Purpose In the present study, effect of earthworm-processed MSW was seen on biochemical, physiological, and yield responses of Abelmoschus esculentus L. Methods Plants were grown on different amendment ratios of municipal solid waste vermicompost (MSWVC). Pot experi - ments were conducted by mixing MSWVC at 0, 20, 40, 60, 80, and 100% ratios to the agricultural soil. Results An increase in photosynthetic rate and stomatal conductance of plants grown at 20 and 40% MSWVC amendment ratios was observed. Total chlorophyll, carotenoid, and protein contents also increased significantly in 20, 40, and 60% amendment ratios at 65 days after germination (DAG). Likewise, proline, peroxidase, and lipid peroxidation increased with increasing levels of MSWVC at both 45 and 65 DAG. Conclusion The study suggests that MSWVC could be used as organic amendment in soil depicted by good yield and anti- oxidative response of lady’s finger ( A. esculentus) at different amendments of MSWVC (up to 60% w/w ratios). Furthermore, agricultural utilization of MSWVC will help in managing dreadful effects of the burgeoning amount of organic solid waste. Keywords Municipal solid waste · Vermicompost · Abelmoschus esculentus L. · Heavy metals · Physiology Introduction the environment with social acceptability. Organic wastes constitute major fraction of municipal solid waste (MSW) Management of the burgeoning amount of organic solid in most of the developing countries. Thus, land application waste is a challenging problem in the contemporary scenario of organic fraction of MSW offers a good option for waste around the globe. The improper handling and unscientific management and its recycling. For example, land application disposal of organic waste has led to countless problems pos- of organic waste such as sewage sludge is a common prac- ing a threat to the ecosystems (soil, air, and water) and envi- tice nowadays (Singh and Agrawal 2008; Lee et al. 2018; ronmental sustainability (Vergara and Tchobanoglous 2012, Sharma et al. 2018). However, there is always a potential Srivastava et al. 2015, 2016). Therefore, it is imperative to threat as it may contain various toxic heavy metals, organic find a solution to this problem that would not only focus compounds such as antibiotics, pharmaceuticals and per- on managing its quantity, but will also help in sustaining sonal care products (PPCPs), and pathogens in traces (Bibby and Peccia 2013, Kang et al. 2013, Prosser and Sibley 2015). The prolonged use of sewage sludge or organic wastes in agricultural field may result in heavy metal accumulation in * Rajeev Pratap Singh email@example.com soil that can be taken up by the crops which may cause many health issues when transferred to higher trophic levels (Sriv- Department of Environment and Sustainable Development, astava et al. 2017).Therefore, composting of organic wastes/ Institute of Environment and Sustainable Development, biosolids is a more interesting option for recycling of wastes. Banaras Hindu University, Varanasi 221005, India 2 Nowadays, composting/vermicomposting of organic fraction Environmental Engineering, Department of Civil of municipal solid waste is gaining attention among work- Engineering, Indian Institute of Technology, Delhi, India 3 ers as it decreases the stabilization time of organic wastes Society for Higher Education & Practical Applications (Fernández et al. 2014, Weber et al. 2014). The quality of the (SHEPA), Varanasi, India Vol.:(0123456789) 1 3 International Journal of Recycling of Organic Waste in Agriculture compost largely depends on factors such as feedstock source, noticed in 100% vermicompost and the largest marketable presence of toxic contaminants/heavy metals, stabilization yield was noticed in 20% pig manure vermicompost. time, and composting design (Hargreaves et al. 2008, Sriv- Plenty of literature are available on the effect of organic astava et al. 2016). waste vermicompost on growth and yield of different plants Cherif et