WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Ghetiya et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 7.632

Volume 9, Issue 2, XXX-XXX Research Article ISSN 2278 – 4357

NUTRIENT AND MICROBIAL STATUS OF CULTIVATED SOIL OF

POPCORN (ZEA MAYS L. EVERTA) AS INFLUENCED BY
APPLICATION OF PHOSPHATE AND POTASH SOLUBILIZING
BACTERIA
1
*Krupa P. Ghetiya, 2Dr. V. B. Bhalu, 3Dr. P. K. Chovatia, 4Dr. R. K. Mathukia and
5
H. B. Sojitra

1
Ph. D. Research Scholar, College of Agriculture, Junagadh Agricultural University,
Junagadh-362 001, Gujarat.
2
Assistant Professor, Department of Agronomy College of Agriculture JAU, Junagadh-362
001, Gujarat.
3
Associate Professor, Department of Agronomy College of Agriculture JAU, Junagadh-362
001, Gujarat.
4
Associate Research Scientist, Department of Agronomy College of Agriculture JAU,
Junagadh-362 001, Gujarat.
5
M. Sc. Agronomist, Department of Agronomy College of Agriculture JAU, Junagadh-
362 001, Gujarat.

ABSTRACT
Article Received on
27 Nov. 2019, To studied the effect of different levels of PSB and KSB on microbial
Revised on 17 Dec. 2019, and nutrient status of cultivated soil of popcorn and their economics
Accepted on 07 Jan. 2020
DOI: 10.20959/wjpps20202-15447
for that an experiment was carried out during rabi season of 2016-17 at
Junagadh Agricultural University. The experimental results revealed
that application of 45 kg P2O5/ha + PSB seed inoculation + PSB soil
*Corresponding Author
application gave significant result on chemical parameters viz., grain P
Krupa P. Ghetiya
Department of Agronomy, content, fodder N and P content, grain and fodder N, P and K uptake, P
College of Agriculture, and K use efficiency; post harvest available P and organic C status of
Junagadh Agricultural soil and microbial parameters of soil viz., total bacterial, total fungal
University, Junagadh -
and PSB count at 30 DAS and harvest, also gave highest yield, net
362001, Gujarat.
return and B:C ratio. Application of 45 kg K2O/ha + KSB seed
krupaghetiya218@gmail.com
pkchovatia@jau.in inoculation + KSB soil application enhanced chemical parameters viz.,
harmishabsojitra33@gmail.c grain P and K content, fodder N and K content, grain and fodder N, P
om
and K uptake, P and K use efficiency; post harvest available K and organic C status of soil
and microbial parameters of soil viz., total bacterial, total fungal and KSB count at 30 DAS
and harvest also gave highest net return and B:C ratio. The joint effect of different levels of

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Ghetiya et al. World Journal of Pharmacy and Pharmaceutical Sciences

PSB and KSB was found significant in case of nitrogen and phosphorus content in fodder and
also in nitrogen and phosphous uptake by fodder.

KEYWORDS: Popcorn, (Zea mays L. Var. Everta), Phosphate and potash solubilizing
bacteria, Microbial status, Nutrient status, Economis.

INTRODUCTION
Maize is considered as the “Queen of Cereals”. Being a C4 plant, it is capable to utilize solar
radiation more efficiently even at higher radiation intensity. Maize (Zea mays L.) ranks third
in total world production after wheat and rice and it is principal staple food in many
countries, particularly in the tropics and subtropics. To satisfy crop nutritional requirements,
P is usually added to soil as chemical P fertilizer, however synthesis of chemical P fertilizer
is highly energy intensive processes, and has long term impacts on the environment in terms
of eutrophication, soil fertility depletion carbon. More over plants can use only a small
amount of this P since 75–90% of added P is precipitated by metal cation complexes, and
rapidly becomes fixed in soils. Such environmental concerns have led to the search for
sustainable way of P nutrition of crops. In this regards phosphate-solubilizing
microorganisms (PSM) have been seen as best eco-friendly means for P nutrition.
Phosphorus is the second important key element after nitrogen as a mineral nutrient in terms
of quantitative plant requirement. Although abundant in soils, in both organic and inorganic
forms, its availability is restricted as it occurs mostly insoluble forms. Potassium though
present as abundant element in soil or is applied to fields as natural or synthetic fertilizers,
only one to two percent of this is available to plants, the rest being bound with other minerals
and therefore unavailable to plants. The most common soil components of potassium, 90 to
98%, are feldspar and mica.

