Eeffect of biofertilizers and foliar application of organic acids on yield, nutrient uptake and soil microbial activity in soybean

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1 Legume Research, 39 (2) 2016 : Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Eeffect of biofertilizers and foliar application of organic acids on yield, nutrient uptake and soil microbial activity in soybean N.N. Lingaraju*, C.S. Hunshal and S.R. Salakinkop University of Agricultural Sciences, Dharwad , Karnataka, India. Received: Accepted: DOI: /lr.v0iOF.6784 ABSTRACT A field experiment was undertaken during kharif 2012 to study effect of biofertilizers and foliar application of organic acids on yield, nutrient uptake and microbial activity of soybean at MARS, Dharwad under rainfed condition. The experiment was laid out in RCBD factorial having twenty treatment combinations and replicated thrice. The treatments comprised of four P-Solubilizers (PSB, VAM, PSB + VAM and Control) and five foliar spray of organic acids (Humic acid, Lecithin, Citric acid, Maleic acid and control). The results revealed that higher seed yield of soybean (35.96 q ha -1 ) was obtained with the treatment combination of dual inoculation of PSB+VAM with foliar spray of 0.1% humic acid at flower initiation and was higher to an extent 22.5 per cent compared to control (27.90 q ha -1 ). Similar trend was observed on microbial activity and nutrient uptake (N, P 2 and K O) of the soybean crop. 2 Key words: Nutrient uptake, Organic acids, PSB, Soybean, VAM. INTRODUCTION Phosphorus (P) is one of the most important elements for plant growth and metabolism. It plays a key role in many plant processes such as energy metabolism, synthesis of nucleic acids and membranes, photosynthesis, respiration, nitrogen fixation and enzyme regulation. Adequate phosphorus nutrition enhances many aspects of plant development including flowering, fruiting and root growth. In many soil types, P is the most limiting nutrient for the production of crops. Maintaining a proper P supplying level at the root zone can maximize the efficiency of plant roots to mobilize and acquire P from the rhizosphere by an integration of root morphological and physiological adoptive strategies. Soil type, ph range, fertilizer (rate and type), cropping system etc., influence the fate of P in soil and plant. The use of phosphate solubilizing bacteria and VAM as biofertilizers and organic acids was suggested as a sustainable solution to improve plant nutrient and production. Soil microorganisms play a key role in soil P dynamics and subsequent availability of phosphate to plants. Co-inoculation of PSM and arbuscular mycorrhizas (AM) may enhance plant acquisition of P from insoluble P sources. Phosphate solubilizing bacteria (PSB) have been used for the crop production since PSB like Bacillus megaterium, Bacillus polymyxa, Pseudomonas striata, Pseudomonas flouresence are important in phosphorus nutrition by enhancing its availability to plants through release from inorganic and organic soil P pools by solubilization and mineralization. Principal mechanism in soil for mineral phosphate solubilization including lowering of soil ph by microbial production of organic acids and mineralization of organic P by acid Phosphatase. Use of phosphorus solubilizing bacteria as inoculants increases P uptake. Greater efficiency of P solubilizing bacteria has been shown through co-inoculation with other beneficial bacteria and mycorrhiza. On the other hand use of organic-mineral fertilizers like humic acid has increased with increasing the agricultural production and the most economical. Humic acid is almost applied directly to the soil or as a foliar application to the plants. It is one of the major components of humic substances. Humic matter is formed through the chemical and biological humification of plant and animal matter and through the biological activities of microorganisms. The effects of humic substances on plant growth depend on the source and concentration, as well as on the molecular fraction weight of humus. MATERIALS AND METHODS A field experiment was conducted at Main Agricultural Research Station Dharwad during Kharif The ph of the soil was neutral (7.3) and low in available *Corresponding author s yadavv39@gmail.com.

