Nano pyrite root treatment in conjunction with soil application of goat droppings boost onion yield and anthocyanin and flavonoids content: A nano-organic farming model towards sustainability

doi:10.21203/rs.3.rs-1630965/v1

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Nano pyrite root treatment in conjunction with soil application of goat droppings boost onion yield and anthocyanin and flavonoids content: A nano-organic farming model towards sustainability

https://doi.org/10.21203/rs.3.rs-1630965/v1

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Agrochemical-intensive farming uses synthetic fertilizers, insecticides, pesticides, and weedicides. It holds the most prominent place in modern agriculture This reliance on synthetic agrochemicals has resulted in the declining health of the environment and living species. To restore the ecological balance, we need to explore strategies to minimize agrochemicals’ use without compromising agricultural production. Nano-formulations help lower dosages of agrochemicals, leading to the emergence of nano-agriculture. Yet the critical challenge is how we could exploit the power of nanomaterials to selectively improve the metabolic performance of crop plants. Here we have achieved the same by root treatment of the onion crop and then growing them in the presence of organic goat manure. In two field trials in consecutive years, we report an increase in yield through root treatment with nano-pyrite (FeS₂) plus soil application of goat dropping (Test) as compared to the use of goat dropping alone (Control). The total bio-mass (bulb + leaf) weight of the test sample was 4.75 kg (n = 86), while control samples weighed 3.5 kg (n = 86). The total bulb weight for the control and test was 2.6 kg and 3.6 kg, respectively. We observed a yield-boosting effect of root treatment with nano-pyrite in onion crops. There is a significant increase in the anthocyanin content in test bulbs and flavonol content in test leaves. We have integrated two types of farming systems: organic farming and nano-agriculture, resulting in a hybrid form; nano-organic farming to bolster the metabolic fitness of the onion (Allium cepa) to achieve sustainable food production.

Nano fertilizer

Sustainable agriculture

environment

nano-agriculture

Through evolution, humankind has developed various strategies for food production. When we as a race shifted from food gatherers to food producers, the first form of farming emerged as natural farming, relying on minimal interference to the ecosystem (Gopal Chandra De. 1989; Loukas et al. 2009; Douglas et al. 2018). Even today, many parts of the world follow natural agriculture. Wisdom-driven advancement of human skills led to the observation that organic matter in the soil helped improve crop productivity, resulting in the emergence of organic farming. One of the salient organic farming features is that essential plant nutrients are entrapped in a mass of carbon, helping sustain soil health and fertility (Rigby and Cáceres 2001; Andreas et al. 2012; Yadav et al. 2013; W. and P. 2015). Various studies conducted in the early 19th century by scientists, namely J B Boussingault, Justus Von Liebig, Jon Bennet Lawes, and JH Gilbert, explored soil, plant, and plant and organic matter which influence soil fertility. They discovered the major plant nutrients, specifically nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, and iron (Gopal Chandra De. 1989).

These discoveries marked the onset of agrochemical-intensive farming. One of the hallmark discoveries was the ammonia synthesis by the Haber-Bosch process at the beginning of the twentieth century (Delwiche 1970). There was a meteoric rise in agrochemicals in the agricultural production system following the green revolution, around the second half of the twentieth century (Gignoux et al. 2011; Pingali 2012; Kravchenko et al. 2017).

The intensive use of agrochemicals in agriculture during the last five decades has resulted in the degradation of soil health and water quality, eventually affecting various life forms of the ecosystem. Reduction in the use of chemical fertilizers is a significant challenge, with the global demand for fertilizer nutrients (N + P2O5 + K2O) being 198107 thousand tonnes in 2019 and forecasted to be 201663 thousand tonnes by 2020 (Nations 2017). There is a need to develop sustainable strategies to restore the ecological balance and maintain agricultural production. The current thrust is to reduce agrochemicals' consumption and increase their efficacy.

Several sustainable solutions, such as the use of bio-fertilizers (Rao 1982), organic manure (Gaur et al. 1984), nano-fertilizers (Tripathi et al. 2011; Sonkar et al. 2012; Tripathi and Sarkar 2015; Raliya et al. 2017; Bhati et al. 2018; Boutchuen et al. 2019), and nano-encapsulations, are currently being explored to reduce the use of chemical fertilizers and prevent disease infestation (Ahuja et al. 2019, 2020; Shukla et al. 2019; Gahukar and Das 2020; El-Moneim et al. 2021). All the strategies of fertilizer reduction target achieving healthy soil dynamics. The area of nano-agriculture holds the potential to achieve sustainable growth through smart farming.

