Reuse of agricultural waste as an alternative substrate in the production of eggplant (Solanum melongena L.) seedlings Received for publication: 19 August, 2019. Accepted for publication: 13 April, 2020 Doi: 10.15446/agron.colomb.v38n1.79497 1 Department of Plant Science, Federal University of Viçosa, Viçosa (Brazil). 2 Department of Plant Science, Federal Institute of Education, Science and Technology of Espírito Santo, Santa Teresa campus, Santa Teresa (Brazil). 3 Department of Plant Science, Darcy Ribeiro North Fluminense State University, Campos dos Goytacazes (Brazil). * Corresponding author: carol.merlo@hotmail.com Agronomía Colombiana 38(1), 29-35, 2020 ABSTRACT RESUMEN Seedling formation is one of the most important phases for the eggplant crop cycle. One of the decisive factors for obtaining quality seedlings and the consequent increase in productivity is the type of substrate used. The objective of this research was to evaluate the growth of eggplant seedlings grown in alternative substrates with increasing levels of “moinha” (residue from the coffee dry milling process) replacing the commercial substrate. The experiment was carried out in a completely randomized design, with six treatments and ten replicates. The treatments were: T1: commercial substrate (control); T2: 0% moinha (MO) + 40% burnt rice husk (BRH) + 15% coconut fiber (CF) + 5% eggshell (ES) + 40% commercial substrate (CS); T3: 10% MO + 40% BRH + 15% CF + 5% ES+ 30% CS; T4: 20% MO + 40% BRH + 15% CF+ 5% ES + 20% CS ; T5: 30% MO + 40% BRH + 15% CF + 5% ES + 10% CS; T6: 40% MO + 40% BRH + 15% CF + 5% ES + 0% CS. The electrical conductivity of the sub- strates and the seedling total dry mass, plant height and stem diameter were evaluated. It is recommended to use the substrate containing 20% MO + 40% BRH + 15% CF + 5% ES + 20% CS. La formación de plántulas es una de las fases más importantes para el ciclo de cultivo de la berenjena. El tipo de sustrato uti- lizado es uno de los factores decisivos para obtener plántulas de calidad y el consiguiente aumento de la productividad. El objetivo de este estudio fue evaluar el crecimiento de las plántu- las de berenjena cultivadas en sustratos alternativos con niveles crecientes de “moinha” (residuo del proceso de molienda en seco de café), en sustitución del sustrato comercial. El experi- mento se realizó en un diseño completamente al azar, con seis tratamientos y diez repeticiones. Los tratamientos fueron: T1: sustrato comercial (control); T2: 0% de moinha (MO) + 40% de cáscara de arroz quemada (CAQ) + 15% de fibra de coco (FC) + 5% de cáscara de huevo (CH) + 40% de sustrato comercial (SC); T3: 10% MO + 40% CAQ + 15% FC + 5% CH + 30% SC; T4: 20% MO + 40% CAQ + 15% FC + 5% CH + 20% SC; T5: 30% MO + 40% CAQ + 15% FC + 5% CH + 10% SC; T6: 40% MO + 40% CAQ + 15% FC + 5% CH + 0% SC. Se evaluó la conductividad eléctrica de los sustratos y la masa seca total de las plántulas, la altura de la planta y el diámetro del tallo. Se recomienda utilizar el sustrato que contiene 20% de MO + 40% de CAQ + 15% de FC + 5% de CH + 20% de SC. Key words: growth, moinha, Solanum melongena, seedling quality, horticulture. Palabras clave: crecimiento, moinha, Solanum melongena, plántulas de calidad, horticultura. Reuse of agricultural waste as an alternative substrate in the production of eggplant (Solanum melongena L.) seedlings Reutilización de residuos agrícolas como sustrato alternativo en la producción de plántulas de berenjena (Solanum melongena L.) Karoline Matiello Almeida1, Paola Alfonsa Vieira Lo Monaco2, Marcelo Rodrigo Krause1, Dandara Lyone Silva de Oliveira3, Lorena Aparecida Merlo Meneghelli1, Caroline Merlo Meneghelli1*, and Louise Pinto Guisolfi1 Introduction In the production of quality vegetables, seedling forma- tion is one of the most important phases for the crop cycle (Zaccheo et al., 2013), and the type of substrate used for growing them is one of the crucial factors for obtaining high-quality seedlings and, therefore, high-yield plants. The substrate should provide an adequate supply of air and water to the root system, besides physically support- ing plant growth (Júnior et al., 2014). In addition, growth media or substrates must be free of plant pathogens, easy to handle, long-lasting, highly available and of low cost (Fernandes et al., 2006; Mesquita et al., 2012). Several studies have been developed to reuse agricultural and agro-industrial residues in the composition of sub- strates to minimize the environmental impact that would be caused by inadequate disposal, and reduce production costs by replacing commercial substrates (Krause et al., 2017; Almeida et al., 2018; Guisolfi et al., 2018; Meneghelli et al., 2018). The reuse of residues to compose substrates http:// 30 Agron. Colomb. 