224

DOI: 10.1002/jpln.200622032

J. Plant Nutr. Soil Sci. 2007, 170, 224233

Long-term effects of fertilization on some soil properties under rainfed soybean-wheat cropping in the Indian Himalayas
Ved Prakash1, Ranjan Bhattacharyya1, Govindan Selvakumar1*, Samaresh Kundu1, and Hari Shanker Gupta1
1

Vivekananda Institute of Hill Agriculture (Indian Council of Agricultural Research), Almora 263 601, Uttaranchal, India

Accepted July 12, 2006

Summary
This study aims to examine the effects of long-term fertilization and cropping on some chemical and microbiological properties of the soil in a 32 y old long-term fertility experiment at Almora (Himalayan region, India) under rainfed soybean-wheat rotation. Continuous annual application of recommended doses of chemical fertilizer and 10 Mg ha1 FYM on fresh-weight basis (NPK + FYM) to soybean (Glycine max L.) sustained not only higher productivity of soybean and residual wheat (Triticum aestivum L.) crop, but also resulted in build-up of total soil organic C (SOC), total soil N, P, and K. Concentration of SOC increased by 40% and 70% in the NPK + FYMtreated plots as compared to NPK (43.1 Mg C ha1) and unfertilized control plots (35.5 Mg C ha1), respectively. Average annual contribution of C input from soybean was 29% and that from wheat was 24% of the harvestable aboveground biomass yield. Annual gross C input and annual

rate of total SOC enrichment from initial soil in the 015 cm layer were 4362 and 333 kg C ha1, respectively, for the plots under NPK + FYM. It was observed that the soils under the unfertilized control, NK and N + FYM treatments, suffered a net annual loss of 5.1, 5.2, and 15.8 kg P ha1, respectively, whereas the soils under NP, NPK, and NPK + FYM had net annual gains of 25.3, 18.8, and 16.4 kg P ha1, respectively. There was net negative K balance in all the treatments ranging from 6.9 kg ha1 y1 in NK to 82.4 kg ha1 y1 in N + FYMtreated plots. The application of NPK + FYM also recorded the highest levels of soil microbial-biomass C, soil microbial-biomass N, populations of viable and culturable soil microbes.
Key words: long-term experiment / soil organic C / soil microbialbiomass C / soil microorganisms / FYM / inorganic fertilizers / sandy-loam soil

1 Introduction
Agricultural practices that improve soil quality and agricultural sustainability have received much attention from researchers and farmers. In any agro-ecosystem, soil receives considerable annual carbon (C) inputs through left-over biomass of leaf-fall, stubble, roots, and root exudates as well as through external sources like farmyard manure (FYM) and compost. Among the agricultural practices, in particular cultivation and organic amendments regulate the soil microbial biomass; this affects C mineralization, nutrient cycling, and turnover of organic matter (OM) (McGill et al., 1986). Seasonal crop growth may influence soil microbial dynamics by altering the temporal and spatial distribution of organic inputs (Franzluebbers et al., 1995), and a decline in the microbial biomass occurs after the introduction of arable agriculture (Saviozzi et al., 1994). Nutrient fluxes through microbial biomass are of at least one order of magnitude faster than in the remaining OM (Dalal, 1998) leading to the suggestion that microbial biomass could be used as an important indicator of changes of soil health and soil quality as influenced by agricultural management practices (Sparling, 1997). Hence, influence of long-term application of chemical fertilizer and manures on soil biological health is an important area of investigation for assessing sustainability. The biological degradation of soil is a less understood area, and it is caused due to the decline in diversity and activity of soil microflora and fauna. Often it is observed that increased inputs compensate for and mask losses in productivity associated with soil degradation (Eivazi et al., 2003).
* Correspondence: Dr. G. Selvakumar; e-mail: gselva74@rediffmail.com

Microorganisms, that comprise the soil microbial-biomass (SMB) pool, control the flow of C and cycling of nutrients in soil. The SMB is regarded as a major nutrient sink during C immobilization (growth) and a source during mineralization (decay). This fundamental process is important to the longterm conservation of SOC through the production of precursors of humic substances. The formation and stabilization of macro-aggregates in FYM-treated soils is likely to be a key mechanism for the protection and maintenance of SOM (Beare et al. 1997) and microbial habitat (Dick, 1992). Soil microbial properties such as microbial biomass, distribution and activity of microbial populations have been used to predict soil biological status and the effects of farm management practices on soil quality (Kaur et al., 2005). The critical cause-effect relationship between soil management and soil quality can only be established through long-term experiments (Lal and Stewart, 1995). Incorporation of OM in the form of FYM enhances the organic-C level of the soil (Kundu et al., 2002) and has direct and indirect effects on soil properties and processes. Longterm experiments (LTEs) are valuable for evaluating the effects of continuous cropping on the capacity of a system to sustain nutrient supply and productivity (Gami et al., 2001). Several LTEs conducted in S Asia, show yield trends and build ups of SOC of varying degrees in treatments which received annual carbon inputs through external application of FYM, while in some locations, annual application of NPK only showed marginal build up of SOC (Nambiar, 1994; Gami et al., 2001; Bhandari et al., 2002). However, most studies were restricted under irrigated conditions in rice-wheat systems. We found no report of yield trends, annual C input, and soil chemical and microbiological properties under a long1436-8730/07/0204-224