On an optimistic note, if all these solutions mentioned above successfully improve soil health, ironically, the benefit will be enjoyed by all the existing flora and the crop. Thus it is imperative that unless the crop plant has higher metabolic fitness to compete with the surroundings, eventually, it will fall short in the competition. Thus the challenge is could we develop a nano-strategy to bolster the metabolic fitness of the crop plant so that it can compete better with the biotic and abiotic stress of the environment. In the present work, we have exploited the role of nano iron pyrite in offering better metabolic fitness to the root system to offer an improved yield. As a case study, we have experimented on onion (Allium cepa) since it has a shallow root system and needs a supply of nutrients close to the soil surface for healthy growth. Goat manure provides the nutrient at the soil surface. So here, we have applied an iron-sulfur nanomaterial as a root treatment agent to boost the root metabolism and used organic manure to supply nutrients- thus following a nano-organic farming approach.

Philosophically, this work draws inspiration from hydrothermal vents' extreme ecosystem, where anaerobic microbes receive their energy from nano pyrite and other metal sulfides. Earlier Srivastava et al. discovered that a 12 h seed treatment of the chickpea in an aqueous nano pyrite suspension resulted in increased biomass (Srivastava et al. 2014a). In the follow-up field trials on chickpea (Jangir et al. 2020b), spinach (Srivastava et al. 2014b), beetroot, carrot, fenugreek, alfalfa, mustard, and sesamum (Das et al. 2016a), we observed higher productivity following pyrite seed treatment. In pyrite-treated rice, an NPK equivalent yield was reported (Das et al. 2018). Jangir et al. recently showed an improved result in wheat crops following nano pyrite seed treatment (Jangir et al. 2020a). The nano pyrite seed treatment gets nullified by co-treatment with an antioxidant nanomaterial, viz., cerium oxide (Das et al. 2016b). Recently Jangir et al. discovered that upon treating the roots of chili, marigold, cabbage, cauliflower, and tomato in an aqueous suspension of nano pyrite for three hours, a significant increase in yield was observed (Jangir et al. 2019b, a; Jangir 2020; Jangir and Das 2021). Further seed and root treatment with nano pyrite improve the plants' root foraging ability (Jangir et al. 2019c, a, 2020b; Jangir 2020; Jangir et al. 2020a; Jangir and Das 2021).

The current study addresses the synergistic effect of nano pyrite root treatment and organic manure (goat droppings) on onion production (Allium cepa). It has tremendous commercial significance. It is a temperate crop, and India is the highest contributor to onion production after China. It has multiple uses- as a spice, salad, and vegetable. The allyl propyl disulfide contributes to onion pungency. The soil has to be rich in organic matter for optimal onion production. Recommended fertilizer dosage is 125:60:100 kg/ha of Nitrogen (N): Phosphorus (P): and Potassium (K) (Gupta 2019; Choudhury 2020).

2.1. Nano pyrite synthesis: Nano pyrite synthesis is a two-step reaction. In the first step, we synthesize fresh polysulfide. We take an aqueous sodium hydroxide solution (0.8 M, 100 ml) divided into two equal parts. In one part, we purged hydrogen sulfide gas until it turned yellow-green, then the remaining half of sodium hydroxide (0.8 M, 100 ml) and 0.04 moles of sulfur were added. The solution was stirred in the presence of hydrogen sulfide for the next two h at room temperature to obtain yellow-orange polysulfide suspension. The suspension was filtered and stored at 4˚C. For the second step, fresh aqueous ferric chloride (0.04 M, 100 ml) was prepared in a round bottom flask and maintained at a pH of 5.6 using a buffer of 91 ml sodium acetate (0.2 M, 100 ml) and 9 ml acetic acid (0.2 M, 100 ml). The flask was placed in an oil bath with continuous argon purging and stirred continuously for one h at room temperature. Then to which, we slowly add 15 ml of polysulfide. The oil bath temperature remains at 120˚C for two h, and a dark grey suspension will appear. Then, the oil bath temperature was switched to 180˚C for the next 10 minutes to obtain a grey-colored suspension. The nano pyrite suspension settles at the bottom of the flask. We wash the suspension with hydrochloric acid, toluene, and acetone. We store the nano pyrite particles in a moisture-free chamber (Srivastava et al. 2014a, b; Das et al. 2016a, b, 2018; Jangir et al. 2019c, a, 2020b; Jangir 2020; Jangir et al. 2020a; Jangir and Das 2021).

