Scientia Horticulturae 226 (2017) 1–9

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Research Paper

Optimization of organic and bio-organic fertilizers on soil properties and MARK

growth of pigeon pea
⁎
Rizwan Ali Ansari , Irshad Mahmood
Section of Plant Pathology and Nematology, Department of Botany, Aligarh Muslim University, Aligarh 202002, India

A R T I C L E I N F O A B S T R A C T

Keywords: Present study was undertaken in order to discover the best organic management system under field conditions.
Bio-organics Consortium of various applied organics and bio-organics exhibited notable improvement in growth and yield of
Fertilization pigeon pea over untreated control (Uc). Rhizobium sp. (Rh) + Municipal waste (Mw) applied field produced
Microbial load more crop growth and yield than Uc. Likewise, Rh + Mw applied plots also enhanced the Peroxidase Activity
Organics
(PA), Nitrate Reductase Activity (NRA) and Chlorophyll (Chl) contents over Uc. Moreover, application of Rh
Pigeon pea
+ Mw also enhanced the plant nutrients (N, P and K) status over Uc. The physico-chemical properties of soil
Soil enzyme
such as pH, Electrical Conductivity (EC), Porosity, Organic Carbon (OC), Bulk Density (BD) and Water Holding
Capacity (WHC) were recorded in treated and untreated plots. The EC and BD were drastically reduced while
porosity, OC and WHC were significantly higher in Rh + Mw treated plots. There was no considerable record of
pH at significance level. Besides, Rh + Mw applied plots registered higher agronomic (N, P and K) variables in
soil. Moreover, Soil Microbial Biomass Carbon (SMBC) and microbial population of fungi, bacteria and acti-
nomycetes were significantly higher in Rh + Mw treated plots as compared to Uc. β-glucosidase (β-glu) were
estimated and found significantly influenced in organically sound plots. β-glu exhibited maximum activities in
Rh + Mw treated plots suggesting to be a good soil indicator. In addition, Rh + Goat manure (Gm) and Rh
+ Poultry manure (Pm) were also found statistically significant over Uc, however, significance quantum was
lesser than Rh + Mw. The fertilization regime of the Rh + Mw was determined as optimal as it offered the
higher growth, yields and ameliorated soil physico-chemical properties ensuring sustainability. This finding has
suggested that Rh + Mw should be further disseminated among the growers in order to maximise crop yield.
Thus, our study is expected to provide a technology in making sustainable crop production a more profitable
income generating and successful industry.

1. Introduction sustainable agriculture, especially in the region where organic nutrients

Intensification of agriculture has accelerated indiscriminate use of
pesticides, chemical fertilizers which has not only negatively affected
the soil biota, quality of produce and human health but also put an
unbearable burden on growers (Ramesh et al., 2005; Littlefield-Wyer
et al., 2008; Yasmin and D'Souza, 2010; Geiger et al., 2010; Damalas
and Eleftherohorinos, 2011; Rivera-Becerril et al., 2017). Henceforth,
emphasis in search of proxy of hazardous fertilizers have been war-
ranted, widened and still on to advance agriculture by curbing the use
of inorganic fertilizers (Zaidi et al., 2009; Ansari et al., 2017a). Judi-
cious use of organics and bio-organics are cost effective in sustained
crop productivity, soil health and provide sufficient nutrient to sup-
plement the parts of chemical fertilizers (Worthington, 2001; Bahadur
et al., 2009a, 2009b; Sumbul et al., 2017). Organic fertilization through
use of animal manure has now been one of the main strategies in
in soils are poor (Chiti et al., 2012). In addition, Mw is undoubtedly
made-up of household and other yard waste and being used in agri-
culture by many developed countries (Hargreaves et al., 2008). Mw
minimizes the waste, kills pathogens, reduces weeds germination in
cultivated lands, and devastates malodorous compound leading to en-
hanced crop productivity (Jakobsen, 1995). Application of Mw in or-
ganic agriculture is gaining popularity due to its affirmative effect on
biological, physico-chemical properties of soil (Anikwe and Nwobodo,
2002; Hansen et al., 2006; Yoshida et al., 2016).
Moreover, plant growth promoting rhizobacteria (PGPR) assists well
in mineralization and channelization of nutrients leading to enhanced
plant productivity (van Loon, 2007; Fischer et al., 2007; Ansari et al.,
2017b). PGPR adopt various possible ways to accelerate the rate of crop
production (Chen, 2006; Rizvi et al., 2015; Ansari et al., 2017b).
Likewise, Rh fix 15–20 kg N/ha and enhance crop attributes to the tune

⁎
Corresponding author.
E-mail address: rizwans.ansari@gmail.com (R.A. Ansari).