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J. Plant Nutr. Soil Sci. 2007, 170, 224233 term rainfed experiment, especially under subtemperate climate of the Indian Himalayas, where low-external-input agriculture is practiced. Hence, the objectives of this study were to examine the impacts of the continuous use of inorganic fertilizers with and without FYM on the yield trends of soybean and wheat, nutrient balances, and soil fertility (total soil organic C, total N, P, and K content, soil microbial-biomass C (SMB-C), soil microbial-biomass N (SMB-N), and populations of major groups of soil microorganisms).

Long-term effects of fertilization on soil properties 225 10 Mg ha1 (N + FYM), and (NPK) + FYM at 10 Mg ha1 (NPK + FYM). The treatments were distributed in a randomized block design with six replications over three uniformly leveled terraces. The net plot size was 5.4 m 2.0 m. Fertilizers used were urea for N, single superphosphate for P, and murate of potash for K. Based on the chemical analysis, the applied FYM had 320.0330.8 g kg 1 moisture and contained 231.4235.6 g C kg1, 7.17.5 g N kg1, 2.12.4 g P kg1, and 5.35.8 g K kg1 on oven-dry basis.

2 Materials and methods

2.1 Experimental site
This study was carried out in a long-term field experiment initiated in June 1973 on a Typic Haplaquept at the experimental farm of Vivekananda Institute of Hill Agriculture, located in the Indian Himalayan region at Hawalbagh (2936 N and 7940 E at an altitude of 1250 m a.s.l.) near Almora (State of Uttaranchal, India). Soil characteristics based on analysis of soil samples taken in 1973 are given in Tab.1. The climate of the mid-hill region of Uttaranchal is subtemperate. Total rainfall, maximum and minimum temperatures during growing periods of both the crops are given in Fig.1. The field was newly reclaimed for cultivation in 1973. Prior to 1973, the field was native grassland, which was yearly cut and grazed.

2.3 Crop management

Farmyard manure was applied on a fresh-weight basis before sowing of soybean crop. Soybean (80 kg seed ha1) was sown in the month of June each year except during 19731975 when it was sown in the first week of July. Before seeding, the land was tilled two times with a spade up to a depth of 15 cm. Soybean cultivar Bragg was used from 1973 to 1998, and thereafter, it was replaced with the cultivar VLS 2. The crop was sown in rows 40 cm apart to a depth of 45 cm by hand. As per the local recommendation, a full dose of N, P, and K was incorporated before sowing. Allachlor [2,3,6-Tetrahydro-N-(1,1,2,2-tetrachloroethylthio) phthalimide] was sprayed 2 d after sowing at 1.5 kg active ingredient ha1 in all the plots to control weeds. Plant-protection measures were applied as and when required to control diseases and pests. After soybean harvest, wheat was sown in the first week of November during 197374 to 197576, in the second fortnight of October during 197677 to 198687 and thereafter (198788 to 200405) in the first fortnight of October. Wheat (cultivar Sonalika from 197374 to 198889, VL 421 from 198990 to 199899, and VL 616 from 19992000 to 200203) was sown (100 kg seed ha1) in rows 22.5 cm apart to a depth of about 56 cm by hand.

2.2 Experimental design and treatments

The experiment included two crops per year, soybean and wheat, with six treatment combinations viz., no fertilizer and manure (unfertilized control); 20 kg N ha1 + 33.2 kg K ha1 (NK); 20 kg N ha1 + 35.2 kg P ha1 (NP); 20 kg N ha1 + 35.2 kg P ha1 + 33.2 kg K ha1 (NPK), 20 kg N ha1 + FYM at

Table 1: Initial properties of the soil as recorded in the year 1973 during the start of the experiment. Parameters pH (soil : water, 1:2) EC (dS m1) CEC [c mol
+

Values in 015 cm soil layer 6.2 0.08 (kg soil)1] 8.7 1.32 5.1 12.8 0.46 0.127 0.24 0.012 0.64 0.065 78.1 16.1 5.8

Methods followed Jackson (1973) Jackson (1973) Jackson (1973) Core Sampler Method Walkley and Black (1934)

Bulk density (Mg m3) Oxidizable soil organic C (g (kg soil)1) C : N ratio Total N (g (kg soil)1) Available N (g (kg soil)1)

Wet Digestion method Alkaline KMnO4 method Page et al. (1982) Olsen et al. (1954) Wood and De Turk (1940) Standford and English (1949) Hydrometer Method

Total P (g (kg soil)1) Olsen-P (g (kg soil)1)

Nonexchangeable K (g (kg soil)1) NH4OAc-K (g (kg soil)1) Sand (%) Silt (%) Clay (%) EC = Electrical conductivity, CEC = Cation-exchange capacity

