Nanofungicides With Selenium And Silicon Can Boost The Growth And Yield Of Common Bean (Phaseolus Vulgaris L.) And Control Alternaria Leaf Spot Disease

There is an urgent need to reduce the intensive use of chemical fungicides due to their potential damage to human health and the environment. The current study investigated whether nano-selenium (nano-Se) and nano-silica (Nano Fungicide-SiO2) could be used against the leaf spot disease caused by Alternaria alternata in a common bean (Phaseolus vulgaris L.). The engineered Se and SiO2 nanoparticles were compared to a traditional fungicide and a negative control with no treatment, and experiments were repeated during two successive seasons in fields and insecticide Fungicide，bactericide Fungicide，Nanoencapsulated pesticides in vitro. The in vitro study showed that 100 ppm nano-Se had an efficacy rate of 85.1% on A. alternata mycelial growth, followed by the combined applications (Se + SiO2 at half doses) with an efficacy rate of 77.8%. The field study showed that nano-Se and the combined application of nano-Se and nano-SiO2 significantly decreased the disease severity of A. alternata. There were no significant differences among nano-Se, the combined application, and the fungicide treatment (positive control). As compared to the negative control (no treatment), leaf weight increased by 38.3%, the number of leaves per plant by 25.7%, chlorophyll A by 24%, chlorophyll B by 17.5%, and total dry seed yield by 30%. In addition, nano-Se significantly increased the enzymatic capacity (i.e., CAT, POX, PPO) and antioxidant activity in the leaves. Our current study is the first to report that the selected nano-minerals are real alternatives to chemical fungicides for controlling A. alternata in common beans. This work suggests the potential of nanoparticles as alternatives to fungicides. Further studies are needed to better understand the mechanisms and how different nano-materials could be used against phytopathogens.

Keywords: biotic stress, phytopathogen, nanofungicide, antioxidants, electrolyte leakage, selenium, silica
1. Introduction

The common bean Phaseolus vulgaris L. is one of the superior leguminous vegetables worldwide and is used for green pod and dry seed consumption. Brazil and Mexico are the largest producers with productions of 495,100 and 373,750 MT per year, respectively [1]. The common bean is the third most important food legume worldwide, after soybean and peanut. Total harvested area is 34.8 million ha year−1, which produces about 27.5 million tons year−1 [2]. The common bean is rich in protein (22%), dietary fiber, fat, and carbohydrates (62%), and the plant contains valuable phytochemicals and antioxidants, as well as acceptable levels of various vitamins and minerals [3]. Growing beans can improve the soil properties and reduce the nitrogen fertilization due to the plant’s characteristic of N-fixation, which increases the soil fertility [4]. However, its production faces many challenges, including pests and diseases, that are related to various bacteria [5,6], fungi [1], nematodes and insects [7], as well as abiotic stresses, e.g., salinity [8], drought [9], and heat [10]. Losses in productivity and quality have commonly reached as high as 60% due to biotic and abiotic stresses



/>Fungal phytopathogens have caused severe symptoms, including leaf and pod spots, leaf blight, rust, and root-rot [11]. Alternaria spp. are destructive to crops such as the apple Malus domestica Borkh. [12], the tomato Solanum lycopersicum L. [13], the potato Solanum tuberosum L. [14], and the common bean [11]. Alternaria alternata caused leaf spot and leaf blight diseases [15]. Other small-spored Alternaria species include A. arborescens E.G. Simmons and A. tenuissima (Kunze) Wiltshire, which both caused brown leaf spots, as well as A. solani Sorauer, which caused early blight [16]. To control these phytopathogens, chemical fungicides have been applied [17]; however, the overuse of these chemicals may represent a real threat to the environment, as well as a serious risk to human health [18]. Although biological control strategies have been developed, they are expensive and, often, take a long time to be effective, so many farmers regard them as infeasible alternatives [19]. Consequently, research has continued for additional alternatives with low environmental and health risks [20]. One such alternative is nano-minerals [21,22,



/>Searching for alternatives to traditional pesticides is an emerging research topic. Much research has been focused on developing novel "sustainable pesticides", which include nutrient-fungicides such as CuO-NPs [24]. The protective role of many elements in nano-form has been confirmed against different plant pathogens, including bacteria, fungi, actinomycetes, and nematodes [25]. These nano-metals/metalloids have included titanium (TiO2-NPs) [26,27], silver (Ag-NPs) [28], magnesium (MgO-NPs) [29], silicon (SiO2-NPs) [30], copper (Cu-NPs) [31], zinc (ZnO-NPs) [32], and other nanomaterials

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/>Certain nanoparticles (NPs) of Se and SiO2 are considered nanofungicides due to their effect against a number of phytopathogens [22,23,30]. Nanofungicides are interesting to explore due to their low dose requirement; low dose-dependent toxicity; high solubility and permeability; targeted delivery; enhanced bioavailability; and controlled release [23]. Nano-selenium and nano-silica are well-known as anti-stressors for various cultivated plants, as confirmed in many studies, and many studies have been carried out to examine their role in supporting plant cultivation under abiotic stress [34], such as drought on strawberry Fragaria spp. [35], the toxicity of heavy metals on rice Oryza sativa L. [36], and salinity on rice [37]. However, few studies have reported on the combined application of nano-Se and nano-silica to mitigate biotic stress. In one study, their combined application was used against root-rot disease induced by Fusarium spp. on bread wheat Triticum aestivum L. [38]. Furthermore, the use of nano-selenium was reported in different studies, e.g., against tomato leaf blight caused by Alternaria alternata [26] and against tomato late-blight disease [39]. Although nano-silicon has the potential for mitigating biotic stress in plants [40,41], more investigations are required. This includes nano-silicon’s effect against different diseases, such as stem canker; stem and leaf blight; leaf and root wilt; leaf spot; and soft rot [42]. The use of nutrients in nano-form has demonstrated their protective role against phytopathogens. Hence, it is worth exploring nanofungicides and the potential of this kind of fungicide. This could provide new directions for the application of sustainable nanofungicides as a potential replacement for traditional chemical fungicides. Therefore, this work aimed to study the role both of nano-selenium (nano-Se) and nano-silica (nano-SiO2), both individually and combined, in controlling leaf spot disease caused by Alternaia alternata in the common bean. Nano-Se and nano-silica were compared to a commercial fungicide and a negative control with no treatment, during two successive seasons. In addition to yield measurements, the chlorophyll content, the enzymatic activities, and the total antioxidants in plant leaves were studied after the application

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