**1. Introduction**

Crop cultivators suffer from high yield loss caused by various diseases. Biotic stress induced by microbes on crop plants reduces the crop yield and decreases the quality. Biotic stress causes disease in crops, which leads to the suffering of the plant. Diseases of the plant need to be controlled to maintain the abundance of food produced by farmers around the world. The management of crop diseases is very necessary to fulfill the food demand. Potato blight disease caused by plant pathogenic fungus *Phytopthora* caused more than one million deaths in Ireland [1]. Around 20–40% of agricultural crop yield losses occur globally due to various diseases caused by phytopathogenic bacteria, phytopathogenic fungi, pests, and weeds [2].

**Citation:** Khan, M.; Khan, A.U.; Hasan, M.A.; Yadav, K.K.; Pinto, M.M.C.; Malik, N.; Yadav, V.K.; Khan, A.H.; Islam, S.; Sharma, G.K. Agro-Nanotechnology as an Emerging Field: A Novel Sustainable Approach for Improving Plant Growth by Reducing Biotic Stress. *Appl. Sci.* **2021**, *11*, 2282. https:// doi.org/10.3390/app11052282

Academic Editor: Anthony William Coleman

Received: 27 January 2021 Accepted: 23 February 2021 Published: 4 March 2021

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**Copyright:** © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

It is estimated that in 2050 the world's human population will reach around 10 billion, and around 800 million people in the world will be hungry and around 653 million people in the world will be undernourished in 2030, thus fulfilling the food demand will remain a huge challenge. The current research progress and disease management strategies are not enough to fulfill the food demand by 2050 [3]. The first green revolution made a huge difference in yield and food production, but in the last few years' crop production has been stagnant and food demand is increasing sharply, so now we need a second green revolution to fulfill the food demand of the population.

Different approaches are used by farmers to mitigate the impact of plant diseases. The agriculture system mainly relies on chemicals to manage crop diseases and inhibit the growth of phytopathogens, which cause diseases before and after crop harvesting. The excessive use of chemical pesticides, herbicides, and fungicides that are mainly used to control plant diseases causes harmful environmental and human health consequences. Tilman et al. [4] observed that the high use of chemical pesticides increases resistance in pathogens and pests, reduces nitrogen fixation, and the bioaccumulation of toxic pesticides occurs.

An example is the synthetic chemical pesticide DDT, dichlorodiphenyltrichloroethane, which was extensively used in agriculture for controlling plant pathogens and was found to be genotoxic in humans, causing endocrine disorders [5]. Water and soil pollution is also caused by the excessive use and misuse of these chemicals. There is an increasing demand day by day to reduce the use of synthetic chemicals. Consequently, the harmful effects of chemicals on wildlife, the environment, and human health have increased the need for alternative measures in the control of plant pathogens, so that some phytopathologists have focused their research on developing a new alternative that should replace the use of chemicals in controlling plant diseases.

Nanotechnology has revolutionized agriculture and can control plant diseases, although the field of nanotechnology is still in the nascent stage and needs more research analysis [6].The use of nanomaterials in agriculture will reduce the excessive use of toxic chemicals used for plant disease management (Figures 1 and 2).

"Nano" denotes one-billionth part, thus nanotechnology deals with small things. The word nano is used for materials with a size range of 0.1 to 100 nanometers [7,8]. The first time the term nanotechnology was used was by Taniguchi in 1974 to the science that largely deals with particles of nano size (1.0 <sup>×</sup> <sup>10</sup>−<sup>9</sup> m). When a bulk material is reduced to nano size, it has a high surface-to-volume ratio that may increase its reactivity and express some new properties [7,9]. The control of plant diseases and improving plant growth by the use of nanomaterials are some of the possible key applications in the area of plant pathology. Approximately 260,000–309,000 metric tons of nanoparticles were produced in 2010 globally, and the worldwide consumption of nanomaterials was approximately from 225,060 metric tons to 585,000 metric tons in 2014 to 2019 [10,11].