38(1) 2020 in eggplant seedling production has been investigated by several authors (Costa et al., 2012; Bardiviesso et al., 2014; Ferreira et al., 2014). Agricultural wastes, such as coir (or coconut fiber), burnt rice husk, eggshell, and the dry residue of coffee beans, also known as “moinha”, are highly available and potential alternative materials to compose substrates for the produc- tion of eggplant seedlings. Coconut coir dust has a predominance of medium and fine particle size fractions, which makes it an adequate substrate when looking for high porosity and presence of micropores. These are important factors to obtain good aeration and water-holding capacity (Zorzeto et al., 2014). Coconut fiber is a renewable and highly biodegradable substrate with improved mechanical properties and low cost (Machado et al., 2014). Therefore, its high availability, low cost, and physical-chemical properties make coconut fiber an adequate alternative material for the composition of substrates.  Burnt rice husk has a high water-holding capacity and fast and efficient drainage. These characteristics provide good oxygenation to the roots, well-aerated conditions, resistance to decomposition, relative structure stability, low density, and pH close to neutrality (Soares et al., 2012). Due to the aforementioned properties, rice husk has become intensively used as a substrate for plant growth. Eggshells are composed of organic and inorganic substanc- es. Calcium carbonate (CaCO3) is the main component in the inorganic phase of hen eggshells (Medeiros and Alves, 2014) and they can be used as one of the nutritional materi- als for the formulation of a substrate. Moinha is a residue generated during the processing of coffee beans. It is composed of plant residues, such as leaves, branches, inf lorescences, and poorly formed coffee beans, which, when dried together with the coffee beans, are burned and released from the dryer. When chemically analyzed, this material was found to contain a high content of organic matter as well as phosphorus, potassium, and nitrogen (Meneghelli et al., 2016). In addition to its poten- tial use as a fertilizer, the authors found a high electrical conductivity in this residue, which is why they verified that concentrations greater than 10% of moinha in the substrate provided lower values in the analyzed variables in Conilon coffee seedlings. In this sense, different concentrations of moinha in the composition of substrates for different crops should be investigated, considering the hypothesis that this residue is an important substrate material, but in an adequate proportion that requires experimentation. It is believed that partial or even total replacement of commercial substrates by the combination of different agricultural residues may be a sustainable alternative for the production of eggplant seedlings since all of them have appropriate characteristics to seedling germination and development. In addition, it reduces costs when compared to commercial growing media (Mesquita et al., 2012; Fer- reira et al., 2014). However, it is necessary to know the exact proportions of moinha and commercial substrate in the composition of the alternative substrate, which will provide high-quality eggplant seedlings. The objective of this research was to evaluate the growth of eggplant seedlings grown in alternative substrates with increasing levels of moinha, replacing the commercial substrate. Material and methods The experiment was carried out at the plant nursery of the Federal Institute of Espirito Santo, Campus Santa Teresa, located in