2.2. Nano pyrite characterization: Structural features of nano pyrite were observed using X-ray diffraction (XRD) and Scanning electron microscopy (SEM). XRD pattern was obtained to study the characteristic planes of nano pyrite crystal, for which a powdered sample was analyzed using X' Pert powder-PANalytical XRD. Sample for SEM and EDS was prepared by drop-casting a suspension of nano pyrite in methanol on a copper stub. The sample was gold coated and observed using FEI, Quanta 200 SEM. Fourier Transform Infrared Spectroscopy (FTIR) was performed to investigate the bonding characteristics of the nano pyrite particles. The sample was prepared by making a pellet of nano pyrite powdered sample with KBr. Perkin-Elmer FTIR spectrum BX spectrometer was employed to obtain the FTIR spectrum. X-ray Photoelectron Spectroscopy (XPS) was performed using PHI 5000 Versa Prob II, FEI for powdered samples. XPS was used to study the surface characteristics of nano pyrite particles (Srivastava et al. 2014a, b; Das et al. 2016a, b, 2018; Jangir et al. 2019c, a, 2020b; Jangir 2020; Jangir et al. 2020a).

2.3. Field trial: Land preparation: We conducted the field trial in the institute nursery of the Indian Institute of Technology Kanpur, India. We selected a nutritionally deficient patch of land adjacent to the nursery’s greenhouse. We first plowed the ground and prepared three plots. Out of the three plots, we maintained the exact dimensions (3*4 ft²) for two plots, while for the third plot, we doubled the length and breadth of the plot (6*8 ft²). We use the third plot (6*8 ft²) as a nursery to grow the onion seedlings before we perform transplantation. For both seasons, the same protocol was used. In figure 1, we have presented a flow chart of the cropping protocol followed during the two years of field trials.

The nutrient profile of the soil: At the beginning of the field trial, we perform a soil nutrient analysis for Nitrogen (N), Phosphorus (P), and Potassium (K). N was estimated using the Kjeldahl method, and P and K were calculated using ICPMS analysis as described earlier (Jangir et al. 2019b, a).

Sowing, root treatment, transplantation, and manure application: For the season I, the field trial was initiated on November 10, 2018. 50 mg onion seeds were germinated in nursery beds (6*8 ft²). After thirty-three days (December 13, 2018), the germinated seedlings were harvested and were randomly divided into two sets of 100 seedlings each. Next, the root treatment was performed for three h before transplantation. In the control group, the roots of seedlings were soaked in water, while in the test group, the roots were soaked in 100 µg/ml aqueous suspension of nano pyrite. After three h of root treatment, the control and test seedlings were transplanted in their respective assigned plots of individual dimensions of 3*4 ft². Goat droppings were used for manuring both plots (1 kg/plot) on January 19, 2019. The second (season II) cropping season follows a similar calendar.

Crop care: Recommended irrigations were given, and all other parameters were kept the same for both sets. All the other agronomic practices were followed as per the recommendations (Gupta 2019; Choudhury 2020).

Harvest: Harvesting was done manually, and the mud was removed and cleaned before weighing the samples.

2.4. Crop Analysis and growth Parameter: The crop was harvested and analyzed for plant growth parameters such as fresh weight, dry weight, leaf area, specific leaf area, leaf area index, size/area of bulb, and onion bulb weight, etc. The relative anthocyanin and flavonol content was also measured for control and test samples. The anthocyanin content of the bulb is an indicator of the coloration of the bulb and thus marketability.

The onion crop yield obtained at harvest was weighed for both control and test groups. Fresh weight was measured for both the whole plant (leaves and bulb) and bulb. The average yield obtained is reported in grams for bulbs and leaves. The weight of leaves was calculated by subtracting the weight of the bulbs from the weight of the whole plant. Total onion bulbs obtained from control and test groups were counted. Twelve and thirteen representative plants were chosen from the control and test groups, respectively. These plants were placed alongside a ruler, and a snapshot was taken. This picture was used to calculate the area of leaves and bulbs using ImageJ. Leaf area is a direct measurement of photosynthetic activity and transpiration. Standard errors have been reported for image analysis. The Student's t-test was performed to calculate the significance of the results. Leaves and bulbs were kept at 70 degrees Celsius for 6 hours in order to measure the dry weight of the samples. Once we had primary data, other plant growth parameters were calculated as per standard procedure using the following (Sudhakar et al. 2016):

Leaf Area Ratio: It is a measure of the relative size of the assimilatory apparatus. In simpler terms gives an idea of the portion of the plant involved in the photosynthetic activity.

