Use of N and P biofertilizers together with phosphorus
Transcription
Use of N and P biofertilizers together with phosphorus
Vol. 2 (3), pp. 168-174, October, 2014. © Global Science Research Journals http://www.globalscienceresearchjournals.org/ Global Journal of Agriculture and Agricultural Sciences Full Length Research Paper Use of N and P biofertilizers together with phosphorus fertilizer Improves growth and physiological attributes of chickpea Moinuddin1, Tariq Ahmad Dar*2, Sajad Hussain2, M. Masroor Akhtar Khan2, Nadeem Hashmi2, Mohammad Idrees2, Mohammad Naeem2, Akbar Ali2 1 Women’s College, Botany Section, Aligarh Muslim University, Aligarh-202002, U.P., India 2 Department of Botany, Aligarh Muslim University, Aligarh-202002, U.P., India Accepted 10 October, 2014 Abstract Leguminous crops grow poorly in phosphorus deficient soils. A two factor factorial experiment was -1 conducted in the net-house to investigate the effect of graded levels of P fertilizer (0, 30 and 60 kg P ha or P0, P30 and P60, respectively) along with Rhizobium (BNF) and/or phosphate solubilizing bacteria (PSB) on growth and physiological parameters of chickpea. Phosphorus was applied as basal dose, while seeds were treated with respective biofertilizer(s) before sowing according to the treatments [BF 0 (control), BNF, PSB and BNF+PSB]. As per the main effects, P60 proved superior or equivalent to P30, while among the biofertilizer treatments, BNF+PSB gave the greatest values or was equal to PSB regarding most growth and physiological parameters. Generally, interaction between P levels and biofertilizer treatments was significant. 30 kg P ha-1 applied with N and P biofertilizers (P30 × BNF+PSB) was the cost-effective interaction for most of the parameters studied. Compared to P 60 applied alone (P60 × BF0), P30 × BNF+PSB resulted in greater dry matter production (10.36%), PN(22.96%), leghemoglobin content of root nodule (47.53%) and nitrate reductase activity (16.67%). In addition, P30 × BNF+PSB was statistically equal to P60 × BNF+PSB regarding dry matter production, net photosynthesis, leghemoglobin content of root nodule and leaf nitrate reductase and carbonic anhydrase activities. Thus, compared to P60, application of P30 × BNF+PSB proved to be more profitable to attain the enhanced values of crop growth attributes and physiological parameters required for the improved crop yield and quality. Key words: Cicer arietinum, growth and physiological attributes, phosphorus levels, Pseudomonas striata, Rhizobium ciceri, phosphorus-deficient soil INTRODUCTION Chickpea (Cicer arietinum L.) is the third most widely grown grain legume in the world after bean and soybean (Soltani et al., 2006). It occupies an important role in *Corresponding Author: Tariq Ahmad Dar, Plant Physiology Section, Department of Botany, Aligarh Muslim University, Aligarh, 202 002 India E-mail: dartariq21@gmail.com, Tel: +91-8755643233 human nutrition due to its high protein content (17-23%) and because of being a good source of carbohydrates, minerals and trace elements (Namvar and Sharifi, 2011). Millions of people of developing countries, who cannot afford the costlier animal protein, consume chickpea in large amounts in their diet (Huisman and Poel, 1994). It is also used as feed for livestock and has a significant role in farming systems (Singh, 1997). Glob. J. Agric. Agric. Sci. 169 Phosphorus is (P) the second limiting plant nutrient after nitrogen (Rudresh et al., 2005). Soils usually contain a high amount of total P, but its availability to plant is very low. As a result, costly phosphatic fertilizers are applied to soil for leguminous crops, which require large amounts P for proper growth and development (Singh, 1983). Photosynthesis and stomatal conductance are reduced by P deficiency (Guidi et al., 1994). On