al. (2009) examined the effect of municipal (Atiyeh et al. 2001, Arancon et al. 2004, Lim et al. 2015, −1 solid waste compost, MSWC (40 and 80 Mg ha ), on wheat Sangwan et al. 2010). However, studies on antioxidative growth and noticed a significant increase of grain yield in response of plants grown on different organic waste ver - −1 both the amendments (58.96 and 60.21 Mg ha , respec- micompost are still in its infancy. Organic waste vermicom- −1 tively) compared to the control (17.65 Mg ha ). Based on post may contain significant amount of trace metals which −1 the treatment effectiveness index, 40 Mg ha of MSWC could pose phytotoxicity (Gupta and Garg 2008, Roodbergen was recommended for agricultural practices. Similarly, agri- et al. 2008, Mohee and Soobhany 2014, Atiyeh et al. 2000). cultural application of MSWC and its effects on the yield Therefore, the present work was aimed to assess the bio- of various crops has been reported in various studies such chemical, physiological, and yield responses of lady’s finger as winter squash (Warman et al. 2009), lettuce (Fagnano (Abelmoschus esculentus L.) grown on soil amendment with et al. 2011), wheat (Lakhdar et al. 2011), and spring triticale different ratios of MSW vermicompost. Also, the potential (Weber et al. 2014). Though the agricultural application of of MSWVC application in agricultural practices was com- MSWC has immense potential for recycling of such type of prehensively evaluated. organic solid waste; however, the presence of heavy metals and other pollutants in MSWC has always been a matter of concern (Singh and Kalamdhad 2013, Alvarenga et al. Materials and methods 2015). Hence, there is a growing interest in vermicomposting Study area technology which has emerged as a new biotechnological tool for recycling of different kind of organic wastes through The experiments were performed at the experimental field the action of earthworms (Wu et al. 2014, Lim et al. 2016). of Institute of Environment and Sustainable Development, Both MSW compost (MSWC) and vermicompost have many Banaras Hindu University, Varanasi, situated in the eastern advantages when compared to inorganic fertilizers which Gangetic plain of the Indian subcontinent at 25°14′N lati- badly affect the soil’s physico-chemical and microbial prop- tude, 82°3′E longitude, and 76.19 m above the sea level. The erties (Srivastava et al. 2016). Agricultural utilization of experiments were performed during August–October 2016. MSW compost/vermicompost (VC) ameliorates the soil’s This period of the year is characterized by mean monthly nutrient profile, texture, water holding capacity, buffering maximum temperatures between 29.0 and 36 °C and mean capacity, soil microbial response, etc. (Weber et al. 2014, monthly minimum temperatures between 18.0 and 27.0 °C. Bouzaiane et al. 2014). MSWC/VC has good organic mat- The total rainfall was 216.90 mm. Maximum relative humid- ter content, nitrogen, phosphorus, and humic substances ity showed variation from 80.0 to 100% and minimum from which are important for maintaining the soil quality (Har- 27 to 93%. greaves et al. 2008). Moreover, it improves the activity of different soil enzymes such as dehydrogenase, urease, Experimental design and raising of plants phosphatase, protease, phosphodiesterase, arylsulphatase, etc. (Perucci 1990, Bhattacharyya et al. 2003). According All the experiments were carried out in earthen pots of to Sim and Wu (2010), it has been demonstrated that through 30 cm diameter and 30 cm depth. The MSWVC used in the vermicomposting, MSW could be sustainably transformed present study was prepared by mixing organic waste and cow into an organic fertilizer known as vermicompost that pro- dung (1:1 ratio). MSW was collected from secondary waste vides great benefits to the agricultural soil and plants. Ati- collection points of municipal corporation, Varanasi district. yeh et al. (2001) studied the effect of earthworm-processed The vermicomposting of