Leaf Area Ratio = Total Leaf Area/Total Dry weight

Specific Leaf Area (SLA): SLA signifies the leaf density of the plant. It tells us about the amount of leaf area used to produce unit biomass.

Specific Leaf Area = Total Leaf Area/Leaf Dry Mass

Leaf Area Index (LAI): The leaf area index (LAI) of a plant is defined as its leaf area per unit of ground area. The leaf area index (LAI) is one of the most important parameters in plant ecology. It signifies how much foliage (leaf) is present on the plant. It measures the active photosynthetic area, as well as the area available for transpiration. More photosynthetic area signifies that the rate of photosynthesis and food production and accumulation is more. In comparison, higher transpiration rates ensure higher uptake of water and minerals. These two factors ensure better nutrition accumulation in edible parts of the plant.

Leaf Area Index = Total Leaf Area/Total Land Area.

The harvested plant materials were further tested for their anthocyanin and flavonol content in both leaves and bulbs. Flavonol is a class of flavonoids that participate in stress responses. These are the most primitive and widespread flavonoids and have a wide range of plant physiological functions. Anthocyanin is a specific class of plant pigments that belong to flavonoids. These pigments provide bright color to the onion bulb and thus increase the marketability and visual acceptability of the crop produce. The higher content of anthocyanin in onions bulbs also signifies earlier maturity, thus helping the farmers get a good price for their early produce. Anthocyanin is also a well-known antioxidant; thus a higher anthocyanin content ensures better health (Martín 2017).

Preparation of plant samples: Plant samples were finely cut into small pieces and homogenized using a pestle and mortar. 2 gm of this homogenized sample was mixed with 7ml of 80% ethanol (0.1% HCl) and further homogenized. The extract was filtered to obtain an alcoholic extract.

Flavonol measurement: Relative flavonol content was measured for control and test samples (leaves and bulb) by taking absorbance at 510nm.

Anthocyanin measurement: Relative anthocyanin content was measured using the ethanol extract and mixing in equal quantity with buffer (pH 4.5) and taking absorbance at 525nm.

3.1. Nano pyrite synthesis: Iron pyrite nanoparticles were synthesized as described in our previous work via a low-temperature nucleophilic reaction (Srivastava et al. 2014a, b; Das et al. 2016a, b, 2018; Jangir et al. 2019c, a, 2020b, a; Jangir 2020). The reaction involves iron (III) chloride and sodium polysulfide, the chief intermediaries being FeS and FeSH+. These intermediates are further attacked by polysulfide nucleophiles, which leads to the formation of FeS₂. The reaction was carried out in an inert environment to prevent interaction with oxygen.

3.2. Nano pyrite characterization: In figure 2, the nano FeS₂ characterization has been shown. XRD peaks at 2θ= 28.8, 33.1, 37.2, 40.7, 47.5, 56.3, 61.5, 64.2, 79.0 correspond to characteristic crystal planes of iron pyrite with indices (111), (200), (210), (211), (220), (311), (302), (321), (420), respectively (Figure 2a). Nanosized iron pyrite flakes can be seen in the SEM image. These flakes assemble together to form large pitcher-shaped structures ranging from 70nm to more than 500nm. (Figure 2b).

EDS was also performed along with the SEM analysis. Figures 3a and 3b show the EDS analysis. Figure 3a shows the EDS overlay with the SEM image, and figure 3b shows the EDS spectrum for nano iron pyrite.

FTIR peaks at 400-800 cm-1 marked pyrite bonding characteristics, the peak at 1076.5 cm-1 showed S-O bonding characteristics marking surface oxidation, and the presence of moisture was marked by peaks at 1623.8 cm-1 and 3407.6 cm-1 (Figure 4).

Fe2p narrow XPS spectrum marked the presence of Fe2p_3/2 and Fe2p_1/2 spin-orbital coupling at 704.8 eV and 717.8 eV, respectively, along with signatures of stoichiometric defects and FeSO₄ at 705.8 eV and 708.2 eV, respectively (Figure 5a). In the S2p narrow XPS spectrum, peaks at 160.2 eV and 161.4 eV depicted S2p_3/2 and S2p_1/2 spin-orbit coupling, respectively. XPS peaks at 162.0 eV and 166.7 eV marked the presence of surface defects and oxidation causing the formation of FeSO₄, respectively (Figure 5b). In our earlier work, we found similar results (Srivastava et al. 2014a, b; Das et al. 2016a, b, 2018; Jangir et al. 2019c, a, 2020b, a; Jangir 2020; Jangir and Das 2021).

