CC BY-NC-ND
0JSSAS 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
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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.
