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Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 6 (2020) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2020.906.133 Soil Enzymatic Activity under Different INM Practices in Rice-Rice Cropping System P.V. Geetha Sireesha*, G. Padmaja, M. Venkata Ramana and P.C. Rao Dept. of Soil Science and Agricultural Chemistry, College of Agriculture, PJTSAU, Rajendranagar, Hyderabad - 500 030, India *Corresponding author ABSTRACT Keywords Dehydrogenase, INM, Phosphatase and urease activity Article Info Accepted: 18 May 2020 Available Online: 10 June 2020 Studies were conducted to understand the influence of integrated nutrient management on soil enzymatic activity under rice-rice cropping system since 1988 at Agricultural College Farm, Rajendranagar, Hyderabad. The enzyme activities viz., urease (55.8, 54.2 µg of NH4+-N g-1 soil 2h-1), dehydrogenase (488.23, 364.3 µg of TPF g-1 soil day-1) and acid (79.28, 70.50 µg p-nitrophenol g-1 soil h-1) and alkaline phosphatase (149.70, 129.53 µg p-nitrophenol g-1 soil h-1) were significantly higher in treatment T3 (50 % RD of NPK + 50 % N through FYM) in both kharif and rabi seasons. The enzyme activity of soils, which is governed by microbial population is also significantly higher in INM treatments. Our results demonstrate that soil enzymatic activity acted as a useful indicator of soil fertility dynamics. Enzymatic activities were positively and significantly correlated with content of organic carbon. Introduction The role that microbial activity plays in ecosystem processes is significant because approximately 80% to 90% of soil processes are mediated by microorganisms (Nannipieri and Badalucco, 2003). Soil microbial population are the driving force behind regulating soil processes such as organic matter decomposition and nutrient cycling, it is imperative to have a better understanding of the factors that regulate its size, activity, and structure (Masto et al., 2006). Soils containing a high microbial diversity are characteristic of a healthy soil-plant relationship, whereas those with low microbial diversity are characterized as an unhealthy soil that often hardly responds to environmental changes (Tejada et al., 2011). Phosphatases find widely in bacteria to mammals, and indicate their importance in 1073 Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 fundamental biochemical processes. The term phosphatase in soil is used to describe a group of enzymes that are responsible for the hydrolytic cleavage of a variety of esterphosphate bonds of organic phosphates and anhydrides of orthophosphoric acid (H3PO4) into inorganic phosphate. Acid and alkaline phosphatases particularly hydrolyse the ester bonds binding P to C (C-O-P ester bonds) in organic matter. During the process, inorganic P is released from organically bound P such as leaf litter, dead root systems, and other organic debris without concomitant release of C (Harrison, 1983). Phosphatase is concentrated in the surface layer and rhizosphere where most of the fresh and less humified organic matter is prevailing (Tarafdar et al., 2001). Phosphatases play a crucial role in the phosphorous acquisition of plants and microorganisms, and thus in the cycling of it within the soil (Schneider et al., 2001). Information on the nature of urease activity in soil was beneficial to develop and employ strategies for nitrogen management. Urease hydrolysis activity is elevated in aerobic condition, and its hydrolysis varied to the plant growth stage within green manure amendment to the crop (Pattnaik et al., 1999). Urease activity is not well when the bioavailability in the soil is troubled Saliha et al., (2006) confirm