ABSTRACT
Actinomycetes are saprophytic soil inhabitants, the most widely distributed group of microorganism in nature. Plant rhizosphere soil habors various kinds of microorganism. Actinomycetes are aerobic, spore forming Gram-positive bacteria belonging to the order Actinomycetales characaterised with substrate and aerial mycelium growth that form the most abundant organisms that form thread-like filaments in the soil and are responsible for characteristically “earthy” smell of freshly turned healthy soil. The present study was conducted with a view to isolate and identifies Actinomycetes from rhizospheric soil of plant (potato, turmeric and coconut) Obtained from Okwuta Ibeku Umuahia using standard microbiological and biochemical techniques. The soil was pre-treated to eliminate the community known microbes using a hot air oven. The culturing was done using spread method and was subcultured using streak method. Two isolates were identified as Actinomyces spp and Streptomyces spp. The highest number of Actinomycetes were isolated from potato rhizoshere soil (2x10-5) from this study. Actinomycetes can be isolated and identified from different rhizosphere of plants. Further study should be carried out on rhisosphere of plants to exploit beneficial actinomycetes which will be useful to man and its environment.
TABLE OF CONTENTS
Title page i
Certification ii
Dedication iii
Acknowledgements iv
Table of contents v
List of tables vii
Abstract viii
CHAPTER ONE
1.1 Introduction 1
1.2 Actinomycetes 2
1.3 Aims and Objectives 3
CHAPTER TWO
2.0 LITERATURE REVIEW 4
2.1 Actinomycetes 4
2.1.1 Actinomycetes diversity and distribution in
nature 6
2.1.2 Actinomycetes a novel microorganism 6
2.1.3 Actinomycetes -brief on taxonomy 8
2.1.4 Role of Actinomycetes in ecosystem 9
2.1.5 Actinomycetes in plant rhizosphere of soils 9
2.2 Actinomycetes enzymes 10
2.2.1 Production of lytic enzymes 10
2.2.2 Therapeutic enzymes 10
2.2.3 Actinomycetes in biodegradation of toxic
pollutants 11
2.2.4 Actinomycetes as a probiotic supplement 12
2.2.5 Actinomycetes as Biocontrol Tools 12
2.3 Characteristics of Actinomycetes 13
2.3.1 Biochemical and physiological
characteristics
13
2.3.2 Molecular characteristics
14
CHAPTER THREE
MATERIALS AND METHODS
17
3.1 Sample collection
17
3.2 Sterilization of Materials
17
3.3 Pre-treatment of Soil Samples
17
3.4.1 Media Used
17
3.5 Serial
Dilution 17
3.5.1 Culturing
18
3.5.2 Colonial Examination 18
3.5.3 Sub-Culturing
18
3.5.4 Gram Staining
19
3.6 Biochemical Test
19
3.6.1 Starch hydrolysis test
19
3.6.2 Hydrogen sulphide test
20
3.6.3 Gelatin hydrolysis .
20
3.6.4 Catalasetest
21
3.6.5 Oxidasetest .
21
3.6.6 Carbohydrate fermentation test
21
CHAPTER FOUR
4.0 Results 23
CHAPTER FIVE
5.1 Conclusions
26
5.2 Recommendations
26
References
27
LIST OF TABLES
Table 4.1 Descriptions of samples collected from
different plants in Okwuta Ibeku 23
Table 4.2 Morphology and Biochemical
Characterization of Isolated Organism
24
Table 4.3 Description of samples and numbers of
Actinomycetes isolates 25
CHAPTER ONE
1.1 INTRODUCTION
Soil is a very diverse ecosystem which
supports the growth of countless organisms be it plants, animals, humans, or
microbes. The major biotic component of soil comprises of the microbiota (D'
Coasta et al; 2007). It harbors millions of micro-organisms like
bacteria and fungi. They are the integral part of every food chain, as they
decompose the dead and decaying organic matter in the soil, into simple
molecules which they use for their nutrition. Soil provides optimum conditions
such as moisture, temperature, PH and organic matter which are favourable for
the growth of microorganisms. These microorganisms produce primary metabolites
which are required for various cellular functions like growth and metabolism.
They also produce secondary metabolites in the later stage of their growth,
which are needed for their defense and survival.
Numerous
antibiotics have been isolated from a variety of microorganism however, studies
are still being conducted to identify novel antibiotics effective against
pathogenic fungi and bacteria. The number and species of microbes in soil very
directly in response to environmental conditions such as nutrient availability
soil texture, and type of vegetation cover. Filamentous Actinomycetes are known
to have the ability to produce a wide variety of secondary metabolites. Indeed
each strain of Actinomycetes likely has the generic potential for the
production of 10 to 20 secondary metabolites. (Bentley et al. 2002;
Sosio et al, 2000). Actinomycetes are useful tools for the production of
antimicrobials against fungi and bacteria. Antibiotics include a chemically
heterogeneous group of small organic molecules of microbial origin that are at
low concentration are deleterious to the growth or metabolic activities of
other microorganisms (Gebreselema el al; 2013).
Antibiotics are important secondary
metabolites that are produced by certain microorganisms such as Actinomycete.
They have immense importance for us human as they help fight many bacterial
infections and diseases. Predominantly Bacilllus and Actinomycetes are
present in soil. They are known for producing inhibitory substances like
Bacitracin and Actinomycin, respectively (Abebe et al., 2013).
1.2 Actinomycetes
Actinomycetes are prokaryotic having G+C content in their
DNA with extreme metabolic possibilities. They represent a transition between
bacteria and fungi. Actinomycetes are the most widely distributed group of
microorganisms in nature. They are attractive, bodacious and charming
gram-positive bacteria.
A majority of the Actinomycetes are free-
living saprophytes found widely distributed in soil, water and colonizing
plants. Their population forms an important component of the soil micro flora.
They form a very important soil inhabiting microbial group and many of them are
able to degrade complex organic compounds such as Keratin and Lignin which are
resistant substance.
Actinomycete
forms a large and important segment of the microflora of the natural
environment. Soils, fresh water, lake and river bottoms, manures and compost
contain an abundance of these organisms. They participate in many important
biochemical processes in soils. They play an important role in the degradation
of organic matter.it is anticipated that isolation, identification and study of
Actinomycetes can be useful in the discovery of novel antibiotics and species
of Actinomycetes .
1.3 Aims and Objectives
The present study is essentially concerned
with the isolation and identification of Actinomycetes from the rhizosphere
soil samples of three plant varieties with the following objectives.
• Isolation of Actinomycetes from the
rhizosphere soil sample of potatoes and turmeric and coconut.
• Identification of the isolates of
Actinomycetes by biochemical methods.
• Characterization of the isolates.
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