organic fraction of MSW (includ- pig manure on germination, growth, and yield response of ing vegetable wastes, fruit wastes, flower wastes, paper Lycopersicon esculentum under greenhouse environment. wastes, and leaf litter) was performed using Eisenia fetida Plants were grown in the standard potting medium amended (an earthworm spp.). Three replicates of six die ff rent amend - with various ratios (0–100%) of pig manure vermicompost ments i.e., control (unamended soil), 20% MSWVC (w/w), (VC). Germination rates were increased significantly by 40% MSWVC (w/w), 60% MSWVC (w/w), 80% MSWVC 20–40% in the VC amended soil than the control. Similarly, (w/w), and 100% MSWVC (w/w) designated as S, A, B, C, tomato seedlings grown in 50% VC had greater number of D, and E respectively were used. The MSWVC was mixed leaves and biomass in comparison to control. However, a uniformly with soil and left for 10 days in the field before sharp decline in growth, number of leaves, and biomass was filling the pots. Lady’s finger (Abelmoschus esculentus L. 1 3 International Journal of Recycling of Organic Waste in Agriculture var. F1 GS-126, Virgo), a common vegetable consumed in photosynthetic system (LI-6200, LI-COR, Inc. Lincoln, NE, central India was used as a test plant in the present study. USA) directly on intact plants in the pot at ambient climatic Required moisture level (25%) was maintained prior to the conditions. Randomly selected three plants were sampled sowing of lady’s finger seeds and thereafter six seeds were from the each experimental setup for estimation of morpho- sown at equal distances in each pot. After germination, thin- logical parameters at 45 DAG. Morphological parameters ning was performed to maintain three plants in each pot. The such as root and shoot length, number of leaves, and leaf pots were placed in open field condition to provide identical area of each plant were estimated. Measurement of leaf area light and temperature to all the amendments and were irri- was performed using leaf area meter (Systronics 211, India). gated with equal amount of water to maintain identical water Dry weights of root, shoot, and leaves were also estimated regime throughout the growth period of plants. after drying the samples at 80 °C till constant weight was achieved. The biomass yield was calculated as fresh weight Soil sampling and analysis at the time of harvest at 65 DAG. Triplicate soil samples collected from each amendment Chemicals and quality control were air dried, grounded, and sieved through 2-mm mesh size and physico-chemical properties were analyzed. The All the chemicals used in this study were of analytical grades pH of the soil at different amendments was measured in the and used without further purification. The analytical qual- suspension of 1:5 (w/v) with the help of pH meter (Model ity control was guaranteed through the use of laboratory 802, Systronics, India) standardized with pH 4, 7, and 9.2 quality assurance and quality control protocols and stand- reference buffers. Electrical conductivity was measured ards. Standard operating procedures were followed for the by electrical conductivity meter (Model 303, Systronics, calibration with standards, analysis of reagent blanks, and India). Soil organic carbon was determined by Walkley recovery of known spiked samples. The reagent blanks and and Black’s rapid titration method (Allison 1973) and total known standards were used throughout the metal analysis nitrogen content of samples was measured by Pelican auto- and used to correct the analytical results. Precision and accu- matic nitrogen analyzer (Model KEL PLUS India). Total racy of heavy metal analysis were assessed through repeated phosphorous was estimated by method described by Allens analysis of the samples against National Institute of Stand- (1974). For heavy metal analysis, 1 gm of soil and VC ard and Technology, Standard Reference Material (SRM samples were acid digested with 20 ml of triacid mixture 