Effect of microbial inoculants on growth and yield parameters in Sunflower (Helianthus annuus).
Soil microbes play an important role in many critical ecosystem processes, including nutrient cycling and homeostasis, decomposition of organic matter, as well as promoting plant health and growth as bio-fertilization (Han et al., 2007). In recent years, biofertilizers have emerged as a promising component of integrating nutrient supply system in agriculture. Our whole system of agriculture depends in many important ways, on microbial activities and there appears to be a tremendous potential for making use of microorganisms in increasing crop production. Microbiological fertilizers are an important part of environment friendly sustainable agricultural practices (Bloemberg et al., 2000).
MATERIALS AND METHODS
The effect of biofertilizers on growth, yield and availability of nutrients in soil in Sunflower was studied at All India Co-ordinated Research Project (AICRP) on sunflower, at Zonal Agricultural Research Station (ZARS), University of Agricultural Sciences, Gandhi Krishi Vignana Kendra (GKVK) campus, Bengaluru during Kharif season 2012.
The experimental study we selected variety of KBSH-44 and field site was located at 12p 58 North latitude, 77p 35 East longtitude with an altitude of 899 meters above mean sea level. The experimental soil was red sandy loam in texture with initial available nitrogen (165.4 Kg/ ha), available phosphorus (26.6 Kg/ ha) and available potassium (65.7 Kg/ ha) and initial soil population was analyzed using serial dilution plate count method. The field experiment was laid out in Randomized Complete Block Design (RCBD) which had 12 treatments and three replications. Treatments T1: 100 % NPK, T2: 75 % NP +100 % K, T3: T1 + S.A of A. chroococum + PSB + T. viride + F.S of T. viride, T4: T2 + S.A of A. chroococum + PSB + T. viride + F.S of T. viride, T5: T1 + S.A of A. chroococum + PSB + T. viride + F.S of S. griseus T6:T2 + S.A of A. chroococum + PSB + T. viride + F.S of S. griseus, T7: T1 + S.A of A. chroococum + PSB + B. subtilis + F.S of T viride, T8: T2 + S.A of A. chroococum + PSB + B. subtilis + F.S of T. viride, T9: T1 + S.A ofA. chroococum + PSB + B. subtilis + F.S of S. griseus
T10:T2 + S.A of A.chroococum + PSB + B. subtilis + F.S of S. griseus, T11: T1 + Mancozeb spray at 0.3 %, T12: T1 + Propiconozole spray at 0.1 %.
Note: SA- Soil application and FS- Foliar spray
1. FYM is common to all the treatments as basal application at recommended dose (7.5 t [ha.sup.-1]).
2. 50% Urea was applied as basal dose and 50% as top dressing.
3. Biofertilizers as soil treatment: Applied to soil 7 days after sowing.
The microbial inoculants used in the experiment were obtained from B iofertilizer scheme of Department of Agricultural Microbiology. The microbial inoculants used in the study were, Azotobacter chroococcum as N fixer, Bacillus megaterium as P-solubilizer, Trichoderma viride, Bacillus subtilis, Pseudomonas fluorescens and Streptomyces griseus as bioagents.
Microbial inoculants were mass multiplied under laboratory condition using respective media like A. chroococcum on Ashby's broth, Bacillus megaterium on Pikovoskaya's broth, T viride on Trichoderma specific medium and Pseudomonas fluorescens on King's B broth. After attaining desirable population, the culture was mixed aseptically in pre-sterilized charcoal powder which was neutralized with calcium sulphate. The mixing proportion was 1: 0.6: 2.5 (1 litre broth: 600g CaSo4: 2.5kg charcoal), and was used in the field study as soil application at the rate of 10 kg [ha.sup.-1].
The observations on crop growth parameters (per cent germination, plant height, number of leaves, stem girth, days to 50 per cent flowering) were recorded at 30,60, 90 days after sowing and at harvest stage, yield parameter (head diameter, seed yield per plot and hectare, oil content and oil yield) and seed quality parameters (hundred seed weight and volume weight,) were recorded after crop harvest.