2 Volume 39 Issue 2 (2016) 257 nitrogen (238.8 kg N ha -1 ), medium in available phosphorous (35.22 kg P 2 ha -1 ) and high in available potassium (341.3 kg K 2 O ha -1 ). The experiment was laid out in RCBD factorial having twenty treatment combinations and replicated thrice and genotype DSb-21 was used in the study. The treatments comprised of four main factors (PSB, VAM, PSB + VAM and Control) and five sub factors of foliar application of organic acids at flower initiation stage (Humic acid, Lecithin, Citric acid, Maleic acid and control). Recommended fertilizer dose of 40:80:25 kg N, P 2 and K 2 O per hectare in the form of Urea, DAP and MOP was applied as a basal at the time of sowing. PSB (Pseudomonas striata) obtained from the Department of Agricultural Microbiology, UAS, Dharwad was treated at the rate of 6g kg -1 of seeds. Rhizobium (Sb-120) was also applied at the rate of 6 g kg -1 of seeds. 20 kg ha -1 was applied to soil at the time of sowing. Five plants from net plot area were randomly selected and observations on growth and yield parameters were recorded at 30, 60 DAS (days after sowing) and at harvest. At harvest, yield and its components such as number of pods plant -1, number of seeds pod -1, hundred seed weight, seed yield plant - 1, seed yield ha -1 and haulm yield ha -1 were determined at maturity stage. All the data pertaining to the present investigation were statistically analyzed as per the method described by Panse and Sukhatme (1967). The level of significance used in F and T test was P= The mean differences among the treatments were compared by Duncan Multiple Comparison Test (DMRT) at 0.05 level of probability. RESULTS AND DISCUSSION Yield and yield components: Higher seed yield of soybean (35.96 q ha -1 ) to the extent of 28 per cent was obtained with the treatment combination of dual inoculation of PSB+VAM with foliar spray of 0.1% humic acid at flower initiation. Compared to control (27.90 q ha -1 ) (Table 2). The higher seed yield was mainly due to positive association between yield attributing characters viz., Such as higher number of pods plant -1, number of seeds plant -1 and 100 seed weight. Significantly higher number of pods plant -1 (92.10 g plant -1 ), number of seeds plant -1 (3.10) and 100 seed weight (13.10 g plant -1 ) (Table 1) were also recorded by inoculation of PSB + VAM with foliar spray of 0.1% humic acid at flower initiation compared to control (64.9 pod plant and 10.33g plant -1, respectively). This might be due to better nodulation of soybean roots owing to increased availability of phosphorus. The improvement in nodulation with inoculationof P-solubilizer might have resulted in higher nitrogen fixation and consequent increase in vegetative Table 1: Number of pods plant -1, number of seeds pod -1 and 100 seed weight (g) in soybean as influenced by biofertilizers and foliar application of organic acids B3 - PSB+VAM F3- Foliar spray of citric 0.1% at flower initiation stage

