The most commonly studied engineered nanomaterials (ENMs) have attracted scientists’ attention due to their extremely small size and very high surface area to volume ratio, which allow them to exhibit unique physical and chemical properties, with various reports demonstrating significant potential applications in modern agriculture, such as nano fertilizers, nano pesticides and other nanoforms of agricultural chemical inputs, which can significantly improve resource use efficiency and the quality of agricultural products, contributing to the sustainable development of agriculture (Raliya et al., 2018; Kumar et al., 2023; Periakaruppan et al., 2023). At the same time, biological effects on plants have become a challenging issue and have yet to be explored in detail although high amounts of nanoparticles (NPs) are discharged into ecosystems. The exponential increase in NPs applications across a wide range of industries, most prominently including medical, pharmaceutical, cosmetic, and electronic, resulting in the release of thousands of tons of nanomaterials into the agricultural production system each year (Liu et al., 2022). The global market of NPs expected to exceed $125 billion by 2024 (Wiseguyreports, 2019). Therefore, there is an urgent need to study the potential effects of these nanomaterials on highly exposed biological systems.

Numerous researchers have documented the biological (positive or negative) effects of ENMs on plant growth which depends on plant species tested, the properties of NPs, including size, functional groups, concentrations, as well as application methods of NPs (Gui et al., 2015; Bora et al., 2022). Among these, different application methods have been explored for nano fertilization to achieve the best promotion effect in the literature, including root irrigation, foliar application, stem injection, and seed presowing treatment. For instance, the foliar spray method was the best for Cu NPs to improve stem growth and new leaf appearance of avocado plants compared to other methods, along with complete uptake and translocation of Cu NPs (López-Luna et al., 2023). Kohatsu et al. found that the elemental composition in lettuce roots was mostly affected by the soil irrigation exposure for CuO NPs, while CuO NPs through foliar spray resulted in less disturbance (Kohatsu et al., 2021). For corn (Zea mays L.), 500 mg/kg Fe₃O₄ NPs applied by soil irrigation for 4 weeks had no impact on plant photosynthesis and biomass (Yan et al., 2020), while another study proposed the potential benefit of the foliar application of 200 mg/L Fe₃O₄ NPs to improve the yield and seed quality in cucumbers (Cucumis sativus L.) (Gupta et al., 2022). The plant growth-enhancing potential of iron sesquioxide (Fe₂O₃) NPs on soybean root growth and photosynthesis has been investigated after foliar spraying, and the increased intensity was far less than that of soil amendments, which may be related to the excessive precipitation of iron ions (Alidoust and Isoda, 2013). Similarly, titanium dioxide (TiO₂) NPs showed no changes in the germination rate of wheat but exhibited dose dependent effects on wheat when exposed to TiO₂ NPs in field soil, and different responses of wheat cultivars in loam and sandy loam soils were reported (Zahra et al., 2019; Mustafa et al., 2021).

Mo is an indispensable trace element for plant growth and development and is a major component of Mo flavin protease and 20 kinds of enzymes that catalyze the conversion of nitrates to nitrites (hydrogenase aldehyde oxidase, nitrate reductase). Mo deficiency can lead to the accumulation of nitrate in plants (Moussa et al., 2022). Mo NPs, as nontoxic and inexpensive materials, have great potential in agricultural applications and can be used in the form of nano fertilizers as microelement supplements (Sharma et al., 2021; Huang et al., 2022). To date, a few rudimentary studies have been performed to evaluate the biological effects of Mo NPs on plant growth. Taran et al. found 1μL/g Mo NPs promoted seed germination and activate the antioxidant enzyme activity of chickpea plants which enhanced the adaptability of crops when applied as a presowing treatment (Taran et al., 2016). Meanwhile, 8 mg/L Mo NPs mixed in soil increased the diversity of the rhizosphere microbial community by changing the root secretions of chickpeas (Taran et al., 2014; Thomas et al., 2017). Especially, 100 mg/L biogenic Mo NPs promoted the morphological parameters, nutrients content and ionic balance of wheat plants cultivated in arsenic-contaminated soil (Ahmed et al., 2022). Another study showed that after the introduction of 1mM Mo nanoclusters into the culture medium, the growth of rice roots was promoted but rice and rape root extension was significantly inhibited under higher exposure, maybe accompanied by root necrosis (Aubert et al., 2012; Adhikari et al., 2013). The biometric indices of pea irrigated with 100 ppm Mo NPs were significantly improved and the mineral element content in plants was increased by two times compared to the control (Sutulienė et al., 2021). In hydroponics condition, the increased growth and elevated protein levels in rice seedlings exposed to 100 ppm of α-MoO₃ and MoS₂ NPs, indicating hormesis. Meanwhile, tissue-specific distribution of NPs in rice seedlings was observed (Sharma et al., 2021). Nevertheless, these studies have mainly focused on the effects of Mo NPs on the seed germination and root growth of crops under short-term exposure, and more attempts from multiple dimensions need to be made to clarify the biological effects of Mo NPs on plants, including the physiological and molecular responses.

