Scholarly article on topic 'Dry matter yield and forage quality traits of oat (Avena sativa L.) under integrative use of microbial and synthetic source of nitrogen'

Dry matter yield and forage quality traits of oat (Avena sativa L.) under integrative use of microbial and synthetic source of nitrogen Academic research paper on "Agriculture, forestry, and fisheries"

CC BY-NC-ND
0
0
Share paper
Keywords
{"Dry matter" / Nitrogen / Inoculum / "Nutritional value"}

Abstract of research paper on Agriculture, forestry, and fisheries, author of scientific article — M. Bilal, M. Ayub, M. Tariq, M. Tahir, M.A. Nadeem

Abstract The natural microbes are potential contributor to build up soil nitrogen through transformation of molecular nitrogen to plant available forms. Therefore, in the present study, we investigated the contribution of biofertilizer to reduce the synthetic nitrogen application without deteriorating the yield and forage quality. The supplementary nitrogen rates included 0, 40, 80 and 120kgha−1 and the seed inoculation was carried out with the mixture of Azospirillum + Azotobacter spp. The experiment was laid out in randomized complete block design with factorial arrangement. The results indicated that organic matter contents and ether extractable fat were negatively associated with both nitrogen and inoculation factors. The inoculation produced 6.58%, 9.58%, 2.51%, 16.94%, 10.26%, 17.59%, 14.02%, 33.81% and 66.18% more No. tillers, plant height, leaf to stem ratio, dry matter yield, mineral matter contents, crude fibre, crude protein, crude protein yield and total digestible crude protein yield, respectively over uninoculation. The interactive effects indicated that inoculation alone without nitrogen application produced 19.16% and 6.87% more dry matter yield and crude protein (%), respectively. The beneficiary effects of biofertilizers on growth and dry matter of oat were more pronounced at intermediate level of inorganic nitrogen which was gradually decreased at higher nitrogen levels. The CP, CPY and DCPY achieved with inoculation alone were statistically equivalent to plots fertilized with 0 and 40kgNha−1. It is clear that plots sown with inoculated seeds must be fertilized with 80kgN to produce higher dry matter and economic returns. However, the highest protein contents in dry matter were recorded with highest fertilization level along with inoculation. By giving due attention to stimulatory effects of bacterial species in the present study, it is therefore, recommended to integrate the use of biofertilizers with supplemental dose of synthetic nitrogen source to sustain crop production.

Academic research paper on topic "Dry matter yield and forage quality traits of oat (Avena sativa L.) under integrative use of microbial and synthetic source of nitrogen"

JSSAS 169 3 November 2015 ARTICLE IN PRESS No. of Pages 6

Journal of the Saudi Society of Agricultural Sciences (2015) xxx, xxx- -xxx

King Saud University Journal of the Saudi Society of Agricultural Sciences

www.ksu.edu.sa www.sciencedirect.com

n irljJII pq__lflU n IifjoIilB nipp^ll

SAUDI SOCIETY FOB A ULTUItAL SCIENCES

FULL LENGTH ARTICLE

Dry matter yield and forage quality traits of oat (Avena sativa L.) under integrative use of microbial and synthetic source of nitrogen

M. Bilala*, M. Ayuba, M. Tariqb, M. Tahira, M.A. Nadeema

8 a Department of Agronomy, University of Agriculture, Faisalabad, Pakistan

9 b Central Cotton Research Institute, Multan, Pakistan

10 Received 19 May 2015; revised 25 July 2015; accepted 18 August 2015

KEYWORDS

Dry matter; Inorganic nitrogen; Microbial fertilizer; Nutritional value

Abstract The natural microbes are potential contributor to build up soil nitrogen through transformation of molecular nitrogen to plant available forms. Therefore, in the present study, we investigated the contribution of biofertilizer to reduce the synthetic nitrogen application without deteriorating the yield and forage quality. The supplementary nitrogen rates included 0, 40, 80 and 120 kg ha-1 and the seed inoculation was carried out with the mixture of Azospirillum + Azo-tobacter spp. The experiment was laid out in randomized complete block design with factorial arrangement. The results indicated that organic matter contents and ether extractable fat were negatively associated with both nitrogen and inoculation factors. The inoculation produced 6.58%, 9.58%, 2.51%, 16.94%, 10.26%, 17.59%, 14.02%, 33.81% and 66.18% more No. tillers, plant height, leaf to stem ratio, dry matter yield, mineral matter contents, crude fibre, crude protein, crude protein yield and total digestible crude protein yield, respectively over uninoculation. The interactive effects indicated that inoculation alone without nitrogen application produced 19.16% and 6.87% more dry matter yield and crude protein, respectively. The beneficiary effects of biofertilizers on growth and dry matter of oat were more pronounced at intermediate level of inorganic nitrogen which was gradually decreased at higher nitrogen levels. The CP, CPY and DCPY achieved with inoculation alone were statistically equivalent to plots fertilized with 0 and 40 kg N ha-1. It is clear that plots sown with inoculated seeds must be fertilized with 80 kg N to produce higher dry matter and economic returns. However, the highest protein contents in dry matter were recorded with

