MOLECULAR CHARACTERIZATION AND DNA BARCODING OF ARID-LAND OF FAMILY IN

By

OSHINGBOYE, ARAMIDE DOLAPO B.Sc. (Hons.) Microbiology (2008); M.Sc. Botany, UNILAG (2012) Matric No: 030807064

A thesis submitted in partial fulfilment of the requirements for the award of a Doctor of Philosophy (Ph.D.) degree in Botany to the School of Postgraduate Studies, University of Lagos, Lagos Nigeria

March, 2017

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SCHOOL OF POSTGRADUATE STUDIES UNIVERSITY OF LAGOS CERTIFICATION This is to certify that the thesis “Molecular Characterization and DNA Barcoding of Arid- Land Species of Family Fabaceae in Nigeria”

Submitted to the School of Postgraduate Studies, University of Lagos For the award of the degree of

DOCTOR OF PHILOSOPHY (Ph.D.) is a record of original research carried out By Oshingboye, Aramide Dolapo In the Department of Botany

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DEDICATION

This project is dedicated to the Almighty Father and to my loving and supportive parents Mr.

Adewole and L/Evangelist Abimbola Oshingboye.

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ACKNOWLEDGEMENTS

I would like to extend thanks to the many people, who so generously contributed to the work presented in this thesis. Firstly, I express my utmost gratitude to my enthusiastic supervisor and mentor Professor Oluwatoyin Temitayo Ogundipe FLS, for his time, patience, motivation, continuous support and mentorship in bringing out the best in me. It’s a rare privilege to be under his tutelage. His dedication and constructive criticism have developed great skills and confidence in me. His guidance helped me in all the time of research and writing of this thesis,

I could not have imagined having a better advisor and mentor. Thank you very much sir for giving me so many wonderful opportunities and the privilege of being one of your protégés. I wholeheartedly appreciate not only your scientific guidance, encouragement, inspiration, advice but also your material, financial and spiritual support. I attribute the level of my Master’s degree to your encouragement and effort and without you, this thesis too, would not have been completed or written. What more can I ask in this great man of God, I simply could not wish for a better supervisor. I pray the Almighty God will continue to bless you and take you to greater heights.

I would also like to appreciate Professor James Dele Olowokudejo, for his scholarly contribution to the completion of this research. I am grateful for your time in reading and supervising this work.

My special thanks go to Alastair Culham, for accepting to host me during my stint at the

University of Reading and providing a conducive working environment in the molecular laboratoty of the Centre of Plat diversity, University of Reading. I sincerely appreciate your support and scholarly contributions and also in finding time out of your tight schedule to supervise this work. Thank you, Alastair, for making the necessary contacts and recommendations to enable me secure another year extension with Jonathan Clark at the

University of Surrey.

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I am also hugely appreciative to Jonathan Clark, especially for sharing his taxonomic, morphometric and bioinformatics expertise so willingly at the Bioinformatics unit, Department of Computer science, University of Surrey.

My profound gratitude goes to Dr. (Mrs.) Temitope Onuminya for her time, foresight, guidance and contributions towards the success of this project. I am grateful for not only your time to read and correct this thesis but your thought-provoking comments and questions.

I must appreciate the immense effort of Dr. Bayo Ogunkanmi, Dr. Khalid Adekoya and Dr.

Akeem Kadiri, for also taking your time out to read, coach my mock presentations and offer corrections to this thesis. Thank you so much sirs for those pointer questions, deliberations and comments.

I am thankful for the support and constant encouragement given to me by Prof. Bola Oboh,

Prof. Olusoji Ilori, Prof. Oluwole Familoni, Prof. Mopelola Olusakin, Prof. Kehinde Olayinka,

Prof. A.A. Adekunle, Prof. (Mrs.) C.E. Umebese, Prof. (Mrs.) Oluyemisi Obashoro-John, Prof.

A.B. Adeloye, Dr. A. Akinsoji, Dr. V. J. Odjegba, Dr. (Mrs.) O.E. Ade-Ademilua, Dr. (Mrs.)