http://dx.doi.org/10.1016/j.scienta.2017.07.033
Received 18 September 2016; Received in revised form 9 July 2017; Accepted 20 July 2017
0304-4238/ © 2017 Elsevier B.V. All rights reserved.
R.A. Ansari, I. Mahmood Scientia Horticulturae 226 (2017) 1–9

of 20% (Youssef and Eissa, 2014). The best studied example for PGPR’s Table 1
activity is Rhizobium-legume symbiosis (Cao et al., 2017). In this case, Chemical composition of organic amendments.
legume releases some flavonoids that act as signalling molecule for the
Organic fertilizers Moisture (%) g per kg
bacterial population to secrete Nod factors. These Nod factors are
quickly captured by root hairs and function like harmone which help in N P K
the root nodule induction. The bacterium is powered by carbohydrates
Ph 59 4.3 3.3 1.5
which is obtained from host plant and provides fixed N for amino acid
Ac 62 3.8 2.8 1.1
synthesis in return (Gray and Smith, 2005). Moreover, phosphate so- Pm 51 4.0 3.1 1.4
lubilising bacteria (PSB) maximise root nodulation, and thereby crop Gm 54 4.6 3.4 1.7
yield through increased phosphate solubilization and nitrogen fixation Mw 76 5.2 3.9 2.1
(Afzal and Bano, 2008). Conjoint application of Rh and PSB fixes the
Ph = Parthenium hysterophorus, Ac = Ageratum conezoides, Pm = Poultry Manure,
biological N, solubilizes insoluble phosphates in soil, optimize the fer-
Gm = Goat manure, Mw = Municipal waste.
tilizer use efficiency and crop productivity (Tripathi et al., 2009).
In addition, soil enzymes are known to play a substantial role in the
(10) Ab (10 kg applied per ha as soil application).
maintenance of soil health by catalyzing biochemical reactions.
(11) Pf (10 kg applied per ha as soil application).
Currently, β-glu is being used as soil indicators (Tejada et al., 2009; Liu
(12) Ph (110 kg N applied per ha as soil application).
et al., 2017). This enzyme responds more quickly to changes than
(13) Ac (110 kg N applied per ha as soil application).
others and being used in monitoring system of good soil health (Zhang
(14) Pm (8 T applied per ha as soil application).
et al., 2015). In order to combat this dearth of literature on the usage of
(15) Gm (8 T applied per ha as soil application).
organics with or without bio-inoculants on crop attributes. The present
(16) Mw (110 kg N applied per ha as soil application).
study was embarked with an objective to assess the impact of organics
and some bio-organics on plant growth attributes and physico-chemical
Bio-organics (Ab and Pf) and composted organics (Ph, Ac, Pm, Gm
and microbial diversity of soil. Based on the work, attempt was also
and Mw) with or without Rh were spread on the soil surface and in-
made to provide amicable solutions to address the challenges of organic
corporated to a 25 cm depth by ploughing and disking the day after
farming with the help of pigeon pea cultivation.
application. Uc plots received the same ploughing but no organic
treatments. Five days prior to seeds sowing the microbial inoculants of
2. Materials and methods
Ab, Pf at the rate of 10 kg per ha were applied as soil application
(Smitha et al., 2015). Before, qualitative and quantitative measures
2.1. Experimental design and plot installation
were taken out to know the chemical constituents of organic manures
(Table 1). Moreover, heavy metal level of Mw was determined by
A field experiment was conducted during cropping season (summer
Atomic Absorption Spectrometer 373, Perkin-Elmer, Norwak CT, USA
crop) of pigeon pea (Cajanus cajan L. cv. UPAS-120) in randomized
according to the procedure described by USEPA (1995). Some com-
block design in 3 replications in 2 × 6 m2 micro-plots with 36 seeds/
monly occurring metals such as Cd (0.43), Cu (49.12), Cr (68.21), Pb
micro-plot. The maintained spacing between plant to plant and row to
(59.31), Zn (193.21) mg per kg soil were obtained. The metal con-
row was 40 × 30 cm respectively. Half meter wide border was main-
centration were also compared with MEF (2000) standard and found
tained for each replication (Goswami et al., 2007). Performance of bio-
within the acceptable limits.
organics, Azospirillum brasilense (Ab), Pseudomonas fluorescens (Pf), and
organics, Parthenium hysterophorus (Ph), Ageratum conezoides (Ac),
Poultry manure (Pm), Goat manure (Gm) and Municipal waste (Mw) 2.2. Preparation of bio-inoculants
with or without Rhizobium sp. (Rh) were evaluated. Experiments were
carried out at the Agricultural Farm House (27.88 latitude and 78.07 2.2.1. Ab and Pf
longitude) of Aligarh Muslim University, Aligarh, India during two Ab and Pf were isolated from the roots and rhizosphere soil of pi-
consecutive years 2012–2013 and 2013–2014. geon pea. Both bio-organics were cultured and maintained for their
The land was prepared after ploughing twice with bullock drawn subsequent use. Ab was purely cultured and maintained by the methods
mould board followed by harrowing using cultivator-tractor and the of Sarig et al. (1986). Likewise, Pf culture was maintained in erlen-
whole plot was levelled with leveller, individual plots were made meyer flask with King’s B media (King et al., 1954).
manually as per experimental sketch. Sixteen treatments were used
(three replications per treatment). The amendment doses were de- 2.2.2. Rh
termined totally based on the recommendation of local farmers: Rh was isolated from the root nodules of pigeon pea growing in the
same field from previous to this experimentation. Briefly, healthy,
(1) Uc (untreated check; no organic input). pinkish, well developed, undamaged nodules were picked out, surface
(2) Rh (seed treatments of 25 ml per kg seed with the help of sticker). sterilized with 0.1% HgCl2 for 3 min followed by repeated washing with
(3) Rh (25 ml per kg seed) + Ab (10 kg applied per ha as soil appli- sterile water. Rh was further maintained on Yeast Extract Manitol Agar
cation). (YEMA) containing Congo red (205 ml of 1% Congo red in 100 ml
(4) Rh (25 ml per kg seed) + Pf (10 kg applied per ha as soil appli- YEMA). Furthermore, nodulation tests were performed for the char-
cation). acterization (Subba Rao, 1999). For inoculations, physically healthy
(5) Rh (25 ml per kg seed) + Ph (110 kg N applied per ha as soil seeds were first surface sterilized by 0.1% HgCl2 to remove all possible
application). microbes for 5 min 1 kg seeds were suspended in 25 ml of Rh strain
(6) Rh (25 ml per kg seed) + Ac (110 kg N applied per ha as soil containing 2 × 108 cfu/ml for 30 min.
application).
(7) Rh (25 ml per kg seed) + Pm (8 T applied per ha as soil applica- 2.3. Estimation of PA, NRA and Chl in plants
tion).
(8) Rh (25 ml per kg seed) + Gm (8 T applied per ha as soil appli- PA in plant leaves was determined from the first fully expanded
cation). leaves following the standard procedure (Chance and Maehly, 1955).
(9) Rh (25 ml per kg seed) + Mw (110 kg N applied per ha as soil NRA was estimated by the methods described by Jaworski (1971). Si-
application). milarly, Chl content was determined by the standard procedures of