1570) for all the heavy metals. The results were found to be (HNO :H SO :HClO ::5:1:1) following Allen et al. (1986). within ± 2% of the certified value. All the experiments were 3 2 4 4 The filtered acid digests were analyzed by atomic absorption carried out in triplicate. spectrophotometer (AA 240 FS, Varian). Data analysis Estimation of biochemical and physiological parameters Statistical analysis was performed using the SPSS version 16 (Illinois, USA) software for windows program. Treatments For biochemical analysis, fresh leaves were plugged manu- were compared using analysis of variance (ANOVA) and ally at 45 and 65 days after germination (DAG) and stored Duncan’s multiple range test (DMRT) was performed to test in deep-freezer till further analyses. Chlorophyll and carot- the significance of difference between the treatments. The enoid contents were estimated by the method of Maclachlan graphs were drawn using Sigma Plot version 10 software. and Zalik (1963) and Duxbury and Yentsch (1956), respec- −1 tively, and expressed as mg g dry leaf. Protein content in the fresh leaves was analyzed following Lowry et al. (1951). Results and discussion Foliar ascorbic acid and proline contents were measured by the method of Keller and Schwager (1977) and Bates The MSWVC used in the study had almost neutral pH −1 et al. (1973). Peroxidase activity was measured using the (7.12) and high electrical conductivity (2.24 mS cm ), method of Britton and Mehley (1955). Total phenol and thiol total organic carbon (31.46%), total N (1.4%), total −1 −1 contents were measured following Fahey et al. (1978) and P (15.81 g kg ), total K (15.36 g kg ), and total −1 Bray and Thorpe (1954), respectively. The MDA (malon- Fe (984.13 mg kg ) contents (i.e., 100% MSWVC) dialdehyde) level representing the index of lipid peroxida- (Table 1). The concentrations of heavy metals of Cr, Pb, tion was measured by the method proposed by Heath and Ni, Cu, Cd, and Zn were 28.68, 2.40, 10.07, 37.7, 1.07, −1 Packer (1968). Physiological parameters of lady’s finger and 294.6 mg kg , respectively, in ready MSWVC. The plants such as the rate of photosynthesis, stomatal conduct- physico-chemical properties and nutrient profile of ready ance, and transpiration rate were measured using portable vermicompost largely depend on the source of substrates/ 1 3 International Journal of Recycling of Organic Waste in Agriculture Table 1 Selected physico-chemical properties of unamended and municipal solid waste vermicompost amended soil (values ± SE) Parameters Soil A (20%VC) B (40%VC) C (60%VC) D (80%VC) E (100%VC) a b c d e f pH 8.05 ± 0.04 7.94 ± 0.03 7.85 ± 0.01 7.67 ± 0.008 7.50 ± 0.01 7.12 ± 0.01 −1 f e d c b a EC (mS cm ) 0.26 ± 0.01 0.45 ± 0.01 0.56 ± 0.008 0.87 ± 0.01 1.09 ± 0.01 2.24 ± 0.008 e d d c b a TOC (%) 0.63 ± 0.01 6.33 ± 0.36 7.83 ± 0.32 14.68 ± 0.43 22.29 ± 1.27 31.46 ± 0.63 e de d c b a TKN (%) 0.24 ± 0.01 0.26 ± 0.00 0.28 ± 0.01 0.36 ± 0.01 0.41 ± 0.01 1.40 ± 0.01 −1 e e d c b a TP (g kg ) 4.14 ± 0.11 4.82 ± 0.07 6.88 ± 0.26 9.37 ± 0.15 11.09 ± 0.22 15.81 ± 0.50 −1 e de d c b a TK (g kg ) 5.85 ± 0.25 6.54 ± 0.21 6.84 ± 0.85 7.99 ± 0.12 9.60 ± 0.44 15.36 ± 0.30 −1 d d cd bc b a Cr (mg kg ) 6.45 ± 1.51 7.62 ± 0.29 8.4 ± 1.00 13.50 ± 0.40 15.17 ± 1.43 28.68 ± 3.43 −1 b b a a a a Pb (mg kg ) 0.98 ± 0.33 1.00 ± 0.18 1.88 ± 0.16 2.13 ± 0.16 2.05 ± 0.25 2.40 ± 0.12 −1 b b b b b a Ni (mg kg ) 5.20 ± 0.51 5.30 ± 0.43 5.88 ± 0.26 6.38 ± 0.42 7.27 ± 0.32 10.07 ± 1.50 −1 d d d c b a Cu (mg kg ) 13.97 ± 0.68 14.43 ± 0.23 15.82 ± 0.41 18.98 ± 0.68 21.28 ± 0.29 37.70 ± 1.72 −1 d d cd c b a Fe (mg kg ) 432.35 ± 21.66 425.28 ± 4.5 468.17 ± 12.38 493.77 ± 5.51 552.63 ± 7.33 984.13 ± 33.02 −1 a a a a a a Cd (mg kg ) 0.66 ± 0.13 0.67 ± 0.04 0.70 ± 0.19 0.72 ± 0.07 0.90 ± 0.09 1.07 ± 0.15 −1 d d d c b a Zn (mg kg ) 52.45 ± 0.33 51.63 ± 1.03 60.17 ± 2.96 91.42 ± 3.43 114.25 ± 7.60 294.6 ± 8.07 Different letters in each group show significant difference at p < 0.05 waste used in the