Why need for increasing microbial count in soil of popcorn?

Due to high in price of fertilizers, especially those of phosphatic and potassic fertilizers, and
also the shortage of fertilizers, searching of alternate option hold promise for reducing cost of
cultivation. Further, efficiency of phosphatic and potassic fertilizers is low due to fixation in
calcareous soil of saurastra.

Organic manures can replace fertilizers up to 50%, but the availability and cost of organic
manures are further become constraints. It thus becomes urgent to investigate the bio-

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Ghetiya et al. World Journal of Pharmacy and Pharmaceutical Sciences

solubilisation of soil P and K reserves so as to alleviate the phosphatic and potassic fertilizer
shortage. Several strains of bacteria and fungi have been identified for P and K solubilization.

MATERIALS AND METHODS

An experiment was conducted during rabi season of 2016-17 at Junagadh Agricultural
University, Junagadh, Gujarat. Maize variety “Amber” was sown at 60 cm row spacing and
20 cm spacing maintain between two plants within row. Soil of the experimental plot was
clayey in texture and slightly alkaline in reaction (pH 7.9 and EC 0.33 dS/m). The soil was
medium in available nitrogen (269 kg/ha), available phosphorus (30.7 kg/ha), available
potash (209 kg/ha) and organic carbon (0.83%) from 0-15 cm depth. The soil was free from
any kind of salinity or sodicity hazards. 120 kg N/ha in two equal splits at sowing and 30
DAS was applied uniformly to all the plots. Entire dose of phosphorus and potash as per
treatments was applied at sowing. Treatment which include four level of phosphate treatment
viz., Control (P0), 60 kg P2O5/ha (P1), 45 kg P2O5/ha + PSB seed inoculation (P2), 45 kg
P2O5/ha + PSB seed inoculation + PSB soil application (P3) and four level of phosphate
treatment viz., Control (K0), 60 kg K2O/ha (K1), 45 kg K2O/ha + KSB seed inoculation (K2),
45 kg K2O/ha + KSB seed inoculation + KSB soil application (K3) arranged in 16 treatment
combinations with four levels each of PSB and KSB was laid out in Factorial Randomized
Block Design (FRBD) with three replications.

Liquid consortium of PSB (Bacillus subtilis) obtained from Junagadh Agricultural University
and KSB (Frateuriaaurantia) obtained from Navsari Agricutural University were used for
seed treatment as well as for soil application. For seed treatment, seeds were spreaded and
PSB/KSB culture (108 viable cells/g) @ 30 mL/kg of seed was sprinkled on the seeds and
then dried in shade. For soil application, PSB/KSB culture (108 viable cells/g) @ 3000 mL/ha
was mixed with FYM and than applied in furrows just after sowing before irrigation.

Microbial count in soil was done by general Colony Formation Unit (CFU) count per gram.
Fungal growth study was done by using Potato Dextrose Agar (PDA) media and bacterial
study was done by using Nutritional Agar (NA) media. Total count of fungus and bacterial
population in the soil samples were made by the plate count method as described by Ismail
and Yap 1994. CFU is an estimate of viable bacteria or fungal numbers. Unlike direct
microscopic count where all cell, dead and living, are counted, CFU estimates viable cells.
The appearance of a viable colony requires significant growth of the microbes. At the time of
counting the colony it was not possible to determine if the colony arose from cell or 1,000

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Ghetiya et al. World Journal of Pharmacy and Pharmaceutical Sciences

cells. Therefore, the results were given as CFU/ml for liquids and CFU/g for solids to reflect
this uncertainty.