The experimental data was analyzed statistically by Fischer's method of analysis of variance as given by Panse and Sukhatme (1967).
RESULTS AND DISCUSSION
The combined application of organic and inorganic fertilizers in combination with biofertilizers has significantly influenced the growth parameters in Sunflower hybrid KB SH 44. Among the treatments, biofertilizer treated plants showed significantly superior plant growth over plants not treated with biofertilizers at all the stages of plant growth (30, 60, 90 DAS and at harvest stage).
The highest germination percentage was recorded treated with microbial consortia of Azotobacter chroococcum, PSB, Trichoderma viride and foliar spray of Trichoderma viride with 100% NPK (T3) showed maximum germination (96.39%) which was statistically on par with the next best treatment combinations of Azotobacter chroococcum, PSB, Trichoderma viride and foliar spray of Streptomyces griseus with 100% NPK (T5) (96.11%). The treatment [T.sub.2] showed the lowest germination percentage (87.77%) having 75%NP + 100% K (Table 1). The application of microbial consortia resulted in better seed germination and enhanced plant growth. These beneficial microorganisms are known to secrete biologically active, plant growth promoting substances like IAA, gibberellins, cytokinins, vitamins, etc., which enhances seed germination, seedling vigour and root growth of plants, thus improving the yield of crops (Moeinzadeh et al, 2010). Many earlier research studies have also showed that combined use of organic manures and biofertilizers along with mineral fertilizers improve soil fertility and productivity (Reddy et al, 2005; Iraj et al., 2009). The plant height was significantly higher in plants treated with consortial application of Azotobacter chroococcum, PSB, Trichoderma viride and foliar spray of Trichoderma viride with 100% NPK (T3) (25.16 cm, 146.95 cm, 189.33 cm and 189.60 cm at 30, 60, 90 DAS and harvest stage respectively) (Table. 1 & Fig.1). The lowest plant height was recorded in the treatment of 7 5%NP + 100%K (T2) (17.90 cm, 118.40 cm, 163.37cm and 164.15 cm). The plant height was significantly increased due to microbial inoculation at all the stages of plant growth viz., 30, 60, 90 DAS and at harvest, which is due to better supply of nitrogen by biological means and efficient solubilization of unavailable P to available P form. The production of biologically active substances by these microorganisms might have helped in better nutrient uptake for the plant growth. These results are in conformity with the findings of many research workers who reported such increased plant height was due to biofertilizers application in crops like canola (Megawer and Mahfouz, 2010; Khan et al., 2010), soybean (Ramamurthy et al., 2001), black gram (Rathi et al., 2009) and sunflower (Zehra Ekin, 2011).
More number of leaves were obtained in the plants inoculated with Azotobacter chroococcum, PSB, Trichoderma viride and foliar spray of Trichoderma viride with 100% NPK at all the stages of plant growth viz., 30, 60, 90 DAS and at harvest (12.67, 30.13, 24.53 and 16.07 respectively) (Table 1).
The stem girth increased in all the growth stages i.e., 30, 60, 90 DAS and decreased at hatvest (Table 1). The highest stem girth was observed in plants supplemented with Azotobacter chroococcum, PSB, Trichoderma viride and foliar spray of Trichoderma viride with 100% NPK (1.25cm, 1.90 cm, 2.31 cm and 2.07 respectively). These results correlate with the findings of Mohsen Javahery et al. (2011) who studied the effects of biofertilizer application on phenology and growth of sunflower (Helianthus annuus L.).
Seed quality parameters
Good seed is the basis for successful crop production programme. The quality seed are ensured with uniform germination, rapid root and shoot development (seedling vigour) to obtain healthy and robust seedlings for increased yield per unit area. The seed quality parameters like 100 seed weight and volume weight yield differed significantly due to treatments. combined application of microbial inoculants viz., A. chroococcum, B. megaterium, T. viride and foliar spray of T. viride with 100 per cent NPK has recorded significantly higher 100 seed weight (7.19 g) and volume weight (56.08 g 100 [ml.sup.-1]) compared to other treatments (Table 2). The improvement in seed quality parameters may be attributed to the nutrition of sunflower crop due to the accumulation of higher quantities of seed reserves like protein and carbohydrates. These results are in agreement with the findings of Seshasailasree (2005) who reported that the combined inoculation of Azotobacter and Azospirillum could enhance the yield of rainfed sunflower to an extent of 37.9 per cent, compared to control.
