Inorganic Phosphorus Fractions and Phosphorus Availability in a Calcareous Soil Receiving 21-Year Superphosphate Application 1

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1 Pedosphere 20(3): , 2010 ISSN /CN /P c 2010 Soil Science Society of China Published by Elsevier Limited and Science Press Inorganic Phosphorus Fractions and Phosphorus Availability in a Calcareous Soil Receiving 21-Year Superphosphate Application 1 WANG Jun 1,2, LIU Wen-Zhao 1, 2, MU Han-Feng 2 and DANG Ting-Hui 1 1 State Key Laboratory of Soil Erosion and Dryland Farming on the Loess Plateau, Institute of Soil and Water Conservation, Northwest A&F University, Yangling (China) 2 College of Urban and Environment, Northwest University, Xi an (China) (Received December 28, 2009; revised March 25, 2010) ABSTRACT A long-term (21-year) field experiment was performed to study the responses of soil inorganic P fractions and P availability to annual fertilizer P application in a calcareous soil on the Loess Plateau of China. Soil Olsen-P contents increased by 3.7, 5.2, 11.2 and 20.6 mg P kg 1 after 21-year annual fertilizer P application at 20, 39, 59, and 79 kg P ha 1, respectively. Long-term fertilizer P addition also increased soil total P and inorganic P (Pi) contents significantly. The contents of inorganic P fractions were in the order of Ca 10 -P > Ca 8 -P > Fe-P > Al-P > occluded P > Ca 2 -P in the soil receiving annual fertilizer P application. Fertilizer P application increased Ca 8 -P, Al-P and Ca 2 -P contents as well as their percentages relative to Pi. Pi application increased Fe-P and occluded P contents but nor their percentages. Soil Ca 10 - P content remained unchanged after fertilizer P application while its percentage relative to Pi declined with increasing fertilizer P rate. All Pi fractions but Ca 10 -P were correlated with Olsen-P significantly. 90% of variations in Olsen-P could be explained by Pi fractions, and the direct contribution of Ca 8 -P was predominant. Long-term annual superphosphate application would facilitate the accumulation of soil Ca 8 -P, and thus improve soil P availability. Key Words: long-term experiment, Olsen-P, path analysis, Loess Plateau Citation: Wang, J., Liu, W. Z., Mu, H. F. and Dang, T. H Inorganic phosphorus fractions and phosphorus availability in a calcareous soil receiving 21-year superphosphate application. Pedosphere. 20(3): INTRODUCTION Phosphorus (P) is an essential element for plant growth. Therefore, maintenance of an adequate amount of soil P through application of inorganic and/or organic P is critical for the sustainability of cropping systems (Sharpley et al., 1994). The availability and fractions of soil P may change due to long-term continuous P fertilization besides its yield-increasing effect (Fan et al., 2003; Lai et al., 2003). Many researches have found that soil phosphorus availability would increase after long-term fertilizer P application (Halvorson and Black, 1985; Samadi and Gilkes, 1998; Fan et al., 2003; Lai et al., 2003; Zhang et al., 2004). However, such an effect of fertilizer P application varies with climatic condition, soil type, and soil test method employed, as well as the rate of fertilizer P applied (Zhang et al., 2004). In calcareous soils, soil Olsen-P content (Olsen et al., 1954) is a good indicator for estimating soil P availability. Samadi and Gilkes (1998) found that soil Olsen-P increased by 200% in fertilized soil compared with virgin soil in Western Australia. On the Loess Plateau of China, both Fan et al. (2003) and Lai et al. (2003) reported that compared with soil without fertilizer P application, soil Olsen- P increases from about 3 to over 20 mg kg 1 after 26 kg P ha 1 application under a winter wheat 1 Supported by the National Basic Research Program (973 Program) of China (No. 2005CB121102), the Knowledge Innovation Program of the Chinese Academy of Sciences (No. kzcx2-yw-424), and the National Natural Science Foundation of China (No ). 2 Corresponding author. wzliu@ms.iswc.ac.cn.