In this review article, recent research progress and the application of various nanoparticles for the sustainable management of the biotic stress of crop systems and impact on plant growth have been discussed. We try to cover the various problems associated with crop cultivation and plant diseases and the use of different nanomaterials to control phytopathogens and improve plant growth.

**Figure 1.** Schematic presentation of nanomaterials in agriculture. [12] **Figure 1.** Schematic presentation of nanomaterials in agriculture [12]. **Figure 1.** Schematic presentation of nanomaterials in agriculture. [12]

*Appl. Sci.* **2021**, *11*, x FOR PEER REVIEW 3 of 16

**Figure 2.** Various applications of nanotechnology in agriculture taken from [12] **Figure 2.** Various applications of nanotechnology in agriculture taken from [12] **Figure 2.** Various applications of nanotechnology in agriculture taken from [12].

#### **2. Nanomaterials in Improving Plant Growth and Yield 2. Nanomaterials in Improving Plant Growth and Yield 2. Nanomaterials in Improving Plant Growth and Yield**

Currently, around 1300 nanomaterials, with widespread potential applications, are available [13,14]. Nanoparticles can penetrate the cell wall because the cell wall is porous to 3.5–20 nm macromolecules. Nanoparticles can enter through stomatal openings. When stomata are present at the lower surface of leaves, the entry of nanoparticles (NPs) becomes difficult [15]. It is reported that nanoparticles of size ≤43 nm can penetrate and enter into stomata [16,17]. Currently, around 1300 nanomaterials, with widespread potential applications, are available [13,14]. Nanoparticles can penetrate the cell wall because the cell wall is porous to 3.5–20 nm macromolecules. Nanoparticles can enter through stomatal openings. When stomata are present at the lower surface of leaves, the entry of nanoparticles (NPs) becomes difficult [15]. It is reported that nanoparticles of size ≤43 nm can penetrate and Currently, around 1300 nanomaterials, with widespread potential applications, are available [13,14]. Nanoparticles can penetrate the cell wall because the cell wall is porous to 3.5–20 nm macromolecules. Nanoparticles can enter through stomatal openings. When stomata are present at the lower surface of leaves, the entry of nanoparticles (NPs) becomes difficult [15]. It is reported that nanoparticles of size ≤43 nm can penetrate and enter into stomata [16,17].

The effect of nanoparticles on crop plants is concentration-based. Many plant processes such as seed germination and plant growth are affected by NP concentration [18]. Many NPs have been reported to be beneficial for plant growth. Mahmoud et al. [19] enter into stomata [16,17]. The effect of nanoparticles on crop plants is concentration-based. Many plant processes such as seed germination and plant growth are affected by NP concentration [18]. Many NPs have been reported to be beneficial for plant growth. Mahmoud et al. [19] The effect of nanoparticles on crop plants is concentration-based. Many plant processes such as seed germination and plant growth are affected by NP concentration [18]. Many NPs have been reported to be beneficial for plant growth. Mahmoud et al. [19] used

Zn, B, Si, zeolite NPs on a potato plant and found that these nanoparticles have a positive effect on potato plants and they improve the plant growth. Khan and Siddiqui [20] treated eggplant with ZnONPs and found a foliar spray of ZnONPs causes the highest improvement in eggplant growth. Awasthi et al. [21] reported that ZnONPs have a positive effect on seed germination in the *Triticum aestivum* plant. Zinc oxide nanoparticles (ZnONPs) can enhance plant biomass and agriculture production [22]. Sabir et al. [23] also showed that nanocalcite (CaCO3) application with Fe2O3, nano SiO2, and MgO improved the uptake of Mg, Ca, and Fe, and also notably enhanced the intake of P with micronutrients Zn and Mn. Venkatachalam et al. [24] found that ZnONPs increase in photosynthetic pigment in the *Leucaena leucocephala* plant. Narendhran et al. [25] reported high chlorophyll-a', chlorophyll-'b' and total chlorophyll content in the *Sesamum indicum* plant when treated with ZnO NPs. Taheri et al. [26] observed that treatment of ZnONPs increases the increase in shoot dry matter in *Zea mays.* Tarafdar et al. [27] found that ZnONPs enhanced shoot and grain yield in the *Pennisetum glaucum* plant.