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Prakash, Bhattacharyya, Selvakumar, Kundu, Gupta

Rainfall ( Soybean growing period) Max. Temp. ( Soybean growing period) Max. Temp. ( Wheat growing period) Rainfall (Wheat growing period) Min. Temp. (Soybean growing period) Min. Temp . ( Wheat growing period) 35

J. Plant Nutr. Soil Sci. 2007, 170, 224233

1200

1000

30

25 800 20 600 15 400 10 200 Temperature ( C) Rainfall (mm)

O

0 1973-74

0 1978-79 1983-84 1988-89 Years 1993-94 1998-99 2003-04

Figure 1: Rainfall and temperature distribution over the years during soybean and wheat growing seasons at Hawalbagh, Almora.

Wheat crop was grown on residual fertility. Wheat was harvested at 5 cm above soil surface in the fourth week of April, and aboveground biomass was removed from the field. The grain yield of soybean and wheat was adjusted to 90 and 120 g water kg1, respectively.

sieve, and the treatments were replicated in the pots. After harvest of aboveground biomass (shoot), soil was removed gently by placing the pot on a sieve and washed slowly using a water jet. Root biomass with entangled nodules was recovered, dried, and dry weights of root and nodules recorded. From dry weights, root-to-shoot and nodule-to-root ratios were estimated. During 19992002, soybean leaves were collected after senescence, nodules were excavated at 65 d after sowing, and roots were excavated at 85 d after sowing. Samples were analyzed for total C and total N content using a CHN analyzer for calculation of C inputs. Rhizodeposition of C from root turnover and exudates was assumed to be 10% of the harvestable aboveground biomass of soybean (Shamoot et al., 1968). Organic C of FYM was 22% C on oven-dryweight basis. For soybean crop, annual N inputs to soil was calculated based on the N inputs by leaf-fall, roots, and nodule. We could not estimate the N2-fixation component and calculated apparent N balance. The addition of wheat biomass to soil was through stubble (3%4% of the straw yield) and roots (30% of the harvestable aboveground biomass as observed by Chander et al., 1997). Samples of stubble and roots of wheat were collected after harvest (19992000 to 20022003) from all the plots and C and N inputs through stubble and roots were calculated based on C and N determination with a CHN analyzer. The contribution of C and N through rhizodeposition from wheat was estimated by multiplying the values of total root C and N inputs, respectively, with a factor of 1.4 as observed by Regmi (1994). During growth of soybean and wheat, weeds were removed so that C and N inputs from roots and rhizodeposition by the weeds were not considered. www.plant-soil.com

2.4 Estimation of apparent nutrient balance in soil

During the period 19952005, samples of soybean and wheat (grain and straw) were collected at maturity from each plot, oven-dried at 65C to constant weight, and ground to pass through a 0.5 mm sieve. The samples were digested in a mixture of HNO3 and HClO4 (3:1). Concentrations of P and K in the digested samples were estimated by the ammonium molybdate (Olsen and Sommers, 1982) and by flame-photometry methods, respectively. For calculation of P and K removal by soybean and wheat, the average yields from 1995 to 2005 were used. However, N removal was estimated using average yields of soybean and wheat from 1973 to 2005.

2.5 Estimation of annual C input to soil

Using biomass yield of soybean and wheat, annual inputs of C added to the soil were computed. In case of soybean, added biomass was through leaf-fall, roots, and nodules. Leaf-fall from all treatments was manually collected during the last 5 y of experimentation from 45 d after sowing until harvest, dried, and dry-weight recorded. Root and nodule biomass of soybean were calculated using the root-to-shoot and noduletoroot biomass ratios recorded from a pot experiment (Kundu et al., 2001). A pot-culture experiment was conducted in earthern pots of 10 kg capacity during the period 19992002. The treatment-wise soil samples collected from the field experiment were processed to pass through 2.0 mm 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

J. Plant Nutr. Soil Sci. 2007, 170, 224233

Long-term effects of fertilization on soil properties 227 ter rainfall, were not considered for the trend analysis. Linearregression analysis was also performed to determine the relationship between initial soybean and wheat yields and yield decline (for first four treatments) across the 32 y period. Statistical analyses were done using Tukeys test with SAS program. Treatment means were compared at 5% level of significance. Technical expertise was provided by Indian Agricultural Statistical Research Institute, New Delhi.

