Original Paper Pradhan et al Int. J. Biotech Biosci, ISSN 2231-0304 Vol 4 (3) : 248-255 (2014) EFFICACY STUDIES OF ORGANIC ACID BASED CHELATED PLANT NUTRIENTS FOR GROWTH AND GRAIN QUALITY UNDER CONTROLLED AND FIELD CONDITION Pradhan A 1 *, Gangurde N S 1, Sairam KVSS 1, Verma V 2 1 Prathista R&D Center, Prathista Industries Limited, 10-170/23, Bharathi Nagar Colony, Temple Alwal, Secunderabad 500 010 2 School of Biotechnology, SMVD University, Jammu Email: *amitbiotechnology@yahoo.co.in Abstract How much of a positive impact on plants can organic acid based chelated agri-inputs have? Research conducted in Prathista R&D center, Prathista Industries limited,choutuppal,telangana, India illustrated and reported fermentative chelated nutrients for improve in qualitative and quantitative crop productivity. Trail conducted on Biozinc (Lactic Acid chelates) and Suryamin (Amino Acid) in controlled environment and field condition on sorghum plant. The results have indicated the improvement in the photosynthesis rate, transpiration rate, stomatal conductance, chlorophyll content and photochemical efficiency. Collectively these chelated agri-inputs improves yield improves the grain quality i.e. protein content 11-37%. INTRODUCTION Agriculture is the backbone of Indian economy. In many parts of the India where poor soil conservation and management methods prevail, long-term productivity is projected to decline substantially unless soil management practices improve. Improving soil fertility is one of the most common tactics to increase agricultural and forest production. Maintaining high levels of available major nutrient nitrogen (N) and minor nutrient zinc (Z), the two most limiting nutrients in soils, remains a major challenge to ecologists and land managers. Application of organic based nutrient supplements have shown great potential as a supplementary, renewable and environmental friendly source of plant nutrients and are an important component of integrated nutrient management (INM) and integrated plant nutrition system (IPNS). INM s goal is to integrate the use of all natural and man-made sources of plant nutrients, so that crop productivity increases in an efficient and environmentally benign manner, without sacrificing soil productivity of future generations. Zinc is one of the essential micronutrients required for optimum crop growth. It is required in relatively small concentrations in plant tissues (5 100 mg/kg). Root cell membrane permeability is increased under Zn deficiency, which might be related to the function of Zn in cell membranes (McInyre et al 2009). Zn deficiency is well reported in the soils of much of the world. Cereals play role in satisfying daily calorie intake in the developing world, but the Zn concentration in the grain is inherently very low, particularly when grown on Zn-deficient soils. The major reason for the widespread occurrence of Zn deficiency problems in crop plants is the low solubility of Zn in soils rather than a low total amount of Zn (Welch and Graham 2004). Zinc plays an important role in many biochemical reactions within the plants. Plants such as sorghum, maize and sugarcane shows reduced photosynthetic carbon metabolism due to zinc defi- 248 International Journal of Biotechnology and Biosciences