O.H. Adekanmbi, Dr. (Mrs.) O.O. Shonubi, Dr. (Mrs.). T.A. Adesalu, Dr. P.A. Adeonipekun,

Dr. (Mrs.) E.M. Adongbede, Dr. (Mrs.) Tope Samuel, Dr. C. Isanbor, Dr. (Mrs.) Ronke

Oyeyiola, Dr. (Mrs.) Temilola Oluseyi, Dr. Sunday Adebusoye, Dr. Andrew Akala, Dr.

Lukmon Adeoti, amongst others.

During my stint at the University of Reading, I was blessed with a friendly group of fellow students “the Culham Ph.D. group”. Thank you Azi for taking me through a fantastic lab training, Kalman for your patience, time and always proffering answers to my endless questions regarding phylogenetic analysis. I appreciate Ahmed senior and Widad for taking out time to put me through TAXONDNA and LUCID software. Jordan, thanks for the MrBayes class,

Maria for the statistical pointers, Oli, Anas, Amal, Ahmed junior, and Andrew. I say a big thank you to you all!

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Special mention goes to the “great family of God” at the RCCG City of Peace, Reading Parish.

Pastors Joseph and Solape Bamidele, for your continuous encouragement, prayers, devotion, love, and counsel; thank you sir and ma for believing in me and permitting me the opportunity to serve and be served. I have very fond memories of my time there, it was indeed a “home away from home” experience.

To my wonderful friends who sacrificed much to make this a reality I say a big thank you.

Many thanks to George Nodza for the motivation, support and providing logistics during my sample collection, Linus Ajikah, Seun Ajani, Bisola Akomolafe, Yinka, Chidi Nnamdi, Paul

Terewase, Dr. Andrew Iloh, Ani Emmanuel, Mosun Ojifo, Dr. Mosun Akinwunmi, Dr. Amii

Usese, Dr. Sandra Akagha, Dr. Nicolas Dibal, Femi Orotope, Dr. Gbenga Adeogun (Elder),

Ojisola Aina, Agboola Oludare, Kunrunmi Kunbi (Dr. K.K.), my Aunty (Dr. Mrs. Queen

Omoregie), Michelle Fasona, Dr. Debola Ba-wallah, Motunrayo Sholola, Gloria Anuroe, Kemi

Owolabi, Sadat Babalola, Salametu Saibu, Akpan, Femi Amusa, Joseph and Naissa Milkiah,

Samuel, Abena, Lydia, Samuel, Peace, Laura, Euphemia, Robert Oboch, Mariola, Hannah,

Eliah, Tosin, Patrick, Ngozi, Blessing of Reading, Izzy, Iz, Ojogu, Femi Adesina, Amen, Ben,

Kate, Temitayo dear, Teniola, and Yemisi. Would like to specially thank Promise for his motivation, devotion, emotional and financial support. I couldn’t forget to appreciate the inner caucus ladies Bolaji Folorunsho, Uju Nwadairo, Mr. and Mrs. Toluwalashe Ogunlade,

Opeyemi Omojola, thank you ladies for believing in me, for the love, prayers, faith, patience and understanding despite my tight schedules. I pray the Almighty God will perfect all that concerns you and yours.

I am grateful to the School of Postgraduate Studies, University of Lagos for the financial support given to me in form of Graduate Fellowship. The Competitive Agricultural Research

Grant Scheme (CARGS) project of DNA Barcoding of arid in Nigeria for birthing this research work (sampling and field work). The UNESCO-L’Oreal for Women in Science

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International Fellowship for funding the bench work and other logistics. It is a privilege to be one of the fellows of this prestigious award, I thank you for finding me worthy of this fellowship and even extending the sponsorship for one more year. Mallam Musa and Nnamadi of the Ahmadu Bello University Herbarium, Zaria, Azila of the Nigerian Forestry, Jos, and Mr.

Tola Oyebanji of the University of Lagos Herbarium for their support in identifying and collection of my samples.