Early seedling stages are the important stages in the plant life cycle, and are preferred for toxicological studies due to their tolerance or sensitivity to external stress (Aslam et al., 1993). Reportedly, vital physiological indicators related to plant health, such as the photosynthetic efficiency, relative water content, and malondialdehyde (MDA) content, and morphological parameters, such as shoot and root lengths and shoot and root weights, can be significantly affected by NPs (Yang et al., 2020b). Photosynthesis is an important energy transfer process in plants and is closely related to stomatal conductance, the photosynthetic rate, the intercellular CO₂ (Ci) content and the transpiration rate (Tr) (Hoshika et al., 2022). Moreover, metal and metal oxide NPs cause oxidative stress in plants by producing reactive oxygen species (ROS), forcing the plants to evolve an antioxidant defense system as the first line of defense, increasing or decreasing antioxidant enzyme activities (Venzhik and Deryabin, 2023). Although many studies have been performed, plant response varies with the dose applied, plant species, application modes, experimental conditions and the exposure duration of NPs in the growth medium. We can still investigate changes in these important indicators to elucidate the Mo NPs-mediated promotion effect mechanisms, which have been very rarely reported, to our knowledge.

Flue-cured tobacco is an economically important crop in China with a planting area that exceeded 1 million/hm² in 2018, making it an indispensable part of the agricultural ecosystem. In recent years, the application of nanotechnology has made remarkable progress in tobacco disease and pest management, high-quality variety cultivation, soil fertilization, and yield improvement (Frazier et al., 2014). Studies on the biological effects of nanomaterials on the growth and development of tobacco seedlings have not been conducted. This study examines alterations in the various agronomic attributes, biomass, physiology and morphology of tobacco seedlings in a growth matrix under greenhouse conditions, and Mo accumulation in plants and the expression of several genes involved in physiological processes were analyzed in response to foliar applications and root irrigation with Mo NPs. The present study aimed to clarify the ecological behavior of Mo NPs. Most importantly, it is expected to provide a reference for the rational and efficient application of nanomaterials in agricultural production.

Notably, in this study, Mo NPs more significantly promoted tobacco seedling growth with root-irrigation application than with the foliar spraying method after 25 d. These results are in accordance with previous reports where strong positive effects on rice and chickpea were observed after Mo NPs treatment. The chickpea seed germination rate was significantly promoted after presowing treatment with Mo NPs, even at the low dosage of 5 mg/L (Adhikari et al., 2013; Taran et al., 2014; Taran et al., 2016). Similarly, the foliar application of Mo sulfide (MoS₂) NPs to rice obviously promoted rice growth through the enhancement of photosynthesis and the acceleration of cell division and expansion (Li et al., 2018). It is speculated that Mo NPs strengthen the water adsorption capability of soil amendments due to the role of Mo, which contributes to enhancing the water utilization efficiency and osmotic-adjustment ability, probably facilitating the translocation efficiency of a variety of nutrient elements. This finding is consistent with the results of Wu et al.’s study that Mo effectively improved plant water retention and drought resistance (Wu et al., 2014).