EFFICACY STUDIES OF ORGANIC ACID BASED CHELATED PLANT NUTRIENTS FOR GROWTH AND GRAIN QUALITY UNDER CONTROLLED AND FIELD CONDITION ciency. Zinc modifies and/or regulates the activity of cabonic anhydrase, an enzyme that regulates the conversion of carbon dioxide to reactive bicarbonate species for fixation to carbohydrates in these plants. Nitrogen is a major limiting nutrient for the plant growth. Nitrate and ammonium are common forms of N available for plants. Plants require substantial energy for nitrogen uptake and assimilation (Hotz and Brown 2004). Nitrate must be reduced to nitrite and then ammonium, which is incorporated into amino acid biosynthesis (White and Zasoske 1999). Amino acids are building blocks of proteins, enzymes, nucleic acids, antioxidants, and other secondary compounds (Hotz and Brown 2004). The use of bio-fertilizer and bioenhancer such as Bio-zinc that contain lacate based zinc supplements and Suryamin that contain different type of essential amino acids produced by enzymatic hydrolysis of protein based nutrients can reduce chemical fertilizer applications and consequently lower production cost. Sorghum (Sorghum bicolor) is one of the major cereal crops consumed in India after rice (Oryza sativa) and wheat (Triticum aestivum). The crop is produced in Maharashtra and southern states of Karnataka and Andhra Pradesh. These three states together account for close to 80% of the all-india production. Sorghum is already the most profitable grain crop in the cooler high-rainfall areas, and is becoming more attractive in the hotter, western areas. Good fertilizer management and weed and pest control are important for good sorghum yields. Nitrogen often limits yields in good seasons. There are opportunities for research on sorghum varieties with adaptation to heat. The main aim of the study is to check the bioeficacy of organic nutrient supplements Bio-zinc and Suryamin to the sorghum crop and also to compare the morphological and physiological effects of sequential foliar treatments relative to Urea and Zinc sulfate fertilizer and control on grain sorghum. MATERIALS AND METHODS Experiments was conducted in controlled environment at green house and in field condition at agricultural farm of Prathista Industries Ltd. Sorghum Dekalb 28E were used as test plant for comparative study, the crop was showed in summer season (April 2011) in green house and agricultural farm. Plant nutrient such as Biozinc T and suryamin T which was manufactured at Prathista Industries Ltd, Hyderabad were used for efficacy testing. Efficacy trials in controlled condition The pots (15-L) were prepared by adding soil and metromix in the ratio of 2:1 by weight. The pots were fully soaked with water and left for 1 day to drain and then five seeds per pot were sown at a 5-cm depth. After emergence (two-leaf stage) at 15 days of growth, seedlings makes thinned to two per pot. The temperature regime was of 32/22 C ±3 C day/night, photoperiod of 12 h and photosynthetic photon flux density of 800-1400 µmolm 2 s 1 provided from natural solar radiation and supplemental fluorescent lights respectively. The relative humidity was 80% in the greenhouse. The seeds were applied in five treatments. The treatment are as follows: T1 : Control, T2 : BioZinc (0.3%), T3 : Zinc sulfate (0.5%), T4 : Suryamin (0.3%), T5 : Urea (4%) (Table 1). Table 1. Randomized block of control and treatment plot Control (T1R1) Biozinc (T2R1) ZnSO4 (T3R1) Suryamin (T4R1) Urea (T5R1) ZnSO4 (T3R2) Biozinc (T2R2) Control (T1R2) Suryamin (T4R2) Urea (T5R2) Urea (T5R3) Suryamin (T4R3) ZnSO4 (T3R3) Biozinc (T2R3) Control (T1R3) Vol. 4 (3) July - September 2014 249
All the experiments were done in triplicate. The nutrient supplements were applied on preflowering, flowering and physiological maturity stage. Physiological traits like stomatal conductance, chlorophyll content, rate of photosynthesis, transpiration rate and inter cellular CO- 2 concentration was measured according to the standard methods. After harvesting dry weight, number of seeds per plant, yield, total protein and total nitrogen content was measured (Fig 1). A). Physical appearance of grains B). Preparation of spray nutrients C).Green house test plot D). fully drenching the plant E). Different treatments F). Measuring photosynthesis Fig 1. Treatment at controlled conditions 250 International Journal of Biotechnology and Biosciences