The application of titanium dioxide (TiO2) on crops promotes plant growth parameters and can enhance the photosynthetic rate. Siddiqui et al. [28] usedTiO<sup>2</sup> and ZnONPs on beet root plants. They found that both NPs increased chlorophyll and carotenoid content, improved plant growth, and also improved super oxide dismutase (SOD), catalase (CAT), H2O2, and proline content in plants. ZnONPs were found to be better than TiO2NPs on beetroot plants. Raliya et al. [29] reported that TiO2NPs treatment improved shoots in the *Vigna radiate* plant. Lawre and Raskar [30] observed that TiO2NPs at a lower concentration enhanced seed germination and seedling growth in onion plants. Rafique et al. [31] found a positive effect of TiO2NPs on the *Triticum aestivum* plant. Mahmoodzadeh et al. [32] found a positive effect of TiO2NPs on the seed germination of the *Brassica napus* plant. Qi et al. [33] reported that treatment of TiO2NPs promotes photosynthetic rate in tomato plants.

Silicon is an important element that plays a key role in several metabolic and physiological activities in plants [34]. SiO2nanoparticles have the potential to enhance the germination and seedling growth of *Agropyron elongatum* [35]. Nano-SiO<sup>2</sup> can be used to produce effective fertilizers for crops and to minimize the loss of fertilizer through slow and controlled release, allowing for regulated, responsive, and timely delivery [36]. Siddiqui et al. [37] found improved seed germination in the *Cucurbita pepo* plant after treatment with Nano SiO2. Haghighi and Pessarakli [38] reported that Nano Si treatment on the tomato plant improves photosynthetic rate in treated plants.

Copper is an essential element for plant growth and development. Copper plays a key role in the activity of many plant enzymes. Copper nanoparticles (Cu NP) are used as antimicrobial agents, gas sensors, catalysts, electronics, etc. [39]. Wang et al. [40] found that CuO NPs improved photosynthesis in the *Spinacia oleracea* plant. Zhao et al. [41] reported that Cu(OH)2NPs improved the antioxidant system of the *Lactuca sativa* plant. Shinde et al. [42] found that Mg(OH)2NP treatment promotes seed germination and seedling growth in the *Zea mays* plant. Hussain et al. [43] reported that MgO NPs improve the antioxidant system in *Raphanus sativus* plants. Cai et al. [44] observed that MgO NPs can promote the plant growth of the Tobacco plant. Imada et al. [45] found that MgO NPs can induce resistance in the tomato plant.

Iqbal et al. [46] reported that AgNP treatment improved plant growth and tolerance to heat stress in the *Triticum aestivum* plant. Mehta et al. [47] found that AgNPs' foliar application enhanced growth and biomass in the *Vigna sinensis* plant. Pilon et al. [48] observed that chitosan NPs protect apple plants after post-harvest. Van et al. [49] found that chitosan NPs improve plant growth in Robusta coffee.

Das et al. [50] found that FeS<sup>2</sup> NPs improved seed germination in *Cicer arietinum, Daucus carota, pinacia oleracea, Brassica juncea,* and *Sesamum indicum* crops. The effects of various nanomaterials have been summarized in the following table (Table 1).