the municipality of Santa Teresa, Espirito Santo State (18°48’ S, 40°40’ W, 130 m a.s.l.), in Brazil. According to Köppen and Geiger (1928), the climate of the region, is Cwa (subtropical dry-winter climate), with an average an- nual temperature of 24.6°C and average annual precipita- tion ranging from 700 to 1200 mm. The temperature and relative humidity of the air during the experimental period ranged from 19.9 to 38.2°C and 47.5 to 69.5%, respectively. The plant nursery was covered with a shading screen, which provided a reduction of solar radiation by 50%. The residues used in the alternative substrate composition for the production of eggplant seedlings were: residues from the coffee dry milling process (moinha), eggshells, dry coconut fiber (coir), and burnt rice husk. Eggshells were collected from local restaurants and dried in the sun for 3 d. Subsequently, they were crushed and ground into a fine powder. Burnt rice husk and coir were donated by the company Fibria, located in Aracruz-ES, Brazil. Moinha was collected from a private property (Sítio da Saudade), located near the experimental area in Santa Teresa, Espirito Santo, Brazil. Before being used, moinha was sieved in a stainless-steel sieve with a 4 mm mesh. The chemical and physical-chemical characterization of the residues used in the composition of the substrates was carried out at the Soil and Solid Waste Laboratory of the 31Matiello Almeida, Vieira Lo Monaco, Krause, Silva de Oliveira, Merlo Meneghelli, Merlo Meneghelli, and Pinto Guisolfi: Reuse of agricultural waste as an alternative substrate in the production of eggplant (Solanum melongena L.) seedlings Agricultural Engineering Department of the Federal Uni- versity of Viçosa. The physical-chemical analysis consisted of the determination of the electrical conductivity (EC), through a laboratory conductivity meter (model P613, Shanghai Yoke Instrument Co. Ltd., Shangai, China). The chemical analysis consisted in the determination of the pH, using a bench pH meter (model HMCDB-150, Highmed Solutions in Measurement Technology Ltd- ME Tatuape, São Paulo, Brazil) and the quantification of organic matter (OM), total nitrogen (TN), phosphorus (P) and potassium (K), according to the methodology described by de Matos (2015). The chemical and physical-chemical attributes (EC) of coconut fiber, moinha, eggshells and burnt rice husk used in the experiment are shown in Table 1. TABLE 1. Chemical and physical chemical attributes (EC) of eggshells (ES), coconut fiber (CF), moinha (MO) and burnt rice husk (BRH) used in the experiment. Residues pH(a) EC OCro OCT OM NT P K Ca dS m-1 g kg-1 ES 9.37 0.37 2.8 3.6 6.2 0.87 0.08 0.06 31.9 CF 7.15 0.09 57.1 74.1 127.7 0.66 0.05 0.14 0.0 MO 5.60 6.49 45.3 58.9 101.5 3.7 0.14 0.71 0.0 BRH 5.9 1.15 31.5 40.9 70.5 0.59 0.08 0.03 0.0 pH(a): potential of hydrogen in water; EC: electrical conductivity; OCro: readily oxidizable organic carbon; OCT: total organic carbon; OM: organic matter; NT: total nitrogen; P: phosphorus; K: potassium; Ca: calcium. A completely randomized experimental design (CRD) was used, with six treatments and ten replicates. Each experi- mental unit consisted of 20 seedlings, totaling 1,200 seed- lings used in the experiment. Six plants were considered useful for each experimental unit. The treatments: T1: commercial substrate (control); T2: 0% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshells + 40% commercial substrate; T3: 10% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshell + 30% commercial substrate; T4: 20% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshell + 20% com- mercial substrate; T5: 30% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshell + 10% commercial substrate; T6: 40% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshell + 0% commercial substrate were evaluated. Chemical attributes of the commercial Bioplant® substrate, obtained by Paixão et al. (2012) are shown in Table 2. TABLE 2. Chemical analysis of the commercial substrate Bioplant®. pH(a) NT P K Ca OCT OM g kg-1 5.62 0.62 1.55 0.44 1.84 21.00 52.21 pH(a) in CaCl2(CaCl2 pH); NT: total nitrogen; P: phosphorus; K: potassium; Ca: calcium; OCT: total organic