EFFICACY STUDIES OF ORGANIC ACID BASED CHELATED PLANT NUTRIENTS FOR GROWTH AND GRAIN QUALITY UNDER CONTROLLED AND FIELD CONDITION Efficacy testing in field condition The field experiments were carried out by selecting random uniform fifteen plots (12 feet). All the plots were grown in rain fielded conditions from sowing to till maturation of grains. The treatment are as same mentioned in the pot condition (Table 1). The rate of photosynthesis was measured by using CI-340 Handled photosynthesis system (CID Bioscience Inc., USA). The physiological parameters like stomatal conductance was measured by using leaf porometer (SC-1 Leaf porometer, Decagon Devices, Inc. Pullman, WA, USA), chlorophyll a fluorescence by chlorophyll fluorometer (OS 30, Opti-Science, Hudson, NH, USA), and chlorophyll content by SPAD (Model 502, Spectrum Technologies, Plainfield, IL, USA). After harvesting root length, shoot length, fresh weight, dry weight, seed weight and seed size were measured. The total nitrogen and protein content of the seeds was also done according to the standard method (Fig 2). A). Sorghum plot B). Hand spraying C). Taking measurements D). Machine harvesting E). Taking measurements F). Measuring photosynthesis Fig 2. Treatment at field conditions Vol. 4 (3) July - September 2014 251
The data were subjected to the statistical analysis of variance for each trait according to the general linear model by statistical analysis software 9.1 (SAS, 2003). Differences among treatment means were compared by least significant differences (LSD) at 5% level of probability. RESULTS Physiological traits in controlled condition Probability level from ANOVA suggests that the leaf chlorophyll content values, PS II activity, photosynthetic rate, stomatal conductance, transpiration rate are highly significant among treatments. Biozinc and suryamin was greatly maintained leaf chlorophyll content had a value 53.8 and 53 SPAD respectively whereas the control contain the chlorophyll content 47.1 SPAD, Urea also improved leaf chlorophyll content had value 48.1 SPAD. Similarly biozinc, suryamin and urea treated pots had similar Fv/Fm (PS II activity) 0.740, 0.742 and 0.742 respectively where as control showed 0.745 Fv/Fm PS II activity. ZnSO 4 treated plant showed lowest PS II activity (0.698) As regrads photosynthetic rate Biozinc and suryamin again maintained good photosynthetic rate 42.6 and 38.2µmol m 2 s -1 respectively. In comparison control showed 35.9 µmol m 2 s -1 photosynthetic rate. Biozinc and suryamin showed stomatal conductance 147.3 and 111.7 mmol m -2 s -1 respectively, in comparison control showed 114.1 mmol m -2 s -1 stomatal conductance. Urea also showed high stomatal conductance 146.7mmol m -2 s -1. Biozinc, suryamin and urea maintained good transpiration rate 2.4, 2.3 and 2.6mmol m -2 s -1 respectively in comparison control showed 2.3mmol m -2 s -1 transpiration rate. Probability level from ANOVA suggests that the Inter-cellular CO 2 conc among the treatments are significant. But the biozinc and suryamin treated plot had less inter cellular concentration than the control, which indicated more CO 2 utilization. (Table 2 and Fig.3). Table 2. Effect of various treatments i.e. T1: control, T2: Biozinc, T3: ZnSo4, T4:Suryamin, T5:Urea on various physiological traits in controlled environment Treatments SPAD PS II activity (Fv/Fm) Photosynthetic rate (µmol m -2 s -1 ) Stomatalconduct ance (mmol m -2 s -1 ) Transpiration rate (mmol m -2 s -1 ) Inter-cellular CO 2 conc. (Ci) (ppm) T 1 47.1 b 0.745 a 35.9 b 114.1 b 2.3 a 143.8 bc T 2 53.8 a 0.740 a 42.6 a 147.3 a 2.4 a 125.8 c T 3 42.2 c 0.698 b 30.7 c 76.7 c 1.6 b 195.1 a T 4 53.0 a 0.742 a 38.2 b 111.7 b 2.3 a 119.4 c T 5 48.1 b 0.742 a 34.2 b 146.7 a 2.6 a 176.8 ab LSD(P=0.05) 1.3 0.01 4.3 26.9 0.4 50.0 Probability level (p) from ANOVA P value < 0.0001 < 0.0001 < 0.0001 < 0.0001 0.007 0.0169 Physiological traits in field condition Probability level from ANOVA suggests that leaf chlorophyll content, PS II activity, photosynthetic rate, stomatal conductance, intercellular concentration are not significantly difference among the treatments but there are difference between treatments. Transpiration rate are significantly different among treatments. Biozinc and suryamin maintained leaf chlorophyll 252 International Journal of Biotechnology and Biosciences