the contrary, increased P supply has been reported to increase photosynthesis (Gao et al., 1989). In leguminous crops, P promotes nodulation, dinitrogen fixation and efficient partitioning of photosynthates between source and sink (Giaquenta and Quebedeaux, 1980). Biofertilizers are living microorganisms, which when applied through seed or soil treatment, promote growth by increasing the supply or availability of nutrients to the host plant (Stephens and Rask, 2000; Moin Uddin et al., 2014). In plants, they also increase the content of growth hormones such as IAA and GA, leading to enhancement in the growth of plants (Asad et al., 2004; Selvakumar et al., 2009). These biofertilizers include the N2 fixing, phosphate solubilizing and plant growth promoting microorganisms (Mahdi et al., 2010). Rhizobium bacteria (BNF) through biological N2 fixation, meet about 80-90% of total N requirements of legumes (Verma, 1993). Phosphate solubilizing microorganisms play a key role in the plant metabolism and crop productivity. They are known to increase P uptake and overall P-use efficiency resulting in better growth and higher yield of crop plants (Alagawadi and Gaur, 1988; Rudresh et al., 2005). The combined inoculation of Rhizobium and phosphate solubilizing bacteria has been reported to increase the nodulation, growth and yield parameters in chickpea (Sattar and Gaur, 1987; Alagawadi and Gaur, 1988; Rudresh et al., 2005). As a matter of fact, supply of N and P biofertilizers along with inorganic P fertilizer could play an important role in manifestation of improved nutrient uptake and enhanced crop yield and quality of chickpea in a cost-effective manner (Moin Uddin et al., 2014). The present results are the part of this experiment aimed at exploring the effect of graded levels of P applied with N and P biofertilizers on growth attributes, physiological parameters, nutrient uptake, yield and quality of chickpea. MATERIAL AND METHODS Pot culture The pot experiment was conducted on chickpea in a nethouse under natural conditions at the Botany Department, Aligarh Muslim University, Aligarh (India) using N and P deficient soil. Climatic conditions, soil characteristics and seed treatment with N and P biofertilizers regarding the pot experiment have earlier been reported (Moin Uddin et al., 2014). Soil was basally dressed with three P levels viz. 0, 30 and 60 kg P ha-1 (P0, P30 and P60, respectively), while four biofertilizer treatments (0, BNF, PSB, and BNF+PSB) were applied to the seeds prior to planting. Each treatment was replicated four times. Each experimental pot carried six healthy plants. Growth attributes Growth attributes were determined at 90 days after planting (DAP). Two randomly selected plants were uprooted carefully from each treatment-replicate to measure growth attributes (plant height, number of branches and leaves per plant, and fresh and dry weight per plant) and leghemoglobin content in the root-nodules. The uprooted plants were washed thoroughly with tap water to remove the adhering dust (from the shoot) and soil particles (from the root). The root-nodules of the plants were again washed with distilled water and stored fresh in polythene bags for the estimation of leghemoglobin content. The clean plants (above ground shoots) were blot-dried and o measured for plant fresh weight. They were dried at 80 C for 24 h, recording the dry weight of the plants thereafter. Photosynthetic Parameters and Transpiration Rate Photosynthetic parameters net photosynthesis (PN), stomatal conductance (gs) and internal CO2 concentration) and transpiration rate (E) were measured at 90 DAP on cloudless day at 1000-1100 hours using the youngest fully expanded leaves with the help of an Infra Red Gas Analyzer (IRGA, Li-Cor 6400 Portable Photosynthesis System, Lincoln, Nebraska, USA). Activity