Abbreviations: DMY, dry matter yield; MMC, mineral matter contents; CF, crude fibre; CP, crude protein; CPY, crude protein yield; TDCPY, total digestible crude protein yield. * Corresponding author.

E-mail address: agronomist2413@gmail.com (M. Bilal). Peer review under responsibility of King Saud University.

http://dx.doi.org/10.1016/j.jssas.2015.08.002

1658-077X © 2015 Production and hosting by Elsevier B.V. on behalf of King Saud University.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

JSSAS 169 3 November 2015

2 M. Bilal et al.

19 highest fertilization level along with inoculation. By giving due attention to stimulatory effects of

20 bacterial species in the present study, it is therefore, recommended to integrate the use of biofertil-

21 izers with supplemental dose of synthetic nitrogen source to sustain crop production.

22 © 2015 Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an open access 24 article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

26 1. Introduction

27 The livestock is dominant segment of agriculture in Pakistan

28 with a share of 11.53% in national GDP (Economic Survey

29 of Pakistan, 2012). The success and prosperity of livestock

30 farming is determined by adequate and timely availability of

31 feed. The green forages are major and the most economical

32 source to fulfil the dietary needs of livestock. The insufficient

33 fodder supply is characterized as major constrain of low ani-

34 mal performance for milk and meat production (Rana et al.,

35 2014; Ahmad et al., 2014). Therefore, a significant proportion

36 of livestock is underfed. On the other hand, the continuous

37 and long term feeding with poor quality forage results in mal-

38 nutrition in animals. In the present scenario, we are annually

39 producing 55.06 million tons fresh forage from 2.46 million

40 hectares with an average yield of 22.38 t ha-1 (Economic

41 Survey of Pakistan, 2012). The oat is fast growing and pro-

42 duces a significant amount of fresh fodder within short period

43 (60-70 days) with adequate nutritional facts. The forage scar-

44 city period during winter months could be managed by bring-

45 ing more area under oat crop.

46 Conventionally, the forage crops are cultivated on marginal

47 land characterized by low nutrient supply. Nitrogen is the

48 major nutrient for plants and becoming deficient in soils which

49 is being supplemented by inorganic nitrogen fertilizers. The

50 molecular forms of nitrogen can be made available to crops

51 through industrial and biological process (biological nitrogen

52 fixation). The former process consumes the fossils fuels,

53 degrades the soil and environment health through CO2 and

54 NO2 enrichment and later is naturally eco-friendly which is

55 carried out by prokaryotic micro-organisms. It has been esti-

56 mated that half of the applied nitrogen is lost in various pro-

57 cesses (Pindi, 2012). It is obvious that synthetic fertilizers

58 cannot be put out from agriculture without compromising

59 the low yield but these must be integrated with biofertilizers.

60 The microbial biomass is associated with rhizosphere which

61 plays an important role in the crop growth and development

62 through secretion of growth promoting metabolites and nutri-

63 ent supply. This indicates that rhizosphere research may be of

64 great agricultural value for crop nutrition rather to rely on syn-

65 thetic sources. In addition to nitrogen fixation, these species

66 release plant growth promoting substances and hormones

67 and improve nutrient and water uptake (Damir et al., 2011;

68 Shridhar, 2012; Glick, 2012). The Azotobacter and Azospiril-

69 lum spp. are nonsymbiotic microbes which have been catego-

70 rized as significant contributor toward cereal yield

71 improvement (Aon et al., 2015; Aazadi et al., 2014).