I would like to thank every member of my family for their continual support, prayers, and understanding during the course of this research. To my uncle, Alhaji Giwa, my aunts, cousins, nephews and nieces, Mr. Olojede, Mr. and Mrs. Oloyede, Mr. and Mrs. Owolabi, Iya Arafat,

Ven. Asoore, Barrister Tosin Ogwezzy, Ven. Soyele, thank you all for your support. I am also grateful to my siblings, big sis, cake flair and my wonderful brother Adeniyi, thank you for the understanding, patience and goodwill during the course of the project. Special thanks to my parents Mr. and Mrs. Oshingboye for their understanding, financial, moral and spiritual support. I wholeheartedly appreciate you mum, my number one cheer leader for being the vessel and anchor God has used to achieve this. Thank you for the prayers, and believing in me; even when I lost faith, your faith propelled me to soar higher, I couldn’t have done this without you mum! You are the most important people in my world and I dedicate this thesis to you.

Finally, I return all the glory to the Alpha and Omega, the Almighty God who calls things which are not as though they were. I exalt my maker, the horn of my salvation for bringing me thus far and for granting me the strength, wisdom, favour, and grace to bring this to successful completion.

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TABLE OF CONTENTS

Caption Page

Title Page i

Certification ii

Dedication iii

Acknowledgements iv

Table of Contents viii

List of Tables xii

List of Figures xiii

List of Plates xvi

Appendices xx

Abstract xxi

Chapter One

1.0 Introduction 1

1.1 Background of Study 1

1.2 Economic Importance of Leguminosae 5

1.3 Statement of Problems 7

1.4 Aim and Objectives 8

1.5 Significance of Study 9

1.6 Research Questions 10

1.7 Operational definition of terms 11

1.7.1 Abbreviations and Acronyms 13

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Chapter Two

2.0 Literature Review 15

2.1 The Family Fabaceae 15

2.1.1 Ecology of the Leguminosae 17

2.1.2 Phytochemistry of the Leguminosae 18

2.1.3 Where and when did legumes originate? 20

2.1.4 Evolution of Fabaceae 23

2.1.5 Classification of Leguminosae 26

2.1.6. Phylogeny of Leguminosae 30

2.2 Legumes: Morphological Characterization and the identification of arid-land

species 37

2.2.1 Morphological Characters 38

2.2.2 Taxonomic Keys 38

2.3 Molecular Characterization 40

2.3.1 Molecular Characters 42

2.3.2 DNA Barcoding 44

2.3.2.1 Barcode data analysis 46

2.3.3 Phylogeny Reconstruction 49

2.3.3.1 Methods of Phylogeny Reconstruction 50

2.3.3.2 Assessing of the reliability of a 52

2.3.3.3 Gene Regions for Phylogeny Reconstruction 53

2.4. Conservation Assessments 54

2.4.1 IUCN Categories 55

2.4.2. IUCN Criteria 56

2.4.3 Components of a red list assessment 60

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Chapter Three

3.0 Materials and Methods 61

3.1 Sample Collection 61

3.2 Morphological Characterization 64

3.3 Molecular Characterization 66

3.3.1 Source of Plant material 66

3.3.2 DNA Isolation 66

3.3.3 Quantification of DNA samples 67

3.3.4 Gel Electrophoresis 67

3.3.5 Loading of samples and running of gel 67

3.3.6 Polymerase Chain Reaction (PCR) 68

3.3.7 Gel Electrophoresis of PCR product 70

3.3.8 Cleaning and quantification of PCR product 70

3.3.9 Sequence Contiging, Editing and Alignment 71

3.4 Data Analysis 71

3.4.1 DNA Barcoding analysis 71

3.4.2 Phylogenetic analysis 73

3.5 Conservation Assessment 74

Chapter Four

4.0 Results 75

4.1 Sample collection 75

4.2 Morphological Characterization 78

4.2.1 Systematic Descriptions 80

4.2.2 Keys 273

4.2.2.1 Bracketed keys to Sub-families 273

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4.2.2.2 Electronic multi-access key 279