of nitrate reductase (NR) and carbonic anhydrase (CA) The NR (E.C. 1.6.6.1) activity in the leaves was determined by the intact tissue assay method (Jaworski 1971). The enzyme activity was expressed as nM NO2 g-1 leaf FW h-1. The activity of CA (E.C. 4.2.1.1) was measured in the leaves using the method described by Dwivedi and Randhawa (1974). The enzyme activity was -1 -1 expressed as μM CO2 kg leaf FW s . Leghemoglobin content Leghemoglobin (Lb) content in the fresh-stored nodules was determined as described by Sadasivam and Manickam (2008). The Lb content in the fresh nodules was calculated using the following formula: Lb content (mM) = [(OD 556 – OD 539)/23.4] × 2D. Where OD 556 and OD 539 represent absorbance values recorded at 556, 539 nm, respectively, and D is the initial dilution. Statistical Analysis Statistical analyses of the data were carried out according to randomized block design. All the parameters were subjected to analysis of variance (ANOVA), using Tariq et al. Table 1: Effect of graded levels of phosphorus applied with N and P biofertilizer treatments on growth parameters of chickpea Treatments Height per plant Number of Number of leaves Fresh weight per Dry weight per (cm) branches per plant per plant plant (g) plant (g) Main effects of phosphorus (P) b b b c c P0 24.84 4.0 54.4 2.54 0.835 a a a b b P30 25.91 4.3 56.1 2.86 0.870 a a a a a P60 26.52 4.3 56.1 2.99 0.905 Main effects of biofertilizers (BF) c b c BF0 23.83 4.0 52.7 2.43 0.760 b b b BNF 25.85 4.3 55.2 2.47 0.867 b a a BPF 26.34 4.3 55.4 3.14 0.926 a a a BNF+BPF 27.00 4.3 58.8 3.14 0.927 Effects of interaction (P × BF) bc e e d P0 × BF0 23.00 4.0 52.1 2.30 0.679 b de e d P30 × BF0 23.50 4.0 53.0 2.40 0.752 ab de de c P60 × BF0 25.00 4.0 53.0 2.60 0.849 b d e c P0 × BNF 24.95 4.0 54.3 2.30 0.851 a d e c P30 × BNF 26.40 4.5 55.0 2.35 0.855 a c d b P60 × BNF 26.20 4.5 56.3 2.75 0.896 ab d b P0 × BPF 25.50 4.0 54.3 2.65d 0.901 a cd a a P30 × BPF 26.13 4.5 55.3 3.35 0.937 a c a a P60 × BPF 27.40 4.5 56.7 3.43 0.939 ab c c b P0 × BNF+BPF 25.90 4.0 57.0 2.90 0.908 a a a a P30 × BNF+BPF 27.60 4.5 61.0 3.35 0.937 a b b a P60 × BNF+BPF 27.50 4.5 58.5 3.17 0.937 Values followed by the same letter in a column section are not significantly different according to Fisher’s Least Significant Difference -1 -1 (LSD) at p<0.05. P0: no phosphorus application (control), P30: 30 kg P ha , P60: 60 kg P ha , BF0: no biofertilizer application (control), BNF: biological N fertilizer (Rhizobium ciceri); BPF: biological phosphorus fertilizer (Pseudomonas striata). LSD (P<0.05) Height per plant Number of branches per plant Number of leaves per plant Fresh weight per plant Dry weight per plant Phosphorus (P) 0.829 0.235 0.753 0.071 0.014 Biofertilizer NS NS 0.870 0.082 0.016 P × Biofertilizer 1.657 NS 1.507 0.142 0.028 Table 2. Effect of three P levels with respect to four biofertilizer treatments on photosynthetic parameters of chickpea Treatments PN(µM -2 -1 CO2 m s ) Internal CO2 concentration (ppm) Stomatal conductance -2 -1 (µM m s ) Transpiration -2 -1 mol m s ) rate (m Main effects of phosphorus (P) b b b b P0 5.23 313 237 6.48 a a a a P30 5.88 320 254 7.08 a a a a P60 5.62 326 256 7.02 Main effects of biofertilizers (BF) c c d BF0 4.86 320 236 5.26 b c c BNF 5.54 320 239 7.18 a b b BPF 5.95 320 257 7.41 a a a BNF+BPF 5.95 321 264 7.60 Effects of interaction (P × BF) bc c f P0 × BF0 4.54 305 227 5.19 b c f P30 × BF0 4.89 320 237 5.23 b bc e P60 × BF0 5.14 334 243 5.37 b cd d P0 × BNF 5.10 314 220 6.54 a bc b P30 × BNF 5.95 320 240 7.48 ab b b P60 × BNF 5.58 325 258 7.52 ab b d P0 × BPF 5.67 315 253 7.05 a b a P30 × BPF 6.35 320 255 7.80 ab b c P60 × BPF 5.82 325 262 7.37 ab b d P0 × BNF+BPF 5.60 320 249 7.15 a a a P30 × BNF+BPF 6.32 321 283 7.82 a b a P60 × BNF+BPF 5.93 322 260 7.83 -1 -1 P0: no phosphorus application (control); P30: 30 kg P ha ; P60: 60 kg P ha . BF0: No biofertilizer application (control); BNF: Biological N fertilizer (Rhizobium ciceri); BPF: Biological phosphorus fertilizer (Pseudomonas striata). LSD (P<0.05) Net photosynthesis Internal CO2 Conc. Stomatal conductance Transpiration rate Phosphorus (P) 0.239 6.742 5.000 0.060 Biofertilizer 0.276 NS 6.000 0.069 P × Biofertilizer 0.479 NS 11.000 0.120 170 Glob. J. Agric. Agric. Sci. 171 Table 3. Effect of three phosphorus levels with respect to four biofertilizer treatments on biochemical parameters of chickpea. Treatments Nitrate reductase activity Carbonic anhydrase activity Leghemoglobin -1 -1 -1 -1 -1 (n mol NO2 g FW h ) (m mol CO2 Kg FW s ) content (mg g ) Main effects of phosphorus (P) b b c P0 247 1105 2.51 a a b P30 272 1133 2.93 a a a P60 276 1143 3.03 Main effects of biofertilizers (BF) b ab d BF0 238 1106 2.14 a a c BNF 273 1131 2.86 a a a BPF 273 1136 3.02 a a a BNF+BPF 277 1135 3.28 Effects of interaction (P × BF) c ab d P0 × BF0 231 1100 2.06 bc a cd P30 × BF0 237 1102 2.12 b a c P60 × BF0 246 1117 2.23 b ab c P0 × BNF 252 1100 2.36 a a b P30 × BNF 278 1142 3.00 a a a P60 × BNF 289 1152 3.23 b a c P0 × BPF 250 1110 2.38 a a a P30 × BPF 287 1142 3.30 a a a P60 × BPF 283 1155 3.37 b a a P0 × BNF+BPF 257 1110 3.24 a a a P30 × BNF+BPF 287 1146 3.29 a a a P60 × BNF+BPF 286 1150 3.31 -1 -1 P0: no phosphorus application (control); P30: 30 kg P ha ; P60: 60 kg P ha . BF0: No biofertilizer application (control); BNF: Biological N fertilizer (Rhizobium ciceri); BPF: Biological phosphorus fertilizer (Pseudomonas striata). LSD (P<0.05) Nitrate reductase activity Carbonic anhydrase activity Leghemoglobin content Phosphorus (P) 6.913 27 0.090 two-factor factorial procedure. Fisher’s least significant difference (LSD) was used to test the significance at the 5% probability level. The data were analyzed using SPSS-17 (SPSS Inc., Chicago, IL, USA). RESULTS Growth Parameters As per the main effects of P, progressive application of P enhanced the plant fresh and dry weight gradually, with P 60 proving the best. For rest of the growth attributes, P 30 and P60 gave statistically equal values. However, P30 as well as P60 proved invariably better than P0 (control) for all the growth characteristics studied (Table 1). Considering the main effects of biofertilizer treatments, the maximal values of the growth attributes were generally shown by BNF+BPF and/or BPF compared to the control (BF0), which always gave the poorest results. Interaction of inorganic P fertilizer and biofertilizer was significant for most of the growth parameters. In general, P30 × BPF+BNF resulted in the highest values. However, for fresh and dry weight per plant, the maximum values were attained with P60 × BPF, which was statistically at par with P30 × BPF and/or P30 × BPF+BNF. Photosynthetic Parameters and Transpiration Rate Main effects of P application resulted in enhanced values of net photosynthetic rate (PN), stomatal conductance (gs) , Biofertilizer 7.983 29 0.110 P × Biofertilizer 13.826 53 0.180 internal CO2 concentration and transpiration rate (E) compared to the control (P0) (Table 2). Level P30 resulted in the highest extent of PN; while P60, being statistically at par with P30, gave the highest stomatal conductance, internal CO2 concentration and transpiration rate. Treatment P30 surpassed the P0 by 12.42, 2.24, 7.17, and 9.26% in PN, internal CO2 concentration, gs and E respectively. As per the main effects of biofertilizer treatments, BNF+BPF resulted in the highest PN, gs, and E, with BNF+BPF excelling the control (BF 0) by 22.42, 11.86, and 44.49%, respectively. Interaction P30 × BPF and P30 × BNF+BPF enhanced the PN to the highest extent. In addition, P30 × BNF+BPF was most advantageous interaction