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R.A. Ansari, I. Mahmood Scientia Horticulturae 226 (2017) 1–9

Table 2 2.6. Observations

Physico-chemical characteristics of the experimental field before installation of
experiment.
At the end of experiment plant height, fresh and dry weight, pollen
Soil Characteristics fertility, nodule density, pods and primary branches/plant, NRA, PA,
Chl, N, P and K (plant and residual soil) were monitored. Moreover,
Sand (%) 63.10 data of SMBC, microbial load of fungi, actinomycetes, bacteria, and soil
Silt (%) 19.20 physico-chemical and β-glu were also recorded.
Clay (%) 17.70
Texture class Sandy loam
Electrical Conductivity (dS m−1) 0.914 2.7. Data analysis
pH 6.5
Organic Carbon (%) 0.93
Available Nitrogen (kg per ha) 133 The data of two years were pooled and means were taken out since
Available Phosphorus (kg per ha) 21 there was no significant variation between the values of two con-
Available Potassium (kg per ha) 40 secutive years. In this way, all data were analysed with the help of ‘R’
Available Zinc (kg per ha) 5
software (R i386 3.2.3-version) to determine the Least Significant
Difference (LSD, P ≤ 0.05) and Duncan’s Multiple Range Test (DMRT).
Hiscox and Israelstam (1979). Multivariate Analysis, Principal Component Analysis (PCA) were car-
ried out with MS office XLSTAT software and drawn by Adobe
Illustrator. Some other complemental calculations were carried out
2.4. Estimation of N, P and K in plants and soil using MS excel 2007.

N content of plants and residual soil were estimated as per proce-

dure of IITA (1975). Similarly, P and K contents of plants were de-
termined by the described by (Lindner, 1944). Moreover, procedures of
(Olsen et al., 1954) for P and (Hanway and Heidel, 1952) for K esti-
mation in residual soil were adopted.
2.5. Soil analysis
The soil of the experimental site was tested before the experi-
mentation in order to know the soil nature (Table 2). Furthermore, soil
samples were collected periodically from the root zone of pigeon pea.
The soil samples were air dried and sieved through 2 mm sieve to
analyse soil chemical properties such as pH by pH meter, OC (Walkley
and Black, 1934), available nitrogen (Page et al., 1982), available
phosphorus (Olsen et al., 1954), available potassium (Knudsen et al.,
1982). Soil microbial biomass carbon (SMBC) was estimated by the
chloroform fumigation extraction method described by Vance et al.
(1987) using a kc = 0.45 conversion factor (Jenkinson and Ladd,
1981). Soil enzyme, β- glu (Eivazi and Tabatabai, 1988), microbial
loads (Dhingra and Sinclair, 1995) were also determined accordingly.
3. Results
3.1. Plant growth, yield attributes, enzymes and agronomic variables
The maximum improvement in the pulse crop with Rh + Mw was
reflected in the yield over Uc. Combined application of Rh + Mw
markedly improved the growth and yield attributes of pigeon pea crops
over Uc. Other treatments such as Rh + Gm and Rh + Pm were also
significantly effective over Uc, however, their levels of effectiveness
were lesser than Rh + Mw. Similarly, maximum number of nodules
was recorded in beds that received the consortia of Rh + Mw as com-
pared to Uc. Incorporation of Rh + Gm and Rh + Pm also registered
improved nodulations over Uc, but their improvement quantum was
lesser than the Rh + Mw. Likewise, plant enzymes such as NRA and PA
were greatly influenced in all scheduled treatments. Application of Rh
+ Mw improved markedly the status of NRA and PA as compared to Uc.
Besides, Rh + Gm and Rh + Pm exhibited the appreciable improve-
ment in both enzymes, but their improvements were once at par among
the other treatments except Rh + Mw. Similarly, higher Chl content
were recorded in Rh + Mw treated plants. Similar trends in Chl

Table 3
Status of plant growth, yield and yield like variables of pigeonpea under different fertilization regime.