2 SOIL PHOSPHORUS AND SUPERPHOSPHATE APPLICATION 305 monoculture system. Shen et al. (2004) found that fertilizer P treatments significantly increases soil Olsen-P in the first few years and then it remains stable under a rice monoculture system. However, how soil Olsen-P can increase with fertilization rates after long-term fertilization in calcareous soils has not been well understood. The knowledge of soil P fractions is important for investigating soil P availability, and several kinds of fractionation methods are available for different soil types (Chang and Jackson, 1957; Hedley et al., 1982; Jiang and Gu, 1989). In calcareous soils, soil inorganic P (Pi) represents the dominant component of the soil P pool, accounting for about 75% 85% of soil total P in calcareous soils in China (Jiang and Gu, 1989). Soil Pi was divided into various fractions such as Ca-P (HCl extractable P), Fe- and Al-P (non-occluded Fe- and Al-bound P), and occluded P (Chang and Jackson, 1957; Solis and Torrent, 1989). However, in calcareous soils, the majority of Pi exists in the various Ca-bound forms and there is a great difference in P availability among the Ca-P fractions. To better characterize Pi transformations, Jiang and Gu (1989) proposed a fractionation scheme for calcareous soils based on the methods described by Chang and Jackson (1957) and Hedley et al. (1982), where soil Pi was divided into six fractions (Ca 2 -P, Ca 8 -P, Al-P, Fe-P, occluded P, and Ca 10 -P) using a sequential extraction procedure, based on their availability and solubility. This has been confirmed by Gu and Qin (1997), Samadi and Gilkes (1998), Lai et al. (2003), and Shen et al. (2004) based on the simple correlation coefficients between Olsen-P and Pi fractions. Such sequential extraction has been widely used for the calcareous soils in Northern China (Gu and Jiang, 1990; Fan et al., 2003; Lai et al., 2003; Shen et al., 2004) and Western Australia (Samadi and Gilkes, 1998) to study the response and availability of soil P fractions to fertilizer application. Samadi and Gilkes (1998) found that in Western Australia the agricultural development of soils resulted in increases in all Pi fractions compared with their virgin analogues, and soil Ca 2 -P and Ca 10 -P were major contributors to available P as determined by the Olsen and Colwell soil tests. On the Loess Plateau, Lai et al. (2003) found that the contents of all Pi fractions but Ca 10 -P increased after soil received annual application of chemical fertilizer or manure for 17 years, and long-term fertilization also raised the percentages of Ca 2 -P, Ca 8 -P, and Al-P. However, how the soil Pi fractions transformed due to long-term fertilization is little known, which is important to understand the impact of fertilization on soil phosphorus availability in calcareous soils. In the present study, a long-term fertilization experiment was performed under a winter wheat monoculture system in calcareous cultivated soils on the Loess Plateau of China, where chemical fertilizer has been annually added before crop sowing to maintain a high crop yield since 1980s (Hao et al., 2007). The study aimed 1) to investigate how much the soil available P (Olsen-P) and total P can accumulate after long-term fertilization, 2) to characterize the changes in Pi fractions under different fertilizer P application rates, and 3) to clarify the correlation and transformation in Pi fractions in such a calcareous soil. MATERIALS AND METHODS A long-term field experiment on winter wheat (Triticum aestivum L., cv. QM4 from 1984 to 1985, cv. CW131 from 1986 to 1995, and cv. CW134 from 1996 to 2005) was carried out to examine yield responses and soil fertility response to fertilizer applications under winter wheat monoculture