for ‘gs’ and ‘E’. It exceeded the control (P0 × BF0) by 29.24, 28.64, and 50.67% for ‘PN’, ‘gs’ and ‘E’, respectively. The effect of biofertilizer treatments and that of interaction between biofertilizer treatments and P levels was not significant regarding the internal CO2 concentration. Activity of Nitrate Reductase (NR) and Carbonic Anhydrase (CA) In relation to the main effects of phosphorus, P60 as well as P30 accounted for the greatest extent of NR and CA activities. Level P30 exceeded P0 by 10.12 and 2.53% in NR and CA activity, respectively (Table 3). As for the main effects of biofertilizer treatments, BNF+BPF was Tariq et al. equal to BNF and BPF, registering the highest levels of NR and CA activities compared to BF0. BNF+BPF resulted in 16.38 and 2.62% increase in the NR and CA activity, respectively, over no biofertilizer application (BF0). Several of the P × biofertilizer interactions, including P30 × BNF+BPF and P60 × BNF+BPF, resulted in statistically equal values for NR and CA activities, exceeding the lowest interaction (P0 × BF0) significantly. Leghemoglobin Content This study revealed a significant effect of phosphorus and N & P biofertilizers on Leghemoglobin content of root nodules (Table 3). Regarding the main effects of P, level P30, equaled with P60, exhibited the highest leghemoglobin content in the root nodules. It excelled the control (P0) by 16.73%. In consideration with the main effects of biofertilizer treatments, BNF+BPF resulted in the maximum content of leghemoglobin, exceeding the control (BF0) by 53.27%. Interaction P60 × BPF, which was statistically equal to P30 × BNF+BPF and several other interactions, resulted in the highest leghemoglobin content. P60 × BPF and P30 × BNF+BPF exceeded the lowest interaction (P0 × BF0) by 63.59 and 59.71%, respectively. DISCUSSION Growth Parameters Growth of plant organs depend on proper supply of mineral nutrients, including N and P that play important role in growth and many other physiological processes of plants (Moorby and Besford, 1983). Of the macronutrients, P is required in large amounts specifically by legumes for their proper growth and development (Wan et al., 1991). Hence, the role of P in promoting the chickpea growth attributes, as observed in this study (Table 1), was expected. In this connection, the present results resemble with those obtained by different workers regarding chickpea (Yahiya and Samiullah, 1995; Bahadur et al., 2002; Pathak et al., 2003). In line with our investigation, there was observed a positive effect of dual inoculation (BNF+BPF) on the growth of chickpea (Sonoboir and Sarawgi, 2000) Soybean (Fatima et al., 2006), Lentil (Kumar and Chandra, 2008) and black gram (Vigna mungo L.) (Selvakumar et al., 2009). The investigations employing biofertilizers to improve growth attributes of chickpea (Saraf et al., 1997; Dutta and Prohit, 2009; Nishita and Joshi, 2010; Namvar and Sharifi, 2011) and other leguminous crops (Naeem and Khan, 2005; Fatima et al., 2006; Selvakumar et al., 2009; Selvakumar et al., 2012) also substantiate the present findings in this regard. 172 Photosynthetic Parameters and Transpiration Rate In this study, P30 proved to be the optimum P level, exhibiting the greatest values for all the photosynthetic parameters as well as for transpiration rate (E) (Table 2). These results are confirmed by the findings of other researchers, who observed a positive effect of P application on several of these parameters regarding soybean (Fredeen et al., 1990), groundnut (Hossaini and Hamid, 2007) and cluster bean (Burman et al., 2009). Increased PN (due to P application could be due to the prompt and adequate supply of carbon dioxide to the mesophyll cells of the leaves that is evident by the Penhanced internal CO2 concentration and stomatal