NA media for bacterial growth

Put the weight amount of 5 gm of peptone and 3 gm beef extract in 500 ml of water. Heated
with agitation to dissolve all constitute. Add more distilled water to make final volume 1 lit.
Add 20 gm of agar-agar powder for solidification of media. Adjust the pH of medium 7.0 by
adding either acid or alkali (1 N Hcl or 1 N NaOH) with the help of pH meter or pH paper.
Fill the test tube or flasks with hot melted media. Plug them with cotton and sterilize in
autoclave at 1210C, 15 pound pressure for 15 min. After completion the autoclaving this
media allow to pour in sterilized petri plates under laminar condition. Then, allowed the
media to solidify for at least 4 to 5 hours. Stored media in flask or in test tube at room
temperature for further use.

PDA media for fungal growth

To prepare potato infusion, boil 200 g sliced, unpeeled potatoes in 500 ml distilled water for
30 minutes filter through cheesecloth, saving effluent, which is potato infusion (or use
commercial dehydrated form). Mix with 20 gm of dextrose, 20 gm of agar powder and made
final volume 1 litre of water and boil to dissolve, then after autoclave at 121°C temperature,
15 psi for 15 minute. PDA plates were prepared by pouring 20 ml of these media into each of
the petri plates. The plates were allowed to solidify, and later used for estimation of microbial
count.

Steps for serial dilution of bacteria and fungi

Each of the experimental soil was first homogenized, and then 1 g was taken and added to 9
ml of sterile distilled water to form a 10-1 dilution. This was then further serially diluted to
form 10-9 dilution. 1 ml of the 10-4 to 10-8 dilution from each of the experimental soil was
taken and spreader on both, PDA and NA plates. The plates were then incubated at room
temperature. The NA plates were observed bacterial colony growth after 24 hr while the PDA
plates were observed for fungal growth after 4-5 days. The colonies were counted and noted
for further calculation of CFU/g, as per the formula given below:

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PSB and KSB Population (CFU/g)

The soil microbial analysis was carried out for total PSB count and total KSB count by serial
dilution method (Aneja, 2008). The microbial study was carried out for initial soil sample as
well at 30, 60 DAS and at harvest. Microbial count of soil was done by general Colony
Formation Unit (CFU) count per gram. PSB count study was done using Pikovskaya’s Agar
media (Yeast extract 0.50 g/L, Dextrose 10 g/L, Calcium phosphate 5g/L, Ammonium
sulphate 0.50 g/L, KCL 0.20 g/L, MgSO4 0.1 g/L, MnSO4 0.0001 g/L, FeSO4 0.0001 g/L,
Agar 15 g/L and distilled water 1 L) and KSB count study was done using glucose, yeast
extract, calcium carbonate (GYC) media (Glucose 20 g/L, Yeast extract 3 g/L, Calcium
carbonate 5 g/L, Agar-Agar 18.5 g/L and distilled water 1 L).

Pikovaskaya’s Agar and GYC plates were prepared by pouring 20 ml of the media into each
of the plates. The plates were then allowed to solidify and later used for estimation of
microbial count. This was then further serially diluted to form 10-6 dilution as given above
steps and noted for further calculation of CFU/g, as per abov formula.

The effectiveness of either seed and/or soil inoculation with biofertilizer was assessed in the
nutrient content, nutrient uptake and microbial count in cultivated soil of popcorn, also
different treatment combinations was worked out and studied economics of all different
tretments.

