Efficacy of certain agrochemicals application at field rates on

microbial count of rhizospheric soil in Maize

M.G. Patil, V.S. Kolhe and R.R. Tamboli, Directorate of Weed Science Research
Centre, MAU, Parbhani.

E-mail: kolhevinaykumar@gmail.com

ABSTRACT:
The efficacy of three different agrochemicals (two herbicides and one
fungicide) and bioinoculant was carried out to determine their effects on the
physicochemical and biological properties of rhizospheric soil in Maize field.
Here, the representative soil samples for every treatment plots were collected for
estimation of total fungal and bacterial count. The colony forming units (CFU)
and total viable counts of total soil fungi and total soil bacteria were determined
by means of serial dilution technique and pore plant method after 30, 50 days of
sowing and at harvest of agrochemicals application in Maize field. The estimated
data indicated that agrochemicals used affected the microbial count of
rhizospheric soil in treated plots compared with the untreated soil. The total
fungal count was minimized in herbicide treated plots as compared to control
treatment. On the other hand, the plots in which seeds were treated with
Azospirillum showed maximum bacterial count as compared with control plot.

KEYWORDS: Agrochemicals, bioinoculant, microbial count, Azospirillum,

Rhizosphere and Maize.

INTRODUCTION:
If the global Maize production is examined, it is inferred that USA
has the lion’s share in total global Maize production accounting almost half of
the world’s harvest (~42.5 percent), other top producing countries includes The
Peoples Republic Of China, Brazil, Mexico, Argentina, India and France.
Worldwide production was around 800 million tonnes in 2007. In this year
(2007), over 150 million hectare of Maize was planted worldwide with a yield of
4970.9 kilogram per hectare. India is the fifth largest producer of Maize in the
world contributing 3 percent of the global production. In the last five decades,
India’s Maize production has increased from less than three million tonnes to 15
million tonnes today (2006-07). Maize, like any other cereal is grown across all
the states in India and in many states it is one of the important staple foods.
Karnataka and Uttar Pradesh are the leading producers of Maize in India while
Andhra Pradesh, Bihar, Madhya Pradesh, Maharashtra and Rajasthan are the
other important producers. According to the Union Ministry of Agriculture, as
on 10th July 2006, area brought under Maize in India was 35.22 lakh hectares up
from 27.36 lakh hectares planted during the corresponding period last year.
Maharashtra State holds prominent rank consisting total area under Maize to be
3,71,000 hectares with annual production of 7,44,000 tonnes and productivity of
2,000 Kg/ha in India (FAO, 2004). Microbial activity i s t h e g e n e r a l term
use d to indicate the vast range of activities carried out by soil
microorganisms Landi et al., (2000). It is markedly influenced by changes in
environmental factors and soil conditions Bloem et al., (2006). Biological
parameters may rapidly reflect even small changes that occur in soil,
providing accurate data about soil quality Smith and Papendick, ( 1 9 9 3 ) .
Pascual et al., ( 2 0 0 0 ) . Soil microorganisms, particularly soil fungi,
represent a biogeosphere component determining the level of their real
toxic effect, since they take part in their detoxification and mineralization,
using them as carbon and energy sources Dukia and Mandia (1998),
Nunez e t a l., (2001). Fungicides applied perform inhibition of soil fungi
development during the first two months following their application
Mandia et al. (2005). Recently, Zhao et al (2008) stated that the application
of elemental sulphur (30 mg/kg 2) increased the amount of soil
microorganism (bacteria, fungi and actinomycetes), and enhance soil
microbial activity. Das and Mukherjee (1998)., stated that, the
Hexachlorocyclohexane (BHC) followed by phorate significantly
stimulated the populations of (total) bacteria, actinomycetes, fungi and
aerobic non-sym biotic N 2 -fixing bacteria in soil.

MATERIAL AND METHOD:

 The present investigations were carried out in field as well as in
laboratory at DWSR Centre, MAU, Parbhani. The seeds of Maize (Karveer)
used for sowing in field experiment. The agrochemicals like fungicide
(Mancozeb), herbicide (Atrazine) and Azospirillum were used with different
dosage. The rhizospheric soil was used for laboratory experiment at different
observation time.