conductance. Similarly, the P-improved ‘E’ recorded in the plants supplied with P30/P60 could also be expected because P application also enhanced the ‘gs’ in the present study (Table 2). In line with our investigation, significant decrease in ‘PN’ and ‘gs’ was reported due to P deficiency in sunflower and soybean (Guidi et al., 1994). Besides, there was observed enhanced photosynthesis as a result of P supply to tobacco (Gao et al., 1989). In this study, BNF+BPF proved to be the most beneficial biofertilizer treatment for ‘PN’, ‘gs’ and transpiration rate. Not many references are available on the effect of biofertilizers on photosynthesis and the related parameters. However, enhancement in photosynthetic efficiency of green gram (Vigna radiata) due to N biofertilizer, as noted by Sharma (2001), could be considered in line with the present results in this regard. Interaction P30 × BNF+BPF, equaled by certain other interactions, enhanced the net photosynthesis, ‘gs’ and ‘E’ to the greatest extent. Accordingly, this interaction resulted into the highest yield of chickpea reported elsewhere (Moin Uddin et al., 2014). Activity of Nitrate Reductase (NR) and Carbonic Anhydrase (CA) The activity of NR in plants is influenced by different growth conditions including not only the environmental factors such as light and temperature, but also by the application of mineral fertilizers, particularly P (Oaks, 1985). The presence of P in the nutrient solution has earlier been reported to induce greater nitrate assimilation in corn (de Magalhaes et al., 1998) and Phaseolus vulgaris L. (Gniazdowaska et al., 1999). In this regard, our results are in agreement with those of Naeem and Khan (2005) in the case of Cassia tora. Carbonic anhydrase is known to have its important role in photosynthesis, which is obvious by its presence in all photosynthesizing tissues (Taiz and Zeiger, 2006). It catalyzes the reversible hydration of CO2, thereby increasing its availability to the photosynthetic enzyme RuBisCO (Badger and Price, 1994). The improvement in CA activity in this study (Table 3) could be as a result of Glob. J. Agric. Agric. Sci. 173 adequate availability of N and P at the site of their metabolism, owing to the application of P and the N and P biofertilizers. A plausible cause for the enhancement of CA activity due to application of inorganic P and P- biofertilizer might the positive influence of P availability to plants or the de novo synthesis of CA as argued by (Okabe et al. 1980). Leghemoglobin Content In accordance with this investigation (Table 3), the increase in leghemoglobin content of the root nodules might be due to the improved availability of P to the root nodules due to combined application of inorganic P fertilizer and N and P biofertilizers (P30 × BNF+BPF). Similar to our results, there was observed a positive effect of P application on nodule leghemoglobin content in Lablab purpureus (Santhaguru and Hariram, 1998) and Cassia tora (Naeem and Khan, 2005). In conformity with our study, there was noted beneficial effect of inorganic P fertilizer as well as that of N and P biofertilizers on leghemoglobin content in chickpea by Dutta and Prohit (2009). The greatest content of leghemoglobin in root nodules (N2 fixation) and maximum level of PN and the activities of CA and NR in the leaves in this study might be the reason for the enhanced yield and quality of chickpea reported elsewhere (Moin Uddin et al., 2014). CONCLUSION Phosphorus application improved all the growth and physiological attributes studied compared to no P application (P0), with P30 and P60 being statistically equal in most cases. Application of N and P biofertilizers increased the values of most of the parameters studied significantly compared to no biofertilizer application (BF0). Of the biofertilizer