4.2.3 UPGMA relationship based on morphological data 282

4.3 Molecular Characterization 286

4.3.1 DNA Isolation 286

4.3.2 DNA Amplification 287

4.3.3 DNA Barcoding Analysis 287

4.3.4 Phylogenetic Analysis 308

4.4 Conservation Assessment of species 315

Chapter Five

5.0 Discussion 321

Chapter Six

6.1 Summary of Findings 335

6.2 Contributions to Knowledge 337

7.0 References 338

Appendices 377

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LIST OF TABLES

Title Page

Table 1 Desertification frontline states of Nigeria 4

Table 2 Summary of Criteria (A) in the International Union for Conservation of

Nature (IUCN) criteria and categories 57

Table 3 Summary of Criteria (B) in the International Union for Conservation of

Nature (IUCN) criteria and categories 58

Table 4 Summary of Criteria (C) in the International Union for Conservation of

Nature (IUCN) criteria and categories 58

Table 5 Summary of Criteria (D) in the International Union for Conservation of

Nature (IUCN) criteria and categories 59

Table 6 Summary of Criteria (E) in the International Union for Conservation of

Nature (IUCN) criteria and categories 59

Table 7 Gene regions and primers screened 69

Table 8 Gene regions and primer sequences employed in the study 69

Table 9 Profiles of PCR reactions 69

Table 10 Analysis of inter-specific divergence between congeneric species and intra-

specific variation of candidate barcodes 288

Table 11 Wilcoxon signed rank tests for inter-specific divergence 288

Table 12 Wilcoxon signed rank tests for intra-specific divergence 288

Table 13 Identification success of the 10 barcodes based on the best match and best

close match program in TAXONDNA 295

Table 14 Conservation status of species 315

Table 15: Summary of Findings 335

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LIST OF FIGURES

Title Page

Figure 1.1 Map of Nigeria depicting desertification frontline states 3

Figure 2.1 Gondwanan biogeographic hypothesis scenario for legumes, showing a

phylogenetic tree superimposed on continental positions from 80 mya 22

Figure 2.2 Boreotropics biogeographic hypothesis scenario for legumes, showing a

phylogenetic tree superimposed on continental positions from the Eocene

50 mya 22

Figure 2.3 Relationships of legumes and allies (, eurosid I clade) to other major

clades of flowering plants 25

Figure 2.4 Phylogenetic relationships at the base of Leguminosae based on parsimony

analyses of trnL intron data 32

Figure 2.5 Schematic consensus phylogeny of Mimosoideae based on molecular data 34

Figure 2.6 Schematic phylogeny of Papilionoideae compiled as a super tree based upon

phylogenetic analyses 36

Figure 2.7 DNA double helix structure 44

Figure 2.8 DNA Barcoding pipeline 47

Figure 2.9 Phylogenetic workflow 50

Figure 2.10 The International Union for Conservation Nature (IUCN) Red List

Categories at the regional level (IUCN, 2014) 55

Figure 3.1 Map depicting different ecological zones and sampling location in Nigeria 63

Figure 3.2 Empty Lucid Builder v3.3 65

Figure 3.3 Spreadsheet being scored 65

Figure 4.1 Lucid player when no feature was selected 280

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Figure 4.2 Lucid player when elliptic was selected, only four entities were left in the

upper right panel 280

Figure 4.3 Lucid player when round base was further selected, E. africana was

identified in two steps 281

Figure 4.4 Lucid player when falcate leaf shape was selected, A. auriculiformis was

identified just in one step 281

Figure 4.5 Unweighted pair group method of arithmetic averages (UPGMA) dendogram

of Mimosoideae species based on morphological characters 282

Figure 4.6 Unweighted pair group method of arithmetic averages (UPGMA) dendogram

of species based on morphological characters 282

Figure 4.7 Unweighted pair group method of arithmetic averages (UPGMA) dendogram

of Papilionoideae species based on morphological characters 284

Figure 4.8 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for ITS region 290

Figure 4.9 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for ITS+matK region 290