Treatments Plant length Fresh weight Dry weight Pollen Number NRA† PAΨ Chl® mg per kg plant
(cm) (g) (g) fertility (%)
Pods Nodules Primary N P K
branches

Uc 118.30i 47.42j 13.62h 67.21i 48.21j 20.33h 13.21h 26.22g 0.317j 1.953k 48.60k 37.26i 26.32j
Rh 138.67h 57.41i 17.22g 70.66hi 74.35i 33.95g 19.96g 31.47f 0.323i 2.155j 71.06j 78.63h 54.81hi
Rh + Ab 173.50bc 72.22b−d 21.82a−e 85.14c−e 86.40b−e 40.30de 23.15a−d 36.56b−e 0.353e 2.465d 89.53e−g 94.72de 60.81de
Rh + Pf 176.53a−c 73.31a−d 22.20a−d 87.61b−d 88.33a−d 41.92bc 23.67a−c 37.29b−d 0.360d 2.515c 93.90c−e 97.48b−d 63.04cd
Rh+ Ph 163.38de 69.52de 20.73b−f 79.33e−g 82.73d−g 37.88ef 22.09c−g 34.40d−f 0.346f 2.357f 83.61hi 89.13fg 56.34f−i
Rh + Ac 168.23c−e 70.47cd 21.29b−e 81.44d−f 84.18c−f 38.88d−f 22.47b−f 35.69c−e 0.346f 2.403e 87.40f−h 91.74ef 59.54ef
Rh + Pm 180.08a−c 75.21a−c 22.69a−c 90.21a−c 89.92a−c 44.42a−c 24.06a−c 38.86a−c 0.367bc 2.563b 100.38b 99.60bc 66.06bc
Rh + Gm 184.93ab 77.40ab 23.32ab 94.11ab 91.89ab 45.21ab 24.60ab 39.60ab 0.372b 2.585ab 105.14ab 101.43b 68.34ab
Rh + Mw 188.36a 77.58a 24.33a 96.33a 93.63a 48.23a 25.13a 41.22a 0.378a 2.602 a 109.17a 105.61a 70.36a
Ab 146.94f−h 61.96g−i 18.97e−g 79.22e−g 78.64f−i 38.92d−f 21.05d−g 33.46ef 0.340g 2.312g 87.14f−h 89.81fg 57.70e−h
Pf 150.73fg 63.17f−h 19.37d−g 81.51d−f 81.31e−h 40.09de 21.75c−g 34.48d−f 0.346f 2.368f 89.45e−g 92.11ef 58.18e−g
Ph 143.92gh 59.59g−i 17.74g 72.33gh 75.77hi 37.02f 20.30fg 31.78f 0.327i 2.213i 81.47i 85.78g 53.49i
Ac 139.85gh 58.43hi 18.32fg 75.61f−h 77.24g−i 38.06ef 20.64e−g 32.03f 0.333h 2.269h 83.75hi 89.13fg 56.12g−i
Pm 156.35ef 64.59e−g 20.05c−g 82.66d−f 82.68d−g 41.31cd 22.28b−g 35.71c−e 0.353e 2.428e 92.44d−f 93.97de 60.55de
Gm 163.74de 68.10d−f 21.14b−f 84.22c−e 83.94c−f 43.39bc 22.50b−f 36.55b−e 0.360d 2.477d 95.36b−d 96.92 cd 62.96cd
Mw 169.42cd 70.76cd 21.96a−d 88.51b−d 85.97b−e 43.42bc 23.01a−d 37.55b−d 0.365cd 2.536c 98.76bc 99.10bc 65.59bc

Uc = Untreated Check, Rh = Rhizobium, Ab = Azospirillum brasilense, Pf = Pseudomonas fluorescens, Ph = Parthenium hysterophorus, Ac = Ageratum conezoides, Pm = Poultry Manure,
Gm = Goat manure, Mw = Municipal waste, NRA = Nitrate Reductase Activity, PA = Peroxidase Activity, Chl = Chlorophyl, N = Nitrogen, P = Phosphorus, K = Potassium
† = μmoles NO2−g−h, Ψ = purpurogallin−mg−min, ® = μg−g fresh leaf tissue. Data are mean of n = 3. Different superscript letters indicate significant differences among fertilizer
treatments at P ≤ 0.05.

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R.A. Ansari, I. Mahmood Scientia Horticulturae 226 (2017) 1–9

Fig. 1. Principal Component Analysis (PCA) of variability for different bio-organics and organics fertilization regimes. Each fertilizer (both bio-organics and organics) was designated
using the initials of the first and second components. Uc = Untreated Check, Rh = Rhizobium, Ab = Azospirillum brasilense, Pf = Pseudomonas fluorescens, Ph = Parthenium hysterophorus,
Ac = Ageratum conezoides, Pm = Poultry Manure, Gm = Goat manure, Mw = Municipal waste. 1 = Uc, 2 = Rh, 3 = Rh + Ab, 4 = Rh + Pf, 5 = Rh+ Ph, 6 = Rh + Ac, 7 = Rh
+ Pm, 8 = Rh + Gm, 9 = Rh + Mw, 10 = Ab, 11 = Pf, 12 = Ph, 13 = Ac, 14 = Pm, 15 = Gm and16 = Mw.

