RESEARCH IN PROGRESS

TITLE: Effects of soil textural class on the soil microbial activity and soil N cycling
enzymes

STATUS: Part of the laboratory work completed, and enzymes

analysis in progress

INTRODUCTION

The quality of soil is related to its physical, chemical, and biological properties, which are affected by
management, land use type as well as the textural class. Soil quality has been extensively measured by
indicators such as the physical and chemical properties. Microbiological indicators are however more
sensitive to soil alterations (Doran & Parkin, 1994; Baretta et al.,(2010). The soil microbial community
plays an important role in agro ecosystem function and are on the field scale essential for plant nutrition
and health (Amann et al., (1995). The texture of the soil determines the distribution of soil
microorganisms and hence their activity. Soil enzymes are used as soil quality indicators due to their
high sensitivity and fast response to soil management and use, as well as the ease of determination
(Tabatabai, (1994). The N cycling enzymes are sensitive to variations in soil texture owing to the effects
of textural class on the distribution of the organisms producing the enzymes (Bastida et al., 2006).

OBJECTIVES

The main objective of this study is to determine the soil microbial status of the soil as influenced
by percentages of clay, silt and sand.

MATERIALS AND METHODS

The study was conducted in the laboratory of the Department of Crop, Soil and Pest
management of the Federal University of Technology, Akure, Ondo State (FUTA) located in the
rainforest vegetation zone of Nigeria. The soil sample was sampled from an area in the field
having no herbicide treatment during the last two years.

Soil Sample Preparation and Incubation

Soil samples were collected from a depth of 0-15 cm in the field and brought to the laboratory in
sealed polyethylene bags. In the laboratory, the samples were sieved using (2mm) to remove
plant materials, soil microfauna, debris and stones. After sieving, the soil samples were
homogenized and brought to a water holding capacity (WHC) of 60% by first wetting the soil to
field capacity after which the moisture content was adjusted to 60% by oven-drying. A portion of
the soil (200g) was weighed and transferred into plastic jars. The experiment was laid down in
completely randomized design (CRD) and the soils in the jars were incubated for 8weeks at
room temperature.

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Measurement of Soil Basal Respiration (SBR)
Basal respiration (μg CO2-C g-1 soil) was determined by the alkali sorption and titration method
described by Anderson and Domsch (1990). Three day prior to sampling, a 10 mL solution of
0.5M NaOH was dispensed into a 50mL beaker and placed inside the glass jars containing the
treated soil to trap CO2 evolved from the soil. On the third day, 5mL of 1.0M BaCl2 was added
to the NaOH solutions from the jars to precipitate carbonate (as BaCO3), thus facilitating the
determination of CO2 evolution (as μg CO2-C g-1 soil) from the treated soil. The evolved CO2-
C was then determined by titration. NaOH in solution was titrated against 0.5 M HCl using
phenolphthalein indicator. Two blanks without soil were prepared to assess the amount of CO2
trapped without respiratory activity.

Determination of Organic Matter

Soil sample was grinded to fine powder from which 1g was weighed out and then transferred
into 250ml conical flask. 10ml of 1M of K 2Cr2O7 (Potassium dichromate) was added and the flask
was swirled to mix. 20ml of Concentrated sulphuric acid H 2SO4 was then added rapidly and the
flask immediately swirled gently until soil and reagents are mixed and allowed to stand for
30minutes. 100ml of distilled water was added after standing for 30minutes and 3 drops of
ferroin indicator added. It was then titrated with 0.5M of ferrous sulphate (FeSO 4).

Determination of pH of the soil

10g of air dried soil was sieved with 2mm sieve and placed into 50ml beaker, 20ml of distilled
water was then added, stirred and allowed to stand for 30 minutes with occasional stirring. The
pH meter was then calibrated with pH 7.0 and pH 4.0 buffers. The electrode was inserted into
partly settled suspension during measurement. It was rinsed with distilled water and wiped dry
with a clean tissue paper after each reading.

Enumeration of Soil Microbial Population

Numbers of fungal spores were estimated by soil dilution technique on yeast extract and Potato
Dextrose Agars as isolation media for yeast and mold respectively. To achieve serial dilution, 5
grams of soil was suspended in 150 ml Erlenmeyer flask containing 95ml of sterilized distilled
water to obtain a 10-1 dilution and was kept under shaking conditions at 120 rpm for 15 minutes.
From the flask 1 ml of suspension was transferred to 9 ml water blank to make 10-2 dilution. The
water blank was vortexed and then again 1 ml of the suspension was transferred to a new water
blank (9 ml) tube to obtain 10-3 dilution. In the similar manner dilutions were made up to 10-5.
The constituents of the Potato Dextrose Agar (gL-1) were peptone 5.0, potato extract 5.0,
dextrose 10.0, Agar 20.0, and Distilled water 1000.0 ml at pH 6.5. Potato extract was replaced
with yeast extract to formulate the yeast agar. A mixture of 1g soil and 10mL of saline solution
was shaken on a mechanical shaker for 10 minutes to dislodge fungal propagules into the
solution. This was followed by serial dilutions to the concentrations of 10-5. 0.5 mL of the
aliquot was spread on Potato dextrose or yeast extract agars to isolate mold and yeast spores
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respectively, and this was incubated at 280C for 4 days. Dilution factors of 5 was used to
determine the yeast and mold spore forming units (SFU).

Activities of soil N cycling enzymes will be determined using methods by Tabatabai (1994)

Analysis of Data

Data collected will be subjected to Analysis of Variance (ANOVA) and means separation will be
done using the Duncan Multiple Range Test (DMRT).