72 The biofertilizers can serve a potential tool for sustaining

73 crop production without deteriorating the soil and environ-

74 ment (Pindi, 2012; Alimadadi et al., 2010). Therefore, integra-

75 tion of inorganic and microbial fertilizers sources seems to be

76 sustainable approach in agricultural systems. The inoculation

77 of wheat with Azotobacter and Azospirillum could save 25%

or 50% of recommended mineral nitrogen (Abd El-Lattief, 78

2012). Therefore, the study was designed to find most suitable 79

dose of nitrogen and microbial combinations to improve the 80

forage production and quality of oat. 81

2. Materials and methods 82

The field study was planned to scrutinize the optimum level of 83

nitrogen for bacterium inoculated and un-inoculated oat seed 84

in terms of forage yield and quality. Azotobacter + Azospiril- 85

lum spp. mixture was used for inoculation purpose and nitro- 86

gen levels included 0, 40, 80 and 120 kg N ha-1. The 87

biofertilizer was obtained from Microbiological Section of 88

Ayub Agricultural Research Institute, Faisalabad. The seeds 89

were dipped in 10% sugar solution and biofertilizer was spread 90

over the seeds and was thoroughly mixed. The seeds were left 91

open to dry for a period of one night at room temperature. The 92

seeds were subjected to inoculation just before sowing. The 93

experiment was conducted at Research Farm of Agronomy, 94

University of Agriculture, Faisalabad, Pakistan, during rabi 95

season 2011-12. The soil samples were randomly collected 96

from experimental site prior to sowing and composite soil sam- 97

ple was subjected to physio-chemical analysis. The soil was 98

clay loam in texture, alkaline in reaction and moderately fertile 99

having 0.72% organic matter, 0.022% total nitrogen, 3.37 ppm 100

available phosphorus and 370 ppm available potassium. The 101

treatments were compared in randomized complete block 102

design (RCBD) under factorial arrangement with three replica- 103

tions. The seeds of variety S-2000 were drilled manually @ 104

75 kg ha-1 in well prepared soils at field capacity level on 105

November 28, 2011 in 20 cm apart rows. The plot size mea- 106

sured 1.8 x 6 m and each plot included 9 rows. The nitroge- 107

nous and phosphatic fertilizers were applied in the form of 108

urea (46% N) and single super phosphate (18% P2O5), respec- 109

tively. The nitrogen was applied as per treatment and phos- 110

phorus was given @ 75 kg ha-1. Half of the nitrogen and 111

full dose of phosphorus were thoroughly mixed in soil during 112

seed bed preparation. The remaining portion of nitrogen was 113

top dressed at first irrigation. All the plots were uniformly trea- 114

ted in terms of cultural operations carried out during experi- 115

mentation period. The plant height was recorded in standing 116

crop immediately before harvesting from average of ten ran- 117

domly selected plants. The crop growth rate was estimated at 118

ten days interval from 40th to 90th days of sowing by the fol- 119

lowing equation: 120

CGR =(W2 - W0/(T2 - T1) (Yaduraju and Ahuja, 1996) 123

where 124

W1 = Dry weight m-2 land area at first harvest, 125

W2 = Dry weight m-2 land area at second harvest. 127

T1 = Time corresponding to first harvest, T2 = Time 128

corresponding to second harvest. 129

Dry matter yield and forage quality traits of oat (Avena sativa L.)

The crop was harvested at 50% heading stage and weighted to get the fresh mass. The dry matter contents in% age were determined from subsample (10 g) of fresh forage which was taken in moisture free aluminium containers and was oven dried at 105 C till no further weight reduction. The resulted value was multiplied with fresh mass to calculate the dry matter yield (DMY) from respective plot. The forage quality was assessed in terms of crude protein (CP), crude protein yield (CPY), total digestible crude protein yield (TDCPY), crude fibre (CF) and mineral matter contents (MMC) and analysis protocol was followed as described by Association of Official Analytical Chemists (1984). The organic matter (OM) contents in dry matter were determined by subtracting the value of mineral matter contents. The CP contents were multiplied with total dry matter to calculate the crude protein yield (CPY). The total digestible crude protein yield (TDCPY) was calculated by equation adopted by Iqbal et al. (2013):

TDCPY = [0.97 x crude protein yield] - 0.67

The economic analysis was carried out following the procedures devised by CIMMYT (1988). The data collected on various parameters were statistically analysed by using Fisher's analysis of variance technique and the significance of treatment means was tested at 5% probability level using Least Significance Difference (LSD) test (Steel et al., 1997).