improvement were also recorded in Rh + Gm and Rh + Pm treated Table 4

plants. Moreover, application of Rh + Mw increased tested agronomic Biological properties of soil after addition of different organic and bio-organic fertilizers.
parameters like N, P and K manifold over Uc. Briefly, Rh + Gm and Rh
Treatments SMBC (log μg g−1 soil) Microbial load (log CFU g−1 soil)
+ Pm were also effective in the growth and nutrient promotion of pi-
geon pea but, their values were lesser than Rh + Mw (Table 3). Bacteria Fungi Actinomycetes
Precisely, PCA explained that first two principal components (PCA 1
h l h
and PCA 2) accounted for ∼95% of total variability between the dif- Uc 2.66 12.56 7.84 9.86j
Rh 2.67h 12.60jk 7.90fg 9.91hi
ferent organic and bio-organic fertilizers, out of which 92% variability Rh + Ab 2.76bc 12.72cd 8.02b−d 10.03b−d
was contributed by PCA 1 (Fig. 1). Rh + Pf 2.76bc 12.73bc 8.04bc 10.05bc
Rh + Ph 2.74c−e 12.69ef 7.99c−e 9.99de
Rh + Ac 2.75cd 12.70de 8.00c−e 9.99de
3.2. Biological properties Rh + Pm 2.78ab 12.75ab 8.04a−c 10.06a−c
Rh + Gm 2.79a 12.76a 8.07ab 10.07ab
3.2.1. Soil Microbial Biomass Carbon (SMBC) Rh + Mw 2.79a 12.77a 8.10a 10.10a
The amendments of different organic and bio-organic with or Ab 2.72e−g 12.65gh 7.95ef 9.96e−g
Pf 2.72e−g 12.67fg 7.97de 9.96e−g
without Rh significantly improved the status of microbial counts re-
Ph 2.71fg 12.64hi 7.94ef 9.93f−h
sulting manifold improvement in SMBC. Higher values of SMBC were Ac 2.70g 12.62ij 7.94ef 9.92gh
observed in Rh + Mw as compared to Uc. Other parallel treatments like Pm 2.73d−f 12.68ef 7.97de 9.97ef
Rh + Gm and Rh + Pm also attained a significance level in SMBC Gm 2.73d−f 12.68ef 7.97c−e 9.97ef
improvement but ranked after Rh + Mw (Table 4). Mw 2.75cd 12.70d 8.00c−e 10.02cd

Uc = Untreated check, Rh = Rhizobium, Ab = Azospirillum brasilense, Pf = Pseudomonas

3.2.2. Microbial population fluorescens, Ph = Parthenium hysterophorus, Ac = Ageratum conezoides, Pm = Poultry
Table 4 revealed that amending soil with different organic and bio- manure, Gm = Goat manure, Mw = Municipal waste, SMBC = Soil Microbial Biomass C,
organics significantly improved the total fungi, actinomycetes and Each value is a mean of three replicates.
Note: Log (x + 1) were portrayed to normalizing observations’ values.
bacterial population as compared to Uc. Maximum improvement in
microbial population was recorded in Rh + Mw sound plots as com-
3.4. Soil physico-chemical properties
pared to Uc. Rh + Gm and Rh + Pm treated plots also gained con-
siderable improvement but their levels were ranked after Rh + Mw. An
Integration of organics and bio-organics improved the soil pH, EC,
initial fungi population in the soil rose from log 7.84 to log 8.10 cfu per
BD, porosity, OC and WHC. An increased soil pH were recorded,
g soil after soil amendments of different organic and bio-organic. Si-
however, improvement were statistically non-significant over Uc. The
milarly, log 9.86 to log 10.10 in actinomycetes and 12.56 to log 12.77
lowest soil BD while higher EC, OC, porosity, WHC were seen in Rh
bacteria were recorded.
+ Mw compared with that of Uc. Rh + Gm and Rh + Pm also assumed
the significance level but lesser than Rh + Mw (Table 5).
3.3. Soil enzymes

Soil enzymes are known to play a substantial role in sustenance of 3.5. Correlation
soil ecosystem. Application of Rh + Mw significantly enhanced the
activity of β-glu over Uc. A wide range of other treatments especially A positive correlation between available N and β-glu were obtained
Rh + Gm and Rh + Pm also assured marked enhancement in this en- suggesting that there is a role of soil N in the enhanced activity of β-glu
zymes, but lesser than the Rh + Mw (Fig. 2). (Fig. 3).

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R.A. Ansari, I. Mahmood Scientia Horticulturae 226 (2017) 1–9