EFFICACY STUDIES OF ORGANIC ACID BASED CHELATED PLANT NUTRIENTS FOR GROWTH AND GRAIN QUALITY UNDER CONTROLLED AND FIELD CONDITION Fig 3. Effect of various treatments i.e. T1: control, T2: Bioznc, T3: ZnSO 4, T4:Suryamin, T5:Urea on leaf chlorophyll and PS II activity in controlled environment. content 48.5 and 47.8 SPAD respectively in comparison control showed chlorophyll content 48.5 SPAD. Urea recorded highest leaf chlorophyll content 49.9 SPAD amongst all the treatments. Similarly biozinc, suryamin and urea treated plots had similar Fv/Fm PS II activity, 0.711, 0.718, and 0.731 whereas control showed 0.690 PS II activity. Biozinc and suryamin again maintained good photosynthetic rate 47.8 and 49 µmolm 2 s -1 respectively. In contrast urea and ZnSO 4 had photosynthetic rate 48 µmol m 2 s -1 and 45.9 µmol m 2 s -1 respectively in comparison showed only 40.8 µmol m 2 s -1. Biozinc and suryamin maintained stomatal conductance 287.4 and 299.4 mmol m -2 s -1 respectively and control showed stomatal conductance 237 mmol m -2 s -1. Table 3. Effect of various treatments i.e. T1: control, T2: Biozinc, T3: ZnSo4, T4:Suryamin, T5:Urea on various physiological traits in field condition Treatments SPAD PS II activity (Fv/Fm) Photosynthetic rate (µmol m -2 s -1 ) Stomatal conductance (mmol m -2 s -1 ) Transpiration rate (mmol m -2 s -1 ) Inter-cellular CO 2 conc. (Ci) (ppm) T 1 48.53 a 0.690 bc 40.76 b 237.09 b 6.79 c 112.69 a T 2 48.49 a 0.711 ab 47.79 a 287.43 a 7.98 b 101.83 a T 3 45.02 b 0.669 c 45.92 ab 264.71 ab 8.27 ab 91.73 a T 4 47.81 ab 0.718 ab 49.03 a 299.41 a 9.22 a 103.36 a T 5 49.92 a 0.731 a 48.13 a 289.74 a 8.68 ab 104.53 a LSD(P=0.05) 3.32 0.03 5.84 41.19 1.11 40.60 Probability level (p) from ANOVA P value 0.0755 0.0250 0.0625 0.0472 0.0074 0.8462 Vol. 4 (3) July - September 2014 253
Transpiration rate was significantaly different amongst the treatments Biozinc, suryamin, ZnSO 4 and urea, they maintained good transpiration rate 8, 9.2, 8.3 and 8.7 mmol m -2 s -1 respectively, in comparison control showed only 6.8 mmol m -2 s -1 transpiration rate. Probability level from ANOVA also suggests that the Inter-cellular CO 2 concentrations among the treatments are not significant. All the treatements had less inter cellular concentration than the control, which indicated more CO 2 utilization. (Table 3). Growth, grain yield and quality traits of controlled environment. Probability level from ANOVA, P value suggested significant difference in plant height, Panicle length, and number of seeds per panicle, 100 seed weight, grain yield, protein content and total nitrogen among the treatments. Biozinc amended treatment showed highest growth in height of the plant. It is 104.8 cm in comparison control showed only 91.8 cm plant height. Suryamin, urea and ZnSO 4 having plant height 89.6 cm, 91.1 cm and 87.7 cm respectively. Similarly the panicle length was also highest in biozinc treatment it showed 24.3 cm of panicle length in comparison control showed 20.7cm of panicle length. Highest number of seeds per panicle was again highest in biozinc that is 922 seeds per panicle in comparison control showed 866 seeds per panicle. Weight of 100 seeds was highest in suryamin among all the treatments (Fig 4). Grain yield per plant was 28 g per plant in case of biozinc