3. Results and discussion

The data pertaining to plant morphological parameters, DMY and forage quality indicators (Table 1) clearly indicated that

plant height was most responsive to nitrogen application and each successive increase in nitrogen dose significantly produced taller plants. It was realized from positive relation of leaf to stem ratio (on fresh weight basis) with nitrogen levels, the leaf mass taken more advantage of surplus nitrogen supply over stems. The plants supplied with highest dose of nitrogen bear more leaves and therefore, produced the highest leaf to stem ratio. Fertilizing crop with 80 kg N ha-1 was the optimum rate for the highest DMY. So, it can be concluded that soil of the experimental site was poor in supplying the nitrogen requirement of the oat. It is concluded that nitrogen enhances the merismetic and photosynthetic activity by regulating up the cell elongation and division and chlorophyll contents of leaves and it reflects the higher DMY. It appears that likewise plant height, DMY must be at the maximum with 120 kg N ha-1. But actually it did not happen because plants might have greater tendency of lodging and hence cannot contribute effectively to yield.

Nitrogen not only raised the CP concentration but at the same time it produced similar trend for MMC. Although, outstanding forage quality in terms of CP and MMC was attained from 120 kg N ha-1, at the same time its dry matter was fibrous. The CPY and TDCPY were increased with subsequent increase in nitrogen but these parameters were at par with 80 and 120 kg ha-1. CPY, being the function of DMY and CP percentage was also significantly improved with successive increase in nitrogen. The increase in MMC was accompanied with decreased OMC at higher nitrogen level due to negative association between these parameters. Beside nitrogen role in synthesis of amino acids, the higher leaf to stem ratio at

Table 1 Effect of nitrogen application and seed inoculation on agronomic attributes, dry matter yield and its forage quality.

No. of tillers Plant height Leaf to stem Dry matter yield Organic Mineral matter Ether extractable

(m-2) (cm) ratio (tha-1) matter (%) contents (%) fat (%)

Nitrogen levels (kg ha ')

0 512.00 c 99.9 d 0.378 c 11.50 d 90.93 a 9.08 d 4.24 a

40 559.00 b 104.7 c 0.400 b 15.85 c 89.24 b 10.76 c 3.99 b

80 620.50 a 118.2 b 0.410 b 23.07 a 87.06 c 12.95 b 3.50 c

120 627.00 a 123.8 a 0.425 a 20.78 b 85.66 d 14.34 a 3.22 d

LSD-value 27.037 4.0243 0.011 1.6523 0.3794 0.3794 0.1508

Seed inoculation

Inoculated 598.09 a 116.7 a 0.408 a 19.19 a 87.65 b 12.36 a 3.54 b

Un- 561.17 b 106.5 b 0.398 b 16.41 b 88.80 a 11.21 b 3.93 a

inoculated

LSD-value 19.118 2.8456 0.007 1.1684 0.2683 0.2683 0.1066

Interaction

IqN0 488.33 97.2 0.370 10.49 e 91.49 8.52 4.36

IQNI 545.67 100.6 0.394 14.32 d 89.86 10.14 4.15

ZoN2 592.33 111.7 0.407 20.29 b 87.75 12.25 3.73

I0N3 618.33 116.6 0.420 20.52 b 86.09 13.91 3.49

I,N0 535.67 102.6 0.386 12.50 de 90.37 9.63 4.11

IN 572.33 108.7 0.405 17.39 c 88.62 11.38 3.82

IN2 648.67 124.7 0.413 25.84 a 86.36 13.64 3.26

IiN 635.67 130.9 0.429 21.03 b 85.23 14.77 2.95

LSD-value NS NS NS 2.3368 NS NS NS

Means not sharing the same letter differ significantly at 5% level of probability.

I0 = un-inoculation, I1, inoculation, N0 = 0 kg N ha ', N1 = 40 kg N ha-1, N2 = 80 kg N ha- ', N3 = 120 kg N ha- 1, NS = Non-significant.

JSSAS 169 3 November 2015 ARTICLE IN PRESS No. of Pages 6

4 M. Bilal et al.

Table 2 Effect of nitrogen application and seed inoculation

on forage quality.

Treatments Crude fibre CP CPY TDCPY

(%) (%) (tha-1) (tha-1)

Nitrogen levels (kg ha -1)

0 23.35 d 7.08 d 0.81 c 0.12 c

40 25.48 c 7.90 c 1.26 b 0.56 b

80 29.76 b 9.47 b 2.21 a 1.47 a

120 31.77 a 10.66 a 2.22 a 1.48 a

LSD-value 1.4779 0.3497 0.1964 0.1912

Seed inoculation

Inoculated seed 29.82 a 9.35 a 1.86 a 1.13 a

Un-inoculated seed 25.36 b 8.20 b 1.39 b 0.68 b

LSD-value 1.0450 0.2472 0.1389 0.1352

Interaction

I0N0 21.55 6.84 e 0.72 e 0.02 e

I0N1 23.33 7.74 d 1.07 d 0.37 d

I0N2 27.15 8.75 c 1.78 b 1.05 b

I0N3 29.42 9.73 b 2.00 b 1.27 b

I1N0 25.15 7.31 de 0.92 de 0.22 de

IiNi 27.62 8.33 c 1.45 c 0.74 c

IN2 32.38 10.19 b 2.63 a 1.88 a

IN 34.12 11.58 a 2.44 a 1.69 a

LSD-value NS 0.4945 0.2778 0.2704

Means not sharing the same letter in columns differ significantly at

5% level of probability.