**Table 1.** Effect of various nanomaterials on plant physiology and growth parameters.


**Table 1.** *Cont.*

#### **3. Nanomaterials in Various Diseases Management**

Nanomaterials have antimicrobial activity. Silver nanoparticles have anti-bacterial and anti-fungal properties. Kim et al. [70] have reported the fungicidal effects of nanosilver against *Alternaria alternata, A. brassicicola, A. solani, Botrytis cinerea, Cladosporium cucumerinum, Corynespora cassiicola, Cylindrocarpon destructans, Didymella bryoniae, Fusarium oxysporum f. sp. cucumerinum, F. oxysporum f. sp. lycopersici, F. oxysporum, F. solani, Fusarium sp., Glomerella cingulata* and a few other fungi. Gautam et al. [71] showed the antifungal and antibacterial activity of AgNPs against *Erwinia sp., Bacillus megaterium, Pseudomonas syringe, Fusarium graminearum, F. avenaceum,* and *F. culmorum* fungi. Rodríguez-Serrano et al. [72] reported the antibacterial activity of AgNPs against *E. coli*. Husseinet al. [73] reported the antibacterial activity of AgNPs against *Staphylococcus aureus* and *Klebsiella pneumonia.* Shehzad et al. [74] reported that AgNPs have antibacterial activity against Gram-positive (*Bacillus subtilis*) and Gram-negative (*Escherichia coli*) bacteria. Mohanta et al. [75] reported that AgNPs have antibacterial activity against food borne pathogens *Pseudomonas aeruginosa*, *Escherichia coli,* and *Bacillus subtilis*. Abdelmale and Salaheldin [76] reported that AgNPs show antifungal activity against *Alternaria alternata, A. citri,* and *Penicillium digitatum* fungi. Krishnaraj et al. [77] found theantifungal activity of AgNPs against *Alternaria alternata, Macrophomina phaseolina, Botrytis cinerea, Sclerotinia sclerotiorum, Curvularia lunata,* and *Rhizoctonia solani* fungi. Jo et al. [78] described the antifungal activity of AgNPs against *Bipolaris sorokiniana* and *Magnaporthe grisea* fungi.

Shahryari et al. [79] reported that AgNPs and a silver–chitoson composite show antibacterial activity against *Pseudomonas syringae* pv. *syringae* bacteria. Divya et al. [80] reported that chitoson NPs have antifungal activity against *Macrophomia phaseolina* and *Alternaria alterneta* fungi. Xing et al. [81] reported that chitoson NPs have antifungal activity against *Fusarium solani* and *Aspergillus niger* fungi. Dang et al. [82] reported that AuNPs have antibacterial activity against *E. coli* bacteria. Attar and Yapaoz [83] observed that ZnO and AuNPs have antibacterial activity against *E. coli* bacteria. The gold nanoparticles showed toxic effect on bacteria, *Salmonella typhimurium*, in which the macro gold did not exhibit. Jayaseelana et al. [84] synthesized gold nanoparticles from *Abelmoschus esculentus* and reported their antifungal activity. The antifungal activity of AuNPs was tested against *Puccinia graministritci, Aspergillus niger, Aspergillus flavus* and *Candida albicans* using the standard well diffusion method. The maximum zone of inhibition was observed in the Au NPs against *P. graminis* and *C. albicans*.

Fan et al. [85] observed the antibacterial activity of Cu composites against *Xanthomonas euvesicatoria.* Huang et al. [86] showed the antifungal activity of CuO NPs against *Botrytis cinerea, Colletotrichum graminicola, Rhizoctonia solani, Colletotrichum musae, Magnaporthe oryzae, Penicillium digitatum,* and *Sclerotium rolfsii.* Giannousiet al. [87] showed the antifungal activity of CuO and Cu2O NPs against *Phytophthora infestans.* Sharmaet al. [88] reported the antifungal and antibacterial activity of MgONPs against *Ralstonia solanacearum* bacteria and *Phomopsis vexans* fungus. Imada et al. [45] found the antibacterial activity of MgONPs against *Ralstonia solanacearum*. Derbalah et al. [89] observed the antifungal property of silica NPs against *Alternaria solani* fungus. Akpinar et al. [90] found that SiO<sup>2</sup> NPs possess antifungal properties against *Fusarium oxysporum* f. sp. *lycopersici* and *F. oxysporum* f. sp. *radicislycopersici.* Park et al. [91] showed the antifungal activity of Nano Si-Ag against *Pythium ultimum, Magnaporthe grisea, Colletotrichum gloeosporioides, Botrytis cineria, Rhizoctonia solani, Pseudomonas syringae*, *Xanthomonas compestris* pv. *vesicatoria*.