Yield and yield parameters
The highest head diameter (24.33 cm) was recorded (Table.2, Fig.2&3) in plants treated with (T3) 100% NPK + Azotobacter chroococcum + PSB + T.viride + Foliar spray of T.viride which was superior over all other treatments and the lowest head diameter was found in (T2) 75%NP + 100%K (18.67 cm). These findings are in accordance with the findings of Sanaz Shoghi Kalkhoran et al., (2010) who studied integrated use of chemical, organic and biofertilizer (Azotobacter and Azospirillum) and with the results earlier obtained in sunflower by Mallikarjuna et al., (2000) and Ram et al., (1992).
The combined inoculation of A. chroococcum, B. megaterium, T. viride and foliar spray of T. viride with 100 per cent NPK has resulted in higher seed yield of sunflower per plot (2.04 kg/plot), seed yield per hectare (2276.64 kg [ha.sup.-1]), oil yield (844.69 kg [ha.sup.-1]) and oil content (37.64%). These results are in accordance with the results obtained by earlier research workers in sunflower (Nanjundappa et al., 2001; Ramamurthy and Shivashankar, 1995 and Mallikarjuna et al., 2000) and Ahmed et al., (2011) who reported the effect of bio-N-P fertilizer on the growth, yield and some biochemical components of two sunflower cultivars i.e., Vedock and Hy-sun 35.
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M.E. Shilpa , B. Latha , G. Dakshayini , H.M. Vikas  and M. Srikantaiah 
 Department of Agricultural Microbiology UAS, GKVK, Bangalore India.
 Department of Horticulture, UAHS, Bagalkot, India.
(Received: 12 April 2015; accepted: 06 May 2015)
* To whom all correspondence should be addressed. E-mail: firstname.lastname@example.org
Table 1. Effect of microbial inoculants on Growth parameters of sunflower hybrid KBSE1 44 Treatments Germination Plant height (cm) (%) at 10 DAS 30 60 90 At DAS DAS DAS Harvest [T.sub.1] 90.94 19.81 120.71 168.80 168.97 [T.sub.2] 87.77 17.90 118.40 163.37 164.15 [T.sub.3] 96.39 25.16 146.95 189.33 189.60 [T.sub.4] 94.39 23.60 140.07 182.60 182.92 [T.sub.5] 96.11 24.90 145.07 187.13 187.39 [T.sub.6] 94.33 23.47 139.40 180.07 180.37 [T.sub.7] 92.33 21.63 133.73 173.60 173.87 [T.sub.8] 90.33 20.99 130.67 171.24 171.31 [T.sub.9] 92.00 21.45 133.20 173.57 173.78 [T.sub.10] 90.67 20.63 130.36 170.93 170.99 [T.sub.11] 88.50 19.12 119.61 164.93 164.59 [T.sub.12] 89.33 19.53 120.77 165.67 165.91 F-test * * * * * S.Em[+ or -] 1.15 0.93 4.42 5.25 5.45 CD at 5% 3.36 2.71 12.97 15.39 15.60 Treatments Number of leaves/plants 30 60 90 At DAS DAS DAS Harvest [T.sub.1] 12.60 22.13 11.80 10.13 [T.sub.2] 11.33 20.67 10.87 9.80 [T.sub.3] 12.67 