carbon; OM: organic matter. Seeds of eggplant (Solanum melongena L.) cultivar Embu were sown in a 200-cell Styrofoam propagation tray, plac- ing two seeds per cell. The seedling production system was arranged in suspended trays, placed in masonry raised beds. Seedlings were manually irrigated twice a day, once in the morning and once in the afternoon without fertil- izer application. Fifteen days after sowing (DAS) thinning was performed leaving only one seedling per cell, the most vigorous one. The variables plant height, stem diameter and total dry mass were evaluated at 31 DAS. Plant height was obtained using a millimeter ruler, measuring from the stem base to the apical bud that originated the last leaf. The stem diameter was measured using a precision digital caliper (Zaas Precision). To obtain the total dry mass, the roots were carefully washed in running water, over a sieve. After- ward, the seedlings were packed in paper bags and placed in a forced air circulation drying oven (model WGL-30B, Westtune, Suzhou Beiyin medical equipment Co., Ltd., Zhejiang, China). The seedlings were maintained at 65°C until a constant weight was achieved. Subsequently, the materials were weighed in an electronic precision scale (1 g) (reference LS5, Scientific Mars, São Paulo, Brazil). In addition to the biometric and gravimetric analysis, the electrical conductivity of the substrates was determined using a conductivity meter (model P613, Shanghai Yoke Instrument Co., Ltd, Shangai, China) following the meth- odology proposed by de Matos (2015). All the evaluated variables were submitted to the tests of normality (Lilliefors) and homoscedasticity (Bartlett), to validate and assess their variance. Due to the qualitative difference between the commercial substrate treatment (T1) and the other substrates (T2 to T6), the analysis of variance for qualitative treatments was performed only considering the contrast between T1 and the other treat- ments in the decomposition of the sums of squares of the treatments. In addition, for the comparisons between treat- ments T2, T3, T4, T5, and T6, related to the moinha level, their adjustments were adopted in regression models using the orthogonal polynomial method. For all procedures, an “α” equal to 0.05 was adopted. All the statistical analyzes were performed through the SAEG 9.1 software (2007). 32 Agron. Colomb. 38(1) 2020 Results and discussion For the variables stem diameter (SD), plant height (PH), and total dry mass (TDM), superiority (P<0.05) was ob- served for the group of treatments containing 0, 10, 20, 30 and 40% of moinha, in comparison to the commercial substrate treatment (Tab. 3). This result may be associ- ated with the greater amount of nutrients contained in this residue, especially nitrogen. Nitrogen can favor leaf expansion and provide greater plant growth (Eichler et al., 2008). Increases in shoot dry mass are expected due to the application of nitrogen doses, mainly because this nutrient contributes to vegetative growth, leaf expansion, and stem growth rate (Aleman and Chaves, 2016) Thus, the higher content of nitrogen in the treatment group containing moinha may have provided greater vegetative growth in the seedlings. The use of eggshells and coconut fiber may have positively contributed to seedling development, even because no moinha was added (0% moinha) in T1. The eggshell min- eralized in a short period of time could have contributed to the availability of nutrients. Studying the use of eggshell as a soil acidity corrective, Lo Monaco et al. (2015) concluded that in a period of 2 weeks, the residue added to the soil in the form of powder was mineralized and favored the increase of pH in the soil. Likewise, Niezer and Silveira (2014), using eggshell to recompose the pH of the soil, found a decrease in acidity after 3 weeks of incubation of the soil with the residue. Although it has the lowest organic matter content compared to the other residues used, eggshells provide nutrients, especially calcium (Tab. 1), in greater amounts than the commercial substrate (Paixão et al., 2012). Calcium is extremely important in the primary growth phase of plants, since it is involved