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Effects of zeolite and cattle manure on growth, yield and yield components ofsoybean grown under water deficit stressR EZA N OZARI,H AMID R EZA T OHIDI-M OGHADAM*,M ASOUD M ASHHADI-A KBAR-B OOJAR1Department of AgronomyVaramin-Pishva Branch, Islamic Azad University, Varamin, Iran*(e-mail : hamid_tohidi2008@)A BSTRACTIn order to study the effects of cattle manure and zeolite on growth, yield and yield components of soybean grown under conditions of water deficit stress, an experiment was conducted in research field of Islamic Azad University, Varamin Branch during 2011 growing season. The experimental design was carried out in a randomized complete block with a split factorial arrangement of treatments in three replications. Main factor was water stress (normal irrigation and irrigation withholding after 50% flowering) and sub factors were included cattle manure (0, 15 and 30 t/ha) and zeolite application (with and without). The results showed that water stress significantly decreased plant growth and production. Conversely, cattle manure and zeolite application led to increase in growth and improved yield and yield components. Zeolite application dramatically increased pod number in main branches. The highest pod number in sub-branches was obtained from those plots which were treated with 30 t/ha cattle manure and zeolite grown under normal irrigation conditions. These results indicated that zeolite and cattle manure application (30 t/ha) increased seed yield when plants were irrigated completely. It is interesting to remark that zeolite application was more effective under stress conditions than normal irrigation condition. Biological yield increased on account of cattle manure and zeolite application. In addition, the highest oil yield was achieved when soybean plants were treated with 30 t/ha cattle manure and zeolite under normal irrigation conditions. In sum cattle manure and zeolite application could improve soybean growth and seed production even under drought stress conditions.Key words : Cattle manure, water stress, yield and yield components, zeolite, Soybean1Department of Biochemistry, Tarbiat Moallem, Tehran University, Tehran.INTRODUCTIONWater stress is still a serious agronomic problem and one of the most important factors contributing to crop yield loss. Plant morphological and biochemical responses to drought stress vary with stress intensity. Drought stress significantly limits plant growth and crop productivity. In Iran, water is a scarce resource, due to the high rainfall variability. The water stress effects depend on deficit timing, duration and magnitude (Pandey et al., 2001). The identification of the critical irrigation timing and scheduling based on a timely and accurate basis to the crop is the key to conserving water and improving the irrigation performance and sustainability of irrigated agriculture (Ngouajio et al.,2007). Igbadun (2006) showed that the crop yield response was very much dependent on the amount of water applied at different crop development stages than the overall seasonal water applied. This approach may save water with little or no negative impact on the final crop yield. In arid and semi-arid environments, both efficient use of available water and a higher safflower yield and quality are in demand (Lovelli et al., 2007; Dordas and Sioulas, 2008; Koutroubas et al., 2008).A decrease in biological yield and pod dry matter have been observed in two canola cultivars, under water deficit conditions (Thomas, 2004). Porous ceramics, diatomaceous earth and zeolites are just a few of the more commonly used inorganic soil amendments. Some characteristics of these products, that make them desirable for improving soils’properties, are a large internal porosity, which results in water retention; a uniform particle size distribution, that allows them to be easily incorporated; and a high cation exchange capacity, which retains nutrients (Ok et al. 2003). The unique physical and chemical properties of natural zeolites, in combination with their abundance in sedimentary deposits and in rocks derived from volcanic parent materials, have made them useful for many industrial applications. These properties have spurred their use in agronomic and horticultural applications (Dwairi1998). Zeolites are hydrated aluminosilicates, characterized