Figure 4.10 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for matK region 291

Figure 4.11 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for matK+trnL-F region 291

Figure 4.12 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for matK+rbcL region 292

Figure 4.13 Distribution of inter-specific divergence between congeneric species and intra-

specific variation for rbcL region 292

Figure 4.14 Distribution of inter-specific divergence between congeneric species and

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intra-specific variation for rbcL+ITS region 293

Figure 4.15 Distribution of inter-specific divergence between congeneric species and

intra-specific variation for rbcL+trnL-F region 293

Figure 4.16 Distribution of inter-specific divergence between congeneric species and

intra-specific variation for trnL-F region 294

Figure 4.17 Distribution of inter-specific divergence between congeneric species and

intra-specific variation of COMBINED dataset 294

Figure 4.18 UPGMA analysis of combined regions sequence data 297

Figure 4.19 Bayesian analysis of combined regions sequence data 301

Figure 4.20 Neigbour joining analysis of combined regions sequence data 304

Figure 4.21 Maximum parsimony analysis based on ITS sequence data 309

Figure 4.22 Maximum Bayesian analysis based on ITS sequence data 310

Figure 4.23 Parsimony analysis based on combined chloroplast regions sequence data 311

Figure 4.24 Maximum Bayesian analysis based on combined chloroplast regions

sequence data 312

Figure 4.25 Parsimony analysis based on combined regions sequence data 313

Figure 4.26 Maximum Bayesian analysis combined regions sequence data 314

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LIST OF PLATES Title Page

Plate 1 (A-D) Photographs of some members of the Fabaceae 75

Plate 2 (E-J) Photographs of some members of the Fabaceae 76

Plate 3 (K-P) Photographs of some members of the Fabaceae 77

Plate 4 Acacia ataxacantha, arrow showing (a) claw shaped prickles and

pod with pointed tip (b) foliage and 82

Plate 5 Acacia auriculiformis reduced to falcate phyllode 84

Plate 6 Foliage and inflorescence of Faidherbia albida 86

Plate 7 Foliage of Acacia sieberiana, arrow showing long pointed thorns 88

Plate 8 Acacia pod, arrow showing paired spines 90

Plate 9 Acacia nilotica flowers 92

Plate 10 Foliage of Albizia lebbeck, arrow showing pod 95

Plate 11 Foliage of Albizia zygia 97

Plate 12 Pithecellobium dulce leaves, arrow showing spiral pods 99

Plate 13 africana leaves 101

Plate 14 Dichrostachys cinerea leaves, arrow showing tightly twisted pod 103

Plate 15 Entada abyssinica leaves 105

Plate 16 Leucaena leucocephala leaves, arrow showing globose inflorescence 108

Plate 17 Mimosa pigra leaves, arrow showing oblong clustered pod 110

Plate 18 Mimosa pudica leaves 112

Plate 19 biglobosa leaves 114

Plate 20 monandra leaves, arrowing showing flowers 117

Plate 21 Bauhinia purpurea leaves, arrow showing flowers 119

Plate 22 Bauhinia rufescens leaves, arrow showing narrow, twisted pods 121

Plate 23 Bauhinia tomentosa leaves 123

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Plate 24 Bauhinia vahlii leaves, arrow showing flowers 125