Jamdagni et al. [92] found that ZnO NPs have promising antifungal activity against *Alternaria alternate Botrytis cinerea, Aspergillus niger, Fusarium oxysporum,* and *Penicillium expansum* fungi. Navale et al. [93] found the promising antifungal activity of ZnO NPs against *Aspergillus flavus* and *Aspergillus fumigates* fungi. Rajiv et al. [94] reported the antifungal activity of ZnO NPs against *Aspergillus flavus*, *A. niger*, *A. fumigates, Fusarium culmorum,* and *F. oxysporium.* Gunalan et al. [95] found that ZnO NPs have promising antifungal activity against *Aspergillus flavus, Trichoderma harzianum, A. nidulans,* and *Rhizopus stolonifer.* Dimkpa et al. [96] have shown the antifungal activity of ZnO nanoparticles on *Fusarium graminearum* fungus. Jayaseelan et al. [97] synthesized ZnO nanoparticles using *Aeromonas hydrophila* and screened their activity against pathogenic bacteria *P. aeruginosa*, and fungi, *C. albicans, A. flavus,* and *A. niger*. Sar et al. [98] reported the antifungal activity of TiO<sup>2</sup> NPs against *Fusarium oxysporum* f. sp. *radicislycopersici* and *Fusarium oxysporum* f. sp. *Lycopersici.* Hamza et al. [99] found the antifungal activity of TiO<sup>2</sup> NPs against *Cercospora beticola.* Ardakani [100] found the nematicidal activity of TiO<sup>2</sup> NPs against *Meloidogyne incognita* nematode. Kasemets et al. [101] reported the antifungal activity of ZnO and TiO<sup>2</sup> NPs against *Saccharomyces cerevisiae.* Cui et al. [102] found that TiO<sup>2</sup> NPs have antibacterial against *P. syringae pv. lachrymans and P. cubensis* (Table 2, Figure 3).


**Table 2.** Various nanomaterials in plant disease management


#### **Table 2.** *Cont.*


**Table 2.** *Cont.*

**Figure 3.** (**A**) Different types of nanoparticles. (**B**) Schematic presentation of delivery methods of different nanoparticles and translocation in plants. (**C**) Various applications of nanoparticles (Taken from Sanzari et al. [116]). **Figure 3.** (**A**) Different types of nanoparticles. (**B**) Schematic presentation of delivery methods of different nanoparticles and translocation in plants. (**C**) Various applications of nanoparticles (Taken from Sanzari et al. [116]).

**5. Conclusions**

Nanomaterials' effect on organisms is largely dependent on the dose, size, and shape, the types of NPs, concentration, and the duration of exposure to NPs and the plant/animal species [117,118]. Nanoparticles at optimum concentration augment the plant's growth, but high concentrations of nanoparticles could be toxic for plants. Kushwah and Patel [119] observed that the optimum concentration of nano TiO<sup>2</sup> in the *Vicia faba* plant ranged from 5–50 mg/L. Other studies proved that TiO<sup>2</sup> NPs may induce stress in plants such as tomato, cucumber and spinach at high concentration [120]. Silver nanoparticles cause chromosomal aberrations in *Vicia faba* [121]. Lopez-Moreno et al.

In summary, the literature shows that food demands will increase with time, and to fulfill the demand of people, the present agricultural practices are not sufficient and chemicals used in agriculture as pesticides have a severe toxic effect on the environment. Thus, we need to develop an alternative approach that has a less toxic effect on the environment and that could help in fulfilling food demands. According to estimates, around 192.8 Mt chemical fertilizers were used in 2016–2017 in the whole world. The use of toxic chemicals and pesticides causes environmental pollution, which affects fauna and flora. Pathogens and pests induce resistance against fungicides and pesticides.

[122] reported that CeO<sup>2</sup> nanoparticles can induce DNA damage in soybean.