treatments, BNF+BPF resulted in the highest values almost invariably. P30 × BNF+BPF was the most profitable interaction between inorganic P levels and biofertilizer treatments for most growth and physiological attributes as it gave the greatest values; hence, it could be realized as the optimum combination of inorganic P level and biofertilizer treatment for chickpea growth and metabolism. REFERENCES Alagawadi AR, Gaur AC (1988). Associative effect of Rhizobium and phosphate-solubilizing bacteria on the yield and nutrient uptake of chickpea. Plant Soil 105, 241-246. Asad SA, Bano A, Farooq M, Aslam M, Afzal A (2004). Comparative study of the effects of biofertilizers on nodulation and yield characteristics of mung bean (Phaseolus vulgaris l.). Int J Agric Biol 6, 837-843. Badger MR, Price GD (1994). The role of carbonic anhydrase in photosynthesis. Ann Rev Plant Physiol Plant Mol Biol 45, 369-392. Bahadur MM, Ashrafuzzaman M, Kabir MA, Choudhary MF, Majumdar AN (2002). Response of chickpea (Cicer arietinum L.) varieties to different levels of phosphorus. Crop Res 23, 293-299. Burman U, Garg BK, Kathju S (2009). Effect of phosphorus application on clusterbean under different intensities of water stress. J Plant Nutr 32, 668-680. de Magalhaes JV, Alves VMC, de Novais RF, Mosquim PR, Magalhaes JR, Bahia AFC, Huber DMF (1998). Nitrate uptake by corn under increasing periods of phosphorus starvation. J Plant Nutr 21, 17531763. Dutta D, Prohit B (2009). Performance of chickpea (Cicer arietinum L.) to application of phosphorus and biofertilizer in laterite soil. Arch Agron Soil Sci 55, 147-155. Dwivedi RS, Randhawa NS (1974). Evaluation of rapid test for the hidden hunger of zinc in plants. Plant Soil 40, 445-451. Fatima Z, Zia M, Chaudhary MF (2006). Effect of Rhizobium strains and phosphorus on growth of soybean (Glycine max) and survival of Rhizobium and P solubilizing bacteria. Pak J Bot 38, 459-464. Fredeen AL, Raab TK, Rao IM, Terry N (1990). Effects of phosphorus nutrition on photosynthesis in Glycine max (L.) Planta 181, 399-405. Gao SJ, Chen SS, Li MQ (1989). Effects of phosphorus nutrition on photosynthesis and photorespiration in tobacco leaves. Acta Phytophysio Sinica 15, 281-287. Giaquenta RT, Quebedeaux B (1980). Phosphate induced changes in assimilate partitioning in soybean leaves during pod filling. Plant Physiol 65, Suppl. 119. Gniazdowaska A, Krawczak A, Mikulska M, Rychter AM (1999). Low phosphate nutrition alters bean plants ability to assimilate and translocate nitrate. J Plant Nutr 22, 551-563. Guidi L, Pallini M, Soldatini GF (1994). Influence of phosphorus deficiency on photosynthesis in sunflower and soybean plants. Agrochimica 38, 211-223. Hossaini MA, Hamid A (2007). Influence of N and P fertilizer application on root growth, leaf photosynthesis and yield performance of groundnut. J Agric Res 32, 369-374. Huisman J, Vander Poel AF (1994). Aspects of the nutritional quality and use of cool season food legumes in animal feed. In: Expanding the Production and Use of Cool Season Food Legumes (Eds. FJ Muehlbauer, WJ Kaiser). Kluwer Academic Publishers, Dordrecht, pp. 53-76. Jaworski, E.G., 1971. Nitrate reductase assay in intact plant tissues. Biochem Biophys Res Commun 43, 1247-1279. Kumar R, Chandra R (2008). Influence of PGPR and PSB on Rhizobium leguminosarum Bv. viciae strain competition and symbiotic performance in lentil. World J Agric Sci 4, 297-301. Mahdi SS, Hassan GI, Samoon SA, Rather HA, Dar SA, Zehra B (2010). Bio-fertilizers in organic agriculture. J Phytol 2, 42-54. Moin Uddin, Hussain S, Khan MMA, Hashmi N, Idrees M, Naeem M, Dar T (2014). Use of N and P biofertilizers reduces inorganic phosphorus application and increases nutrient uptake, yield, and seed quality of chickpea. Turk J Agric For 38, 47-54. Moorby J, Besford RT (1983). Mineral