70
Rh+Pm Rh+Gm Rh+Mw Ab Pf Ph
Ac Pm Gm Mw
70 60

60 50

50 40

40 30

30 20

20 10
135 155 175 195 215 235 255 275

10
Fig. 3. A relationship between available N and β-glucosidase.

0
Fig. 2. Status of soil enzymes β-glucosidase under different fertilization regimes of or-
ganics and bio-organics. Each value is a mean of three replicates.
4. Discussion
4.1. Plant growth, yield, enzymatic and agronomic variables
The results of this study has revealed that organics and bio-organics
supplied plots improved the pigeon pea yield, physico-chemical prop-
erties of soil leading to enhanced soil enzyme and microbiome. The
improvement tune by the different fertilizers with or without Rh on
various yield variables might be the results of good source of nutrients.
Application of beneficial microbes to crop plants enhanced the fixation
of atmospheric N, organic decomposition and augmented the soil
properties. These organics are considered to produce bioactive com-
pounds like wide range of vitamins, phytohormones which accelerated
plant growth and yield (Bahadur et al., 2009a, 2009b; Upadhyay et al.,
2012). Besides, organic matters like paddy straw are naturally available
in plenty and play a crucial role in the sustenance of crop plants. Such
organics have the potential to replenish soil OC and essential micro-
nutrients leading to ameliorated soil health offering more crop produce
(Surekha et al., 2003). Rh + Mw registered maximum improvement
among all treatments suggesting good source of nutrients leading to
improved plant growth and yield traits.
Moreover, NRA was also increased, however, at varying extent. This
might be explained as due to ease in availability of biodegradable
materials and large nutrients status in Mw and other supplied organics.
Our studies are in consistent with Ruiz et al. (2000) where, increased
NRA was directly proportional to greater synthesis of protein, amino
acids and N assimilation. These facts might also be the responsible for
higher nodulation. Villora et al. (2002) reported that NRA could be
altered by optimizing the concentration of NO3− and Mo essential in
the acceleration of enzymatic reaction and utilization of N. However,
reproductive parts such as flowers and fruits accelerated the significant
improvement in NRA (Carelli et al., 2006; Márquez-Quiroz et al., 2014).
Moreover, these findings corroborated the results of Amal et al. (2010)
who reported an increased PA in plants due to certain biotic and abiotic
factors (Ansari and Mahmood, 2017).
The use of all organic fertilizers along with Rh enhanced the plants’
physiological parameters like Chl which might be due to enhanced
nutrients absorption and assimilation. Ondiek et al. (2011) found
higher Chl a and b in organically amended plots than did in non-
amended control in African nightshade species. Besides, decomposition
of organic inputs releases wide spectrum of nutrients which accelerate
the rate of protein, amino acids and photosynthetic activity leading to
enhanced root development and eventually increased potassium

Table 5
Effect of different organic and bio-organics amendments on soil physico-chemical properties.

Treatments pH EC (dS m−1) Porosity (%) OC (%) BD (g cm−3) WHC (%) Available in kg per hectare

N P K

NS a f m a f k m
Uc 6.04 0.996 49.03 0.96 1.39 39.62 140.52 23.62 42.21ij
Rh 6.36NS 0.736b 52.97ef 1.71l 1.22b 49.19e 175.96j 29.56l 46.26i
Rh + Ab 6.44NS 0.496f−i 58.47a−e 2.03f 1.11b−e 49.98c−e 225.45d−f 58.96e−g 52.37e−g
Rh + Pf 6.47NS 0.457h−j 59.49a−d 2.09e 1.08c−f 51.51be 233.56c−e 60.52c−e 54.68d−f
Rh+ Ph 6.40NS 0.546e−g 56.49a−e 1.87h 1.15b−d 48.80e 206.06gh 56.70g−i 50.66e−i
Rh + Ac 6.42NS 0.526f−h 57.66a−e 1.93g 1.13b−e 49.57d−e 213.74fg 58.39e−h 51.56e−h
Rh + Pm 6.49NS 0.402j−l 60.39a−c 2.18c 1.03d−f 53.97b−d 242.17bc 62.43bc 59.61bc
Rh + Gm 6.52NS 0.348lm 61.45ab 2.22b 0.97fg 55.95ab 249.03ab 64.02ab 61.06ab
Rh + Mw 6.56NS 0.299m 62.57a 2.31a 0.87g 59.30a 262.38a 65.95a 64.86a
Ab 6.46NS 0.605de 54.91c−f 1.85i 1.15b−d 49.94c−e 205.19gh 54.34ij 49.90f−i
Pf 6.47NS 0.563ef 55.02c−f 1.94g 1.11b−e 50.40c−e 212.21fg 56.02h−j 52.57e−g
Ph 6.40NS 0.695bc 53.56d−f 1.73k 1.22b 49.53de 184.94ij 53.05k 47.36hi
Ac 6.42NS 0.654cd 54.38c−f 1.77j 1.19bc 49.73c−e 196.19hi 53.67jk 48.21g−i
Pm 6.49NS 0.477g−i 56.22b−e 2.02f 1.05d−f 51.99b−e 223.75ef 57.68f−h 55.20c−e
Gm 6.51NS 0.426i−k 57.67a−e 2.12d 1.01ef 54.34bc 233.57c−e 59.09d−f 57.92b−d
Mw 6.52NS 0.374kl 59.64a−d 2.21b 0.96fg 55.56ab 239.97b−d 61.46cd 58.11b−d

Uc = Untreated Check, Rh = Rhizobium, Ab = Azospirillum brasilense, Pf = Pseudomonas fluorescens, Ph = Parthenium hysterophorus, Ac = Ageratum conezoides, Pm = Poultry Manure,
Gm = Goat manure, Mw = Municipal waste, EC = Electrical Conductivity, OC = Organic C, BD = Bulk Density, WHC = Water Holding Capacity, N = Nitrogen, P = Phosphorus,
K = Potassium, NS = Not Significant. Data are mean of n = 3. Different superscript letters indicate significant differences among fertilizer treatments at P ≤ 0.05.