Io = un-inoculation, I1, inoculation, N0 = 0 kg N ha-1,

N1 = 40 kg N ha-1, N2 = 80 kg N ha-1, N3 = 120 kg N ha-1,

NS = Non-significant.

190 highest level of nitrogen is another support for higher protein

191 concentration in dry matter as leaves contain more protein

192 than stems. Nitrogen, being an essential component of chloro-

193 phyll, hormones, enzymes and amino acid improved the

194 growth, dry matter yield and protein concentration. The ether

195 extractable fat showed a negative relation with nitrogen appli-

196 cation rates and its concentration in dry matter was dropped

197 significantly at each successive increase in nitrogen. The

198 growth and forage quality promoting effects of nitrogen are

199 confirmation of findings of Tariq et al. (2011), Afzal et al.

200 (2012), Iqbal et al. (2013) (see Tables 2 and 3).

201 The co-inoculation of Azotobacter and Azospirillum pro-

202 duced significantly higher values for plant growth and dry

matter with improved forage quality traits over plots treated as 203

control. The plant roots secrete exudates which attract and 204

encourage the multiplication of microbes in rhizosphere which 205

in turn benefits the plants with improved germination and 206

healthy seedlings, nitrogen fixation, synthesis of growth 207

promoting hormones, phosphorus solubilization and improved 208

nutrients and water uptake (Kumar et al., 2001; Yasmin et al., 209

2004; Asghar et al., 2002; Steenhoudt and Vanderleyden, 2000; 210

El-Komy, 2004; Vessey 2003). The inoculants increased the 211

root surface through lateral root formation for higher nutrient 212

and water uptake and changed the rhizosphere composition 213

to facilitate the nutrient resource acquisition. The CGR 214

value was comparatively higher in inoculum treatments over 215

un-inoculated seeds irrespective of nitrogen rates (Fig. 1). It 216

was also realized that CGR was increased up to 60-70 and 217

70-80 DAS in inoculation and control treatment, respectively. 218

It is suggested that inoculation resulted relatively faster growth 219

and reduced decline in growth which would lead to higher 220

forage supply at early and greenish fodder at later growth 221

stages, respectively. Our results are in confirmation of those 222

of El-Toukhy and Abdel Azeem (2000), Naserirad et al. (2011), 223

Naseri et al. (2013) where inoculation with Azotobacter 224

+ Azospirillum spp. also resulted higher values for leaf to stem 225

ratio and yielded attributes of barley and maize, respectively. 226

The microbes modify the soil environment conducive for the 227

release of nutrients. The inoculation alone produces 9.58%, 228

2.51%, 16.94%, 14.02%, 17.59%, 10.26%, 33.81% and 229

66.18% higher plant height, leaf to stem ratio, dry matter, 230

CP, CF, MMC, CPY and TDCPY, respectively than untreated 231

seeds. Both the factors of the present study had promoting 232

effects on forage yield and its protein value and it is highly 233

desirable to increase protein concentrations in fodder crops 234

to reduce the complete reliance on protein supplements 235

(Eskandari et al., 2009). The improvement in growth and yield 236

parameters from inoculum treatments was also reported in ear- 237

lier studies conducted on various crops (Bashan et al., 2004; 238

Shaalan, 2005; Abd El-Ghany et al., 2010). 239

The interactive effects were non-significant except for 240

DMY, CP, CPY and TDCPY. The response of crop was mod- 241

ified with inoculum treatment and their use in crop fertilization 242

programme seems to be promising. The benefits of inoculums 243

were also clear for DMY without supplemental nitrogen fertil- 244

ization where inoculum produced 2.01 tons ha-1 more DMY 245

over uninoculated plots. The results have also been confirmed 246

by Narula et al. (2005) who observed 20% more yield in wheat 247

Table 3 Economic interpretation of the study.