2.6 Soil chemical and microbiological analysis

Surface soils (015 cm) were sampled with an auger ( 5 cm) during the peak-growth stages of soybean (August 2004) and wheat (March 2005), respectively. Ten soil cores were taken from the plots in the space between the rows of soybean and wheat crop and bulked separately for each plot. All samples were sieved through 2 mm sieve, adjusted to 40% of water-holding capacity, and pre-incubated for 10 d to permit uniform rewetting and to allow microbial activity to equilibrate after the initial disturbances. Such equilibrated samples were used for the estimation of the soil microbiological properties. Subsamples of each sample were air-dried and ground for chemical analysis. The pH of the soil was measured with a glass electrode using a soil-to-water ratio of 1:2.5. Representative subsamples were ground to pass through a 0.2 mm sieve to determine total soil organic C and total soil N using a CHN analyzer (Foss Heraeuss CHN-O Rapid) by the dry-combustion method. Total P in soil samples was determined by the method of Olsen and Sommers (1982), and total K was determined following the method of Prat (1967). The chloroform fumigation-incubation method (Jenkinson and Powlson, 1976) was used to estimate the soil microbialbiomass C (SMB-C), while the fumigation-extraction method as described by Jrgensen (1995) was used to estimate the soil microbial-biomass N (SMB-N). The viable and culturable microbial population was determined by standard serialdilution plate technique (Subba Rao, 1977) using selective media. Soil-extract agar, Martins rose bengal agar, and Kenknight & Munaiers agar obtained from Hi-Media, India, were used for the enumeration of total bacteria, fungi, and actinomycetes, respectively. Jensens N-free agar and Pseudomonas agar (Hi-Media, India) were used for the enumeration of heterotrophic free-living N2-fixing bacteria and fluorescent pseudomonads, respectively. Colonies that showed fluorescence under an ultraviolet source were designated as fluorescent pseudomonads.

3 Results and discussion

3.1 Yield of soybean and wheat
All the fertilized treatments significantly (p < 0.05) improved soybean yields relative to unfertilized control (Tab. 2). The mean yield of soybean and wheat observed under unfertilized control treatment suggests that the soil of our experimental field is capable of supporting average yield of soybean and wheat to the tune of 0.56 and 0.72 Mg ha1, respectively, without any external application of nutrient inputs. Soybean responded to inorganic-nutrient application (NPK), increasing average yield of soybean and wheat to 0.86 and 0.40 Mg ha1, respectively (Tab. 2). The highest mean grain yield of soybean was observed in the plots under NPK + FYM and the lowest under unfertilized control. Wheat grain yields were significantly affected by residual effect of the treatments. Due to residual effect of FYM, the productivity of succeeding wheat crop was also enhanced by 43% and 71% in N + FYM and NPK + FYM treatments, respectively, over NPK treatment. The yield data obtained over the period 19732005 clearly demonstrates benefits of organics apart from N, P, and K supply, such as improvements in microbial activities, better supply of macro- and micronutrients such as S, Zn, Cu, and B, which are not supplied by inorganic fertilizers, and less losses of nutrients from the soil (Yadav et al., 2000). The higher wheat yields obtained on FYM-amended plots were possibly caused by the better supply pattern of N, P, and K and improved soil physical conditions (YadvinderSingh et al., 2004).

2.7 Statistical analysis

Simple linear-regression analyses of grain yields across years were performed to determine yield trends (slopes). Low wheat yields in 1977 and 1984, associated with very low win-

3.2 Nutrient balance in soil

Following harvest of soybean, considerable amount of N is generally added to the soil through the residual biomass consisting of leaf-fall, nodules, and rhizodeposition. The annually

Table 2: Soybean and wheat yields (19732005) as affected by the long-term application of farmyard manure and fertilizers. Treatments Control NK NP NPK N + FYM NPK + FYM Soybean yield (Mg ha1) 0.56 f* 0.63 e 0.88 d 1.42 c 2.40 b 2.90 a Soybean yield change (kg ha1 y1) 17 19 22 21 +28 +27 Wheat yield (Mg ha1) 0.72 f 0.81 e 0.92 d 1.12 c 1.60 b 1.90 a Wheat yield change (kg ha1 y1) 2 1 7 6 +6 +5

* Means followed by the same lower-case letter within a parameter indicate that they are not significantly different (at p < 0.05 according to Tukeys multiple-range test).

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Prakash, Bhattacharyya, Selvakumar, Kundu, Gupta

J. Plant Nutr. Soil Sci. 2007, 170, 224233 wheat and fertilizers and in all the treatments (Tab. 4), thus showing a net negative N balance ranging from 27.0 kg ha1 y1 in NK- to 67.1 kg ha1 y1 in NPK-treated plots. Our results are of similar trends with that of Kundu et al. (1996). It was observed that the soils under the unfertilized control, NK, and N + FYM treatments suffered a net annual loss of 5.1, 5.2, and 15.8 kg P ha1, respectively, whereas the soils under NP, NPK, and NPK + FYM treatments had net annual gains of 25.3, 18.8, and 16.4 kg P ha1, respectively (Tab. 3). In plots under N + FYM and NPK + FYM treatments, total P uptake by soybean and wheat crops increased by 93% and

added N of soybean and wheat ranged from 16.7 to 101.4 kg ha1 and from 12.1 to 32.8 kg ha1, respectively (Tab. 3), depending upon the biomass-yield responses to applied fertilizer and FYM. The estimated N contributed through leaffall, root, nodules, and rhizodeposition varied largely depending on the biomass yield of soybean. Buresh and DeDatta (1991) found that leaf-fall during crop-legume development and the nodulated roots could each supply up to 40 kg N ha1. Removal of N by the system was 73% higher in the plots under NPK + FYM treatment than that under NPK treatment (151.9 kg N ha1). It was found that annual removal of N by the crops surpassed the amount of annual N applied to the soil through addition of residual biomass of soybean and