30.13 24.53 16.07 [T.sub.4] 11.93 26.87 21.87 13.20 [T.sub.5] 12.53 29.07 23.87 14.40 [T.sub.6] 11.80 26.47 21.47 13.07 [T.sub.7] 12.53 28.13 23.73 14.40 [T.sub.8] 11.80 26.27 20.00 12.07 [T.sub.9] 12.53 26.73 22.10 14.13 [T.sub.10] 11.67 25.33 19.73 11.27 [T.sub.11] 11.33 21.67 10.93 9.67 [T.sub.12] 12.47 21.73 11.20 10.20 F-test NS * * * S.Em[+ or -] -- 1.05 0.82 0.53 CD at 5% -- 3.09 2.42 1.57 Treatments Stem Girth (cm) 30 60 90 DAS DAS DAS [T.sub.1] 1.15 1.65 1.75 [T.sub.2] 1.10 1.49 1.71 [T.sub.3] 1.25 1.90 2.31 [T.sub.4] 1.21 1.63 2.01 [T.sub.5] 1.23 1.81 2.25 [T.sub.6] 1.19 1.60 2.07 [T.sub.7] 1.23 1.67 2.13 [T.sub.8] 1.22 1.65 2.02 [T.sub.9] 1.21 1.63 2.04 [T.sub.10] 1.19 1.65 1.91 [T.sub.11] 1.11 1.58 1.68 [T.sub.12] 1.13 1.62 1.73 F-test NS * * S.Em[+ or -] -- 0.06 0.08 CD at 5% -- 0.17 0.23 Treatments Stem Girth Days to (cm) 50% At Harvest [T.sub.1] 1.70 66.67 [T.sub.2] 1.57 67.00 [T.sub.3] 2.07 64.33 [T.sub.4] 1.87 65.00 [T.sub.5] 2.01 64.67 [T.sub.6] 1.81 65.33 [T.sub.7] 1.99 64.67 [T.sub.8] 1.79 66.00 [T.sub.9] 1.81 64.67 [T.sub.10] 1.80 66.33 [T.sub.11] 1.60 66.00 [T.sub.12] 1.65 66.33 F-test * NS S.Em[+ or -] 0.09 -- CD at 5% 0.26 -- Table 2. Effect of microbial inoculants on yield parameters of sunflower hybrid KBSH 44 Treatments Head Seed yield Seed yield 100 seed diameter per plot (kg Weight (cm) (kg) [ha.sup.-1]) (g) [T.sub.1] 20.07 1.44 1603.32 5.81 [T.sub.2] 18.67 1.31 1458.24 5.61 [T.sub.3] 24.33 2.04 2276.64 7.19 [T.sub.4] 22.07 1.72 1919.52 6.10 [T.sub.5] 24.20 2.00 2235.72 7.10 [T.sub.6] 21.47 1.74 1941.84 6.30 [T.sub.7] 22.87 1.75 1953.00 7.09 [T.sub.8] 21.93 1.71 1908.36 5.94 [T.sub.9] 22.20 1.74 1938.12 6.19 [T.sub.10] 21.60 1.73 1926.96 5.84 [T.sub.11] 19.80 1.37 1525.20 5.66 [T.sub.12] 20.11 1.40 1562.40 5.78 F-test * * * NS S.Em[+ or -] 0.68 0.09 100.79 -- CD at 5% 1.99 0.26 295.61 -- Treatments Volume seed Oil Oil yield weight content (kg (g/100 ml) (%) [ha.sup.-1]) [T.sub.1] 53.52 37.18 596.88 [T.sub.2] 53.41 37.00 540.92 [T.sub.3] 56.80 37.64 844.69 [T.sub.4] 55.05 37.26 722.43 [T.sub.5] 56.05 37.50 840.22 [T.sub.6] 54.15 37.29 729.54 [T.sub.7] 55.72 37.43 734.01 [T.sub.8] 54.91 37.16 715.64 [T.sub.9] 55.37 37.32 721.94 [T.sub.10] 54.84 37.14 714.63 [T.sub.11] 53.58 37.07 564.30 [T.sub.12] 53.94 37.10 567.97 F-test NS NS * S.Em[+ or -] -- -- 38.04 CD at 5% -- -- 111.57
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|Author:||Shilpa, M.E.; Latha, B.; Dakshayini, G.; Vikas, H.M.; Srikantaiah, M.|
|Publication:||Journal of Pure and Applied Microbiology|
|Date:||Sep 1, 2015|
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