3.3. Field trial: Nutrient profile of the soil: In the present study, a nutritionally deficient patch of land (total nitrogen (159 kg/ha), total phosphorus (0.3 kg/ha), total potassium (0.4 kg/ha)) was selected in the Institute Nursery of Indian Institute of Technology Kanpur, India. In earlier work, it has been shown that the above-mentioned nitrogen, phosphorous, and potassium level is considered nutritionally deficient (Jangir et al. 2019b, a, 2020a).

3.4. Crop Analysis (2018-2019): In figure 6a, representative pictures of the control (n=12) and the test (n=13) bulbs obtained at the time of harvest are shown (2018-2019 season). Visual observations show comparatively denser foliage in test plants as compared to the control. In figure 6b, the total bulb harvested from the control (n=83) and the test (n=86) crop is shown. Initially, a total of 100 seedlings were sown for both the control and test group.

A detailed crop analysis is shown in figure 7 a-f. The test samples resulted in higher productivity in terms of fresh weight in comparison to untreated plants. The average fresh weight of the whole plant (leaves and bulb) for control samples was found to be 42.16 grams while the test group had an average fresh weight of 55.23 grams (Figure 7a). In order to investigate the distribution of the weight between foliage and the economic yield, the leaves and the bulbs were weighed separately. The average bulb weight for test samples was significantly higher (41.86 gm) as compared to control (31.32 gm) (Figure 7e). Similarly, the average fresh weight of test leaves (13.37 gm) was higher in comparison to control leaves (10.84 gm) (Figure 7c).

Average water accumulation in both control and test plants was approximately equal. Since the fresh weight of the test samples was high, the average dry weight of the test plants (8.13 gm) was also more as compared to the control (6.158) untreated plants (Figure 7b). An approximately equal amount of moister content was found in both test and control sample leaves and bulbs. The average dry weight of control leaves was found to be 0.889 gm while the test leaves had an average dry weight of 1.096 gm (Figure 7d). Similar to the leaves the bulbs also accumulated the approximately same amount of water for both control and test samples. Therefore, the average dry weight of test bulbs (7.04 gm) was higher as compared to the control onion bulb samples (5.26 gm) (Figure 7f).

The average bulb area for test samples (2.33 cm²) was significantly higher as compared to control samples (1.59 cm²) (Figure 8a). Significantly denser leaf foliage was observed in test samples with an average leaf area of (11.53 cm²) as compared to control (6.99 cm²) (Figure 8b). The increase in leaf area resulted in significantly higher specific leaf area (10.51) (Figure 8c), leaf area index (0.08) (Figure 8d), and leaf area ratio (Figure 8e) in test samples as compared to control (7.86, 0.05, and respectively).

The relative anthocyanin content in control leaves (0.052) was considerably higher as compared to test leaves (0.03) (Figure 9a). However, there is a highly significant increase in the anthocyanin content in test bulbs (0.069) as compared to control bulbs (0.02) (Figure 9b). The results indicate that there is a selective partitioning of the pigment in leaves and bulbs. The test plants tend to accumulate significantly higher amounts of pigment in the food storage/edible parts. The relative flavonol content in test leaves (0.253) (Figure 9c) was found significantly higher with respect to control samples (0.086). On the other hand, the flavonol content in the bulb (Figure 9d) was approximately equal in both control and test samples.

As mentioned earlier that philosophically, this work is inspired by an extreme ecosystem viz., hydrothermal vents in the deep ocean rifts of the pacific, where microbes like archeobacteria and thiobacillus receive their energy from nano pyrite and other metal sulfides. The fundamental hypothesis for the use of nano pyrite pivots around an exothermic process, whereby metal sulfides are converted to sulfates to liberate 200 kcal/mol of energy (Tributsch 1984). This kinetics is further accelerated when metal sulfides are covered with a thin film of water approximately 4% of their weight in the presence of oxygen (Tributsch 1984). Subsequent reduction of oxygen leads to the release of trace amounts of H₂O₂, which further attacks additional sulfide molecules resulting in the generation of Fe²⁺, Fe³⁺, SO₄^2-, and S⁰. So, when plant roots are treated in aqueous suspension of pyrite, it is briefly exposed to iron, sulfur, pyrite, peroxide, and proton-rich environment. These species orchestrate a cascade of metabolic changes that enhance plant metabolism (Jangir et al. 2019b, a; Jangir 2020; Jangir and Das 2021). In other words, this treatment results in metabolically smarter plants. Here, we tested the feasibility of this root treatment in plants with shallow root system to draw nutrients efficiently from organic matter. The overall premise of the work is summarized in figure 10.

