S, Umar A, Ameen F, Gancarz M, Eldin Darwish DB. Influence of silicon nano-particles on Avena sativa L. to alleviate the biotic stress of Rhizoctonia solani. 2023, 13(1):15191.

  54. Feigl G, Kumar D, Lehotai N, Pető A, Molnár Á, Rácz É, Ördög A, Erdei L, Kolbert Z, Laskay GJABH. Comparing the effects of excess copper in the leaves of Brassica juncea (L. Czern) and Brassica napus (L.) seedlings: growth inhibition, oxidative stress and photosynthetic damage. 2015, 66(2):205–21.

  55. Dagari M, Umar GJCJ. Impacts of EDTA on uptake and accumulation of Cu2 + by spinach (Spinacia oleracea L.) seedlings replanted in hydroponic solutions. 2017, 8(1):74–80.

  56. Ambrosini VG, Rosa DJ, de Melo GWB, Zalamena J, Cella C, Simão DG, da Silva LS, Dos Santos HP, Toselli M, Tiecher TLJPP et al. High copper content in vineyard soils promotes modifications in photosynthetic parameters and morphological changes in the root system of ‘Red Niagara’plantlets. 2018, 128:89–98.

  57. Ahanger MA, Agarwal R, Tomar NS, Shrivastava MJJ. Potassium induces positive changes in nitrogen metabolism and antioxidant system of oat (Avena sativa L cultivar Kent). J Plant Interact. 2015;10(1):211–23.

  58. Guo TR, Zhang GP, Zhang YH. Physiological changes in barley plants under combined toxicity of aluminum, copper and cadmium. Colloids Surf B Biointerfaces. 2007;57(2):182–8.

  59. Huang WL, Wu FL, Huang HY, Huang WT, Deng CL, Yang LT, et al. Excess copper-induced alterations of protein profiles and related physiological parameters in citrus leaves. Plants. 2020;9(3):291.

  60. Younis U, Malik SA, Rizwan M, Qayyum MF, Ok YS, Shah MHR, et al. Biochar enhances the cadmium tolerance in spinach (Spinacia oleracea) through modification of cd uptake and physiological and biochemical attributes. Environ Sci Pollut Res. 2016;23:21385–94.

  61. Zengin FK, Kirbag SJ. Effects of copper on chlorophyll, proline, protein and abscisic acid level of sunflower (Helianthus annuus L.) seedlings. J Environ Biol. 2007;28(3):561.

  62. Ferreira IC, Barros L, Soares ME, Bastos ML, Pereira JA. Antioxidant activity and phenolic contents of Olea europaea L. leaves sprayed with different copper formulations. Food Chem. 2007;103(1):188–195.

  63. Şahin S, Bilgin, MJJotSoF. Agriculture: Olive tree (Olea europaea L.) leaf as a waste by-product of table olive and olive oil industry: a review. 2018;98(4):1271–9.

  64. Batool A, Azizullah A, Ullah K, Shad S, Khan FU, Seleiman MF, et al. Green synthesis of Zn-doped TIO2 nanoparticles from Zanthoxylum armatum. BMC Plant Biol. 2024;24(1):820.

Download references

Acknowledgements

The authors expend their appreciation to the Researchers Support Project number (RSP-2024R374) of King Saud University, Riyadh, Saudi Arabia.

Funding

This work was supported by funding from Natural Science Foundation of China (32172187, 31771961), Chinese Postdoctoral Science Foundation (2023M661761) and Science & Technology Platform Construction Program of Jiangxi Province. The authors expend their appreciation to the Researchers Support Project number (RSP-2024R374) of King Saud University, Riyadh, Saudi Arabia.

Author information

Authors and Affiliations

  1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China

    Umar Zeb, Asif Ali Khan & Feng-Jie Cui

  2. Department of Botany, Bacha Khan University Charsadda, Charsadda, Khyber Pakhtunkhwa, 24420, Pakistan

    Fazli Rahim

  3. Department of Biology, Faculty of Biological and Biomedical Science, The University of Haripur, Haripur, Khyber Pakhtunkhwa, 22620, Pakistan

    Umar Zeb & Azizullah Azizullah

  4. Faculty of Science, Zarqa University, Zarqa, 13110, Jordan

    Ibrahim A. Saleh

  5. Department of Botany, Islamia College University Peshawar, Peshawar, Khyber Pakhtunkhwa, 25120, Pakistan

    Sher Wali

  6. School of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China

    Hanif Khan

  7. Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore, 54590, Pakistan

    Sajid Fiaz

  8. Department of Botany and Microbiology, Faculty of Science, Beni-Suef University, Beni-Suef, 65541, Egypt

    Hamada AbdElgawad

  9. School of Environment and Safety Engineering, Jiangsu University, Zhenjiang, 212000, China

    Babar Iqbal

  10. Botany and Microbiology Department, College of Science, King Saud University, P.O. Box 2455, Riyadh, 11451, Saudi Arabia

    Mohammad K. Okla

  11. Department of Agronomy, Abdul Wali Khan University Mardan, Mardan, 23200, Khyber Pakhtunkhwa, Pakistan

    Shah Fahad

Authors
  1. Umar Zeb
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fazli Rahim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Azizullah Azizullah
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ibrahim A. Saleh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sher Wali
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Asif Ali Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hanif Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sajid Fiaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hamada AbdElgawad
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Babar Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Mohammad K. Okla
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Shah Fahad
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Feng-Jie Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

UZ: Investigation; Conceptualization; writing - original draft; FR: Methodology, writing-original draft, review & editing; IAS and AA: Investigation; methodology; H.A.E and SW: Writing-review & editing, methodology; A.A. K: Writing-review & editing; H.K: Writing-review & editing; S.F: Investigation, resources; MKO and BI: Conceptualization, writing-original draft, review & editing, project administration, funding acquisition; SF and FC: Conceptualization, funding acquisition. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Shah Fahad or Feng-Jie Cui.

Ethics declarations

Ethics approval and consent to participate

Not Applicable.

Consent for publication

Not Applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zeb, U., Rahim, F., Azizullah, A. et al. Effects of copper sulphate stress on the morphological and biochemical characteristics of Spinacia oleracea and Avena sativa. BMC Plant Biol 24, 889 (2024). https://doi.org/10.1186/s12870-024-05566-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12870-024-05566-8

Keywords

BMC Plant Biology

ISSN: 1471-2229

Contact us