which was highest among the treatments. ZnSO 4, Suryamin and Urea had 18, 24 and 22 gm per plant in comparison control showed 26 gm per plant yield. Protein content and total nitrogen was also highest in case of suryamin and biozinc among all the treatments. Growth, grain yield and quality traits of field condition Probability level from ANOVA, P value suggested there are no significant differences in harvest index, grain yield, number of seeds per panicle, 100 seed weight. Biozinc had highest harvest index 0.46 whereas control showed 0.44 harvest index. Suryamin and Fig 4. Effect of various treatments i.e. T1: control, T2: Bioznc, T3: ZnSO 4, T4:Suryamin, T5:Urea on 100 seed weight and Protein content biozinc had highest number of seeds per panicle; it showed 2863 and 2488 seeds per panicle respectively whereas control showed 2130 seeds per panicle. Weight of 100 seeds was also highest in Biozinc it having 2.80 gm and control showed 2.49 gm (Table 4, 5). Probability level from ANOVA, P value suggested that there are significant differences in the protein content and total nitrogen among the treatments. The protein content in suryamin treatment was highest among all the treatments it showed 9.0 % of protein and control showed 6.6 % of protein, biozinc also increased protein content it showed 8.79 % of protein content. 254 International Journal of Biotechnology and Biosciences
EFFICACY STUDIES OF ORGANIC ACID BASED CHELATED PLANT NUTRIENTS FOR GROWTH AND GRAIN QUALITY UNDER CONTROLLED AND FIELD CONDITION Table 4: Effect of various treatments i.e. T1: control, T2: Biozinc, T3: ZnSo4, T4:Suryamin, T5:Urea on growth, grain yield and quality traits in controlled environment Treatments Plant (cm) height Panicle length (cm) No. of seeds 100 seed panicle -1 weight Grain yield (g plant -1 ) Protein % TN T 1 91.8 b 20.7 b 866.6 a 2.98 b 26.3 ab 10.05b 1.61 b T 2 104.8 a 24.3 a 922.4 a 3.23 ab 28.0 a 10.74ab 1.72 ab T 3 87.7 b 21.9 b 688.2 b 2.58 c 18.4 c 9.95b 1.59 b T 4 89.6 b 22.7 ab 759.1 ab 3.49 a 23.6 abc 11.92a 1.91 a T 5 91.1 b 20.6 b 682.8 b 3.28 ab 21.7 bc 9.83b 1.61 b LSD (P=0.05) 10.7 2.4 173.4 0.35 5.8 1.22 0.23 Probability level (p) from ANOVA P value 0.0355 0.0355 0.0395 0.0017 0.0294 0.0174 0.0498 Table 5. Effect of various treatments i.e. T1: control, T2: Biozinc, T3: ZnSo4, T4:Suryamin, T5: Urea on, grain yield and quality traits in field conditions. Treatments Harvest index No. of seeds Grain yield panicle -1 (kg ha -1 ) 100 seed weight (g) Protein % % Increase TN T 1 0.44 a 2131 b 7102 ab 2.49 b 6.59 b 1.05b T 2 o.46 a 2489 ab 7809 a 2.80 a a 8.79 33 1.40a T 3 0.42 a 2205 b 5944 b 2.63 a b 6.75 03 1.08b T 4 0.43 a 2863 a 7394 a 2.75 a a 9.04 37 1.45a T 5 0.44 a 2256 ab 6685 ab 2.76 a ab 7.85 19 1.25ab LSD (P=0.05) 0.064 640.0 1160 0.242 1.378 0.221 Probability level (p) from ANOVA P value 0.7683 0.1508 0.0409 0.0869 0.0071 0.0076 CONCLUSION Experiment suggests that application of biozinc and suraymin in sorghum improves the photosynthesis rate transpiration rate, stomatal conductance, leaf chlorophyll content and photochemical efficiency results more yield and most importantly it improves the grain quality by increasing the protein content 11-37% against absolute control. REFERENCES Hotz C and Brown KH (2004) Assessment of the risk of Vol. 4 (3) July - September 2014 zinc deficiency in populations and options for its control. Food Nutr Bull., 25,94 204 McIntyre BD., Herren HR., Wakhungu J and Watson RT (2009). International Assessment of Agricultural Knowledge, Science and Technology for Development: Global Report http://www. agassessment.org/. White JG and Zasoski RJ (1999) Mapping soil micronutrients. Field Crop Res., 60,11 26 Welch RM and Graham RD (2004) Breeding for micronutrients in staple food crops from a human nutrition perspective. J Exp Bot., 55,353 364 255