Bulk Mo fertilizer, commonly derived from ammonium molybdate and sodium molybdate, is considered an effective strategy to improve crop physiological traits. When wheat is chronically exposed to ammonium molybdate (0.41 kg ha⁻¹) in the field, the biomass, grain yield and uptake of P from soil are significantly increased (Rana et al., 2020). In a short-term treatment, seed priming with Mo (0.5 g/L of sodium molybdate) for 8 h increased chickpea yield in a pot study and in field study by 27% and 20%, respectively (Farooq et al., 2012). Other findings indicated that sodium molybdate positively affected leguminous plant (alfalfa and hairy vetch) growth, whether foliar application or soil addition, especially in low-pH soil (5.2), resulting in a significant increase in nodule counts and size, thus accelerating the biological N fixation process (Adhikari and Missaoui, 2017; Alam et al., 2015). However, higher Mo doses (1.0 mg kg⁻¹) led to the deterioration of nodule structure (Alam et al., 2015). In contrast, in this study, the concentration of Mo NPs used was far greater than that in the above studies without any phytotoxicity, indicating better biocompatibility. In addition, 50–70% of traditional fertilizers can be lost by means of evaporation, drift, hydrolysis, leaching, runoff, and microbial or photolytic degradation of nutrients, causing the very low use efficiency (Marchiol, 2019). It is worth highlighting that, as nano fertilizer, the physicochemical peculiarities of NPs contribute to the improvement of nutrient delivery efficiency in unique ways, such as specific release, controlled release, moisture release and rapid diffusion (Mikkelsen, 2018).

Some nanomaterials can change the ability to absorb water and regulate the absorption of metal elements in plants due to their hydrophilic properties (Hu et al., 2017). He et al. reported that GO can act as a “water transporter” in the soil, accelerating water absorption, and ultimately promoting the germination of spinach and chive seeds, but GO sheets are not absorbed by the seeds (He et al., 2018). This is slightly different from the mechanism by which MWCNTs puncture the seed coat and promote vegetable seed germination and plant growth by accelerating water uptake by means of enhanced aquaporin gene expression, whereafter they are delivered and accumulate in the plant (Villagarcia et al., 2012; Lahiani et al., 2013).

Chlorophyll, a photoreceptor, channels the energy of sunlight into chemical energy, converting it through the process of photosynthesis. Mo NPs increased the chlorophyll content of tobacco, especially under root irrigation, which is in agreement with the findings of Taran et al., who reported stimulating effects of Mo NPs on the chlorophyll content under soil treatment (Taran et al., 2014), while Yang et al. showed that exposure to all tested concentrations less than 100 mg/kg of Mo nanomaterials had no significant effect on the chlorophyll content in soybean leaves and therefore did not affect the net photosynthetic rate (Yang et al., 2020a). The growth promotion effect of Mo NPs on tobacco after root irrigation treatment is better than that of spraying treatment, which may be due to the better NPs absorption by plant roots. A higher chlorophyll content can improve light absorption and conversion efficiency, thus Mo NPs promote the photosynthesis process of tobacco plants (Hideaki, 2006). Our speculation was verified in this research. It is interesting to note the multiple effects of NPs on the chlorophyll content in a wide range of species (peanut, cotton, corn) in previous studies, including positive and negative effects, but they rarely involved Mo NPs (Adhikari et al., 2013; Thomas et al., 2017; Li et al., 2018; Chen et al., 2022). We speculated that the improved chlorophyll content of tobacco seedlings under Mo NPs exposure may result from the absorption of Mo, which is usually present in the active center of several plant enzymes that catalyze the key steps of nitrogen, carbon, and sulfur metabolism, participate in the chloroplast structure and regulate the production and balance of H₂O₂ in plants, promoting the synthesis of chlorophyll in leaves at the seedling stage. The activated mechanisms can be similar to those of MgO NPs (Cai et al., 2018). MgO NPs can be absorbed by plants, and participate in the synthesis and metabolism of magnesium porphyrin compounds in chlorophyll, thereby accelerating the photosynthesis of watermelon and tobacco plants, and even enhancing the resistance of plants to soilborne diseases (Wang et al., 2013).