RESULTS AND DISCUSSION

Table 1: Effect of different trtments on microbial count.
Total bacterial Total fungal PSB KSB
7 4 6
Treatment count (1 × 10 ) at count (1 × 10 ) at Count (1 × 10 ) at Count (1 × 106) at
30 DAS Harvest 30 DAS Harvest 30 DAS Harvest 30 DAS Harvest
PSB
P0 77.74 62.99 33.34 29.63 4.49 3.52 6.89 4.00
P1 79.61 65.36 34.68 31.34 3.77 2.38 6.42 4.25
P2 84.69 67.88 35.98 32.77 8.76 6.84 6.83 4.26
P3 86.36 70.50 37.27 33.89 9.48 7.30 7.39 4.63
S.Em.± 1.24 1.07 0.55 0.47 0.26 0.23 0.34 0.21
C.D. at 5% 3.58 3.08 1.58 1.35 0.74 0.67 NS NS
KSB
K0 77.29 64.18 33.87 30.09 6.23 4.58 5.79 3.28
K1 80.46 65.56 34.75 31.52 5.93 4.49 5.45 3.06
K2 83.79 67.65 35.87 32.43 6.35 5.06 7.69 5.48
K3 86.86 69.34 36.78 33.60 6.66 5.32 8.60 6.05
S.Em.± 1.24 1.07 0.55 0.47 0.26 0.23 0.34 0.21
C.D. at 5% 3.58 3.08 1.58 1.35 NS NS 0.99 0.61

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Note: Before sowing no observed much more microbial activity and interaction between PSB
and KSB observed non significant.

Table 2: Effect of different treatments on nutrient status.

Nutrient content (%) Available nutrient (kg/ha) in
Treat-
Nitrogen Phosphorus Potassium soil at 0-15 cm depth
Ment
Grain Fodder Grain Fodder Grain Fodder N P K OC
P0 1.67 0.34 0.426 0.108 0.46 1.08 244.48 26.44 266.60 0.65
P1 1.77 0.44 0.439 0.140 0.44 1.08 249.78 29.08 258.04 0.70
P2 1.75 0.46 0.459 0.158 0.45 1.12 260.38 37.78 273.98 0.76
P3 1.86 0.54 0.473 0.163 0.49 1.16 267.06 40.10 275.66 0.80
S.Em.± 0.05 0.02 0.007 0.004 0.01 0.03 6.61 0.81 5.82 0.02
C.D. at 5% NS 0.05 0.020 0.012 NS NS NS 2.45 NS 0.05
K0 1.74 0.36 0.431 0.134 0.40 1.03 243.85 28.69 253.61 0.65
K1 1.71 0.43 0.438 0.141 0.44 1.06 250.22 32.50 269.04 0.71
K2 1.75 0.48 0.454 0.144 0.48 1.16 260.14 33.50 274.78 0.74
K3 1.85 0.51 0.475 0.149 0.51 1.18 267.48 35.62 288.85 0.81
S.Em.± 0.05 0.02 0.007 0.004 0.01 0.03 6.61 0.86 5.82 0.02
C.D. at 5% NS 0.05 0.020 NS 0.04 0.09 NS NS 16.80 0.05
PxK
S.Em.± 0.10 0.03 0.014 0.008 0.03 0.06 13.22 1.62 11.63 0.03
C.D. at 5% NS 0.09 NS 0.024 NS NS NS NS NS NS
C.V.% 9.67 12.23 5.366 10.12 10.46 9.53 8.96 8.56 7.5 7.6

Table 3: Interaction effect of PSB and KSB on nitrogen and phosphorus content in
fodder.
Nitrogen content in fodder Phosphorus content in fodder
Treatments (%) (%)
K0 K1 K2 K3 K0 K1 K2 K3
P0 0.26 0.33 0.34 0.43 0.11 0.13 0.14 0.15
P1 0.44 0.44 0.42 0.45 0.08 0.15 0.15 0.14
P2 0.24 0.46 0.60 0.54 0.14 0.12 0.14 0.16
P3 0.50 0.49 0.56 0.62 0.10 0.15 0.18 0.17
S.Em.± 0.03 0.01
C.D. at 5 % 0.09 0.02

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Table 4: Effect of different treatments on nutrient uptake, yield and economics.