Treatments Green fodder Gross Total fixed Cost of Cost of TVC Total Net BCR

yield income cost inoculation nitrogen (Rs ha-1) expenditure benefit (Rs h

(kg ha-1) (Rs ha-1) (Rs ha-1) (Rsha-1) (Rs ha-1) (Rs ha-1) (Rs ha-1)

T1 = I0N0 54,430 163,290 54,400 0 0 0 54,400 108,890 3.00

T2 = I0N1 61,230 183,690 54,400 0 3043.60 3043.60 57443.6 126246.4 3.20

T3 = I0N2 75,770 227,310 54,400 0 6086.96 6086.96 60486.96 166,823 3.76

T4 = I0N3 79,220 237,660 54,400 0 9130.43 9130.43 63530.43 174129.6 3.74

T5 = I1N0 57,420 172,260 54,400 250 0 250 54,650 117,610 3.15

T6 = IIN1 68,150 204,450 54,400 250 3043.60 3293.60 57693.6 146756.4 3.54

T7 = IiN2 91,830 275,490 54,400 250 6086.96 6336.96 60736.96 214,753 4.54

Tg = I1N3 84,750 254,250 54,400 250 9130.43 9380.43 63780.43 190469.6 3.99

TVC: Total Variable Cost, BCR: Benefit-Cost Ratio.

Dry matter yield and forage quality traits of oat (Avena sativa L.)

□ 80

□ 120

18 16 14 12 10 8 6 4 2 0

Inoultion Un-inoultion

Days after sowing (DAS)

Figure 1 Effect of nitrogen rates (kg ha-1) on periodic crop growth rate (g m-2 day-1) with and without inoculation.

nitrogen was applied at the rate of 80 kg ha-1.The minimum 271

benefit was recorded in T1 where seeds were not inoculated 272

and no nitrogen was applied. 273

5. Conclusion 274

The use of inoculums alone as well as in combination with sup- 275

plementary nitrogen application increased the DMY and its 276

forage quality constituents over un-inoculated seeds. However, 277

the application of nitrogen at the rate of 80 kg ha-1 and the 278

crop raised from inoculated seed with Azotobacter + Azospir- 279

ilium spp. seems to be the optimum dose for obtaining maxi- 280

mum yield of forage oat. 281

6. Uncited references 282

Dobbelaere et al. (2002), Fayez et al. (1985), Gholami et al. 283

(2009), Manske et al. (2000); Saikia and Jain (2007). 284

248 with inoculation of Azotobacter and Azospirillum strains. The

249 performance of inoculum for DMY was significant up to

250 80 kg N ha-1 where it improved the DMY from 20.29 to

251 25.84 tons ha-1. The similar trend has been observed in wheat

252 where Azotobacter and Azospirillum performance was gradu-

253 ally declined with successive increase in nitrogen Narula

254 et al. (2002).

255 The plots sown with inoculated seeds and received

256 120 kg N ha-1 produced 1.85% more crude protein over

257 untreated seed. Furthermore, the protein contents obtained

258 at 120 kg N ha- can be achieved with 80 kg N ha-1 and

259 inoculums which saved 40 kg N. Similar results were reported

260 by Kader et al. (2002) who stated that inoculating the wheat

261 seeds with biofertilizer helped to reduce the use of nitrogenous

262 fertilizer up to 20%. Fig. 2 shows that the contribution of inoc-

263 ulation to improve the DMY, CPY and TDCPY was signifi-

264 cantly with addition of surplus nitrogen and it reached at the

265 best with 80 kg N ha-1. However, for CP (%) this rate was

266 1 20 kg N ha-1 and it suggested that further treatments may

267 be designed to find the maximum rate.

268 4. Economic analysis

269 It is revealed from the data that maximum benefit cost ratio

270 (4.54) was obtained in T7 where seeds were inoculated and

nitrogen levels (kg ha-1)

Figure 2 Effect of inoculation (%) on various levels of supplementary nitrogen for dry matter and crude protein.