Photosynthesis is an essential process by which green plants and certain other organisms transform light energy into chemical energy stored in carbohydrate molecules such as sugars and other organic compounds, which are synthesized from carbon dioxide and water. Chlorophyll, the most important class of pigments, assists in this process by trapping solar energy. Confirmatively, we next verified that the improved chlorophyll levels in Mo NPs-exposed tobacco resulted in an increased photosynthetic rate. A previous study demonstrated that CeO₂ NPs, MoS₂, ZnO, Fe₂O₃, and TiO₂ NPs stimulated the photosynthetic rates of crops at relatively low exposure levels (Zheng et al., 2005; Alidoust and Isoda, 2013; Tarafdar et al., 2014; Cao et al., 2017; Li et al., 2018; Sun et al., 2020). It was also reported that other engineering nanomaterials, such as CuO, Cu, MoO₃ and CeO₂ NPs, could exert a negative impact on photosynthesis at high concentrations by reducing the chlorophyll and carotenoid contents in rice, soybean, corn, and tomato (Du et al., 2015; Yang et al., 2020b; Rai et al., 2021; Sutulienė et al., 2023), or as by damaging photosynthesis-related genes (Wang et al., 2016). The underlying mechanism of this facilitating effect may be attributed to the enhancement of photosynthetic pigments and the function of NPs in regulating biological processes such as stomatal aperture, the synthesis of photosynthesis-related enzymes, enzymatic reactions, and water uptake.

When plants are stimulated by exogenous biotic or abiotic sources, for example, environmental factors, such as plant hormones, metal ions, relative reactive oxygen species (oxygen ions, peroxides, oxygen-containing free radicals, etc.) will be produced in various parts of the plant, resulting in the oxidation of biomolecules such as lipids, proteins, and DNA, and causing cell death (Taran et al., 2016). Hence, ROS-mediated oxidative pressure in plants is usually an effective indicator of phytotoxicity induced by ENMs. Subsequently, plants perceive that their tissues are threatened, which induces the activation of important defense enzymes, such as SOD, POD, and CAT, to eliminate ROS species in plants (Taran et al., 2016). SOD is highly efficient in eliminating superoxide radicals (O₂ ⁻) to produce O₂ and H₂O₂ in plants. CAT and POD can directly convert H₂O₂ into H₂O and O₂ through disproportionation and hydrolysis reactions, respectively (Imada et al., 2016). As has been reported previously, plants exposed to ENMs exhibit significantly increased defense enzyme activities compared to water-sprayed control plants but reduced MDA contents, especially under abiotic stress conditions, such as UV-B radiation, well-watered and drought stress (Aghdam et al., 2016; Imada et al., 2016; Venkatachalam et al., 2017; Sun et al., 2020). Similar changes were observed in other studies showing higher antioxidant enzyme activity of pea at 50 and100 ppm Mo NPs but lower levels of H₂O₂ and MDA (Taran et al., 2016; Sutulienė et al., 2021) The results of this study show that after Mo NPs treatment, the activities of three antioxidant enzymes in tobacco seedlings were enhanced in a concentration dependent manner to different degrees, especially after root irrigation treatment. Combining the results in Figure 3 ., it can be observed that Mo NPs not only display strong promotion effects on tobacco plants, but can also serve as an elicitor to stimulate the plant defense system against ambient stress.

Similar results have been reported in previous studies where ZnO NPs significantly improved the leaf total soluble protein contents in green peas and cotton even at lower concentrations (Mukherjee et al., 2016; Venkatachalam et al., 2017). Activating the expression level of stress proteins could possibly explain these results, which is also a response mechanism in plants for protecting their cells from potential oxidative damage caused by NPs (Zhao et al., 2012). Similarly, soluble sugars, especially sucrose and glucose, might also play central roles in higher plant defense responses under biotic and abiotic stresses (Couée et al., 2006). Most importantly, sugar signaling and sugar-modulated gene expression are related to maintaining reactive oxygen species balance in various metabolic reactions, such as mitochondrial respiration or photosynthesis regulation (Koch, 1996; Koch, 2004; Couée et al., 2006). Therefore, it can be speculated that the increase in the soluble sugar levels in tobacco plants after treatment with Mo NPs could be emerging as a signaling molecule that is, conducive to the strength of plant innate immunity.