Nutrient uptake (kg/ha) NUE
Grain Fodder Net
(kg grain/kg B:C
Treatment Nitrogen Phosphorus Potassium yield yield return
Fertilizer applied) Ratio
(kg/ha) (kg/ha) (₹/ha)
Grain Fodder Grain Fodder Grain Fodder P K
P0 47.34 16.98 12.07 4.71 13.06 47.63 0.00 4.88 2819 4380 28931 1.87
P1 55.31 27.17 13.71 6.78 13.80 52.36 4.81 6.44 3109 4840 32366 1.90
P2 57.31 31.49 15.03 8.48 14.78 57.66 10.14 6.85 3276 5128 36531 2.02
P3 64.78 38.80 16.39 8.66 17.22 61.34 13.88 9.74 3452 5301 40018 2.11
S.Em.± 2.61 1.23 0.49 0.27 0.64 1.99 - - 98.35 125.32 - -
C.D. at 5% 7.54 3.55 1.40 0.79 1.86 5.76 - - 284.04 361.95 - -
K0 49.87 19.46 12.27 5.90 11.58 45.32 5.07 0 2839 4376 28628 1.84
K1 51.65 26.48 13.38 7.00 13.39 51.88 8.52 3.01 3015 4858 30749 1.86
K2 58.48 31.89 15.11 7.44 16.14 59.46 8.96 10.84 3325 5121 37770 2.06
K3 64.86 36.60 16.55 7.94 17.74 62.33 6.51 14.15 3476 5295 40695 2.14
S.Em.± 2.61 1.23 0.49 0.27 0.64 1.99 - - 98.35 125.32 - -
C.D. at 5% 7.54 3.55 1.40 0.79 1.86 5.76 - - 284.04 361.95 - -
PxK
S.Em.± 5.22 2.45 0.97 0.55 1.29 3.99 - - 196.69 250.64 - -
C.D. at 5% NS 7.09 NS 1.58 NS NS - - NS NS - -
C.V.% 16.08 14.86 11.76 13.43 15.16 12.62 - - 10.77 8.84 - -

Table 5: Interaction effect of PSB and KSB on nitrogen and phosphorus uptake by
fodder.
Nitrogen uptake by fodder (kg/ha) Phosphorus uptake in fodder (kg/ha)
Treatments
K0 K1 K2 K3 K0 K1 K2 K3
P0 10.72 23.66 13.96 29.50 4.29 6.13 6.45 6.73
P1 14.89 26.67 30.37 34.00 3.35 7.32 7.94 8.31
P2 16.85 27.74 40.63 42.34 6.48 5.82 8.20 9.27
P3 25.45 30.60 42.30 49.35 4.73 7.88 9.92 9.22
S.Em.± 2.45 0.55
C.D. at 5 % 7.09 1.58

PSB – P0K0 PSB – P2K2 PSB – P3K3

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KSB – P0K0 KSB – P2K2 KSB – P3K3

Plate (A): Different PSB and KSB colony isolated from soil and their growth.

EFFECT OF PSB
Effect of PSB on nutrient and micronial status, yield and economics of popcorn
Maize crop fertilized with 45 kg P2O5/ha + PSB seed inoculation + PSB soil application (P3)
produced significantly highest nitrogen content in fodder, phosphorus content in grain and
fodder, available phosphorus (P) and organic carban (OC) content in soil (Table 2) which
remained statistically at par with application of 45 kg P2O5/ha + PSB seed inoculation (P2).
Also highest nitrogen, phosphorus and potassium uptake by grain and fodder, grain yield
(3452 kg/ha) and fodder yield (5301 kg/ha), which was 22.45 and 21.01% higher over the
control treatment (K0) respectively and also gave maximum net return (₹ 40018/ha) and B:C
ratio (2.11) (Table 4) in P3 Treatments which was on par with application of 45 kg P2O5/ha +
PSB seed inoculation (P2). An increase in grain and fodder yields with these treatments might
be due to PSB solubilized the fixed soil phosphorus and readily hydrolysed the
organophosphate and degraded them in the soil and increase the availability of fixed P and
applied P to the plant owing to its favourable effects on division and multiplication of cells.
Moreover, phosphorus is the important constituent of co-enzymes which are important for
photosynthesis and protein synthesis. One of the main roles of the phosphorus in plant is in
transfer of energy through ATP and also involved in root development and in metabolic
activities especially in synthesis of protein. The results are in conformity with those reported
by Kaur and Reddy (2014) maize.