References 285

Aazadi, M.S., Siyadat, S.A., Syahbidi, M.M.P., Younesi, E., 2014. The 286

study effect of nitrogen, Azotobacter spp. and Azospirillum spp. on 287

phonological and morphological traits of durum wheat cultivars in 288

Dehloran region, Iran. Cercetari Agronomice in Moldova 47 (157), 289

15-21. 290

Abd El-Ghany, Bouthaina F., Arafa, Rhawhia A.M., El-Rahmany, 291

Tomader A., El-Shazly, Morsy M., 2010. Effect of some soil 292

microorganisms on soil properties and wheat production under 293

north sinai conditions. J. App. Sci. Res. 6 (5), 559-579. 294

El-Lattief, E.A., 2012. Improving Bread Wheat Productivity and 295

Reduce Use of Mineral Nitrogen by Inoculation with Azotobacter 296

and Azospirillum Under Arid Environment in Upper Egypt. In: 297

Int. Conf. on App. Life Sciences (ICALS2012) Turkey, September 298

10-12, 2012. 299

Afzal, M., Ahmad, A., Ahmad, A.H., 2012. Effect of nitrogen on 300

growth and yield of sorghum forage (Sorghum bicolor (L.) Moench 301

cv.) under three cuttings system. Cercetari agronomice in Moldova 302

45 (4), 57-64 . 303

Ahmad, S., Jabar, M.A., Khalique, A., Saima, Shahzad, F., Ahmad, 304

N., Fiaz, M., Younas, U., 2014. Effect of different levels of ndf on 305

voluntary feed intake, dry matter digestibility and nutrients 306

utilization in dry Nili Ravi buffaloes. J. Anim. Pl. Sci. 24 (6), 307

1602-1605 . 308

Alimadadi, A., Jahansouz, M.R., Besharati, H., Afshari, R.T., 309

Tavakkoli, M., 2010. Evaluating the effects of phosphate solubi- 310

lizing microorganisms, mycorrhizal fungi and seed priming on 311

nodulation of chickpea. Iranian J. Soil Res. (Formerly Soil Water 312

Sci.) 24 (1), 44-53. 313

Aon, M., Khalid, M., Hussain, S., Naveed, M., Akhtar, M.J., 2015. 314

Diazotrophic inoculation supplemented nitrogen demand of 315

flooded rice under field conditions. Pak. J. Agri. Sci. 52 (1), 145- 316

150. 317

Asghar, H.N., Zahir, Z.A., Arshad, M., Khaliq, A., 2002. Relationship 318

between in vitro production of auxins by rhizobacteria and their 319

growth promoting activities in Brassica juncea L. Bio. Fertil. Soil. 320

35, 231-237 . 321

Association of Official Analytical Chemists, 1984. Official Methods of 322

Analysis, fourteenth ed. Arlington Virginia, USA. 323

Bashan, Y., Holguin, G., De-Bashan, L.E., 2004. Azospirillum- plant 324

relationships: physiological, molecular, agricultural, and environ- 325

mental advances. Can. J. Microbiol. 50 , 521-577 . 326

CIMMYT, 1988. From agronomic data to farmer recommendation: 327

an economic training manual. Mexico, D.F: 25-33. 328

Damir, O., Mladen, P., Bozidar, S., Srnan, N., 2011. Cultivation of the bacterium Azotobacter chroococcum for preparation of biofertiliz-ers. Afric. J. Biotech. 10 (16), 3104-3111.

Dobbelaere, S., Croonenborghs, A., Thys, A., Ptacek, D., Okon, Y., Vanderleyden, J., 2002. Effect of inoculation with wild type Azospirillum brasilense and Azospirillum irakense strains on development and nitrogen uptake of spring wheat and grain maize. Biol. Fert. Soil. 36, 284-297.

Economic survey of Pakistan, 2011-12. Ministry of Food, Agriculture and Livestock Division (Economic Wing), Islamabad, Pakistan, pp. 17-37.

El-Komy, H., 2004. Coimmobilization of Azospirillum lipoferum and Bacillus megaterium for successful phosphorus and nitrogen nutrition of wheat plants. Food Techn. Biotech. 43 (1), 19-27.

El-Toukhy, S.A., Abdel-Azeem, H.H., 2000. Response of barley (Hordeum vulgare) to biofertilization technology. Annal. Agric. Sci. 2, 539-559.

Eskandari, H., Ghanbari, A., Javanmard, A., 2009. Intercropping of cereals and legumes for forage production. Not Sci. Biol. 1 (1), 7-13.

Fayez, M., Emam, N.F., Makboul, H.E., 1985. The possible use of nitrogen fixing Azospirilum as biofertilizer for wheat plants. Egypt. J. Microbiol. 20 (2), 199-206.

Gholami, A., Shahsavani, S., Nezarat, S., 2009. The effect of plant growth promoting Rhizobacteria (PGPR) on germination, seedling growth and yield of maize. Int. J. Biol. Life Sci. (Suppl 1), 35-40

Glick, B.R., 2012. Plant Growth-Promoting Bacteria: Mechanisms and Applications, Scientifica, ID 963401, p. 15, <http://dx.doi.org/10. 6064/2012/963401 >.

Iqbal, M.F., Iqbal, Z., Farooq, M., Ali, L., Fiaz, M., 2013. Impact nitrogenous fertilizer on yield and quality of oat. Pak. J. Sci. 65 (1), 1-4.