system. The experiment was performed at the Changwu Agro-ecological Experimental Station ( E, N) in Shaanxi Province, China. With an average annual temperature of 9.1 C, an annual frost-free period of about 171 d, and an annual precipitation of 585 mm, rainfed agriculture is the dominant production system. The soil is a calcareous soil, and referred to as a Heilu soil following the Chinese Soil Classification System, which corresponds to a Calcarid Regosol according to the FAO/UNESCO Classification System (FAO-ISRIC, 1988). It had a ph value of 8.3, organic matter of 10.5 g kg 1, total nitrogen of 0.80 g kg 1, available nitrogen of 37.0 mg kg 1, total phosphorus of 0.66 g kg 1, available phosphorus of 3.0 mg kg 1 (extracted with 0.5 mol L 1 NaHCO 3 ), available potassium of mg

3 306 J. WANG et al. kg 1 (extracted with 1 mol L 1 NH 4 CH 3 COO), and CaCO 3 of g kg 1 based on tests carried out in Soil loss by water and wind is rather low. The long-term experiment had seventeen fertilization treatments arranged in a randomized complete block design with three replications (Hao et al., 2007). Each plot area was 22 m 2. Urea and superphosphate were used as the sources of N and P fertilizers, respectively, and applied prior to wheat sowing for each year. Five phosphorus fertilization rates were applied (i.e., 0, 20, 39, 59 and 79 kg P ha 1 )at the same nitrogen fertilizer level (90 kg N ha 1 ). Soil samples were collected after harvesting on July 15, By using a 5 cm diameter tube auger, five random cores were taken to a depth of 20 cm in each plot. Large pieces of organic matter were removed by hand. Moist subsamples were brought to the laboratory, air-dried, and ground to pass through 1 and 0.25 mm nylon screens for analysis. Total P in soil was measured after the digestion with HClO 4 -H 2 SO 4 (Bao, 2000). Olsen-P was determined by extraction of air-dry soil with 0.5 mol L 1 NaHCO 3 at ph 8.5 (Olsen et al., 1954). Soil Pi fractionation was carried out according to a fractionation scheme for calcareous soils based on methods described by Chang and Jackson (1957) and Hedley et al. (1982). In the Pi fractionation scheme, soil Pi was divided into six fractions as Ca 2 -P, Ca 8 -P,Al-P,Fe-P,OccludedP,andCa 10 -P using a sequential extraction procedure with 1) 0.25 mol L 1 NaHCO 3 solution at ph 7.5 to remove Ca 2 -P, 2) 0.5 mol L 1 NH 4 Ac at ph 4.2 to remove Ca 8 -P, 3) 0.5 mol L 1 NH 4 F at ph 8.2 to remove Al-P, 4) 0.1 mol L 1 NaOH-0.1 mol L 1 Na 2 CO 3 to remove Fe-P, 5) 0.3 mol L 1 Na 3 (citrate)-na 2 S 2 O mol L 1 NaOH solution to remove occluded P, and 6) 0.25 mol L 1 H 2 SO 4 to remove Ca 10 -P. The P pools measured by the scheme are strictly empirical and to some extent overlap, but the fractions roughly correspond to the following P pools. The Ca 2 -P represents monocalcium phosphate [Ca(H 2 PO 4 ) 2 ] and dicalcium phosphate (CaHPO 4 2H 2 O) equivalents. The Ca 2 -P includes water-soluble P, citrate soluble P and partial surface-adsorbed P, and can be readily taken up by plants. The Ca 8 -P represents a group of phosphates with chemical structure similar to Ca 8 H 2 (PO 4 ) 6 5H 2 O. The Ca 8 -P fraction belongs to the sparingly soluble P, but can be partly used by plants. The Al-P (Al phosphates) and Fe-P (Fe phosphates) fractions have very low availability to plants, but can be constrainedly utilized by plants when available soil P is severely depleted. The Ca 10 -P represents a group of phosphates with chemical structure similar to Ca 10 (PO 4 ) 6 (OH) 2, which is difficult to be used by plants (Gu and Jiang, 1990). Variance analysis, correlation analysis, and path analysis were conducted using SAS software PROC ANOVA and PROC REG (SAS Institute, 1989). Path analysis is a statistical technique that partitions