Table 3: Apparent annual N, P, and K balances as affected by the long-term application of farmyard manure and fertilizers. Treatments N Annual addition by soybean crop* (kg ha1) Control NK NP NPK N + FYM 16.7 e 16.9 e 30.4 d 46.2 c 79.5 b Annual addition by wheat crop** (kg ha1) 12.1 d 13.5 d 14.4 d 18.6 c 27.3 b 32.8 a Addition Removal Balance as (kg ha1) (kg ha1) fertilizer/ manure (kg ha1) 0 20 20 20 75.8 75.8 68.8 e 77.4 e 99.5 d 151.9 c 220.3 b 261.8 a 40 c 27 d 34.7 cd 67.1 a 37.7 cd 51.8 b P Addition Removal Balance as (kg ha1) (kg ha1) fertilizer/ manure (kg ha1) 0 0 35.0 35.0 15.4 50.4 5.1 e 5.2 e 9.7 d 16.2 c 31.2 b 35.5 a 5.1 e 5.2 e +25.3 a +18.8 b 15.8 d +16.4 c K Addition Removal Balance as (kg ha1) (kg ha1) fertilizer/ manure (kg ha1) 0 33.2 0 33.2 37.4 70.6 32.4 e 40.1 d 37.0 d 62.9 c 119.8 b 152.2 a 32.4 b 6.9 c 37.0 b 29.7 b 82.4 a 81.6 a

NPK + FYM 101.4 a

* Addition from N2 fixation was not estimated. Annual addition by soybean crop = N added through leaf-fall, roots, and nodule ** Annual N addition by wheat crop = N added through stubble, roots, and rhizodeposition *** Means followed by the same lower-case letter within a parameter indicate that they are not significantly different (at p < 0.05 according to Tukeys multiple-range test). Table 4: Estimated annual C input by the soybean-wheat cropping system as affected by the long-term application of farmyard manure and fertilizers. Treatments Harvestable aboveground biomass (shoot) yield of soybean (kg ha1) Control NP NK NPK N + FYM 1301 f* 2146 d 1605 e 3410 c 5809 b Annual C input (kg ha1) by soybean crop and farmyard manure Leaffall Roots Nodules Rhizo- FYM deposition Total Harvestable Annual C input abovewheat crop ground Stubble Roots biomass (shoot) yield of wheat (kg ha1) 2150 de 2533 d 2396 d 3248 c 4638 b 5504 a 37 c 38 c 41 c 39 c 60 b 75 a 204 e 244 d 236 d 307 c 450 b 514 a (kg ha1)$ by Rhizo- Total deposition Total C input by the system (kg ha1)

47 e 126 d 45 e 289 c 431 b 488 a

173 f 276 d 201 e 400 c 549 b 723 a

18 e 28 d 22 e 57 c 73 b 91 a

130 f 215 d 161 e 341 c 581 b 717 a

1587 1587

368 645 429 1087 3221 3606

286 e 341 d 331 d 430 c 631 b 719 a

527 e

895

623 d 1268 608 d 1037 776 c 1863 1141 b 4362 1308 a 4914

NPK + FYM 7170 a

We assumed that entire amount of biomass through leaf fall and FYM and 90% of the biomass contributed by roots, nodules, and rhizodeposition remained within 045 cm soil depth. $ We assumed that entire amount of biomass through stubbles and 80% of the biomass contributed by roots and exudates remained within 045 cm soil depth. * Means followed by the same lower-case letter within a parameter indicate that they are not significantly different (at p < 0.05 according to Tukeys multiple-range test).

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J. Plant Nutr. Soil Sci. 2007, 170, 224233 119% over that in NPK-treated plots (16.2 kg P ha1 y1). Gami et al. (2001) also observed a positive P balance in fertilizer Pamended plots after 20 years of rice-wheat cropping. It can be inferred that mean annual removal of K by the crops surpassed the amount of annual K applied to the soil in all the treatments (Tab. 3), thus showing a net negative K balance ranging from 6.9 kg ha1 y1 in NK- to 82.4 kg ha1 y1 in N + FYMtreated plots. Both NK- and NPK-treated plots received similar amounts of fertilizer K, but the plots under NPK treatment showed considerably higher negative balance due to significant yield increase of soybean to applied P. Even in NPK + FYM treatment, where 70.6 kg K ha1 were applied annually, the removal of K exceeded the amount of K added, with a negative K balance of about 81.6 kg K ha1 y1. Negative K balances are attributed to labile K either leached from the plough layer or being fixed in less readily available form (Blake et al., 1999; Singh et al., 2002a, b). Results of several other LTEs showed that considerable amounts of K have either leached out of or have been fixed in the soil layers (Srinivasa Rao et al., 1999; Singh et al., 2002a).