by three-dimensional networks of SiO4and AlO4tetrahedral and linked by shared oxygen atoms. Clinoptilolite is not the most well-known zeolite, but it is one of the most useful. The chemical formula of clinoptilolite is (Na3K3) (Al6Si30O72).24H2O. Extensive deposits of clinoptilolite are found in the Western United States, Bulgaria, Hungary, Japan, Australia and Iran. Amendment of clinoptilolite zeolite to sandy soils has been reported to lower nitrogen concentrations in leachate and to increase moisture and nutrients in the soil, due to increased soil surface area and cationexchange capacity (He, 2002). Agronomic operations, which involve heavy application of chemical fertilizers, may cause reduction of nutrients in soil and certain others would generally accumulate in excess resulting in nutrient imbalance which affects the soil productivity. Among the means available to achieve sustainability in agricultural production, organic manure like cattle manure plays an important and key role because it possesses many desirable soil properties and exerts beneficial effect on the physical, chemical and biological characteristics of the soil. Cattle manure is a good source of nutrients for vegetation especially when supplemented with commercial nitrogen fertilizer. Cattle manure application improves physical and chemical properties of soil. In addition, it increases micro organism activity and water retention capacity (Gupta et al.,2004). Crop yield is usually increased by manure application because of the increased nutrient availability and the improved soil structure (Matsi et al., 2003). Eghball and Power (1999) found that application of cattle manure resulted in maize yield similar to that from commercial fertilizer application. Water stress is still a serious agronomic problem and one of the most important factors contributing to crop yield loss. Plant morphological and biochemical responses to drought stress vary with stress intensity. In arid and semi-arid regions like Iran, soils are generally characterized by poor soil structure, low water-holding capacity, lack of organic matter and nutrient deficiency. Thus, the application of zeolite substances and organic fertilizers may be effective on better establishment of plant under water stress conditions. Therefore, this study was undertaken to evaluate the effects of both cattle manure and zeolite on soybean plant under water deficit stress.MATERIALS AND METHODSIn order to study the effects of different amount of organic fertilizer (cattle manure) and zeolite (with and without zeolite) on growth, yield and yield components of soybean grown under normal and limited irrigation, an experiment was conducted in research field of Islamic Azad University, Varamin Branch during 2011 growing season. Site of study was situated at 31°519 E and 20°359 N and 1050 m above sea level. Before beginning of experiment, soil samples were taken in order to determine the physical and chemical properties. A composite soil sample was collected at a depth of 0-30 cm. It was air-dried, crushed and tested for physical and chemical properties. The research field had a clay loam soil. Details of soil properties areshown in Table 1. After plow and disk, plots were prepared. The experimental design was laid out in a randomized complete block with a split factorial arrangement of treatments in three replications. Main factor was water stress (normal irrigation and irrigation withholding after 50% flowering) and sub-factors included cattle manure (0, 15 and 30 t/ha) and zeolite application (with and without). The 16 m2plots were prepared with 4 m length and consisted of five rows, 0.65 m apart. Among all main plots, 2 m alley was kept to eliminate all influence of lateral water movement. Polyethylene pipeline was performed for control of irrigation as dropping irrigation. Cattle manure was applied before seed sowing so that manure was spread on the soil surface and then was mixed into the soil manually. Soybean seeds [Glycine max (L.) Merr. cv. Williams] were purchased from Pars Abad Oil Seed Company and inoculated with Rhizobium japonicum and sown in certain experimental plots with 10 cm apart each other. Irrigation was performed immediately after seed sowing and after seedling establishment irrigation was done every week. At 5-leaf stage plants were thinned to appropriate