nutrition and growth. In: Encyclopedia of Plant Physiology (New Series). Vol. 15B (Eds. A Pearson, MH Zimmerman). Springer Verlag, Berlin, pp. 481-527. Naeem M, Khan MMA (2005). Growth, physiology and seed yield of Cassia tora (syn. Cassia obtusifolia) as affected by phosphorus fertilization. J. Med Aromat Plant Sci 27, 4-6. Namvar A, Sharifi RS (2011). Phenological and morphological response of chickpea (Cicer arietinum L.) to symbiotic and mineral nitrogen fertilization. Žemdirbystė=Agriculture 98, 121-130. Nishita G, Joshi NC (2010). Growth and yield response of chickpea (Cicer arietinum) to seed inoculation with Rhizobium sp. Nature Sci 8, 232-236. Okabe K, Lindlar A, Tsuzuki M, Miyachi S (1980). Carbonic anhydrase and ribulose 1, 5-biphosphate carboxylase and oxygenenase. FEBS Letters 114: 42-144. Oaks A (1985). Nitrogen metabolism in roots. Ann Rev Plant Physiol Plant Mol Biol 36: 407-414. Pathak S, Namdeo KN, Chakrawarti VK, Tiwari RK (2003). Effect of biofertilizers, diammonium phosphate and zinc sulphate on growth and yield of chickpea (Cicer arietinum L.). Crop Res 26, 42-46. Rudresh DL, Shivaprakash MK, Prasad RD (2005). Effect of combined application of Rhizobium, phosphate solubilizing bacterium and Trichoderma spp. on growth, nutrient uptake and yield of chickpea (Cicer aritenium L.). Appl Soil Ecol 28, 139-146. Tariq et al. Sadasivam S, Manickam A (2008). Biochemical methods. 3rd ed, New Age International (P) Ltd. Publishers, New Delhi, India. Santhaguru K, Hariram N (1998). Effect of Glomus mosseae and Rhizobium on growth, nodulation and nitrogen-fixation in Lablab purpureus (L.) under different phosphorus regimes. Indian J Plant Physiol 3, 156-158. Saraf CS, Shivakumar BG, Patil RR (1997). Effect of phosphorus, sulphur and seed inoculation on performance of chickpea (Cicer arietinum L.). Indian J Agron 42, 323-28. Sattar MA, Gaur AC (1987). Production of auxins and gibberellin by phosphate dissolving micro-organisms. Zentralblatt Fiir Mikrobiologie 142, 393-398. Sharma S (2001). Growth, physiological and yield aspects of mungbean (Vigna radiate L.) as affected by inoculation treatment by different strains of Bradyrhizobium culture. Res Crops 2, 112-115. Selvakumar G, Lenin M, Thamizhiniyan P, Ravimycin T (2009). Response of biofertilizers on the growth and yield of blackgram (Vigna mungo L.). Recent Res Sci Tech 1, 169-175. Selvakumar G, Reetha S, Thamizhiniyan P (2012). Response of biofertilizers on growth, yield attributes and associated protein profiling changes of blackgram (Vigna mungo L. Hepper). World App Sci J 16, 1368-1374. Singh KB (1997). Chickpea (Cicer arietinum L.). Field Crops Res 53, 161-170. 174 Singh KC (1983). Response of green gram (Vigna radiata) to phosphorus application in arid zone. Annals Arid Zone Res 21, 275278. Soltani A, Robertson MJ, Mohammad-Nejad Y, Rahemi-Karizaki A (2006). Modeling chickpea growth and development: Leaf production and senescence. Field Crops Res 99, 14-23. Sonoboir HL, Sarawgi SK (2000). Nutrient uptake, growth and yield of chickpea as influenced by phosphorus, Rhizobium and phosphate solubilizing bacteria. Madras Agric J 87, 149-151. Stephens JHG, Rask HM (2000). Inoculants production and formulation. Field Crops Res 65, 249-258. Taiz L, Zeiger E (2006). Plant physiology. 4th ed, Sinauer Associates, Sunderland, Massachusetts. Verma LN (1993). Organic in soil health and crop production. In: Tree Crop Development Foundation, (Ed. PK Thampan), Cochin, India, pp. 151-184. Wan Othman WM, Lie TA, Mannetje LT, Wassink GY (1991). Low level phosphorus supply affecting nodulation, N 2 fixation and growth of cowpea (Vigna unguiculata L. Walp). Plant Soil 135, 67-74. Yahiya M, Samiullah (1995). Effect of phosphorus on nodulation and nitrogen fixation of chickpea (Cicer arietinum L.). Bioved 6, 161-166.