Plate 25 reticulatum leaves 127

Plate 26 leaves, arrow showing pod 129

Plate 27 africana leaves 131

Plate 28 megalophylla (a) showing leaves (b) leaves, arrow showing

corky pod 133

Plate 29 oliveri leaves 135

Plate 30 Detarium macrocarpum leaves and pods 137

Plate 31 Isoberlina tomentosa leaves and pod 139

Plate 32 Burkea africana leaves 143

Plate 33 Caesalpinia pulcherrima leaves, arrow showing flowers 145

Plate 34 Delonix regia leaves 147

Plate 35 arereh leaves 149

Plate 36 tora leaves, arrow showing pod 154

Plate 37 Senna alata leaves and flower 156

Plate 38 Senna hirsuta leaves, arrow showing pod 158

Plate 39 leaves, arrow showing pod 160

Plate 40 Senna occidentalis leaves, arrows showing flowers and pods 162

Plate 41 Senna siamea leaves 164

Plate 42 Senna singueana leaves, arrow showing pod 166

Plate 43 nigricans leaves 168

Plate 44 Chamaecrista mimosoides leaves 170

Plate 45 Chamaecrista rotundifolia leaves 172

Plate 46 Abrus precatorious leaves, arrow showing flowers 176

Plate 47 Adenodolichos paniculatus leaves 178

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Plate 48 Cajanus cajan leaves, arrow showing flowers 180

Plate 49 mucunoides leaves, arrow showing flowers 183

Plate 50 Vigna subterranea leaves 188

Plate 51 pruriens leaves, arrow showing pod 193

Plate 52 senegalensis leaves, arrow showing inflorescence 195

Plate 53 Erythrina sigmoidea leaves 197

Plate 54 glumaceus leaves 199

Plate 55 Alysicarpus rugosus leaves and flowers 201

Plate 56 Desmodium gangeticum leaves 205

Plate 57 Desmodium scorpiurus leaves 207

Plate 58 Desmodium tortuosum leaves (pinnate-trifoliate leaflets) 209

Plate 59 Desmodium velutinum leaves 211

Plate 60 comosa leaves 213

Plate 61 Crotalaria hyssopifolia leaves 215

Plate 62 Crotalaria lachnosema leaves 217

Plate 63 Crotalaria macrocalyx leaves 219

Plate 64 Crotalaria pallida leaves 221

Plate 65 Crotalaria naragutensis (a) leaves (b) arrow showing inflorescence 223

Plate 66 Crotalaria retusa leaves 225

Plate 67 (a) sisso leaves (b) arrow showing inflorescence 229

Plate 68 Pterocarpus erinaceus leaves 231

Plate 69 Stylosanthes erecta leaves 233

Plate 70 Gliricidia sepium leaves, arrow showing pod 238

Plate 71 arrecta leaves, arrow showing flowers 240

Plate 72 Indigofera hirsuta leaves, arrow showing inflorescence 242

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Plate 73 Indigofera nummulariifolia leaves 244

