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Diversity Study of Drumstick (Moringa Oleifera Lam.) Using Morphological Characters and Molecular Markers

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Diversity Study of Drumstick (Moringa Oleifera Lam.) Using Morphological Characters and Molecular Markers.

ABSTRACT  

Germplasm diversity analysis and characterization provide the foundation for a crop improvement program. This work has used morphological and molecular tools to study diversity and characterize Moringa oleifera. The study was undertaken at the Forestry Research Institute of Nigeria (FRIN), Jericho Ibadan arboretum from 2013 to 2016 with the aim of determining accessions for future genetic improvement programs. The experiment was laid out in a randomized complete block design with three replications.

The morphological data were subjected to Principal Component Analysis (PCA), Rank Summation Index (RSI) to determine their pattern of variation, identify major traits responsible for the variation, and suggest appropriate accessions for breeding. Significant (P < 0.05) variations were observed among the accessions for surviving ability, pod diameter, and a number of seed/pod and highly significant (P < 0.01) differences for days to flowering, days to fruiting, pod length, plant height, and secondary branches.

Phenotypic variances and phenotypic coefficients of variation were slightly higher than genotypic variances and genotypic coefficients of variation for all the traits. Positive significant (P<0.05) correlations were observed between survival ability and vegetative characters, as well as between survival ability and yield-related characters. Negative significant correlations were observed between early flowering and yield-related characters; likewise, between early flowering and vegetative characters. 

TABLE OF CONTENTS

TITLE PAGE
DECLARATION……………………………………………………………………………………………………i
CERTIFICATION ………………………………………………………………………………………………. iv
DEDICATION …………………………………………………………………………………………………….. v
ACKNOWLEDGEMENTS ………………………………………………………………………………….. vi
ABSTRACT…………………………………………………………………………………..v
TABLE OF CONTENTS……………………………………………………………………vii
LIST OF FIGURES………………………………………………………………………….x
LIST OF TABLES…………………………………………………………………………..xi
PLATES AND APPENDICES……………………………………………………………..xii
ABBREVIATIONS…………………………………………………………………………..xiii

1.0 INTRODUCTION………………………………………………………………………………………….. 1
Background…………………………………………………………………………………1
Justification for the Study……………………………………………………………………3
Objectives……………………………………………………………………………………5

CHAPTER TWO …………………………………………………………………………………………………. 6
2.0 LITERATURE REVIEW……………………………………………………………………………….. 6
2.1 Origin and Geographical Distribution……………………………………………….6
2.2 Botanical Description of M. oleifera Lam……………………………………………7
2.3 Uses of Moringa oleifera Lam………………………………………………………8
2.4 Tree Improvement………………………………………………………………….12
2.5 Genetic Improvements of M. oleifera Lam…………………………………………14
2.6 Importance of Provenance………………………………………………………….15
2.7 Diversity within Species……………………………………………………………16
2.8 Studying Genetic Diversity within Species…………………………………………17
2.8.1 Morphological characterization……………………………………………………………. 18
2.8.2 Molecular assessment of genetic diversity……………………………………………… 22

3.0 MATERIALS AND METHODS ……………………………………………………………………. 28
3.1 Materials Used for the Study and their Sources.……………………………………28
3.1.1 Nursery establishment………………………………………………………………………… 28
3.1.2 Field establishment ……………………………………………………………………………. 30
3.2 Morphological Variation Study…………………………………………………….31
3.2.1 Data collection………………………………………………………………………………….. 31
3.2.2 Data Analysis……………………………………………………………………………………. 32
3.2.3 Estimation of component of variance ……………………………………………………. 34
3.2.4 Heritability estimates …………………………………………………………………………. 35
3.2.5 Correlations ……………………………………………………………………………………… 35
3.2.6 Rank summation index……………………………………………………………………….. 36
3.2.7 Principal component analysis………………………………………………………………. 37
3.2.8 Cluster analysis…………………………………………………………………………………. 38
3.2.9 Euclidean similarity distance……………………………………………………………….. 38
3.3 Materials and Methods for Molecular Markers Analysis…………………………..38
3.3.1 Plant materials ………………………………………………………………………………….. 39
3.3.2 DNA extraction…………………………………………………………………………………. 39
3.3.3 DNA quantification……………………………………………………………………………. 40
3.3.4 Genotyping using SSR Markers …………………………………………………………… 40
3.3.5 Gel scoring ………………………………………………………………………………………. 43
4.0 RESULTS……………………………………………………………………………………………………. 44
4.1 Morphological Analysis Results……………………………………………………44
4.1.1 Mean performance …………………………………………………………………………….. 44
4.1.2 Analysis of variance…………………………………………………………………………… 49
4.1.3 Variance components and heritability……………………………………………………. 49
4.1.4 Estimates of the correlation coefficients………………………………………………… 53
4.1.5 Rank summation index……………………………………………………………………….. 53
4.1.6 Principal component analysis………………………………………………………………. 56
4.2 Molecular Characterization Results…………………………………………………71
4.2.1 Primers characteristics ……………………………………………………………………….. 71
4.2.2 Cluster analysis from the SSR markers………………………………………………….. 74

5.0 CHAPTER FIVE …………………………………………………………………………………………. 77
5.1 DISCUSSIONS…………………………………………………………………….77

6.0 CHAPTER SIX ……………………………………………………………………………………………. 84
6.1 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS………………….84
6.1.1 Summary …………………………………………………………………………………………. 84
6.1.2 Conclusions ……………………………………………………………………………………… 85
6.1.3 Recommendations……………………………………………………………………………… 87

REFERENCES…………………………………………………………………………………………………… 88

INTRODUCTION  

Moringa oleifera Lam commonly known as a drumstick is the most widely cultivated species of the monogenetic family, Moringaceae (Fuglie, 2013). Drumstick is a fast-growing deciduous perennial tree, the bark is whitish-grey in color and it is surrounded by a thick cork (Parotta, 1993). It is native to India in Asia and was introduced in the 1920s to the lower part of China, southeast to the Philippines, westward to Africa and America (Pandey et al., 2010).

A total of 13 tropical and subtropical species of its genus are known out of which some species such as M. Arborea, M. borziana, M. longituba, M. rivae, M. ruspoliana, and M. stenopetala are endangered (Stephenson and Fahey, 2004). Moringa oleifera Lam. is the only cultivated species in the Moringa genus (Sanchez et al., 2006). The tree comprises four different edible parts: leaves, pod, stem and root (Morton, 1991).

It is also referred to as ‗Mother‘s best friend‘ as it is consumed to increase a woman‘s milk production, and has sometimes been prescribed for treating anemia (Siddhuraju and Becker, 2003; Estrella et al., 2000). It is referred to as the ‗Natural nutrition of the tropics. Different parts of the Moringa tree such as the leaves, fruits, flowers, and immature pods are highly nutritious and have been used as a source of food in many countries like India, Pakistan, the Philippines, Hawaii, and several countries in Africa (Anwar and Bhanger,2003).

M. oleifera leaves have been reported to be a rich source of β-carotene, protein, vitamin C, calcium, and potassium. In the Philippines, in addition to food uses, M. oleifera is used for animal feed (Sanchez et al., 2006). When supplemented in the diet of dairy animals, the leaves improve dry matter intake, digestion, and milk production without affecting 2 the smell, taste, or color of milk produced by the animal (Sanchez et al.,2006). 

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CSN Team.

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