density. Weeds were controlled manually at 5-leaf stage, stem elongation and flowering stage. In order to stress induction, irrigation was stopped at 50% flowering stage in stressed plots till end of growing stage. At physiological maturity stage, crop was completely harvested and plant height, branch number, pod number in main branches, pod number in sub-branches, total pod number, 1000-seed weight, seed yield, biological yield, harvest index, oil and protein yield were measured. Oil and protein percentage were calculated using Soxhlet and Kjeldahl methods, respectively. Oil and protein yield and also harvest index were obtained by following formulae, respectively :Oil yield = Oil percentage × Seed yield /100Protein yield = protein percentage × Protein yield /100HI = Economic yield (seed yield)/ biological yield × 100 All data were analyzed from analysis of variance (ANOVA) using the GLM procedure in SAS (SAS Instituteinc., 2002). The assumptions of variance analysis were tested by ensuring that the residuals were random, homogenous, with a normal distribution about a mean of zero. Duncan's multiple range test was used to measure statistical differences between treatment methods and controls.RESULTS AND DISCUSSIONAnalysis of variance showed that water stress had significant effect on all agronomic traits (Table 2). In addition, effect of cattle manure was significant on alltraits except for harvest index (Table 2). Among all agronomic traits, plant height and harvest index were not affected by zeolite application (Table 2). Interaction of experimental factors (water stress × cattle manure × zeolite) was significant except for biological yield and harvest index (Table 2). As can be seen from Table 3, water stress significantly decreased plant height. Although, cattle manure had not any effect on plant height, zeolite increased plant height under water stress conditions. The most branch number was observed from those plants which were treated with 30 t/ha cattle manure and zeolite application given under normal irrigation conditions (Table 3). By contrast, the lowest branch number was related to stress plants treated with 0 or 15 t/ha cattle manure without zeolite application (Table 3).The results showed that zeolite application dramatically increased pod number in main branches (Table 3). The lowest pod number in main branch was observed when soybean plants were just stressed treated neither by cattle manure nor by zeolite application (Table 3). Similarly, the highest pod number in sub-branches was obtained from those plots which were treated with 30 t/ha cattle manure and zeolite application given under normal irrigation conditions. Conversely, the lowest pod number in sub-branches was achieved in stressed plants without any further treatment (Table 3). In general, the highest and lowest pod number in plant was obtained when plants were treated with 30 t/ha cattle manure alongwith zeolite application given under normal irrigation and stressed plants treated without cattle manure application, respectively (Table 3). Our results indicated that zeolite application and cattle manure application (30 t/ha) significantly increased seed yield when plants were irrigated completely (Table 3). Zeolite application was more effective under stress conditions than normal irrigation conditions (Table 3). According to obtained results the highest oil yield was achieved when soybean plants were treated with 30 t/ha cattle manure and zeolite application both under normal irrigation conditions and drought stress conditions (Table 3).Protein yield significantly decreased due to water stress. Cattle manure had significant effect on protein yield especially under normal irrigation conditions. So, the highest protein yield belonged to treatments treated with 30 t/ha cattle manure and zeolite application both under normal irrigation conditions and drought stress conditions (Table 3). Interaction between water stress and cattle manure application was significant on biological yield (Fig. 1). Biological yield increased on account of cattle manure in drought stress condition (Fig. 1). The heaviest biological yield wasobtained when 30 t/ha cattle manure was applied under normal irrigation conditions, while the lightest biological yield was related to no cattle manure application treatment alongwith irrigation withholding at the end of