correlations into direct and indirect effects, and attempts to differentiate between correlation and causation. This technique also features multiple linear regressions and generates standardized partial regression coefficients (path coefficients) (Wei et al., 2006). RESULTS AND DISCUSSION Crop yield, total P and Olsen-P Table I shows the mean annual wheat yield and the contents of soil total P and Olsen-P. The mean annual yield ( ) increased significantly after long-term fertilizer P application according to the reports by Hao et al. (2007). However, no significant difference was found in crop yield for different fertilizer P application rates, which means that the increasing fertilizer P application would result in the increasing P accumulation in soil because of the similar P uptake by wheat. Consistently, soil total P contents increased by 4.2%, 26.0%, 36.5% and 49.8% in the treatments receiving 20, 39, 59, and 79 kg P ha 1, respectively. Regression analysis in Table I showed that soil total P content was closely correlated to fertilizer P rate, although no significant differences were found among fertilizer P 0, 20, and 39 kg P ha 1, and between fertilizer P 59 and 79 kg P ha 1. Soil Olsen-P content in the soil without fertilizer P application decreased from the initial value of

4 SOIL PHOSPHORUS AND SUPERPHOSPHATE APPLICATION 307 TABLE I Mean annual crop yields, soil total P contents and Olsen-P contents after long-term fertilizer P applications at Changwu Agro-ecological Experimental Station Fertilizer rate Yield a) Total P Olsen-P kg P ha 1 kg ha 1 mg kg b b) 753c 2.3d a 789c 6.0cd a 837b 7.4c a 1 027a 13.5b a 1 127a 22.9a Linear regression with fertilizer P rate (n = 15) Slope Intercept R P - < < a) Cited from Hao et al. (2007). b) Values within one column followed by the same letter are not significantly different at P< to2.3mgkg 1 after 21-year continuous wheat monoculture. This is consistent with the findings of Ma et al. (2009) who reported that in the soil without P added, soil Olsen-P concentration decreased with cultivation time until about 3 mg kg 1, afterwards it remained constant and did not decline although the P balance was negative. The decrease in Olsen-P caused by crop uptake can be replenished by other forms of soil P because of low uptake and crop yield at this range of Olsen-P in soils (Selles et al., 1995). Compared with no fertilizer P application, applications of 20, 39, 59, and 79 kg P ha 1 increased the levels of soil Olsen-P by 3.7, 5.2, 11.2 and 20.6 mg P kg 1 soil, respectively. These values were very consistent with the results reported by Fan et al. (2003) in the same experimental site, where Olsen-P increased by 4.2 mg kg 1 in the soil receiving 26 kg P ha 1. These results were also similar but higher than those reported by Halvorson and Black (1985) on a glacial till soil, where they found that soil Olsen-P content increased by 1, 2, 4, and 8 mg P kg 1 in the treatments receiving fertilizer P application at 22, 45, 90, and 180 kg P ha 1 year 1 for 16 years, respectively. In five long-term wheat-maize crop rotation experiments, Ma et al. (2009) found that the Olsen-P concentration in soils with fertilizers increased with cultivation time, and the accumulation rate of P in soils was affected mainly by P fertilization rate, crop yield and soil ph (Ma et al., 2009). In this study, there was a strong linear correlation between Olsen-P and fertilizer P rate, and fertilizer P rate might be the only factor for affecting the accumulation of Olsen-P due to the similar crop yield and soil ph (data not shown) among different fertilizer P application treatments. The increasing rates of Olsen-P are 0.18, 0.24, 0.53 and 0.98 mg kg 1 year 1 when the fertilizer P rates are 20, 39, 59 and 