NPs can usually be absorbed by the leaves or roots of plants and are mainly transported through the xylem and phloem pathways, followed by localization and accumulation in various plant tissues, potentially regulating plant physiological and biochemical processes (Gui et al., 2015). It is implied that both treatment measures accelerated Mo uptake in tissue, however, it seems that the root irrigation treatment shows weaker transport element capacity and causes Mo retention in the root system ( Figure 6 ). The same phenomenon was found in another study, where a strong accumulation of Mo in pea roots watered with Mo NPs was determined (Sutulienė et al., 2021). The reason for this may be related to the agglomeration of NPs in soil, which may allow them to pass through narrow vascular tissues in the stem with great difficulty (Collin et al., 2014). Yang et al. found that Mo mainly accumulated in soybean leaves when added to soil (Yang et al., 2020a). Additionally, in most described cases of NPs uptake by plants, either under hydroponic conditions or soil amendments, and with either foliar or irrigation application, it was clear that the absorption and distribution of ENMs by plants were very distinct according to the crop species, soil texture, NPs size, surface charge, and morphology, especially the differences in dispersion (Lv et al., 2019). For example, after ZnO NPs were absorbed by cluster bean and mung bean plants, the released Zn²⁺ was used as a cofactor in the catalytic activity center of phosphatase and improve the activities of phosphatase and inositol hexaphosphatase, consequently the absorption rate of soluble phosphorus in plants increased by 84-108%, and the absorption rate of insoluble phosphorus that is difficult for plants to absorb increased by 10.8% (Raliya and Tarafdar, 2013; Raliya et al., 2016). Therefore, we speculate that Mo NPs can also carry macronutrient elements into the plant during the process of accelerated plant water absorption, thus promoting plant growth and development. This is because Mo is an important component of nitrate reductase and dehydrogenase, which are involved in nitrogen assimilation and transport in higher plants (Kovacs et al., 2015). These enzymes are the key enzymes for plants to convert nitrate nitrogen into ammonium for the synthesis of amino acids and proteins. At the same time, Mo participates in the conversion of inorganic phosphorus to organic phosphorus in plants. When tobacco seedlings absorb NPs through roots or stomata, Mo will inevitably be released into the plant, contributing to the absorption of water and nutrient elements, enhancing photosynthesis, and finally promoting the biomass accumulation of tobacco seedlings (Li et al., 2018).

For most plant species, lignification has been regarded as a basal defense mechanism in the plant immune response that can function as a defensive physical/chemical barrier to external environmental and biotic stress, especially in preventing phytopathogen invasion and colonization (Adhikari et al., 2013; Dhoke et al., 2013; Zhang et al., 2019). Importantly, compared to the control, many more lignifying cells in roots treated with irrigation of Mo NPs than with foliar spraying were observed. Most likely, Mo NPs were identified as exogenous substances, thus activating the defense response in tobacco plants. This reflects the observation that tobacco seedlings seem to respond more sensitively to Mo NPs under root irrigation than spraying, which may be due to roots exhibiting higher NP or metal uptake and accumulation, as indicated in the above results ( Figure 6 ). Previous studies have suggested that plant roots are more responsive to ENMs as a result of the higher accumulation of NPs compared with leaves and stems, even under soil-grown conditions (Chen et al., 2022). Our results also showed that the application of Mo, especially irrigation exposure, promoted the differentiation of vascular tissue in stems and leaves and enhanced the leaf vein areas, which will be beneficial to plant growth. Leaf veins have the physiological function of transporting water and nutrients (Hu et al., 2005). Studies have shown that the development of the vascular bundle directly affects the transport of water, inorganic salt via xylem and photosynthates via phloem, and determines the yield of crops (Yu et al., 2010). These changes increase the flow of water and nutrient elements from roots to leaves after Mo NPs irrigation exposure, and further improve the transportation capacity of vascular tissue, significantly promoting the growth of tobacco seedlings.

Further study was conducted with the assumption that the promotional effects on tobacco in the presence of Mo NPs were ultimately associated with gene regulation at the molecular level. AQPs are integral membrane proteins that increase the permeability of membranes to water as well as to other small molecules such as CO₂ and glycerol, which play a key role in maintaining plant water balance and water use efficiency. We found that all PS- and AQP-related genes were upregulated in the tobacco plants exposed to Mo NPs by irrigation in comparison to the control plants. Commonly, the activity of AQPs can be impacted by many stress factors such as drought, heavy metals, salinity, pH, and anoxia (Tyerman et al., 1999). Under NPs stresses, some important stress-signaling pathways can be regulated in response to the uptake of NPs, which could induce physiological processes in plants (Sun et al., 2020). The tested AQP genes encode plasma membrane intrinsic proteins (PIPs). NtAQP1, a stress-induced gene, plays vital roles in controlling the CO₂ and water permeability of mesophyll tissues in tobacco plants, in addition to increasing the photosynthetic rate, stomatal opening, and leaf growth, especially under abiotic stress. NtPIP2;1 has also been reported to have high water transport activity. The increased transcription levels of NtPIP2;1 in tobacco tissues showed that the irrigation application of Mo NPs significantly promoted water uptake, and indicated the moderate increase in the water content in tobacco after foliar spraying with 100 μg/mL Mo NPs. A similar study found that the water-channel protein (LeAqp2) was upregulated in tomato root tissues exposed to MWCNTs (Khodakovskaya et al., 2011).