growing season. In addition, biological yield was affected by water stress and zeolite application (Fig. 2). Zeolite application increased biological yield both under normal irrigation conditions and soybean plants were under stressed conditions. Also the highest biological yield was obtained when 30 t/ha cattle manure was applied in both zeolite application and without zeolite application (Fig. 3). Also the results showed that the harvest index decreased due to water stress, however, it was not affected by cattle manure or zeolite (Fig. 4).The results of this research showed that the water stress decreased growth, yield and yield components of soybean. The decrease in yield and yield components in different safflower genotypes due to water deficiency was also reported by other researchers (Zaman and Das, 1991; Kar et al., 2007; Lovelli et al., 2007). Anyia and Herzog (2004) indicated that water deficit caused between 11 and more than 40% reduction of biomass across the genotypes of cowpea (Vigna unguiculata L.) due to decline in leaf gas exchange and leaf area. Obtained result indicated that the seed filling stage was more sensitive in soybean growth stages. Nielsen and Nelson (1998) and Nuez et al. (2005) also identified the number of pods per plant as the principal cause of yield losses of bean subjected to drought stress, followed by the number of seeds per pod and seed weight. This result corroborated the earlier findings of Castaeda et al. (2006).The diverse effects of water stress on physiological and metabolic responses in plant growth were well documented (Kramer and Boyer, 1995). Application of integrated chemical fertilizer with biological fertilizer caused to produce highest yield compared with application of chemical and biological treatment alone (Rizwan et al., 2008). Aowad and Mohamed (2009) evaluated the effects of manure and mineral fertilizer on sunflower characteristics and found that the highest values of stem diameter and plant height were produced due to combination of nitrogen fertilizer with farm yard manure. Ibeawuchi and Onweremalu (2007), in an experiment, showed that at first of all the highest 1000-kernel weight was in integrated fertilizer treatments and the following was in chemical fertilizer. Majidian and Ghalavand (2006) showed that integrated chemical and biological fertilizer obtained highest kernel number per ear compared with sole application of them. It seems that zeoliteincreases water retention capacity and thus water stress intensity will be decreased. Xiubin and Zhanbin (2001) showed that zeolite improved water retention capacity and cation exchange capacity in arable soils. Leggo (2000) studied the response of wheat to poultry manure amended by zeolite and found out that crop faced a better growth rate when zeolite was applied in poultry manure, and reported that the increase of growth and yield was due to nitrogen availability by zeolite.CONCLUSIONIrrigation deficit reduced growth, biomass and yield in soybean. The flowering stage was the most sensitive to water deficit. Drought stress, during this period, caused a reduction in the number of grains per plant and in time to maturity. Consequently, reduced sink capacity and shorter growing period led to lower seed yield. Under water limited conditions, soil water extraction was a more important component in soybean yield. Efficient management of soil moisture was important for agricultural production in the light of scarce water resources. Soil conditioners contributed significantly to provide a reservoir of soil water to plants on demand in the upper layers of the soil where the root systems normally develop. Zeolite apart from improving the soil physical properties, also served as buffers against temporary drought stress and reduced the risk of plant failure during establishment. This was achieved by means of reduction of evaporation through restricted movement of water from the sub-surface to the surface layer. At the same time, cattle manure application improved physical and chemical properties of soil. In addition, it increased micro- organism activity and water retention capacity. Also cattle manure could be substituted with inorganic fertilizer for the nitrogen and micronutrients supplementation and better growth of soybean. In conclusion, this study showed that application of zeolite and cattle manure, in