Long-term effects of fertilization on soil properties 229 C ha1 y1), an additional gain of 932 kg C ha1 y1 occurred under soybean with NPK + FYM compared to NPK only. Like soybean, roots and rhizodeposition from wheat also contributed the highest amount of C input to soil (Tab. 5). Total C input from unfertilized wheat was 527 kg ha1 y1 and 1308 kg ha1 y1 from wheat grown on residual fertility of NPK + FYM. The residual effect of FYM resulted in an additional gain of 532 kg C ha1 y1 in wheat over that under NPK only. The harvestable aboveground biomass yield of wheat varied due to variation in residual fertility from different fertilizer treatments applied to soybean. Average C input from wheat to soil (Tab. 4) was 24% of the harvestable aboveground biomass (R2 = 0.99*, p < 0.01). Kundu et al. (2001) reported a value of 32%. Kemper et al. (1998) reported a root-to-shoot ratio of 0.25 in wheat. In wheat, considerable amount of C input (on average 445 kg C ha1 y1) to soil originated from rhizodeposition. Study with 14C indicated that rhizodeposition during the growing season of wheat was 30% of the total C accumulated aboveground (Keith et al., 1986).

3.3 Annual C input to soil

The average annual total C input to soil from soybean and wheat (Tab. 4) varied with aboveground yield responses of both crops under different fertilizer application. Average C input from soybean to soil (Tab. 4) was 29% of the harvestable aboveground biomass (R2 = 0.99*, p < 0.01). Roots of soybean crop contributed the highest amount of annual addition of biomass to soil. Kundu et al. (1997) also estimated that root biomass of soybean added the highest amount (31% of the aboveground biomass) to soil. Mean C concentration of soybean leaves, roots, and nodules was 38%, 34%, and 32%, respectively. Without fertilizer, total C input from soybean was 368 kg ha1 y1. Soil organic-C inputs were 1087 kg ha1 y1 with NPK and 3606 kg ha1 y1 with NPK + FYM. Apart from FYMs own contribution (1587 kg

3.4 Soil chemical properties

Continuous cropping over the years has resulted in a marginal reduction in soil pH (Tab. 5). Maximum reduction of 0.61 units in soil pH was observed in NPK treatment in the surface layer. Inspite of build-up of Ca and Mg in soil profiles under N + FYM and NPK + FYM treatment (Bhattacharyya et al., unpublished data), there was reduction in soil pH after 32 y of cropping. The higher plant-biomass production along with greater metabolism under manure-amended soils might have lead to higher soil acidity (Bellakki et al., 1998). Another important factor regulating mineral soil pH is the capacity of the soil to buffer protons produced by plant metabolism, biological transformations in the soil, or imported to the area with decomposition. The decrease in pH in the fertilizer-amended plots was due to decrease of exchangeable base cations like

Table 5: Chemical properties* of the soil (015 cm) as affected by the long-term application of farmyard manure and fertilizers. Treatments Initial soil Control NK NP NPK N + FYM NPK + FYM pH 6.2 5.90 a** 5.81 a 5.72 a 5.59 a 5.86 a 5.82 a Total SOC (g kg1) NA 6.19 e 6.74 d 7.63 d 8.08 c 11.12 b 11.91 a Total N (g kg1) 0.46 0.55 d 0.57 d 0.68 c 0.64 d 0.76 b 0.83 a C : N ratio 12.8 11.5 d 11.9 c 11.0 e 12.5 b 14.1 a 14.2 a Total P (g kg1) 0.24 0.228 d 0.210 e 0.307 b 0.262 c 0.235 d 0.347 a Total K (g kg1) NA (0.64)*** 4.57 b (0.50 c) 4.64 a (0.59 a) 4.50 cd (0.48 cd) 4.52 c (0.52 b) 4.50 cd (0.46 d) 4.56 b (0.49 c )

* Mean value of observations during the growth of 32nd soybean and wheat crops SOC = Soil organic C, NA = Not available. ** Means followed by the same lower-case letter within a parameter indicate that they are not significantly different (at p < 0.05 according to Tukeys multiple-range test). *** Data in parentheses indicate the nonexchangeable soil K.