vermicomposting process (Srivastava et al. et al. 2014). Previous studies have shown positive correla- 2015, 2016). For example, Garg et al. (2006) showed varia- tion between plant heavy metal uptake and total heavy metal tions in pH of different organic wastes vermicompost ranged content in the soil (Singh and Agrawal 2007, 2009, 2010a, between 7.7 (sludge) and 8.3 (kitchen waste). Similarly, EC b). Therefore, MSWVC amendments may pose risk to soil −1 was ranged between 0.7 mS cm (institutional waste) and and plant health at higher doses. −1 2.3 mS cm (kitchen waste) depending on the nature of Compared to the control, the total chlorophyll content waste. of plants at 45 and 65 DAG was increased in MSWVC In the present study, physico-chemical analysis of amended soil (Fig. 1). Total chlorophyll content in A. escu- −1 MSWVC amended soil showed a significant decrease in pH lentus was 1.84, 1.97, 2.51, 2.28, 1.71, and 0.99 mg g dry and increase in EC with increasing levels of VC as com- weight in S, A, B, C, D, and E, respectively, at 65 DAG. pared to unamended soil (Table 1). This trend may be attrib- Maximum increase of 61.25 and 36.41% was observed in uted to lower pH and higher EC of MSWVC used in the total chlorophyll content of plants grown in 40% MSWVC study. The release of humic substances might be the reason (B) at 45 and 65 DAG. However, significant decrease in total for such trends (Suthar et al. 2015). Atiyeh et al. (2001) chlorophyll was noticed in 100% MSWVC (E) treatment and observed similar results in application of pig manure ver- a decrease of 40.96 and 46.20% was recorded at 45 and 65 micompost for horticultural uses. Soil organic carbon, total DAG, respectively. Lakhdar et al. (2012) studied the effect of nitrogen, total phosphorus, and total potassium contents MSW compost on photosynthetic performance of Triticum −1 were increased in VC amended soil (Table 1). Improvement durum, following application up to 300 t ha . An increase −1 in the nutrient profile of soil due to amendment of organic of 14 and 15% was recorded at 40 and 100 t ha applica- waste compost/vermicompost has been reported previously tion rate, followed by a progressive decline in total chloro- −1 (Sangwan et al. 2010, Avramidou et al. 2013). However, phyll content at higher doses (200 and 300 t ha ). Similarly, higher doses of MSWVC treatments led to increased concen- Sangwan et al. (2010) demonstrated increased level of total tration of heavy metals in soil (Table 1). The concentrations chlorophyll content in marigold at different application rates of heavy metals in the MSWVC amended soils were found of cow dung and filter cake (sludge from sugar mill waste in the order of Fe > Zn > Cu > Cr > Ni > Pb > Cd. Zn is an water treatment) + horse dung vermicomposts. Decrease in important micronutrient for different metabolic functions photosynthetic rate may be attributed to elevated levels of in plants; however, it’s toxic levels in soil lead to reduced heavy metal accumulation that can adversely affect electron plant growth, leaf chlorosis, photosynthesis impairment, and transport as well as photosynthetic metabolism (Burzyhski reactive oxygen species (ROS) production that poses harm and Zurek 2007). to membrane integrity and permeability (Cambrollé et al. Carotenoids are non-enzymatic antioxidants, which pro- 2012, Subba et al. 2014, Srivastava et al. 2017). Moreover, tect the photosynthetic pigment against oxidative stress high concentrations of Zn adversely affect dehydrogenase, posed by heavy metal-induced reactive oxygen species β-D-glucosidase, urease, catalase, and invertase activities (ROS) (Halliwell 1987). Carotenoid content increased in soil (Kunito et al. 2001, Yang et al. 2006, Ciarkowska significantly at both the ages and maximum increase of 1 3 Total chlorophyll content Carotenoid -1 -1 (mg g dry wt) (mg g dry wt) International Journal of Recycling of Organic Waste in Agriculture S A(20%) B(40%) C(60%) D(80%) E(100%) 8.0 65 days 45 