soils exposed to drought stress, could maintain soil water content and improve plant growth and yield.REFERENCESAnyia, A. O. and Herzog, H. (2004). Water use efficiency, leaf area and leaf gas exchange of cowpeas under mid-season drought. Eur. J. Agron.20:327-39.Aowad, M. M. and Mohamed, A. A. A. (2009). The effect of bio, organic and mineral fertilization on productivity of sunflower seed and oil yields. J. Agric.Res. Kafr El Sheikh Univ.35 : 1013-27.Castaeda, S. M. C, Cordova, T. L, Gonzalez, H. V. A, Delgado, A. A., Santacruz, V.A. and Garca de los, S. G. (2006). Espuestas fisiol gicas, rendimientoycalidad de semilla en frijol sometido a estrés hdrico. Interciencia.31 :461-66.Dordas, C. A. and Sioulas, C. (2008). Safflower yield, chlorophyll content, photosynthesis, and water use efficiency response to nitrogenfertilization under rainfed conditions. Indust. 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Zeolite application for enhancing infiltration and retention in loess soil. Resource, Conservation and Recycling,Amsterdam34 : 45-52.Zaman, A. and Das, P. K. (1991). Effect of irrigation and nitrogen on yield and quality of safflower. Indian J. Agron.36 : 177-79.1112Table 2. Analysis of variance growth parameters, yield and yield components, harvest index, oil and protein yield affected by water stress, cattle manure and zeoliteSources of variation dfheight number in main branch sub branch number weight yield yield index yield yield Replication 2 ns ns ns ns ns ns ns ns ns ns ns Water stress 1 * ** ** ** ** ** ** ** * ** ** Error (a) 2 Cattle manure 2 ** ** ** ** ** ** ** ** ns ** ** Zeolite1 ns ** ** ** ** ** ** ** ns ** ** Water stress × cattle manure2 ns ** ** ** ** ** ** * ns ** ** Water stress × Zeolite 1 ns ns ** ** ** ** ** * ns ** * Cattle manure × Zeolite2 ns ** ** ** ** ns ** * ns ** ** Water stress × cattle manure × Zeolite 2 * ** ** ** ** * * ns ns ** ** Error (b)20C.V (%)4.3316.854.026.333.896.119.1611.907.338.398.90*,** and ns significant at 0.05, 0.01 and no significantTable 1. Soil properties of the experimental siteDepth EC (ds m -1) pH O.C (%)T.N.V (%) K (ppm) P(ppm) Total N (%) Texture 0-30 cm4.17.40.71<1036825.90/079Clay loamTable 3. Interaction between water stress, cattle manure and Zeolite on growth parameters, yield and yield components, oil yield, Protein yieldWater stress Cattle manure Zeolite Plant height(cm)BranchnumberPod number inmain branchPod number insub branchTotal podnumberSeed yield Oil yieldProteinyieldNo stress 0 ton ha-126.000e 56.667g6870.0d1266.7d 1988.7e+ 141.667bcd 4.000de 44.000c 27.333de 71.333d 8296.7c 1612.0c 2423.0d 15 ton ha-1- 146.000bc 4.000de 40.667d 27.667de 68.000de 7953.3c 1624.3c 2910.0c+ 144.667bc 4.000de 63.333a 48.667b 112.000b 9506.7b 1865.7b 3159.3bc 30 ton ha-1- 148.667b 9.000b 49.000b 38.667c 87.667c 8750.0bc 1840.3b 3528.0b+ 167.000a 13.000a 65.667a 68.667a 134.333a 13533.3a 3024.0a 5528.3aWater stress 0 ton ha-1- 131.333d 3.000e 29.333h 15.000f 44.333h 2770.0f 483.3f 723.0h+ 135.000cd 3.333de 34.667fg 27.667de 60.333fg 3830.0ef 707.7e 1037.0gh 15 ton ha-1- 135.667cd 3.333de 34.667fg 25.667e 60.333ef 3756.7ef 694.0e 1091.7gh+ 141.000bcd 5.000cd 38.000de 31.000d 69.000de 4496.7e 881.7e 1492.3f 30 ton ha-1- 145.333bc 4.333de 35.667ef 29.667de 65.333fg 4153.3e 814.3e 1382.7fg+ 142.333bcd 6.000c 50.667b 42.000c 92.667c 5890.0d 1206.3d 2154.0deTreatment means followed by the same letter within each common are not significantly different (P < 0.05) according to Duncan’s Multiple Range test13Fig. 1.Interaction between water stress and cattle manure application on biological yield. Treatment means followed by the same letter within each common are not significantly different (P < 0.05) according to Duncan’s Multiple Range test14letter within each common are not significantly different (P < 0.05) according to Duncan’s Multiple Range test15Fig. 3. Interaction cattle manure and zeolite application on biological yield. Treatment means followed by the same letter within each common are not significantly different (P < 0.05) according to Duncan’s Multiple Range test1617Fig. 4. Main effect of irrigation on harvest index. Treatment means followed by the same letter withineach common are not significantly different (P < 0.05) according to Duncan’s Multiple Range test5101520253035404550No stress water stressB i o l o g i c a l y i e l d ( k g h a -1 )。