that urease activity increased along with microbial population in soil amended with liquid organic substrate. Materials and Methods Site description: The present investigation was carried out in the on-going AICRP on Integrated Farming Systems which was initiated in kharif, 1988 at the College Farm, College of Agriculture, Rajendranagar, Hyderabad. The monthly mean maximum temperatures during the crop growth period ranged from 28.0 0C to 37.7 0C with an average of 31.3 0C, while the monthly mean minimum temperature ranged from 10.1 0C to 23.9 0C with an average of 19.2 0C. The total rainfall received during the crop growth period was 1052.7 mm distributed throughout the year. Experimental design and treatments: This experiment was laid out in randomized block design with eight treatments and three replications. The treatment details during kharif season were as follows: T1 – Control (No fertilizer, no organic manure) T2 – 100 % RD of NPK T3 – 50 % RD of NPK + 50 % N through FYM T4 – 75 % RD of NPK + 25 % N through FYM T5 – 50 % RD of NPK + 50 % N through Paddy straw T6 – 75 % RD of NPK + 25 % N through Paddy straw T7 – 50 % RD of NPK + 50 % N through Green leaf manure T8 – 75 % RD of NPK + 25 % N through Green leaf manure The treatments in rabi season were as follows: T1 – Control (No fertilizer, no organic manure) T2 – 100 % RD of NPK (120-60-60 kg N, P2O5 and K2O ha-1) T3 – 100 % RD of NPK T4 – 75 % RD of NPK T5 – 100 % RD of NPK T6 – 75 % RD of NPK T7 – 100 % RD of NPK T8 – 75 % RD of NPK The organic sources such as FYM, Paddy straw and Glyricidia (green leaf manure) were applied two weeks before transplanting of paddy as per the treatments. All the PK and 1/3rd of N fertilizer were applied at the time of 1074 Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 transplanting while remaining nitrogen was applied in two equal splits. All cultural practices were performed. Soil sample analysis: The initial soil was sandy clay loam, neutral in reaction pH 8.5 (Jackson, 1973), non saline in nature EC 0.24 dS m-1 (Jackson, 1973) (Table 1), medium in organic carbon OC 0.54 % (Walkley and Black (1934), low in available N 151 kg ha-1 (Subbiah and Asija (1956), medium in available P 24.0 kg ha-1 (Olsen et al., 1954) and medium in available K 224 kg ha-1 (Jackson, 1973). Urease activity was assayed by quantifying the rate of release of NH4+ from the hydrolysis of urea as described by Tabatabai and Bremner (1972). Dehydrogenase activity in the soil was determined by the procedure given by Casida et al., (1964). The method involved spectrophotometric determination of the Tri Phenyl Formazon (TPF) produced when soil is treated with Triphenyl Tetrazolium Chloride (TTC). The acid and alkaline phosphatase activity was assayed by quantifying the amount of p-nitrophenol released and expressed as µg of p-nitrophenol released g-1 soil h-1 as described by Tabatabai and Bremner (1969). Statistical analysis: The data on the observations made were analyzed statistically by applying the technique of analysis of variance for randomized block design as suggested by Panse and Sukhatme (1978). disturbance on their activity, enzymes can serve as sensitive indicators of soil quality (Dick et al., 1994: Nedunchezhiyan et al., 2013). The data pertaining to the effect of different INM treatments on the enzyme activities and their correlations with organic carbon content were presented in the table 2. Urease (µg of NH4+-N g-1 soil 2h-1) Urease activity ranged from 32.50 to 55.76 µg of NH4+ released g-1 soil 2h-1 at harvest of kharif rice (Table 4.4). The highest urease activity (µg of NH4+ released g-1 soil 2h-1) was recorded in the treatment receiving 75 % RD of NPK + 25 % N