days 45 days a 65 days 20 a d 6.0 15 a 4.0 f c e d e 2.0 0.0 1.0 2 45 days a 65 days 45 days 65 days b a a 0.8 b b 0.6 b a 0.4 0.2 0.0 Fig. 1 Protein, ascorbic acid, total chlorophyll, and carotenoid content in lady’s finger plants grown at different MSWVC amendments at two dif- ferent ages (Mean ± 1 SE). Bars with different letters in each group show significant difference at p < 0.05 65.38 and 50.80% was found in 40% MSWVC (B) at 45 salinity due to higher dose of MSWVC might have resulted and 65 days, respectively, as compared to control (Fig. 1). in reduced photosynthetic rate (Mysliwa-Kurdziel et al. Increase in carotenoid content depicted that the plant defense 2002). Similarly, the transpiration rate also increases due to system is actively responding to oxidative stress; however, a adequate nutrient supply (Adamtey et al. 2011). Ascorbic decrease of 32.69% in 100% MSWVC (E) at 45 days showed acid is a powerful antioxidant, which functions as a redox that the plant defense mechanism collapsed and failed to buffer and prevents plants from free oxygen radicals. It also defend the plant against stress. works as a cofactor for enzymes participating in photo- Photosynthetic (Ps) and transpiration rate increased sig- synthesis (Smirnoff and Wheeler 2000). In this study, no nificantly in 40% MSWVC (B), whereas stomatal conduct- significant changes were noticed in ascorbic acid content ance increased significantly in 40 and 60% MSWVC (B and of A. esculentus grown in different MSWVC amendments. C) compared to control (Table 2). An increase of 71.60% in Singh and Agrawal (2010c) found similar kind of trends for photosynthetic rate of plants grown in 40% MSWVC (B) was ascorbic acid content while studying the response of Oryza observed, whereas it declined to 31.49% in 100% MSWVC. sativa L. grown at different sewage sludge application rates Lakhdar et al. (2012) reported increased CO net assimila- (Fig. 1). It is presumed that reactive oxygen free radicals −1 tion in Triticum durum plants grown at 40 and 100 t ha might have oxidized ascorbic acid to dehydroascorbic acid MSW compost, whereas a significant reduction was seen at (DHA) resulting in reduction and a non-significant response −1 300 t ha . Increased photosynthetic rate in plants is ascribed of ascorbic acid content of plants. Similarly, Rao and Sresty to nutrient rich profile of MSWVC (Chen et al. 2005); (2000), during their study on seedlings of Cajanus cajan however, bioavailability of heavy metals to the plants and (L.) Millspaugh under Zn and Ni stresses found negative Table 2 Variation in selected Amendments Photosynthetic rate Transpiration rate Stomatal conduct- Ci/Ca physiological characteristics −2 −1 −2 −1 −1 (µmol m s ) (mmol m s ) ance (cm s ) of A. esculentus plants grown b b cd a in different amendment rates Soil 7.43 ± 0.22 3.55 ± 0.12 0.12 ± 0.00 0.72 ± 0.03 of MSW vermicompost b b c ab A (20%VC) 7.78 ± 0.11 3.62 ± 0.04 0.13 ± 0.01 0.71 ± 0.01 (values ± SE) a a a a B (40%VC) 12.75 ± 0.28 4.78 ± 0.14 0.25 ± 0.01 0.72 ± 0.02 c b b b C (60%VC) 6.49 ± 0.35 3.85 ± 0.11 0.16 ± 0.02 0.64 ± 0.03 d b cd a D (80%VC) 5.24 ± 0.20 3.41 ± 0.23 0.10 ± 0.01 0.75 ± 0.04 d b d b E (100%VC) 5.09 ± 0.37 3.63 ± 0.35 0.09 ± 0.01 0.64 ± 0.02 1 3 Protein Ascorbic acid -1 -1 (mg g fresh wt) (mg g fresh wt ) Peroxidase content Thiol content (μM purpurogalin formed -1 -1 -1 (μΜ g fresh wt) min g fresh wt) International Journal of Recycling of Organic Waste in Agriculture correlation between ascorbic acid content and increasing Increased level of malondialdehyde (MDA) acts as levels of heavy metals. marker of lipid peroxidation. MDA content was increased Protein content increased significantly in plants with significantly with increasing level of MSWVC when com - increasing rate of MSWVC (A, B, C, and D) compared pared with the control plants at both the ages (Fig. 2). The to control; however, a significant decrease was noticed in maximum increase of 125% was noticed in MDA content of 