in the construction of cell walls, in addition to other essential processes such as photosynthesis, cell division, cytoplasmic movements, and increased cell volume (Malavolta et al., 1997). Moreover, calcium plays an important role in ion absorption and when in deficit it affects root growth points (Vitti et al., 2006). Coconut fiber may have contributed to the physical quality of the substrate, providing adequate aeration and water retention. Granulated coconut fiber shows a predominance of intermediate and fine fractions, which may be appropriate when seeking high porosity and presence of micropores, which are responsible for good aeration and water retention in the environment (Zorzeto et al., 2014). In highly weathered tropical and subtropical soils, organic matter is of great importance for crop nutrient supply, cation retention, toxic and micronutrient complexation, structure stability, infiltration and retention, aeration, and microbial activity, constituting a fundamental component of its productive capacity (Bayer and Mielniczuk, 1999). Or- ganic matter added to soil favors numerous microbiological processes related to mineralization and nutrient release to plants, nitrogen fixation (symbiotic to non-symbiotic), and decomposition of organic waste. It also improves the physi- cal qualities of the soil such as structure development and aggregate stability, which are beneficial for plant growth and development (Bento, 1997). In addition to physical factors, the increase in organic matter content promotes improvements in chemical attributes such as increased pH and base saturation, as well as aluminum complexation and soil solution precipitation (Mello and Vitti, 2002). Regarding the variable electrical conductivity (EC), there was no difference (P<0.05) between the two groups of contrasted treatments. TABLE 3. Significance levels, contrast estimators and P value between the conventional substrate treatment and those with different levels of moinha in the composition. Treatments Conventional 0% 10% 20% 30% 40% Estimator P value Contrast coefficients Moinha level 5 -1 -1 -1 -1 -1 - - Means of the treatment Stem diameter (mm) 1.67 1.57 1.88 1.99 2.05 1.87 -1.00 0.0189 Means of the treatment Plant height (cm) 4.00 4.00 5.09 6.04 5.78 4.48 -5.38 0.0082 Means of the treatment Total dry mass (mg) 213.75 298.63 270.25 492.0 454.63 376.63 -823.38 0.0034 Means of the treatment Electrical conductivity (µS cm-1) 48.75 45.00 55.00 55.00 31.25 83.75 -26.25 0.4986 33Matiello Almeida, Vieira Lo Monaco, Krause, Silva de Oliveira, Merlo Meneghelli, Merlo Meneghelli, and Pinto Guisolfi: Reuse of agricultural waste as an alternative substrate in the production of eggplant (Solanum melongena L.) seedlings When the different moinha levels were evaluated (poly- nomial regression), second-degree effects (P<0.05) were observed for the variables electrical conductivity (EC), stem diameter (SD) and plant height (PH). For the total dry mass (TDM), no adjustment was observed in regres- sion models (P<0.05). It was observed that, in general, increasing percentages of moinha in the substrate composition led to an increase of the salinity level in the substrate (greater electrical con- ductivity) (Fig. 1). This result was already expected since moinha showed higher electrical conductivity than the other residues (Tab. 1). The high concentration of salts is a stress factor for plants, because it reduces the osmotic potential and causes the action of ions on the protoplasm (Harter et al., 2014). Water is osmotically retained in the saline solution, so that the increased concentration of salts makes it less and less available to plants (Ribeiro et al., 2001). FIGURE 2. Stem diameter of eggplant seedlings as a function of increa- sing percentages of moinha in the composition of substrates. The smaller amount of nutrients present in the substrate, due to the absence of moinha, may have been the reason for having a smaller stem diameter in T2. In the highest concentrations of moinha, the decrease in this variable can be attributed to the higher salinity of this residue, which impairs plant growth and development, as mentioned earlier. A similar result was