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J. Plant Nutr. Soil Sci. 2007, 170, 224233 treatment and possibly because of that reason, the gain of total P in soil profile was the highest. Although the total P balance at 015 cm soil depth in NPK + FYMtreated plots was highly positive, the same in N + FYM treatment was negative. This might be due to the fact that the major P fraction added through FYM is in the organic pool, which mineralized slowly with time apart from the fact that in the later treatment, no mineral P was added (Yadvinder-Singh et al., 2004). We could not judge the balance of total soil K at the surface soil layer due to unavailability of total soil-K data for the initial soil. The highest concentration of total K in 015 cm soil layer was found in NK-treated plots, followed by the unfertilized control plot (Tab. 5). It was observed that total K content in unfertilized plots was higher than that in plots receiving no fertilizer K (NP) after 32 y of cropping, and other workers found the similar results (Dikshit et al., 1995; Kumari, 2004). Where fertilizer K was limited by NP applied, K was probably more effectively utilized from the soil layer resulting in depletion of soil K reserves. Considerable depletion of nonexchangeable K content was observed throughout the soil profiles after 32 crop cycles for all the treatments. The results show that the surface soil layer (015 cm depth) under all the treatments was in net loss of nonexchangeable K, ranging from 100 kg ha1 in the plots under NK treatment to a maximum of 360 kg ha1 for the N + FYMtreated plots (Tab. 5). For the unfertilized plots, the rate of depletion of nonexchangeable K was 8.8 kg ha1 y1, and this rate of depletion decreased to 7.5 and 3.1 kg ha1 y1 due to application of NPK and NK. This depletion of nonexchangeable K might have caused the negative K balances in the plots under all the treatments. The greater depletion of nonexchangeable K was due to higher crop uptake in comparison to addition of K along with higher release rate (of nonexchangeable K to exchangeable and soil-solution K) in this light-textured soil (Singh et al., 2002b).

Ca+2, Mg2+, and K+ by leaching, runoff losses, and crop uptake (Bellakki et al., 1998). Soil organic-C concentrations remained positively changed from the initial values in plots under unfertilized control (Tab. 5). At the end of 32 y, application of NPK showed significantly higher SOC (8.1 g C kg1) over the unfertilized control (6.2 g C kg1) in 015 cm soil layer. Application of NPK also showed significantly higher SOC concentrations over either NP or NK. There was significant improvement in SOC in NP (7.6 g C kg1) over NK (6.7 g C kg1) at the same depth. Soils under NPK + FYM treatment contained 47% higher SOC in the 015 cm soil layer (11.9 g C kg1) over those observed in NPK-treated plots. Considerably higher build-up of SOC in NPK + FYMtreated plots as compared to the plots under N + FYM treatment indicated the beneficial effect of balanced fertilization in improving SOC status of the soil. Soil organicC concentrations marginally improved from the start of the experiment in the unfertilized plots due to C addition through the roots and crop residues, higher humification rate constant, and lower decay rate (Kundu et al., 2002). The higher concentration of C in the fertilizer + FYM-treated plots was a result of increased yields of roots and plant residues and the application of organic C through FYM (Singh et al., 1998). Differences in C storage were due to greater roots and wheat stubble inputs for higher-yielding treatments. It was found that there were significant improvements of total soil N (TSN) in all the treatments over unfertilized soils (Tab. 5). Total soil N concentrations remained positively changed from the start of the experiment in the unfertilized plots (0.55 g kg1 in the 015 cm soil layer). The highest TSN content in the 015 cm layer was observed under NPK + FYM treatment (0.835 g kg1). Soils under NPK + FYM treatment contained higher TSN by 31% at the 015 cm soil depths over those observed under NPK-treated plots. The C : N ratio under chemically fertilized plots showed marked reduction, while there was an increase in the C : N ratio in the FYMamended plots. Farmyard manure is known to stimulate biological N2 fixation in the soil, which may have been responsible for the increase in TSN (Ladha and Kundu, 1997) in the FYM-treated plots over those NPK-treated plots, apart from FYMs own N contribution. The positive balance of TSN observed under all the treatments could be attributed to higher SOC concentrations and low rate of mineralization under subtemperate climatic conditions. Total N concentrations increased in all the treatments despite the observed negative balances of N in all the treatments. We could not measure the biological N2 fixation in this experiment. Hence, it is inferred that the biological N2 fixation might have contributed largely to compensate the crop uptake and other losses (if any). The status of total P content showed that there was net depletion of total P in the treatments, which did not receive any P dressing as fertilizer (Tab. 5). The apparent P balances were also negative in the plots under the unfertilized control, NK, and N + FYM treatments. In the NP- and NPK-treated plots, the build-ups of total P after 32 y were 67 and 22 mg P (kg soil)1, respectively, in the 015 cm soil layer. About 15.4 kg P ha1 were annually supplied to the soil through FYM apart from fertilizer-P in the plots under NPK + FYM 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

3.5 Soil microbiological properties

The treatments receiving FYM recorded significantly higher levels of SMB-C and SMB-N as compared to the plots that did not receive manure for the past 32 y (Tab. 6). The NPK + FYM treatment recorded the highest SMB-C and SMB-N. The SMB-C content of the surface soil under NPK was 165.6 mg (kg soil)1 which was 73.4% higher than in the unfertilized control plot. The biomass C : N ratio was also the highest in plots under NPK + FYM treatment followed by N + FYM treatment. The increase of SMB under FYM-amended soils could be attributed to several factors, such as higher moisture content, greater soil aggregation, and higher C content. The FYM-amended plots provided a steady source of organic C to support the microbial community compared to NPK-treated plots. Fauci and Dick (1994) suggested that soils with a longterm history of organic amendments, such as animal manure, generally maintain high levels of microbial biomass. It should be noted here that the population of the different groups of soil microbes is only an indicator of the trends in the dynamics of the different groups. In general, it was observed that the application of FYM had a beneficial effect on the population levels of all the soil microorganisms, irrespecwww.plant-soil.com