79 kg P ha 1 year 1, respectively. Pi and its fractions Soil Pi contents ranged from to mg kg 1 (Table II), and accounted for 56.2% to 67.5% of the total P. Compared with no fertilizer P application, soil Pi increased by 19.7%, 16.4%, 64.6%, and 75.0% when fertilizer P rates were 20, 39, 59, and 79 kg P ha 1, respectively, and the percent increases were statistically correlated to fertilizer P rates. The contents of all Pi fractions but Ca 10 -P increased after fertilizer P applications as shown in Table II, although no significant differences were found among two low fertilizer P levels and no fertilizer P treatment for Ca 8 -P, Al-P, Fe-P and occluded P. Table II also showed a high correlations coefficient between fertilizer P rates and all Pi fractions but Ca 10 -P. The slope of linear correlation equations between Pi fractions and fertilizer P rate was in the order of Ca 8 -P > Al-P > Fe-P > Ca 2 -P occluded P, which indicated the extent how Pi fractions increased or affected by fertilizer P applications. Long-term fertilizer P application had a more positive impact on the accumulation of soil Ca 8 -P than the

5 308 J. WANG et al. TABLE II Concentrations of soil Pi fractions after long-term fertilizer P applications at the Changwu Agro-ecological Experimental Station Fertilizer rate Ca 2 -P Ca 8 -P Al-P Fe-P Occluded P Ca 10 -P Total Pi kg P ha 1 mg kg d a) 70.3b 10.2c 26.6c 13.1b 296.7a 421.5b c 88.2b 16.6bc 34.2c 13.4ab 342.0a 504.6b bc 114.8b 21.8bc 37.8bc 17.2ab 288.4a 490.7b ab 200.0a 44.4ab 49.0ab 20.4ab 365.5a 693.8a a 242.2a 56.0a 57.4a 24.0a 341.8a 737.6a Linear regression with fertilizer P rate (n = 15) Slope Intercept R P < < < < < ns b) < a) Values within one column followed by the same letter(s) are not significantly different at P<0.05. b) Not significant. other fractions, which is consistent to the results from Samadi and Gilkes (1998). The contents of Pi fractions after fertilizer P application were in the order of Ca 10 -P > Ca 8 -P > Fe-P > Al-P > occluded P > Ca 2 -P, and only Al-P exceeded occluded P when comparing to that without fertilizer P application (Fig. 1). As the dominant component, soil Ca 10 -P accounted for 70.5% of the total Pi in the soils without fertilizer P application, and with increasing fertilizer P rate the percentage decreased to 42.8% when receiving 79 kg P ha 1. Similar result was also found by Lai et al. (2003) where the percentage of Ca 10 -P declined from 75.1% to 59.5% after receiving 26 kg P ha 1. In contrast, the percentages of Ca 2 -P (1.1% 2.2%), Ca 8 -P (16.6% 32.7%), and Al-P (2.4% 7.7%) relative to total Pi increased with fertilizer P application, while the percentages of Fe-P and occluded P remained stable after fertilizer P applications, being 6.3% 7.8% and 2.7% 3.5%, respectively. Fig. 1 Percentages of Pi fractions under different rates of fertilizer P application. Relationship between Olsen-P and Pi fractions Simple correlation coefficients showed that Olsen-P was positively correlated with all Pi fractions except for Ca 10 -P (Table III). According to Jiang and Gu (1989), Pi fractions had different contributions to plant-available P which could be reflected by the extraction sequence. However, the correlation coefficients did not decrease in the extraction order, and high correlations were also found among these Pi fractions except for Ca 10 -P as shown in Table III, so the transformation among these fractions would be more important to understand their potential of P release and direct availability. Path analysis was used to partition the relationship between Olsen-P and P fractions into direct and