The inhibitory action of nanoparticles on fungi and bacteria includes disruption by pore formation in the cell membrane, disturbance in membrane potential, cell wall damage, direct attachment to the cell surface, DNA damage, cell cycle arrest, the inhibition of enzyme activity and reactive oxygen species (ROS) generation, and this finally leads to death. Nanoparticles generate the ROS, which causes damage to the cellular structures. The different components of reactive oxygen species include free radicals, such as hydrogen peroxide (H2O2), superoxide (O<sup>2</sup> <sup>−</sup>), singlet oxygen (1O2), carbon dioxide radical (CO<sup>2</sup> −), hydroxyl (HO· ), hydroperoxyl (HO2), carbonate (CO<sup>3</sup> −), peroxyl (RO2), and alkoxyl (RO), and nonradicals, such as ozone (O3), nitric oxide (NO), hypobromous acid (HOBr), hypochlorous acid (HOCl), hypochlorite (OCl−), peroxy nitrite (ONOO−), organic peroxides (ROOH), peroxo monocarbonate (HOOCO<sup>2</sup> −), peroxy nitrous acid (ONOOH) and peroxy nitrate (O2NOO−), and these nanoparticles accumulate in the membrane of bacteria or fungi, which leads to change in the permeability of the cell membrane and disturbs the proton motive force (PMF).Oxidative stress due to the higher concentration leads to single- and double-strand breaks and nitrogen base and pentose sugar lesions [103,104].

#### **4. Toxic Effect of Nanoparticles**

Nanomaterials' effect on organisms is largely dependent on the dose, size, and shape, the types of NPs, concentration, and the duration of exposure to NPs and the plant/animal species [117,118]. Nanoparticles at optimum concentration augment the plant's growth, but high concentrations of nanoparticles could be toxic for plants. Kushwah and Patel [119] observed that the optimum concentration of nano TiO<sup>2</sup> in the *Vicia faba* plant ranged from 5–50 mg/L. Other studies proved that TiO<sup>2</sup> NPs may induce stress in plants such as tomato, cucumber and spinach at high concentration [120]. Silver nanoparticles cause chromosomal aberrations in *Vicia faba* [121]. Lopez-Moreno et al. [122] reported that CeO<sup>2</sup> nanoparticles can induce DNA damage in soybean.

#### **5. Conclusions**

In summary, the literature shows that food demands will increase with time, and to fulfill the demand of people, the present agricultural practices are not sufficient and chemicals used in agriculture as pesticides have a severe toxic effect on the environment. Thus, we need to develop an alternative approach that has a less toxic effect on the environment and that could help in fulfilling food demands. According to estimates, around 192.8 Mt chemical fertilizers were used in 2016–2017 in the whole world. The use of toxic chemicals and pesticides causes environmental pollution, which affects fauna and flora. Pathogens and pests induce resistance against fungicides and pesticides. Hence, optimizing of the use of toxic chemical pesticides and fungicides is needed. Nanotechnology is flamboyant and has provided nanostructure materials as pesticide and fertilizer carriers. Nanomaterials can develop smart fertilizers as they can enhance nutrient availability and reduce environmental pollution [123]. Novel nanotechnology can be an alternative that can reduce crop diseases and enhance crop yield. Previous studies reported a significant positive effect of nanomaterials on crop plants. This novel technology can reduce the use of toxic chemicals and pesticides that contaminate soil, the environment, and groundwater. Further research is needed to develop this technology on a large scale (Figure 4).

Hence, optimizing of the use of toxic chemical pesticides and fungicides is needed. Nanotechnology is flamboyant and has provided nanostructure materials as pesticide and fertilizer carriers. Nanomaterials can develop smart fertilizers as they can enhance nutrient availability and reduce environmental pollution [123]. Novel nanotechnology can be an alternative that can reduce crop diseases and enhance crop yield. Previous studies reported a significant positive effect of nanomaterials on crop plants. This novel technology can reduce the use of toxic chemicals and pesticides that contaminate soil, the environment, and groundwater. Further research is needed to develop this technology on

**Figure 4.** Diagram showing general applications of nanoparticles in agriculture **Figure 4.** Diagram showing general applications of nanoparticles in agriculture.