obtained by Almeida et al. (2018), who verified a decrease in the stem diameter of cucumber seedlings with the increase of the residue in the alternative substrate. The plant height increased significantly with the gradual replacement of the commercial substrate by moinha up to the percentage of 21.80%, providing a maximum height of 5.94 cm (Fig. 3). Above this percentage, there is a decrease in the plant height values. However, similarly to what occurred for the stem diameter, the plant height values obtained at higher moinha concentrations were still greater than those registered for the treatment in which no moinha was added (T2). ŷ = 52.0000 - 1.2125x + 0.0437x² R² = 37.48%; P = 0.034 El ec tr ic al c on du ct iv ity ( µ S c m -1 ) Moinha in the substrate (%) 0 10 20 30 40 20 30 40 50 60 70 80 90 ŷ = 1.5701 + 0.0368 - 0.0007x² R² = 98.81%; P = 0.001St em d ia m et er ( m m ) Moinha in the substrate (%) 0 10 20 30 40 1.5 1.6 1.7 1.8 1.9 2.0 2.1 FIGURE 1. Electrical conductivity as a function of increasing percentages of moinha in the composition of the substrates. The stem diameter is a key variable to evaluate the survival potential and growth of seedlings (Souza et al., 2006). These authors emphasize that within the same species, plants with large stem diameters have higher survival rates because they have the capacity to generate and develop roots. In this sense, it can be observed that the substrate containing 26.3% moinha provided the highest stem diameter (2.05 mm), with a decrease in the variable above this percent- age (Fig. 2). However, the stem diameter values obtained at doses higher than 26.3% were still greater when com- pared to the treatment without the addition of moinha in the substrate (0% moinha + 40% burnt rice husk + 15% coconut fiber + 5% eggshell + 40% commercial substrate), demonstrating the beneficial effect of this residue on sub- strate composition. ŷ = 3.89211 + 0.1875 - 0.0043x² R² = 96.48%; P = 0.0001 P la nt h ei gh t ( cm ) Moinha in the substrate (%) 0 10 20 30 40 3.5 4.0 4.5 5.0 5.5 6.0 6.5 FIGURE 3. Plant height of eggplant seedlings as a function of increasing percentages of moinha in the composition of the substrates. 34 Agron. Colomb. 38(1) 2020 In general, it was verified that the highest values obtained for the assessed variables were those in which around 20% moinha was added in the composition of the substrates. When moinha proportion increased above this percent- age, all variables showed decreasing values. It is believed that this result could be associated with the higher salinity levels achieved with increasing moinha percentages in the substrate, evidenced by the greater electrical conductivities (Fig. 1). The high concentration of salts is a stress factor for plants, as it reduces the osmotic potential and provides the action of ions on the protoplasm (Harter et al., 2014). The water is osmotically retained in the saline solution, making it less and less available to the plants, and, consequently, interfering negatively in the growth and productivity of the crop. Studying the use of agricultural residues in the composi- tion of substrates for the production of tomato seedlings, Krause et al. (2017) found that the highest values obtained from the analyzed variables were those in which around 15 to 32% of moinha was used in the composition substrates. With the increase in this proportion, all variables showed decreasing values. The authors also believe that this fact may be associated with higher salinity in the substrate, evidenced by the greater electrical conductivity in the residue. Likewise, Meneghelli et al. (2016), studied moinha as an alternative substrate in conilon coffee seedlings and also found a decrease in the analyzed growth variables and concluded that the high salinity of the residue may have favored this result. The use of about 20% of moinha in the substrate composi- tion promoted the best development of eggplant seedlings, probably because at this proportion the negative effects of salinity did not overcome the beneficial effects of the nutrients, especially nitrogen, present in higher amounts (Tab. 1). 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