ASSESSMENT OF GENETIC (VIA PROTEIN ELECTROPHORESIS) AND NUTRITIONAL VARIABILITIES AMONG TEN ACCESSIONS OF SPHENOSTYLIS STENOCARPA (HOCHT. EX. A. RICH) HARMS

ABSTRACT

The genetic variability among ten (10) accessions of Sphenostylis stenocarpa from three states and National Centre for Genetic Resources and Biotechnology Moor Plantation, Ibadan, Nigeria were assessed using sodium dodecyl polyacrylamide gel electrophoresis (SDS - PAGE). There were variations in the banding patterns of the accessions and polymorphism was also observed. Twelve polypeptide bands were obtained ranging from 9KDA to 219KDa. The Unweighted Pair Group Method with Mean Algorithm (UPGMA) dendrogram grouped the ten accessions into two clusters and six groups. Some accessions appeared in the same group, while others were observed to be in different groups. The proximate and nutritional analysis also detected variations. A strong and positive correlation was observed among some of the attributes studied. The principal component analysis also showed some variation among the accessions. The highest variability was observed in the ash content which shows that ash content could be a major indicator for nutritional variability in these 10 accessions followed by the Vitamin B₂ in the second component.

TABLE OF CONTENTS

Title page

Table of contents

List of plates

List of figures

List of tables

Abstract

CHAPTER ONE: INTRODUCTION

CHAPTER TWO: LITERATURE REVIEW

CHAPTER THREE: MATERIALS AND METHODS

3.1.      SODIUM DODECYL SULPHATE – POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE)

3.1.1. Protein extraction

3.1.2    Preparation of gel

3.1.3    Loading the electrophoresis tank

3.1.4.   Loading the samples

3.1.5    Staining the gel

3.1.6    Destaining

3.1.7    Gel documentation and analysis

3.2.  NUTRITIONAL ANALYSIS

3.2.1. PROXIMATE ANALYSIS

            3.2.1.1.Determination of crude protein content

            3.2.1.2 Determination of carbohydrate content

            3.2.1.3 Determination of crude fat content

            3.2.1.4.Determination of crude fiber content

            3.2.1.5 Determination of ash content

            3.2.1.6 Determination of moisture content

3.3.1    MINERALS AND VITAMIN ANALYSIS

            3.3.1.1.Determination of Vitamin A

            3.3.1.2.Determination of Vitamin B

            3.3.1.3 Determination of Vitamin C

            3.3.1.4 Determination of Zinc

            3.3.1.5 Determination of Lead

            3.3.1.6 Determination of Iron

3.4.      DATA ANALYSIS

CHAPTER FOUR: RESULTS

4.1.      Evaluation of Genetic variability

4.2.      Nutritional Variability

CHAPTER FIVE: DISCUSSION AND CONCLUSION

CONCLUSION

REFERENCES

APPENDICES

CHAPTER ONE

INTRODUCTION

African Yam Bean (AYB) (Sphenostylis stenocarpa Ex. A. Rich, Harms) is an underutilized food leguminous crop of the family Fabaceae, sub family Papilionoideae (Azeke et al., 2005; Moyib et al., 2008). The domestication, cultivation and distribution of the crop are very evident in the tropics and Africa where it had been reported to exhibit very high diversity (Okpara et al., 1997). The distribution of the crop according to Genetic Resources Information Network (GRIN), (2009), includes the following countries within the tropical regions of Africa: Chad and Ethiopia (Northeast tropical Africa); Kenya, Tanzania and Uganda (East tropical Africa); Burundi, Central African Republic and Democratic Republic of Congo (Central tropical Africa); Cote d’voire, Ghana, Guinea, Mali, Niger, Nigeria and Togo (West tropical Africa) etc.

Nigeria is very significant in AYB production where extensive cultivation had been reported in the Eastern, Western and Southern areas of Nigeria (Saka et al., 2004). AYB thrives in weathered soils where the rainfall can be extremely high. It grows well even in acidic and highly leached sandy soil of the humid low land tropics (Uguru et al., 2001).

The plant is an annual, with climbing, vine-like stems that require staking; the stem is often reddish in colour. In Nigeria, it is usually grown in association with yams, maize, cassava, etc., so that the same stake of the yam serves as support for both crops. It is cultivated mainly for home consumption and only about 30% of the dry grain produced is sold. It is also planted for soil restoration (Saka et al., 2004). The seed is planted at the base of yam mounds in June or July. Germination is hypogeal and takes about 5 days after planting. It begins flowering at about 80-120 days and set fruits from late September and October. The large bright coloured (purple) flowers result in long linear pods that could house about 20 seeds and they vary in size, shape, colour, etc. Pods start maturing about 150 days after sowing. The tubers which develop more slowly than the flowers, normally take 5 to 8 months to swell to harvestable size. Most are dug up towards the end of the rainy season. AYB seeds can be brown, white, speckled etc, with a hilum having a dark brown border (Klu et al.,2001)(plate 2).....

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