Kader, M.A., Mian, M.H., Hoque, M.S., 2002. Effects of Azotobacter inoculant on the yield and nitrogen uptake by wheat. Online J. Biol. Sci. 2, 259-261.

Kumar, V., Behl, R.K., Narula, N., 2001. Establishment of phosphate solubilizing strains of Azotobacter chroococcum in the rhizosphere and their effect on wheat cultivars under greenhouse conditions. Microbiol. Res. 156, 87-93.

Manske, G.G.B., Qritz-Monasterio, J.I., Ginklel, M.V., Gozzalez, R. M., Rajaram, S., Molina, E., Vlek, P.L.G., 2000. Traits associated with improved P-uptake efficiency in CIMMYTs semi dwarf springbread wheat grown on an acid soil in Mexico. Pl. Soil 221, 189-204.

Naseri, R., Moghadam, A., Darabi, F., Hatami, A., Tahmasebei, G. R., 2013. The Effect of deficit irrigation and Azotobacter chroococ-cum and Azospirillum brasilense on grain yield, yield components of maize (S.C. 704) as a second cropping in western Iran. Bull. Env. Pharmacol. Life Sci. 2 (10), 104-112.

M. Bilal et al.

Narula, N., Behl, R.K., Dudi, H.R., Suneja, S., Lakshminaryana, K., 2002. Response of wheat genotypes to Azotobacter inoculation under rainfed conditions. Rachis 17, 66-67.

Narula, N., Kumar, V., Singh, B., Bhatia, R., Lakshminarayana, K., 2005. Impact of biofertilizers on grain yield in spring wheat under varying fertilizer conditions and wheat-cotton rotation. Arch. Agron. Soil. Sci. 51 (1), 79-89.

Naserirad, H., Soleymanifard, A., Naseri, R., 2011. Effect of integrated application of bio-fertilizer on grain yield, yield components and associated traits of maize cultivars. American-Eurasian J. Agric. Environ. Sci. 10, 271-277.

Pindi, P.K., 2012. Liquid microbial consortium- a potential tool for sustainable soil health. Pindi J. Biofertil. Biopestic. 3 (4), 1-9.

Rana, A.S., Ahmad, A.U.H., Saleem, N., Nawaz, A., Hussian, T., Saad, M., 2014. Differential response of sorghum cultivars for fodder yield and quality. J. Glob. Innov. Agric. Soc. Sci. 2 (1), 6-10.

Saikia, S.P., Jain, V., 2007. Biological nitrogen fixation with non legumes: an achievable target or a dogma? Curr. Sci. 92 (3), 317-322.

Shaalan, M.N., 2005. Influence of biofertilizers and chicken manure on growth, yield and seeds quality of (Nigella sativa L.) plants. Egypt. J. Agric. Res. 83, 811-828.

Shridhar, B.S., 2012. Review: nitrogen fixing microorganisms. Int. J. Microbiol. Res. 3 (1), 46-52.

Steel, R.G.D., Torrie, J.H., Dicky, D.A., 1997. Principles and Procedures of Statistics: A Biometrical Approach, third ed. McGraw Hill Book Co., Inc., New York, USA, 352-358.

Steenhoudt, O., Vanderleyden, J., 2000. Azospirillum, a free-living nitrogen fixing bacterium closely associated with grasses: genetic, biochemical and ecological aspects. FEMS Microbiol. Rev. 24, 487-506.

Tariq, M., Ayub, M., Elahi, M., Ahmad, A.H., Chaudhary, M.N., Nadeem, M.A., 2011. Forage yield and some quality attributes of millet (Pennisetum americannum L.) hybrid under various regimes of nitrogen fertilization and harvesting dates. Afric. J. Agric. Res. 6, 3883-3890.

Vessey, J.K., 2003. Plant growth promoting rhizo-K. 2003. Plant growth promoting rhizo- K. 2003. Plant growth promoting rhizobacteria as biofertilizers. Plant Soil. 255, 571-586.

Yaduraju, N.T., Ahuja, K.N., 1996. NAR and CGR. In: Yaduraju, N. T., Ahuja, K.N. (Eds.), The Illustrated Dictionary of Agric. Venus Publish House, 11/298 Press Colony, Mayapuri, New Delhi, India, pp. 200/240.

Yasmin, S., Rahman, M., Hafeez, F.Y., 2004. Isolation, characterization and beneficial effects of rice associated plant growth promoting bacteria from Zanzibar soils. J. Basic Microbiol. 44 (2004), 241-252.