through FYM (T3) followed by T4 (53.38), T7 (52.25) and T8 (50.63). However, T4, T7 and T8 were on par with T3 but significantly superior to T2 (49.73), T5 (49.63) and T6 (45.88). Urease activity ranged from 54.23 to 30.14 µg of NH4+ released g-1 soil 2h-1 at harvest of rabi rice. The highest urease activity was recorded in the treatment of 75 % RD of NPK (T3) followed by T4 with urease activity of 53.84 µg of NH4+ released g-1 soil 2h-1 (Table 2). Long-term organic amendments increased the capacity of the small-sized fractions to protect soil microorganisms; urease activity was mainly located in that fraction (Kandeler, 1996), and the activities of total urease significantly correlated with microbial biomass-C (Klose and Tabatabai, 1999). Dehydrogenase (µg of TPF g-1 soil day-1) Results and Discussion Soil enzyme activity is an indirect indication on the activities of microbes which is directly correlated with soil microbial dynamics. Enzyme activity in the soil environment is considered to be a major contributor of overall soil microbial activity (Burns et al., 2013). Due to the effects of external Dehydrogenase is an enzyme that occurs in all intact viable microbial cells. These soil enzymes function as a measure of the metabolic state of soil microorganisms by relating it to the presence of viable microorganisms and their oxidative capacity. 1075 Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 Table.1 Long-term effects of INM on available N, P and K status (kg ha-1) of the soils after harvest of rice-rice system at Rajendranagar Treatments OC (%) Kharif T1 – Control Rabi T1 – Control T2 – 100 % RD of NPK T2 – 100 % RD of NPK T3 – 50 % RD of NPK + 50 % N through FYM T3 – 100 % RD of NPK T4 – 75 % RD of NPK + 25 % N through FYM T4 – 75 % RD of NPK T5 – 50 % RD of NPK + 50 % N through Paddy straw N P2O5 K2O Kharif Rabi Kharif Rabi Kharif Rabi Kharif Rabi 0.57 0.59 125.5 124.7 18.6 16.2 204.5 200.1 0.68 0.72 0.57 0.65 196.5 217.4 208.3 233.1 32.9 44.0 34.8 45.5 362.6 392.9 380.5 394.0 T5 – 100 % RD of NPK 0.67 0.70 0.61 0.68 230.0 179.8 216.0 200.7 40.4 33.9 40.6 32.6 363.3 320.6 355.7 347.2 T6 – 75 % RD of NPK + 25 % N through Paddy straw T7 – 50 % RD of NPK + 50 % N through Green leaf manure T6 – 75 % RD of NPK 0.71 0.70 183.1 198.2 32.3 30.3 306.4 332.6 T7 – 100 % RD of NPK 0.68 0.60 204.0 218.3 38.6 36.2 356.5 387.7 T8 – 75 % RD of NPK + 25 % N through Green leaf manure Initial T8 – 75 % RD of NPK 0.66 0.71 209.1 207.3 37.0 29.8 344.5 370.0 0.54 151.0 24.0 224.0 CD (P=0.05) NS 0.09 28.7 30.00 8.01 2.91 50.08 60.69 SEm + 0.04 0.03 9.70 10.23 2.67 1.03 16.65 20.47 1076 Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 Table.2 Long-term effects of INM on enzyme activities of the soils under rice-rice cropping system at Rajendranagar Treatments Kharif Acid phosphatase T1 – Control T2 – 100 % RD of NPK Rabi T1 – Control T2 – 100 % RD of NPK T3 – 50 % RD of NPK + 50 % N through FYM T4 – 75 % RD of NPK + 25 % N through FYM T3 – 100 % RD of NPK T4 – 75 % RD of NPK T5 – 50 % RD of Paddy straw T6 – 75 % RD of Paddy straw T7 – 50 % RD of Green leaf manure T8 – 75 % RD of Green leaf manure CD (P=0.05) SEm + NPK + 50 % N through Alkaline phosphatase (µg p-nitrophenol g-1 soil h-1) Kharif Rabi Kharif Dehydrogenase (µg of TPF g-1 soil day-1) Kharif Rabi Urease (µg of NH4+-N g-1 soil 2h-1) Kharif Rabi Rabi T5 – 100 % RD of NPK 38.55 44.41 79.28 77.78 65.20 35.60 52.60 70.50 68.43 58.33 98.22 108.54 149.70 142.98 128.93 94.51 101.53 129.53 123.76 105.83 290.06 359.03 488.19 477.43 408.20 255.79 302.34 364.25 358.01 325.61 32.50 49.73 55.76 53.38 49.63 30.14 42.61 54.23 53.84 46.95 NPK + 25 % N through T6 – 75 % RD of NPK 62.73 55.56 121.10 102.63 403.06 321.18 