6 SOIL PHOSPHORUS AND SUPERPHOSPHATE APPLICATION 309 TABLE III Pearson correlation coefficients for soil Olsen-P and Pi fractions Olsen-P Ca 2 -P Ca 8 -P Al-P Fe-P Occluded P Ca 2 -P 0.81*** Ca 8 -P 0.92*** 0.86*** Al-P 0.80*** 0.71** 0.82*** Fe-P 0.75** 0.83*** 0.90*** 0.75** Occluded P 0.83*** 0.84*** 0.93*** 0.66** 0.89*** Ca 10 -P 0.25ns a) 0.20ns 0.02ns 0.03ns 0.03ns 0.31ns **, ***Significant at P<0.01 and P<0.001, respectively. a) Not significant. indirect effects. Direct path coefficients measure the direct effect of a fraction on Olsen-P, while indirect path coefficients specify the effect of a fraction passed through other fractions. A high path coefficient indicates a strong effect on Olsen-P. Inorganic P fertilizer accounted for 90% of variations in Olsen-P (Table IV). The direct path coefficient from path analysis showed that the importance of soil P fractions to Olsen-P was in the order of Ca 8 -P > Fe-P > Ca 2 -P >occluded P > Ca 10 -P > Al-P. Fe-P had a negative direct path coefficient, and its high correlation with Olsen-P was mainly from indirect effect through Ca 8 -P and Ca 2 -P. This indicated that Fe-P might be an intermediate in the transformation of Pi fractions. As the most labile fraction, Ca 2 -P can be easily absorbed by crop or transformed into Ca 8 -P, which resulted in a low direct effect on Olsen-P. Al-P and occluded P both had a very low direct path coefficient to Olsen-P, the high correlation between them and Olsen-P was mainly through the indirect effect with Ca 8 -P (positive) and Fe-P (negative). Being consistent with correlation analysis, Ca 10 -P had a low direct and indirect path coefficient. The transformation mainly occurred among other Pi fractions. TABLE IV Path coefficients a) of soil Pi fractions for Olsen-P Direct path coefficient Indirect path coefficient Ca 2 -P Ca 8 -P Al-P Fe-P Occluded P Ca 10 -P Ca 2 -P Ca 8 -P Al-P Fe-P Occluded P Ca 10 -P a) R 2 = P added to the soil was distributed between labile and non-labile pools. Shen et al. (2004) found that labile P (Ca 2 -P, Ca 8 -P, and Al-P) could be readily fixed into non-labile P (Fe-P, occluded P, and Ca 10 -P) due to the association with hydrous Fe oxides and calcareous compounds in calcareous soils. However, the present study showed as a labile pool, more Ca 8 -P accumulated due to the transformation from other fractions after long-term continuous inorganic fertilizer application, which had a significant contribution to soil P availability. CONCLUSIONS In the Heilu soil or Calcarid Regosol on the Loess Plateau of China, Olsen-P increased linearly with fertilizer P application rates after long-term annual P (superphosphate) application. Compared with no fertilizer P application, applications of 20, 39, 59, and 79 kg P ha 1 increased soil Olsen-P content

7 310 J. WANG et al. by 3.7, 5.2, 11.2 and 20.6 mg P kg 1 soil, respectively. Soil total P and Pi accumulated largely after long-term fertilizer P application and were strongly correlated with fertilizer P rate. In the soil with fertilizer P application, the contents of Pi fractions were in the order of Ca 10 -P > Ca 8 -P > Fe-P > Al-P > occluded P > Ca 2 -P, and soil Pi fractions changed with fertilizer P rate differently. Fertilizer P increased both Ca 8 -P, Al-P and Ca 2 -P contents and their percentages relative to Pi, while only contents for Fe-P and occluded P increased. Soil Ca 10 -P remained stable after long-term fertilizer P application, and its relative content to Pi declined linearly with increasing fertilizer P rate. Strong correlations were found between Olsen-P and the Pi fractions but Ca 10 -P after long-term fertilizer P application. 