Extensive studies have been performed regarding the effects of different nanomaterials on crop growth under different application methods. Some findings are contrary to the results of this study. Adhikari et al. found that the length of roots was obviously inhibited after rice exposure to Mo NPs at concentrations greater than 5 mg/L (Adhikari et al., 2013). Interestingly, our findings demonstrated that exposure to 100 mg/L Mo NPs still promoted tobacco plant growth without any phytotoxicity in either case. For metal oxide NPs, compared to soil amendment, Fe₂O₃ NPs more significantly enhanced the biomass and photosynthetic rates of soybean when foliar sprayed, which was attributed to the increase in stomatal opening (Alidoust and Isoda, 2013). Some studies on plant exposure to ZnO NPs indicated that ZnO NPs suppressed wheat root growth and reduced the biomass under root irrigation in field soil and sandy soil, which mostly depended on the continuous uptake of many zinc ions released from NPs, thus inducing the production of oxidative stress and causing biological toxicity (Du et al., 2011; Dimkpa et al., 2012). However, the foliar spraying of ZnO NPs facilitated the growth and productivity of wheat, tomatoes, and mung beans (Dhoke et al., 2013; Raliya et al., 2015). These different results may be due to differences in experimental methods, NPs properties, plant species, and the nutrient absorption and metabolic ability of plants at different growth stages.

According to our previous investigation, the soil available Mo content in the Chongqing tobacco growing area was generally low (0.05-5 mg/kg), mainly due to nutrient loss caused by long-term soil acidification (He et al., 2005; Kaiser et al., 2005). Unlike other metal ions, Mo usually exists in soil as organic compounds or water-soluble molybdate ions (MoO^4-) and ammonium molybdate ions (NH₄Mo²⁺) and participates in the life activities of plants (Gupta and Lipsett, 1981). At present, the use of nanotechnology to improve the disease resistance and vegetative growth of flue-cured tobacco has achieved initial results that are of great significance for reducing fertilizer input and ecological and environmental protection (Chen et al., 2022). This study was carried out in potting substrates and demonstrated that Mo NPs can promote the growth of tobacco seedlings and the synthesis of chloroplasts and proteins, which is largely dependent on the uptake of Mo (Raven, 1988). Therefore, Mo NPs not only have good biocompatibility but can also be expected to be used as micronutrient fertilizers for plant nutrition. However, the biological effects of Mo NPs in soil and the migration behavior of Mo NPs in plants are still unclear and need to be further explored.

The aim of this study was to clarify the effects of Mo NPs on tobacco growth under two exposure modes. The results demonstrated that the biological effects of Mo NPs applied by irrigation were superior to those obtained in response to foliar exposure, which exerted significantly positive effects on tobacco seedling growth, resulting in a remarked physiological and biochemical changes. After soil application, more Mo accumulated in stems and roots compared to that observed under foliar spraying, especially 100 mg/L Mo NPs exposure. The increase in the defensive enzyme activity and upregulation of eight genes involved in the photosynthesis process and aquaporins indicated enhanced disease resistance. This pattern on the increased Mo content suggests that Mo NPs may be absorbed by tobacco plant, although more experiments are needed to confirm it. Therefore, the transport kinetics of nanoparticles and location in plant tissues still need to be further clarified. And what’s more, the necessity of the Mo NPs environmental risk assessment must be emphasized before wide use in agriculture. Firstly, it is necessary to evaluate the biological effects of Mo NPs based on a wider range of plant species. Secondly, more studies on the effects of Mo NPs on soil microbial communities are also required to consider the use of Mo NPs as potential nano nutritional supplements, especially whether they interfere with soil beneficial microbial composition and function.

The original contributions presented in the study are included in the article/ Supplementary Material . Further inquiries can be directed to the corresponding author.

JC, conceptualization, writing the original draft. YY, KS, and YZ, performed the experiments and statistically analyzed the data. WD, funding acquisition, English editing, revised preparation. All authors contributed to the article and approved the submitted version.