3 258 LEGUME RESEARCH - An International Journal Table 2: Seed yield (q ha -1 ) and haulm yield (q ha -1 ) in soybean as influenced by biofertilizers and foliar application of organic acids Treatment Seed yield (q ha -1 ) Haulm yield (q ha -1 ) Spray of organic acids F1 F2 F3 F4 F5 Mean F1 F2 F3 F4 F5 Mean B bf 29.86ei 31.05cg 30.81fi 29.34be 30.54b 52.39ac 49.41be 50.67bd 49.03be 46.93df 49.68b B di 30.07bf 31.44bf 30.67ch 29.58fi 30.79b 53.05ab 49.69be 50.91bd 50.42bd 47.49cf 50.31b B a 32.71bc 33.81b 32.94bc 32.47bd 33.58a 55.75a 51.52ad 53.01ab 52.35ac 49.80bd 52.48a B di 28.27hi 29.33fi 28.61gi 27.90i 28.88c 46.54df 46.98df 44.82ef 43.07f 43.71f 45.03c Mean 32.53a 30.23c 31.41b 30.76bc 29.82c 51.93a 49.40b 49.85ab 48.72bc 46.98c S.Em+ S.Em+ B F BXF B1 - PSB B2 - VAM B3 - PSB+VAM B4 - Control F1- Foliar spray of Humic 0.1% at flower initiation stage F2- Foliar spray of 0.1% at flower initiation stage F3- Foliar spray of Citric 0.1% at flower initiation stage F4- Foliar spray of Maleic 0.1% at flower initiation stage growth and dry matter production and also mediated by biological process as noticed by increased microbial activity. Biofertilizers play a key role in enhancing the efficiency of utilization of native as well as applied nutrients. The greater yield response due to dual inoculation of VAM fungus and PSB than with PSB alone can be attributed to the activity of the VAM fungus in transporting extra phosphorus solubilized by PSB from and beyond the root zone into the plant roots which in the absence of VAM hyphae gets refixed by soil constituents during the course of slower diffusion towards plant roots. These results are in accordance with the findings of Chitale et al. (2012) in soybean. They reported that higher seed yield of soybean was due to inoculation of PSB + VAM than inoculation of either PSB or VAM alone. Foliar spray of 0.1% humic acid helps in quick absorption of nutrients, which enhanced the growth of root and shoot effectively resulting in higher uptake of nutrients. Spraying of humic acid mainly reduced the flower drop and ultimately enhanced the pod setting and resulted in higher seed yield. The results are in line with the findings of Dixit and Elamathi (2007). Combined application of PSB+VAM with foliar spray of 0.1% humic acid might have helped to enhance the biological activity in the soil, soil characters improvements, better root development, improved transport of nutritional elements, enhanced chlorophyll content, protein synthesis and photosynthesis, solubilization of nutrients resulting in higher nutrients uptake by soybean compared to other treatments (Lee and Bartlett, 1976) suggesting existence of synergetic effect of combined applications of mineral nutrients and humic acid substances. Nutrient uptake in soybean: Significantly higher uptake of nitrogen, phosphorus and potassium by soybean at harvest (223.65, and 240 kg ha -1, respectively) (Table 3) by soybean was recorded with foliar application of 0.1% humic acid at flower initiation as compared to other sprays and control. Higher P uptake might be due to solubilization of fixed phosphorus by P-solubilizer due to secretion of organic acids. Similar findings were made by Sharma and Namdeo (1999). Foliar application of humic acid significantly enhances the presence of various antioxidants in the leaves and stimulates root initiation thus resulting in higher uptake of nitrogen, phosphorous, potassium, magnesium, calcium, zinc, iron and copper (Schmidt and Zhang, 1998). Combined effect from biofertilizers and humic acid might have resulted in the improvement of soil fertility and crop productivity. Microbial activity: Significantly higher microbial load of rhizobium population and Rhizosphere P- Solubilizers population (Number X 10 4 g -1 ) was influenced by combined application of PSB+VAM. At 30, 60 DAS and at harvest, dual inoculation of PSB+VAM recorded significantly higher rhizobium population (18.60, and 25.6 X 10 4 g -1 ) compared to singular inoculation of VAM (18.60, and 22.2 X 10 4 g -1 ) and PSB (16.93, and 22.9 X 10 4 g -1 ) alone over the control (18, and 22.1 X 10 4 g -1 ). Among the foliar sprays, 0.1% humic acid recorded significantly higher rhizobium population at 60DAS (34.23 X 10 4 g -1 ) compared to control (27.54 X 10 4 g -1 ). With respect to interaction, dual inoculation of PSB+VAM and foliar spray of 0.1% humic acid recorded

4 Volume 39 Issue 2 (2016) 259 Table 3: Nutrient uptake (N, P 2 and K 2 O kg ha -1 ) of soybean at harvest as influenced by biofertilizers and foliar application of organic acids B3 - PSB+VAM F3- Foliar spray of Citric 0.1% at flower initiation stage DAS Days after sowing Table 4: Rhizobium colony count (Number X 10 4 g -1 in soybean at different growth stages as influenced by biofertilizers and foliar application of organic acids B3 - PSB+VAM F3- Foliar spray of Citric 0.1% at flower initiation stage