Media used:

Different semi-solid media were used for different microorganisms.

Nutrient agar for estimation of bacteria and Martin Rose Bengal agar media for
estimation of fungi (Pepper et al.., 1995)

Microbial analysis:

After application of agrochemicals at 1 DAS, 30 DAS and at 50 DAS in

treated plots. The total number of fungi and bacteria were counted. The number
of colony forming units (cfu) in the selective media were determined by means
of serial dilution technique and the pour plate method (Salle, 1973). Analysis
were done in three replicates.

Serial Dilution:

First prepared series of dilution blanks consisting of sterile water. For

each series, 250 ml conical flasks containing 90 ml of sterile water and 8-10 test
tubes containing 9 ml distilled water were required. Transfer 1 gm of soil sample
in 9 ml sterile distilled water in test tube (10-1) and shake properly. Transfer 1 ml
of suspension from this tube to another tube containing 9 ml of sterile distilled
water (10-2). Again transfer 1 ml of suspension from this tube to another tube
containing 9 ml sterile distilled water (10 -3). Similar dilution process was
continued as per requirements i.e. 10-4 for bacteria and 10-6 for fungi.

Pour plate method:

 Using fresh sterile micro pipette, pour 1 ml aliquot from the last dilution
into each of the three Petri dishes. After this, poured approximately 20 ml
volume of molten medium in each of the plates. Immediately after pouring,
gently moved the plates in a whirling motion to mix the contents, then allowed
the medium to solidify and incubate in BOD incubator at 30 0 + 2 0C in inverted
position.

Calculation of total viable count:

After proper incubation period, the number of colonies per plate was
recorded by digital colony counter. Total viable counts used are calculated by
formula

Total viable count = Avg. No. of colonies X Size of aliquot X Dilution factor.

RESULTS:

The present investigation is an attempt to check the effect of different

agro-chemicals on microbial count in relation to chickpea rhizosphere. The study
was conducted in field and in laboratory of Directorate of Weed Science
Research Centre, Marathwada Agricultural University, Parbhani. Result of these
experiments compiled, statistically analyzed and is presented in this chapter.

Effect of agrochemicals on microbial count of soil in relation to Maize

rhizosphere.
Effect of agrochemicals on total fungal count in Maize rhizosphere.
Table 1 Total fungal count.

Total Fungal Count

Tr. (cfu x 104)

Treatment
No. 30 50 At
DAS DAS harvest
T1 Weedy Check 35.33 38.66 47.66
T2 Mechanical weeding 25.00 35.00 45.33
T3 Atrazine @ 0.75 Kg/Ha 26.33 33.00 40.00
T4 Atrazine @ 1.50 Kg/Ha 26.66 34.00 43.00
Atrazine @ 0.75 Kg/Ha + 2,4-D @ 0.5
T5 32.00 33.66 42.66
Kg/Ha
T6 Mancozeb @ 0.5 % 24.00 28.33 34.66
T7 Mancozeb @ 0.2 % 27.00 32.00 38.00
Seed Treatment with Azospirillum @
T8 30.66 35.66 39.00
250g/ 10 Kg seeds
SE ± 0.97 1.09 1.09
 CD at 5% 2.95 3.32 3.32

In the above table, maximum fungal count was observed in treatment (T 1)

i.e., Weedy Check at 30 DAS (35.33), 50 DAS (38.66) and at harvest (47.66),
whereas minimum fungal count was noticed in treatment (T 6) i.e. Mancozeb @
0.5 % at 30 DAS (24.00), at 50 DAS (28.33) and at harvest (34.66).

Effect of agrochemicals on total bacterial count in Maize rhizosphere.

Table 2 Total bacterial count.