**Author Contributions:** Writing—original draft: M.K. & A.U.K.; Writing—review & editing: M.K. & A.U.K., V.K.Y., K.K.Y., M.M.C.P.; Conceptualization: G.K.S. & S.I. Data curation: M.A.H. & A.H.K.; Formal analysis: K.K.Y., N.M.,& G.K.S.; Funding acquisition: M.A.H., M.M.C.P., A.H.K. & S.I.; Investigation: N.M., S.I., & G.K.S.; Methodology: M.K., A.U.K., & M.M.C.P.; Project administration: M.K., A.U.K., M.M.C.P.; Resources: M.A.H., K.K.Y., S.I., G.K.S. & A.H.K., Software: K.K.Y., G.K.S. & N.M., Supervision: A.U.K., M.M.C.P., A.H.K., V.K.Y., Validation: V.K.Y., M.A.H., A.H.K., Visualization: V.K.Y., S.I. & N.M. All authors have read and agreed to the published version of the manuscript. **Author Contributions:** Writing—original draft: M.K. and A.U.K.; Writing—review & editing: M.K., A.U.K., V.K.Y., K.K.Y. and M.M.C.P.; Conceptualization: G.K.S. and S.I. Data curation: M.A.H. and A.H.K.; Formal analysis: K.K.Y., N.M. and G.K.S.; Funding acquisition: M.A.H., M.M.C.P., A.H.K. and S.I.; Investigation: N.M., S.I. and G.K.S.; Methodology: M.K., A.U.K. and M.M.C.P.; Project administration: M.K., A.U.K., M.M.C.P.; Resources: M.A.H., K.K.Y., S.I., G.K.S. and A.H.K.; Software: K.K.Y., G.K.S. and N.M.; Supervision: A.U.K., M.M.C.P., A.H.K., V.K.Y., Validation: V.K.Y., M.A.H. and A.H.K.; Visualization: V.K.Y., S.I. and N.M. All authors have read and agreed to the published version of the manuscript.

**Funding:** Funding for this work has been provided by the Deanship of Scientific Research, KKU, Abha, Kingdom of Saudi Arabia, under research grant award number R.G.P2/85/41. **Funding:** Funding for this work has been provided by the Deanship of Scientific Research, KKU, Abha, Kingdom of Saudi Arabia, under research grant award number R.G.P2/85/41.

**Institutional Review Board Statement:** Not Applicable **Institutional Review Board Statement:** Not Applicable.

**Informed Consent Statement:** Not Applicable **Informed Consent Statement:** Not Applicable.

a large scale (Figure 4).

**Data Availability Statement:** The raw data used for this proposed work have been cited in the manuscript. Moreover, the derived data supporting the findings of this study have been graphically depicted and are available with the corresponding author on request. **Data Availability Statement:** The raw data used for this proposed work have been cited in the manuscript. Moreover, the derived data supporting the findings of this study have been graphically depicted and are available with the corresponding author on request.

**Acknowledgments:** The authors thankfully acknowledge the Deanship of Scientific Research, King Khalid University, Abha, for providing administrative and financial support. Funding for this work has been provided by the Deanship of Scientific Research, KKU, Abha, Kingdom of Saudi Arabia, under research grant award number R.G.P2/85/41. The authors also acknowledges the **Acknowledgments:** The authors thankfully acknowledge the Deanship of Scientific Research, King Khalid University, Abha, for providing administrative and financial support. Funding for this work has been provided by the Deanship of Scientific Research, KKU, Abha, Kingdom of Saudi Arabia, under research grant award number R.G.P2/85/41. The authors also acknowledges the contribution and support provided by the University of Aveiro, Portugal. The authors wish to acknowledge the work of all the references used in this study.

**Conflicts of Interest:** The authors declare that there is no conflict of interest regarding the publication of this paper.

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