The experiment was purposefully conducted in a nutritionally deficient patch of land to study the dual effect of root treatment and organic manure, thus, closely simulating a sustainable organic farm environment with a nano intervention. We chose goat dropping as an organic supplement as it has high contents of organic matter, nitrogen, and phosphorus, and indirectly impacts fertilizer mobilization and biological cycling. Also, the total nitrogen mineralization over one-twenty days has been found maximum for goat droppings as compared to that of poultry and cattle (Gichangi et al. 2009, 2010; Azeez and Van Averbeke 2010; Rizk et al. 2012). Further, goat manure is commonly used as a source of liquid fertilizer (Sunaryo et al. 2018).

Earlier, it has been observed that organic manure solely cannot suffice for maximum onion productivity. This could be due to the shallow root system of onion crops with lesser root penetrability. Thereby, onion crop requires nutrient application close of the root system, which was the principal reason that we applied goat manure on the soil surface (Gichangi et al. 2009, 2010; Azeez and Van Averbeke 2010; Rizk et al. 2012). Here, by pre-treating onion seedlings with nano pyrite before manure application, we demonstrated that boosting the metabolism of onion seedlings in the test group lead to better nutrient acquisition and higher yield as compared to the control group. Higher leaf area, specific leaf area, leaf area ratio, and leaf area index indicate a higher photosynthetic rate and higher transpiration. All these factors may have contributed to an increase in the size of onion bulbs of test plants. Bigger bulbs ensure better market and price for the farmer.

An increase in anthocyanin content in bulbs indicates that the economic yield is brighter colored and has matured earlier. It also indicates that the test yield shall have better marketability and may sell at a higher price. Higher flavonol content in test leaves ensures better stress response, particularly to UV-B radiation (van de Staaij et al. 2002). Flavonols play very important role in pollen germination and thus plant fertility (Mahajan et al. 2011). The results indicate that the test plants have significantly higher flavonols and thus are less prone to UV stress, also; the plants may have higher fertility and therefore higher seed production capability.

In summary, on treatment with an aqueous suspension of nano pyrite, the root experiences a brief energy-rich microenvironment, which shapes its subsequent metabolism and growth. These metabolically boosted plants acquire nutrients efficiently from organically supplemented soil and hence have higher competitive fitness. This approach has the feasibility to open up avenues for nano-organic sustainable farming and further boosting the economy of goat farmers of Asian and African continents.

While sustainable farming aims at reducing chemical fertilizer by using organic manure, bio-fertilizers, nano fertilizers; a crop-specific increase in metabolic fitness could positively influence the yield. Here we show that a nano pyrite root treatment during transplantation could boost the metabolic fitness of the plant so that it can capitalize on the maximum benefits derived from organic farming; thus leading to a novel nano-organic farming strategy.

Data availability

All the data are provided in the manuscript and the related references are cited.

Conflict of interest

The authors declare no conflict of interest

Funding statement

HJ is funded by MHRD, GOI. Part of the funding for the project is obtained from the “Scheme for Transformational and Advanced Research in Sciences (STARS)” Ministry of Education, India; project of MD.

Acknowledgments

HJ is funded by MHRD GOI and the work is an integral part of HJ's doctoral thesis. The authors are thankful to Mr. D D Pal for XPS, PGRL, IITK for SEM, Mr. A Tiwari for XRD facility, Ms. V Singh for FTIR at Advance Centre for Material Science, IIT Kanpur; Prof. A Singh for providing the ICPMS facility; Mr. Satish Singh for providing Institute Nursery facility. Part of the funding for the project is obtained from the “Scheme for Transformational and Advanced Research in Sciences (STARS)” Ministry of Education, India; project of MD.

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No competing interests reported.

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Nano pyrite root treatment in conjunction with soil application of goat droppings boost onion yield and anthocyanin and flavonoids content: A nano-organic farming model towards sustainability

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