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Long-term effects of fertilization on soil properties 231

Table 6: Microbial-biomass C and N content and population of different viable and culturable soil microbes during the 32nd cropping cycle. Treatments Microbial biomass C (mg (kg soil)1) Microbial Total Bacteria biomass N (106 cfu (g oven-dry soil)1) (mg (kg soil)1) Soybean Control NK NP NPK N + FYM NPK+ FYM

Fungi (103 cfu (g oven-dry soil)1) Soybean 10.0 d 15.0 c 20.0 b 22.0 b 27.5 a 27.5 a Wheat 30 b 36 b 43 ab 50 a 50 a 56 a

Actinomycetes (102cfu (g oven-dry soil)1) Soybean 10.7 bc 12.5 b 13.2 b 13.3 b 14.0 b 19.2 a Wheat 10.3 bc 11.3 bc 13.3 b 14.3 b 16.0 a 18.3 a

Heterotrophic free-living N2 fixers (103 cfu (g oven-dry soil)1) Soybean 5.2 bc (37 c) 4.0 c (51 bc) 5.7 bc (59 b) 8.7 b (64 b) 10.7 b (96 a) 15.5 a (104 a) Wheat 14 c (32 e) 16 c (42 d) 17 c (47 d) 30 b (63 c) 27 bc (82 b) 40 a (97 a)

Wheat 1.8 c 2.6 bc 5.0 b 6.7 b 10.7 a 12.6 a

95 f** 131 e 148 d 166 c 216 b 248 a

17 f 23 e 25 d 27 c 31 b 35 a

3.4 c 3.5 c 4.5 c 8.2 c 15.5 b 23.0 a

Mean value of observations during the growth of 32nd soybean and wheat crops as recorded in the surface layer (015 cm soil depth). Data in parentheses indicate Fluorescent Pseudomonads (103 cfu (g oven-dry soil)1). * Means followed by the same lower-case letter within a parameter indicate that they are not significantly different (at p < 0.05 according to Tukeys multiple-range test).

tive of the crop that was cultivated. The population levels of soil fungi and heterotrophic free-living N2-fixing bacteria were higher in the soils under wheat as compared to the soils under soybean in all the treatments, while the total soil bacterial population, actinomycetes, and fluorescent pseudomonad populations were higher in the soils under soybean (Tab. 6). The results showed that continuous application of FYM in combination with N or NPK had higher bacterial, fungal populations, and the population of actinomycetes in the surface layers of the soil. In the plots under NPK + FYM treatment, we observed the highest population of free-living N2-fixing bacteria, which was significantly higher than the population levels recorded in the plots under chemical fertilization alone. It was further observed that the plots receiving P fertilization recorded higher levels of heterotrophic free-living N2-fixing bacteria than the plots that did not receive P. The population of fluorescent pseudomonads was also higher in the plots receiving FYM along with N or NPK. The predominance of fungi and heterotrophic free-living N2-fixing bacteria in the wheat soils could be attributed to the nature of the root exudates from wheat crop and the weather conditions that prevail during the wheat-growing season. Wheat being a cereal crop is more responsive to colonization and proliferation of free-living N2-fixing bacteria like Azospirillum (Boddey et al., 1986). The low temperatures that prevailed during the wheat-growing season could have influenced the proliferation of fungi, which require a lower temperature for their growth and proliferation compared to bacteria. Besides the huge amount of biomass in the form of leaf litter and underground stubbles from the previous soybean crop could have influenced the predominance of fungi in the wheat soils.

yield of soybean in the plots under NPK + FYM treatment was 98.7% higher than that observed under NPK treatment (1.43 Mg ha1). The residual effect of NPK + FYM treatment was also considerable, resulting in 71.2% increase in wheat yield over that observed under NPK treatment (1.10 Mg ha1). Addition of FYM with NPK fertilizers increased total soil organic C (SOC), N, and P by 37.8%, 20.8%, and 30.2%, respectively, over NPK-alone treatment, and FYM application along with N or NPK showed significantly higher microbialbiomass C and populations of microorganisms over chemically fertilized plots. Application of inorganic fertilizer resulted in higher SOC over the unfertilized control, due to greater crop residue and root biomass produced. The annual rate of increase in SOC content over initial condition was 74, 328, 711, and 900 kg C ha1 under unfertilized, NPK, N + FYM, and NPK + FYM treatments. Thus, the rainfed soybeanwheat cropping in the subtemperate Indian Himalayas is a system that can sequester SOC. Long-term application of N, P, and K resulted in declining yields of soybean. The results clearly reveal that current fertilizer recommendations are inadequate in the long run, and application of FYM appeared to be important in this experiment. So, a nutrient management strategy is needed to ensure high and stable overall productivity in this rainfed soybean-wheat cropping system.

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4 Conclusions
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