ACTINOMYCETES ASSOCIATED WITH THE RHIZOSPHERE OF SOME SELECTED PLANTS

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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