5 260 LEGUME RESEARCH - An International Journal higher rhizobium count at 60DAS and at harvest (35.0 and X 10 4 g -1 ). Significantly lower rhizobium count was recorded by uninoculated control (16 and 17.0 X 10 4 g -1 ) (Table 4). Microorganisms with phosphate solubilizing potential increase the availability of soluble phosphate and enhance the plant growth by improving biological nitrogen fixation. Pseudomonas spp. enhanced the number of nodules, dry weight of nodules, yield components, seed yield, nutrient availability and uptake in soybean crop as reported by Sharma et al. (2011). Adequate supply of P with Rhizobium strains (mixture inoculation) play an important role in physiological and developmental processes in plant life and the favourable effect on nutrient uptake accelerate the growth processes, which ultimately resulted in increased seed yield of the crop. Increase in N content in seed, shoot and total N uptake due to Rhizobium inoculation was mainly due to significant increase in nodulation, resulting in higher accumulation of N due to atmospheric N 2 fixation. Higher P uptake with Rhizobium inoculation was due to the ability of applied rhizobium to solubilize precipitated P components thereby increased P uptake in plants as reported by Fatima et al. (2007). Higher available P due to the solubilization and inoculated PSB might cause enhancement of nutrient uptake and population of nitrogen fixers. Similarly, at 30, 60 DAS and at harvest, dual inoculation of PSB+VAM recorded significantly higher P- Solubilizers population (19.86, and CFU x 10 4 g -1 soil) than inoculation of PSB (16.10, and CFU x 10 4 g -1 soil) and VAM alone (11.29, and CFU x 10 4 g -1 soil) compared to the uninoculated control (9.90, and CFU x 10 4 g -1 soil) (Table 5). The greater yield response due to dual inoculation of VAM fungus and PSB than with PSB alone can be attributed to the activity of the VAM fungus in transporting extra phosphorus solubilized by PSB from and beyond the root zone into the plant roots which in the absence of VAM hyphae gets refixed by soil constituents during the course of slower diffusion towards plant roots. Increase in the PSB population enhances the phosphorous availability to plants by lowering soil ph and by microbial production of organic acids and mineralization of organic P by acid phosphatases. The phosphate- solubilizing bacteria promoted root colonization when associated with mycorrhizal fungi. These organisms besides providing P also facilitate the growth of plants by improving the uptake of nutrients and stimulating the production of phytohormones. Son et al., (2007) revealed that inoculation with PSB enhanced beneficial bacterial count and their communities in the soils and improved the nutrient uptake. Table 5: Rhizosphere P- Solubilizers population (Number X 10 4 g -1 in soybean at different growth stages as influenced by biofertilizers and foliar application of organic acids B3 - PSB+VAM F3- Foliar spray of Citric 0.1% at flower initiation stage DAS Days after sowing

6 Volume 39 Issue 2 (2016) 261 CONCLUSION Combined application of biofertilizers and organic acids (PSB+VAM in combination with foliar spray of 0.1% humic acid) along with recommended fertilizer proved effective in significantly enhancing the yield, microbial activity and nutrient uptake by soybean. Dual inoculation of PSB+VAM and foliar spray of 0.1% humic acid recorded significantly higher rhizobium and P- solubilizer population in soil. REFERENCES Chitale, S., Sarawgi, S. K. and Sandeep, (2012), Effect of phosphorus application along with PSB, Rhizobium and VAM on P fractionation and productivity of soybean. Indian J. Agron., 57(1): Dixit, R. S. and Elemathi, (2007), Effect of foliar application of DAP, micronutrients and NAA on growth and yield of green gram (Vigna radiata. L). Legume Res., 30(40): Fatima, Z., Zia, M. and Chaudhary, M. F., (2007), Interactive effect of Rhizobium strains and P on soybean yield, nitrogen fixation and soil fertility. Pak. J. Bot., 39: Lee, Y. S. and Bartlett, R. J., (1976), Stimulation of plant growth by humic substances. Soil Sci. Soc. Am. J., 40: Panse, V. G. and Sukhatme, P. V., (1967), Statistical Methods for Agricultural Workers, ICAR., Publication New Delhi, 359. Schmidt, R. E. and Zhang, Z., (1998), How humic substances help turfgrass grow. Golf Course Management. 66(7): Sharma, K. N. and Namdeo. K. N., (1999), Effect of biofertilizer and phosphorus on NPK content, uptake and seed quality of soybean and nutrient status of soil. Crop Res., 17(2): Son, Diep, C. N., Truong, T. M. and Tran, T. A. T., (2007), Effect of co-inoculants (bradyrhizobia and phosphate solubilizing bacteria) liquid on soybean under rice based cropping system in the mekong delta. Omonrice, 15: Sharma, U. C., Datta, M. and Sharma, V., (2011), Effect of phosphorus on the yield and nutrient uptake by soybean cultivars on acidic soil. J. Soil Sci., 1:

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