5
R.A. Ansari, I. Mahmood
content which assist in stoma regulations. Behaviour of stomatal con-
ductance is an important phenomenon for plants due to its participation
in CO2 acquisition and assimilation leading to increased photosynthetic
activity and chlorophyll pigments (Dodd, 2003).
As far as ameliorations in plants nutrient status is concerned, en-
hanced nutrients like N, P and K might be due to enhanced vegetative
growth and improved soil health leading to increased root formation.
Healthy roots acquire more minerals from the soil pumping out towards
the photosynthetic machinery leading to enhanced nutrient pool
(Sarkar et al., 2003; Pernes-Debuyser and Tessier, 2004). Santibáñez
and Varnero (2014) reported that seed cake application enhanced sig-
nificantly the N, P and K constituent of plant which might be attributed
to presence of abundant number of soil microorganisms, microbial re-
spiration and microbial biomass C. In addition, increased level of P
might be due to presence of adequate organic acids which were released
after organic matter decomposition and subsequently help in the solu-
blization of the indigenous phosphates (Bharadwaj and Omanwar,
1994). Similarly, presence of increased pool of K might be due to po-
sitive role of organic inputs on the mitigation of potassium fixation in
the soil. Organics gets interacted with the K clay to liberate K from un-
exchangeable form to available sink (Ramesh et al., 2005).
4.2. Biological properties
In the present study, incorporation of organics and bio-organics
with or without Rh triggered a manifold increase in SMBC and popu-
lation of fungi, bacteria and actinomycetes. Such surprising improve-
ments in microbial status may be the reason behind augmented soil
health.
4.2.1. SMBC
SMBC were significantly higher in Rh + Mw amended plots. This
might be due to enhanced mineralization of organic leading to in-
creased microbial activity (Nagar et al., 2016). Incorporation of or-
ganics supplied easily mineralizeable and hydrolyseable C leading to
enhanced microbial activities ensuring greater SMBC (Manna et al.,
2001; Chandrashekhar, 2012). Increased population of Azotobacter,
PSB, soil enzymes, OC exhibited positive correlation with SMBC
(Majumdar et al., 2014). Moreover, significant improvement in SMBC
might be due to more stimulation in zymogenous microbial populations
(Blagodatsky et al., 2000; Schaffers, 2000). In addition, SMBC and soil
respiration were markedly higher in fulvic acid rich organic due to
presence of large labile fraction of organic matter. The labile fractions
are degraded rapidly leading to increased mineralization and channe-
lization of chaste energy to microbes (Cook and Allan, 1992).
4.2.2. Microbial loads
The rhizosphere is also a niche of beneficial microflora. Addition of
organic and bio-organic with or without Rh exhibited enhanced mi-
crobial activity. It could be explained that higher root exudation, OC, N
and moisture content in amended organics powered the microbes for
their proliferation (Nagar et al., 2016). Decomposition of root tissue
and root nodules provided sufficient energy in terms of C and N for the
multiplication of fungi (Sharma and Guled, 2012), bacteria (Singh
et al., 2012) and actinomycetes (Sharma et al., 2000) leading towards
the building up of heavy microbial population. Moreover, higher po-
pulation in beneficial microbes may be due to balanced water supply,
particulate organic matter and presence of respirable substances
(Rupela et al., 2005; Sharma et al., 2011; Majumdar et al., 2014). Latha
and Sharanappa (2014), reported that significant bacteria (34.58 × 106
cfu/g), fungi (21.77 × 103 cfu/g) and actinomycetes (11.28 × 103 cfu/
g) population were recorded over bacteria (19.77 × 106 cfu/g), fungi
(9.5 × 103 cfu/g) and actinomycetes (5.44 × 103 cfu/g) due to varia-
tion in organic C nutrient status and rate of decomposition of supplied
organics.
6
Scientia Horticulturae 226 (2017) 1–9
4.3. β-glu
In the present study, β-glu was more significant in beds treated with
Rh + Mw. Incorporation of organic inputs to the soil led to increased β-
glu activity. This might be due to presence of higher amount OC in the
soil (Eivazi and Tabatabai, 1990). Addition of organic matter ac-
celerated the microbes towards the production of soil enzymes (Tejada
et al., 2009). Maximum β-glu was observed in Rh + Mw amended soil
suggesting that this consortium provided abundant C sink for microbes
consequently increased β-glu activity (Bandick and Dick, 1999; Tejada
et al., 2009; Zhang et al., 2015; Liu et al., 2017).
4.4. Soil physico-chemical properties
Physico-chemical properties of soil were markedly influenced in the
organically sound plots, especially Rh + Mw as compared to Uc.
Spaccini et al. (2004) and Tejada et al. (2006) propounded that in-
corporation of organic manures acted as cementing materials which
helps in flocculation of soil assisting in soil aggregations. However,
improvement depends on the composition and nutrient status of or-
ganic inputs applied into soil. Another report with respect to this study
suggested that soil aggregation is markedly correlated with the humic
acids but not fulvic acids due to its direct role in clay-organic formation
(Whalen et al., 2003). Mucilage and mucilage like compounds produced
by microbes registered marked improvement in soil aggregation
(Oades, 1993; Six et al., 2004).
The changes in pH varied from 6.04 to 6.56 and higher but re-
mained non-significant statistically in Rh + Mw and other treatments
as compared to Uc. The increased soil pH might be due to the C mi-
neralization and later on production of OH− ions through exchange of
ligand and basic cations like K+, Ca2+, and Mg2+ introduction
(Mkhabela and Warman, 2005).
In addition, EC was found to be increased considerably in organi-
cally sound soil being greater in Rh + Mw. Concentration of dissolved
solutes contents might be the reason of enhanced EC because EC of the
soil solution is related and often used to measure soil salt level (Brady
and Weil, 1996).
Soil BD significantly decreased in the organic amended soil than Uc.