100% MSWVC (E) amendment at both 45 and 65 DAG. plants grown in 100% MSWVC at 65 DAG. Increased bio- The total protein content in A. esculentus plants was 17.31, availability of heavy metals to the plants at higher doses of −1 19.26, 20.97, 19.52, 16.92, and 12.55 mg g fresh weight MSWVC led to ROS production that could have damaged the in S, A, B, C, D, and E, respectively, at 65 DAG. Maxi- plasma membrane (Cuypers et al. 2011). Singh and Agrawal mum increments of 19.96 and 21.14% were observed in 40% (2007) reported increased level of lipid peroxidation in Beta MSWVC (B) at 45 and 65 DAG, respectively. Whereas in vulgaris grown at 20 and 40% sewage sludge amendments. It 100% MSWVC (E), a sharp decline of 30.30, and 27.50% was observed that peroxidase content increased significantly in total protein content was noticed at 45 and 65 DAG. High in plants with increasing doses of MSWVC; the increments organic matter content in MSWVC might be the main rea- were more prominent in 60, 80, and 100% MSWVC in 45 son for increased level of protein content as it slows down and 65 DAG (Fig. 2). Similar findings have been reported the release of N in VC amended soil (Singh and Agrawal earlier in Beta vulgaris (Singh and Agrawal 2007) and in 2010c). Also, heavy metal toxicity may induce heat shock Oryza sativa (Singh and Agrawal, 2010c) grown in dif- proteins (Kim et al. 2014). The elevated level of heavy met- ferent sewage sludge amendments. Phenols, a secondary als at higher doses of MSWVC might be the reason for metabolite and antioxidant, play a crucial role in defense decreased protein content. mechanism of plants against oxidative and abiotic stresses. 12 18.0 65 days 45 days 45 days a 65 days 16.0 c b b a b 14.0 8 12.0 b ab f e e d 10.0 8.0 6.0 4.0 2.0 0 0.0 0.4 45 days 65 days 65 days 45 days a 16 0.3 c b a d e e c e c 0.2 0.1 0.0 2.0 45 days 65 days 1.5 1.0 e e 0.5 de c e d 0.0 Fig. 2 Phenol, proline, thiol, peroxidase, and lipid peroxidase content in lady’s finger plants grown at different MSWVC amendments at two dif- ferent ages (Mean ± 1 SE). Bars with different letters in each group show significant difference at p < 0.05 1 3 Lipid peroxidation Phenol content Proline content -1 -1 -1 (mg g fresh wt) (n mol ml ) (mg g fresh wt ) International Journal of Recycling of Organic Waste in Agriculture Table 3 Morphological characteristics (at 45 DAG) and yield* response (at 65 DAG) of lady’s finger (values ± SE) Amendment Root length Shoot length Leaf area (cm ) No. of leaves Root biomass Shoot biomass Leaf biomass Total biomass Yield* (g −1 −1 −1 −1 −1 −1 (cm) (cm) (plant )(g plant )(g plant )(g plant )(g plant ) plant ) bc b e bc cd b cd bc cd S 6.20 ± 0.62 28.03 ± 1.21 136.17 ± 4.15 5.00 ± 0.00 0.05 ± 0.01 0.32 ± 0.04 0.30 ± 0.02 0.67 ± 0.07 121.33 ± 2.09 b ab c b b b bc b bc A 7.27 ± 0.41 34.87 ± 1.52 206.07 ± 7.39 6.00 ± 0.58 0.10 ± 0.00 0.63 ± 0.07 0.37 ± 0.03 1.05 ± 0.08 138.33 ± 8.44 a a a a a a a a a B 9.77 ± 0.07 43.83 ± 2.33 432.0 ± 7.42 8.0 ± 0.58 0.19 ± 0.03 1.72 ± 0.40 1.08 ± 0.13 2.99 ± 0.55 196.64 ± 7.23 a b b b cd b b b b C 8.8 ± 0.50 30.53 ± 6.08 280.83 ± 5.20 5.67 ± 0.67 0.05 ± 0.01 0.54 ± 0.07 0.58 ± 0.06 1.17 ± 0.11 147.70 ± 4.09 b b d b bc b bc bc d D 7.13 ± 0.15 28.73 ± 3.42 182.97 ± 4.03 5.33 ± 0.33 0.06 ± 0.01 0.31 ± 0.03 0.46 ± 0.06 0.84 ± 0.10 115.46 ± 6.76 c c f c d b d c e E 5.13 ± 0.20 12.73 ± 0.94 41.03 ± 6.89 3.67 ± 0.33 0.01 ± 0.00 0.11 ± 0.03 0.09 ± 0.01 0.23 ± 0.04 72.01 ± 6.85 Different letters in each group show significant difference at p < 0.05 Phenol content increased significantly in plants at different seen in leaf area of B, but it was declined by 69.87% in E amendment rates (20, 40, and 60% MSWVC); however, a amendment compared to control. Likewise, total biomass decrease was noticed in 80 and 100% MSWVC indicating was increased significantly in B, but reduced significantly failure of plant defense system at higher doses at 45 DAG in plants grown at E amendment. Yield response recorded at (Fig. 