Total Bacterial Count

Tr. (cfu x 106)

Treatment
No. 30 50 At
DAS DAS harvest
T1 Weedy Check 49.00 52.33 55.00
T2 Mechanical weeding 47.33 50.00 53.33
T3 Atrazine @ 0.75 Kg/Ha 42.00 45.33 49.00
T4 Atrazine @ 1.50 Kg/Ha 43.33 45.00 49.00
Atrazine @ 0.75 Kg/Ha + 2,4-D @ 0.5
T5 46.00 49.33 51.66
Kg/Ha
T6 Mancozeb @ 0.5 % 44.00 46.00 48.66
T7 Mancozeb @ 0.2 % 42.66 45.66 49.00
Seed Treatment with Azospirillum @
T8 53.33 56.00 57.66
250g/ 10 Kg seeds
SE ± 1.53 1.39 1.71
CD at 5% 4.66 4.22 5.20

In the table mentioned above, total bacterial count found maximum at 30

DAS, 50 DAS and at harvest in treatment (T 8) i.e. seed treatment with
Azospirillum @ 250g/ 10 Kg seeds, which were (53.33), (56.00) and (57.66)
respectively. On the other hand, minimum total bacterial count was noticed at 30
DAS (42.00) in treatment (T3) i.e. Atrazin @ 0.75 Kg/Ha, followed by treatment
(T4) i.e. Atrazin @ 1.50 Kg/Ha at 50 DAS (45.00) and finally treatment (T 6) i.e.
Mancozeb @ 0.5 % at harvest (48.66).

DISCUSSION:

In present investigations, the rhizospheric soil samples in Maize plants

were taken from different agrochemical treatment plots from Maize field. The
maximum fungal population was observed in control plot (T 1) at 30 DAS, 50
DAS and at harvest, while most of the treatments were recovered. Herbicides
treated plots at recommended rates resulted lower fungal count compared to
control plots. Also, lowest fungal count observed in treatment (T6) i.e. Mancozeb
@ 0.5 % followed by (T7) i.e. Mancozeb @ 0.2 % due to fungicidal action.
Fungal count was minimum at 30 DAS followed by 50 DAS.
 Similar results were obtained by several research workers (Houseworth
and Tweedy, 1973; Cole, 1976; Hamed et al.., 1980; Frioni, 1981; Shukla et al..,
1990; Thopate et al.., 1990; Martinez Toledo et al.., 1998; Vazquez et al.., 2000;
Jose et al.., 2002; Khan and Ahmed, 2002; Aggarwal et al.., 2005; Hu et al..,
2005; Jose et al.., 2005; Ayansina and Oso, 2006; Huang et al.., 2006 (a); Huang
et al., 2006 (b); Cycon and Piotrowska, 2007; Jose et al., 2007; Ghany and
Ahmed, 2009)

In case of bacterial count, significantly highest bacterial count was

observed in treatment (T2) i.e. mechanical weeding followed by treatment (T 8)
i.e. seed treatment with Azospirillum @ 250/ 10 Kg seeds. On the other hand,
minimum total bacterial count was noticed at 30 DAS (42.00) in treatment (T 3)
i.e. Atrazin @ 0.75 Kg/Ha, followed by treatment (T 4) i.e. Atrazin @ 1.50 Kg/Ha
at 50 DAS (45.00) and finally treatment (T 6) i.e. Mancozeb @ 0.5 % at harvest
(48.66). Due to higher dose of Mancozeb (0.5 %), the fungal population along
with bacterial population got inhabited. On the other hand, in treatment (T 8) i.e.
seed treatment with Azospirillum @ 250/ 10 Kg seeds was strictly followed
without any interference of other agrochemicals. Hence overall microbial
population enhanced. Also, similar results were also reported by research
workers (Houseworth and Tweedy, 1973; Kapulnik et al.., 1981; Sarig et al..,
1983; Shukla et al.., 1990; Thopte et al.., 1990; Hernandez et al.., 1997; Krol
and Kobus, 1997; Burdman et al., 1997; Jacoud et al.., 1999; Vazquez et al.,
2000; Khan and Ahmed, 2002; Piutti et al.., 2003)

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