This might be due to increased soil porosity which happened due to
joint action of organics and bio-organics. Tejada et al. (2009) reported
that soil BD were decreased due to compost addition to the soil leading
to enhanced humic acid concentration, soil aeration, dilution of the
denser soil mineral fractions, later on porosity and structural soil sta-
bility. Present study corroborated the earlier findings of Kay and
Vanden Bygaart (2002), Shukla and Tyagi (2009) and Nagar et al.
(2016).
Increased OC were recorded in consortium (organics + bio-or-
ganics) applied plots. Oxidation rate of organics by soil microbes are
the main cause behind increased OC (Babhulkar et al., 2000; Ramesh
et al., 2009). Higher WHC were noticed in Rh + Mw due to increased
soil pores. The decomposition magnitude of organic matter has become
a significant factor on increased WHC; controlled by pore size, abun-
dance and surface area (Kladivko and Nelson, 1979). Furthermore, re-
duced BD leads to enhanced alteration of small pores (Khaleel et al.,
1981). Application of organic and bio-organic fertilizers with or
without Rh leads to increased surface area and subsequently higher
surface tension which is very close to wilting point of soil might be the
another reason of higher WHC (Gupta et al., 1977; Khaleel et al., 1981).
4.5. Nutrient status in soil
Incorporation of organic and bio-organic with or without Rh en-
hanced the soil nutrient status such as N, P and K significantly than did
the Uc plots. This might be associated with the availability of higher
quantum of N, P and K in the organic supplied plots. Gopinath et al.
(2008) reported that addition of FYM and Lantana sp. resulted higher
R.A. Ansari, I. Mahmood
yield in growth attributes and increased nutrient status such as avail-
able N, P and K into the soil, might be due to presence of plethora of
nutrients into the organic applied to the field. In present study, other
treatments also responded significantly but Rh + Mw ranked first fol-
lowed by other ones. This could be explained due to slower rate in
release of nutrients to the soil from such organic inputs (MacRae et al.,
1993). Vyas et al. (2003) reported that application of 5 kg Zn + 10 T
FYM/ha enhanced the N status of soil due to addition of organic manure
and inorganic fertilizers to the soil in mint-mustard intercropping. Such
significant improvement might be due to faster release of P and K after
mineralization of organic inputs. Tiwari et al. (2002) reported sig-
nificant improvement in N, P and K status of soil in the organically (15T
per ha) sound plots. Moreover, multivariate analysis showed that con-
sortia of manure and inorganic fertilizers could be a better option in the
maintenance of environmental issues (Liang et al., 2012). Similarly, in
our experiment, Rh + Mw applied plots exhibited significantly max-
imum yield, nutrient (plants and soil), SMBC and microbial activities
suggesting faster turnover of soil nutrients (Liu et al., 2017).
4.6. Correlation
In our results, positive correlation between two important variables
was obtained. Available soil N and β-glu showed a considerably positive
relationship. This might be due to enhanced SMBC, a determinant
factor of the increased β-glu. Moreover, MBC is the important source of
C supplying higher energy to the microbes leading to enhanced enzy-
matic activities (Navas et al., 2011; Liu et al., 2017).
5. Conclusions
The inference could be drawn that conjoint application of Rh + Mw
showed higher impacts on crop produce. Plant nutrients (N, P and K)
enzymes (PA and NRA) and photosynthetic pigments (Chl) registered
manifold improvement in organically sound (Rh + Mw) plots as com-
pared to Uc. Other supplied treatments like Rh + Gm and Rh + Pm
were also gave promising results over Uc. However, their influences
were lesser than the Rh + Mw treated plots. Biological properties of the
soil were determined and found to be significantly influenced. SMBC
was found to be higher in Rh + Mw treated plots as compared to Uc.
Similar trends were recorded in the microbial (fungi, bacteria and ac-
tinomycetes) populations. Likewise, soil physico-chemical properties
were also found to be greatly influenced. Higher values of pH, EC,
porosity, OC and WHC were recorded in Rh + Mw treated plots. A
significant reduction in BD was recorded in the same treatments as
compared to Uc. Rh + Gm and Rh + Pm also exhibited considerable
improvement in soil physico-chemical properties. However, levels of
significance were found lesser than what was seen in Rh + Mw treated
plots.
Therefore, based on these findings it is concluded that not only
chemical fertilizers could produce a good yield and fetch a good re-
muneration but also consortia (Rh + Mw) could also achieve the tar-
geted yield and good returns under better management practices.
Possibly, use of this fertilization regime would offer an opportunity for
energy savings and mitigation of soil pollution. Although prior to use,
analysis for the detection of heavy metals should be deployed which are
supposed to be more pronounced in Mw because presence of heavy
metals beyond the permissible level offer manifold enhancement in the
soil toxicity level. Thus, waste utilization by this way may provide a
solution for the problem associated with the safe disposal of the Mw. It
would also be very enthusiastic to monitor plant quality like taste of
yield and morphology, physiognomy, susceptibility towards biotic pa-
thogens as these are fundamental principle for the consumers/en-
trepreneur before its marketing.
7
Scientia Horticulturae 226 (2017) 1–9
Conflict of interest
Both authors have declared no conflict of interest.
Acknowledgement
The Chairperson of Department of Botany, AMU, Aligarh is grate-
fully acknowledged for permitting us to carry out this research. RAA is
highly grateful to University Grant Commission (UGC), Government of
India for providing Non-NET fellowship during the course of this re-
search. Anonymous reviewers and editorial team are highly thanked for
their kind support during the revision of this article.
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