2). Similarly, increased level of polyphenol content 65 DAG showed significant increase at 40 and 60% MSWVC −1 in Mesembryanthemum edule grown at 40 t ha of MSW soil amendments; however, it was decreased in 80 and 100% compost was reported by Lakhdar et al. (2011). MSWVC amendments. Proline is an osmolyte which plays a pivotal role dur- ing abiotic and oxidative stress in plants and acts as metal chelator, antioxidant, and a signaling molecule (Hayat et al. Conclusion 2012). Proline content was increased significantly in plants with increasing levels of MSWVC at both the ages with The present study clearly suggests that MSWVC showed maximum increase in 100% MSWVC compared to control. a positive effect on soil nutrient profile represented by This could be due to either high salt concentration, plant increased organic carbon and NPK. Higher doses of phytotoxicity in higher dose of VC, and/or heavy metal accu- MSWVC led to either salinity stress, plant phytotoxicity or mulation in amended soil (Arancon et al. 2004, Sangwan bioavailable heavy metal accumulation depicted by increased et al. 2010, Srivastava et al. 2016). Heavy metals in plants level of antioxidative plant response, lipid peroxidation, and may affect the permeability of membranes leading to water peroxidase activity. However, MSWVC showed positive stress environment causing proline accumulation (Singh and effect on biochemical, physiological, and yield responses of Agrawal 2007, 2009). Similarly, thiol content increased at A. esculentus (Lady’s finger) up to 60% MSWVC amended different amendment ratios of MSWVC (A, B, C, and D) soils represented by increased rate of photosynthesis, sto- and was declined at higher doses (E) at both 45 DAG and matal conductance, and improved antioxidative response. 65 DAG (Fig. 2). In addition, the leaf area, total biomass, and yield responses Root length and shoot length increased significantly in increased significantly with respect to control. The present 40% MSWVC whereas it declined in 100% MSWVC at 45 study concludes that up to 60% amendments of MSWVC can DAG (Table 3). Reduction in root and shoot length may be used as manure for improving soil fertility in agricultural be ascribed to increased levels of heavy metals in 100% applications. This could help in the sustainable management MSWVC amendments. Increased levels of heavy metals of the burgeoning amount of organic waste. might have decreased the mitotic activity within the root Acknowledgements RPS is thankful to Department of Science cells leading to suppressed root growth in 100% MSWVC and Technology (P-45/18) for the project grant and to the Director, (Thounaojam et al. 2012). Furthermore, high concentrations Dean and Head, Institute of Environment and Sustainable Develop- of heavy metals caused more ROS production that posed ment, Banaras Hindu University for providing necessary facilities. negative effects on plant physiology resulted in reduced VS is grateful to ICMR for awarding JRF and SRF. Authors extend their sincere thanks to Mr. Kamlesh Kumar for his assistance in field shoot growth (Srivastava et al. 2017). Similarly, Singh and preparation. Agrawal (2007) reported significant decrease in root lengths of Beta vulgaris plants at 20 and 40% (w/w) sewage sludge Open Access This article is distributed under the terms of the Crea- amendments (SSA), whereas shoot lengths were decreased tive Commons Attribution 4.0 International License (http://creat iveco significantly at 40% SSA due to heavy metal stress. Leaf mmons.or g/licenses/b y/4.0/), which permits unrestricted use, distribu- tion, and reproduction in any medium, provided you give appropriate areas were increased significantly with increasing MSWVC credit to the original author(s) and the source, provide a link to the (A, B, C ,and D), while decrease in leaf area was noticed in Creative Commons license, and indicate if changes were made. E amendment at 45 DAG. 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