45.88 45.08 NPK + 50 % N through T7 – 100 % RD of NPK 70.86 64.40 137.26 117.73 451.15 357.23 52.25 49.76 NPK + 25 % N through T8 – 75 % RD of NPK 68.52 62.46 133.47 116.65 449.28 350.00 50.63 47.98 6.93 2.26 6.27 2.04 16.39 5.35 14.69 4.80 53.67 17.53 43.31 14.15 4.05 1.32 4.97 1.62 Table.3 Simple correlations between different organic carbon and soil enzyme activities Variables r-values Kharif Rabi Organic carbon vs Acid phosphatase 0.8567 0.8800 Organic carbon vs Alkaline phosphatase 0.8189 0.8203 Organic carbon Organic carbon vs vs Dehydrogenase Urease 0.7455 0.8233 0.7317 0.8891 1077 Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1073-1080 The results related to dehydrodenase activity revealed that highest activity (488.19 µg of TPF g-1 soil day-1) was recorded in treatment receiving 50 % RD of NPK + 50 % N through FYM (T3) at harvest of kharif crop whereas lowest activity observed in control (290.06 µg of TPF g-1 soil day-1). However, T3 showed on par results with T4 (477.43), T7 (451.15) and T8 (449.28) and significantly increased dehydrogenase activity (µg of TPF g-1 soil day-1) than T2 (359.03), T5 (408.20) and T6 (403.06) (Table 2). Dehydrogenase activity during rabi season indicated that highest activity (364.25 µg of TPF g-1 soil day-1) was observed in treatment 100 % RD of NPK (T3) followed by T4 (100 % RD of NPK) and T7 (100 % RD of NPK) and T8 (75 % RD of NPK) with dehydrogenase activity of 358.01, 357.23 and 350.0 µg of TPF g-1 soil day-1, respectively, and on par with T3. The lowest activity was observed in (255.79 µg of TPF g-1 soil day-1) control (Table 2). Marinari et al., (2000) showed that a higher level of dehydrogenase activity was observed in soil treated with vermicompost and manure compared to soil treated with mineral fertilizer. In this study, the combined effect of organic manure and chemical fertilizer was also better than that of only chemical fertilizers. Because biological energy matter such as organic manure can supply available energy, thus it can accelerate microorganism and enzyme cell multiplication to improve organism and enzyme living environment and then to increase soil organism and enzyme composition and activity (Li et al., 2000). Acid and Alkaline Phosphatase (µg pnitrophenol g-1 soil h-1) Phosphatases are a group of enzymes that are capable of catalyzing hydrolysis of esters and anhydrides of phosphoric acid. Tabatabai and Eivazi (1977) reported that air drying of the soils increased the activity of acid phosphatase and phosphodiesterase but decrease the activity of alkaline phosphatase. In soil ecosystems, these enzymes play critical roles in P cycles (Speir and Ross, 1978) as evidence showed that they were correlated to P stress and plant growth. The data on acid phosphatase activity revealed that T3 showed highest 79.28 and 70.50 µg pnitrophenol released g-1 soil h-1 at harvest of both kharif and rabi rice, respectively. The lowest 38.55 and 35.60 µg p-nitrophenol released g-1 soil h-1 in T1 (control). However, T3 and T4 were on par with each other (Table 2). All the INM treatments showed significant effect on acid phosphatise activity under both kharif and rabi. The results related to alkaline phosphatase activity revealed that T3 (50 % RD of NPK + 50 % N through FYM) showed highest activity 149.70 and 129.53 µg p-nitrophenol released g-1 soil h-1 at harvest of kharif and rabi crops, respectively. The lowest activity of 98.22 and 94.51 µg p-nitrophenol released g-1 soil h-1 was observed in T1 (control) at harvest of kharif and rabi crops, respectively (Table 2). All the treatments did not show any significant effect soil acid phosphatase activity. 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