90% of variations in Olsen-P could be explained by Pi fractions, and the direct contribution was mainly from Ca 8 -P. The high correlations between Olsen-P and Ca 2 -P, Al-P, and occluded P were mainly through their positive indirect effect on Ca 8 -P and negative indirect effect on Fe-P. As a labile pool, more Ca 8 -P accumulated due to the transformation from other fractions after long-term continuous inorganic fertilizer application, which improved soil P availability. REFERENCES Bao, S. D Analysis of Soil and Agrochemistry (in Chinese). 3rd Edition. China Agriculture Press, Beijing. Chang, S. C. and Jackson, M. L Fractionation of soil phosphorus. Soil Sci. 84: Fan, J., Hao, M. D. and Wang, Y. G Effects of rotation and fertilization on soil fertility on upland of Loess Plateau. Res. Soil Water Conserv. (in Chinese). 10(1): Food and Agriculture Organization-International Soil Reference and Information Centre (FAO-ISRIC) FAO- UNESCO Soil Map of the World. Revised Legend. FAO, Rome, Italy. Gu, Y. C. and Jiang, B. F The fraction method for determining soil inorganic P in calcareous soils. Soils (in Chinese). 22: Gu, Y. C. and Qin, S. W Effect of long-term phosphate fertilization on soil phosphorus accumulation, transformation and availability in a fluvo-aquic soil. Soils (in Chinese). 29: Halvorson, A. D. and Black, A. L Long-term dryland crop responses to residual phosphorus fertilizer. Soil Sci. Soc. Am. J. 49: Hao, M. D., Fan, J., Wang, Q. J., Dang, T. H., Guo, S. L. and Wang, J. J Wheat grain yield and yield stability in a long-term fertilization experiment on the Loess Plateau. Pedosphere. 17: Hedley, M. J., Stewart, J. W. B. and Chauhan, B. S Changes in inorganic and organic soil phosphorus fractions induced by cultivation practices and by laboratory incubations. Soil Sci. Soc. Am. J. 46: Jiang, B. F. and Gu, Y. C A suggested fractionation scheme of inorganic phosphorus in calcareous soils. Sci. Agri. Sin. (in Chinese). 22(3): Lai, L., Hao, M. D. and Peng, L. F The variation of soil phosphorus of long-term continuous cropping and management on Loess Plateau. Res. Soil Water Conserv. (in Chinese). 10(1): Ma, Y. B., Li, J. M., Li, X. Y., Tang, X., Liang, Y. C., Huang, S. M., Wang, B. R., Liu, H. and Yang, X. Y Phosphorus accumulation and depletion in soils in wheat-maize cropping systems: Modeling and validation. Field Crop. Res. 110: Olsen, S. R., Cole, C. V., Watanable, F. S. and Dean, L. A Estimation of Available Phosphorus in Soils by Extraction with Sodium Bicarbonate. USDA Circular No U.S. Government Printing Office, Washington, DC. Samadi, A. and Gilkes, R. J Forms of phosphorus in virgin and fertilized calcareous soils of Western Australia. Aust. J. Soil Res. 36: SAS Institute SAS/STAT User s Guide, Version 6. 4th Edition. SAS Institute, Cary, NC. Selles, F., Campbell, C. A. and Zentner, P. R Effect of cropping and fertilization on plant and soil-phosphorus. Soil Sci. Soc. Am. J. 59: Sharpley, A. N., Sims, J. T. and Pierzynski, G. M Innovative soil phosphorus availability indices: Assessing inorganic phosphorus. In Havlin, J. and Jacobsen, J. (eds.) Soil Testing: Prospects for Improving Nutrient Recommendations. SSSA Special. Publication No. 40. Soil Science Society of America, Madison, USA. pp Shen, J., Li, R., Zhang, F., Fan, J., Tang, C. and Rengel, Z Crop yields, soil fertility and phosphorus fractions in response to long-term fertilization under the rice monoculture system on a calcareous soil. Field Crop Res. 86: Solis, P. and Torrent, J Phosphate fractions in calcareous Vertisols and Inceptisols of Spain. Soil Sci. Soc. Am. J. 53: Wei, X. R., Hao, M. D., Shao, M. A. and Gale, W. J Changes in soil properties and the availability of soil micronutrients after 18 years of cropping and fertilization. Soil Till. Res. 91: Zhang, T. Q., MacKenzie, A. F., Liang, B. C. and Drury, C. F Soil test phosphorus and phosphorus fractions with long-term phosphorus addition and depletion. Soil Sci. Soc. Am. J. 68:

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