Total bacterial, total fungal and PSB count at 30 DAS and harvest (Table 1) were
significantly higher with application of 45 kg P2O5/ha + PSB seed inoculation + PSB soil

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application (P3) and on par with application of 45 kg P2O5/ha + PSB seed inoculation (P2).
Application of biofertilizers along with inorganic fertilizers increse microbial count may be
due to the fact that inorganic fertilizer which is use for body building of microorganism, so
that introduces fertilizer produce a high amount of beneficial microflora and phytohormones
in the soil which stimulating the growth of bacterial strains. Also observed significantly high
fungi population, this may be the result of increased nutrient availability in the rhizosphere
due to mineral fertilizers and activities of inoculated species which might have improved
growth of fungal hyphae. These results were in accordance with works Jilani et al. (2007),
Laxman, et al. (2017).

EFFECT OF KSB
Effect of KSB on nutrient and micronial status of soil, yield and economics of popcorn
An application of 45 kg K2O/ha + KSB seed inoculation + KSB soil application (K3)
recorded significantly the highest nitrogen and potassium content in grain and fodder,
phosphorus content in grain and available potassium (K) and OC content in soil (Table 2)
which was found statistically comparable to application of 45 kg K2O/ha + KSB seed
inoculation (K2) except in case of potassium content in fodder. Significantly the highest
nitrogen, phosphorus and potassium uptake by grain and fodder (Table 5) also gave highest
grain (3476) and fodder yield (5295 kg/ha) to the extent of 22.43 and 21.01% over treatment
K0 (control) respectively, net return (₹ 40695/ha) and B:C ratio (2.14) (Table 4) was
registered under the treatment P3 followed by P2 except in case of nitrogen uptake by fodder
and phosphorus uptake by fodder. With help of KSB mobilization of K from soil because of
secretion of organic acids by the bacterial strains, thereby enhanced plant growth and
development, and finally greater acquisition of nutrients. These results are in close
conformity with the finding of Padma and Sukumar (2015) and Yallapa et al.(2016). In case
of micobial status total bacterial and fungal and KSB count at 30 DAS and harvest were
significantly higher with K3 tretments, but it was found statistically at par with application 45
kg K2O/ha + KSB seed inoculation (K2).

INTERACTION EFFECT OF PSB AND KSB

The joint effect of different levels of PSB and KSB was found significant in case of nitogen
and phosphorus content and it’s uptake by fodder (Table 4). Wherein, the treatment
combination P3K2 recorded significantly the highest P content in fodder (0.18%), however it
remained statistically at par with the treatment combinations P3K3 and P2K3 also with help of

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P3K3 recorded significantly the highest N content in fodder (0.62%), however it remained
statistically at par with the treatment combinations P2K2, P2K3 and P3K2. Wherein, the
treatment combination P3K3 observed significantly the highest N uptake by fodder (49.35
kg/ha), however it remained on par with the treatment combinations, P2K3 and P3K2, While,
the treatment combination P0K0 (control) registered significantly the nitrogen uptake by
fodder (10.72 kg/ha). These results are in close conformity with the finding of Savliya et al.
(2014).

CONCLUSION
On the basis of one year field experimentation, it seems quite logical to concluded that seed
inoculation and soil application of PSB and KSB which increse microbial count in soil leads
to enhanced nutrient content in soil of popcorn (var. Amber), helps to increse nutrient uptake
and production can be secured by application of 45 kg P2O5/ha + PSB seed inoculation + PSB
soil application and 45 kg K2O/ha + KSB seed inoculation + KSB soil application at sowing
along with 120 kg N/ha (50% as basal + 50% as top dressing at 30 DAS) on medium black
calcareous clayey soil. It could be concluded that combine application of organic and
inorganic ferilizers enhanced grain yield with higher net return and saving of 25% fertilizer
dose of phosphorus and potash.

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