Plate 74 leaves 246

Plate 75 Indigofera suffruticosa leaves 248

Plate 76 Bobgunnia madagascariensis leaves 254

Plate 77 Tephrosia elegans leaves, arrow showing stipule 256

Plate 78 Tephrosia bracteolata leaves 258

Plate 79 Tephrosia linearis leaves 260

Plate 80 Tephrosia platycarpa leaves 262

Plate 81 sericeus leaves 265

Plate 82 Lonchocarpus cyanescens leaves 267

Plate 83(a-d) Electropherogram of extracted DNA 286

Plate 84(a-d) Electropherogram of PCR product 287

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APPENDICES Title Page

Appendix I List of species, voucher and Genbank numbers 377

Appendix II Morphological characters and states recorded 380

Appendix III Character descriptions scored for each species 386

Appendix IV Items List used in DELTA 395

Appendix V Tokey List used in DELTA 395

Appendix VI DNA Bank number and Spectrophotometric readings 396

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ABSTRACT

As a direct consequence of the appreciable threats confronting drylands, many arid-land species are lost with meagre information and research on their diversity, species composition, characteristics and endemism. It has become necessary to understand, describe and document species relationships and characterize the diversity of dryland species so as to help provide baseline information for efficient conservation strategies and sustain the livelihood of over one billion indigent inhabitants. A total of 324 leaf samples representing 118 species collected randomly across the arid and semi-arid regions of Nigeria were used for both morphological and molecular studies. Morphological features were recorded and used to develop keys for identification and to reveal species relationships. Molecular characterization involved isolation of total genomic DNA according to a modified CTAB protocol, amplification of four selected gene regions and sequencing. Raw sequenced data were contigned, edited and aligned, to be used for both DNA Barcoding and Phylogenetic analysis. Red-listing of species were performed according to IUCN categories and criteria. Samples comprised 22 Mimosoideae, 38 Caesalpinioideae and 58 Papilionoideae species; based on habit, members are either , , climbers or herbs, which are somewhat distinct in each sub-family. Morphological features recorded were mostly compound leaves (paripinnate, imparipinnate, pinnate, bipinnate, trifoliate and pinnate-trifoliate in some Papilionoideae species); flowers are zygomorphic in both Caesalpinioideae and Papilionoideae while actinomorphic in Mimosoideae. Fourty-seven characters and a hundred and eighty-four character states were recorded and used to develop a bracketed key and an electronic multi access key using Lucid 3.3 programme. Pair-wise analysis based on unweighted algorithm depicted various relationships and clustering of species taxonomically. For molecular studies, total genomic DNA isolated were predominantly high molecular weight DNA of 10,000bp and of good quality with A260/280 ratio within 1.75 – 2.01. The barcoding performance of 10 regions comprising three (3) cpDNA regions (rbcL, matK, trnL-F), one (1) nuclear region (ITS), six (6) combinations of regions (matK+rbcL, ITS+matK, ITS+rbcL, ITS+matK+rbcL+trnL-F, matK+trnL and rbcL+trnL-F) revealed the Internal Transcribed Spacer (ITS), exhibited significantly the highest inter-specific discrimination higher than matK and trnL-F while rbcL was the least. Wilcoxon signed rank tests further confirmed that ITS region exhibited the highest intra and inter-specific divergence between congeneric species, whereas rbcL exhibited the lowest. ITS region again exhibited the highest barcoding gap of 2.00, ITS+matK, ITS+rbcL, ITS+matK+rbcL+trnL-F, barcoding gap of 1.00 while other regions revealed no barcoding gap. This implies ITS region can either be used singly or in combination with either matK or rbcL as a potential plant barcode marker. Phylogenetic analysis based on three data matrices (ITS, rbcL+matK+trnL-F and ITS+rbcL+matK+trnL-F) support the monophyly of the family Fabaceae but depict two different topologies. Parsimony results based on ITS sequence data could not clearly resolve relationships among species; it inferred a monophyletic Fabaceae with Papilionoideae nested within a paraphyletic Caesalpinioideae while Mimosoideae is polyphyletic. The parsimony analysis of combined chloroplast regions and all combined regions produced a monophyletic family tree with a monophyletic Papilionoideae and Mimosoideae nested within a paraphyletic Caesalpinioideae; but members of the tribe Detarieae were the basal-most clade instead of Cercidieae. However, Bayesian analysis presented members of Cercidieae as the basal- most clade. IUCN red-listing of species revealed one, two, three and six species in the “Critically Endangered”, “Endangered”, “Vulnerable” and “Least Concerned” categories respectively. This study has explored and characterized the diversity of Nigerian arid land legumes while contributing to the ongoing project of re-constructing the phylogeny of Fabaceae and towards the quest of achieving the United Nation Strategic goals for Biodiversity 2011 - 2020.

Key words: Arid-land, Biodiversity conservation, DNA barcoding, Fabaceae, Phylogeny

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CHAPTER ONE INTRODUCTION

1.1 Background to study

The problem of land degradation, desert encroachment and biodiversity loss in arid and semi- arid regions worldwide have been of major concern amongst the international community, this has prompted the urgent need to protect desert regions and mitigate desert encroachments. Arid and semi-arid lands include the Tropical grassland and Savanna/woodland Savanna, the warm desert and semi-desert, temperate grasslands, tundra communities, and cold deserts biomes

(Udvardy, 1975). According to the FAO 1997 statistics, these regions occupy more than one third of the earth’s land area (out of which 32% is in Africa) and approximately one billion mostly poor people depend on these regions for their survival and livelihood. From these regions are many important food crops that are known for their great resistance to disease and stress, adaptability, endemism and valuable sources for plant breeding (Hassan, 2003). In addition, these florae are notable for their high within-species genetic diversity, rather than between-species variation or species richness; yet they contain a significant endowment of plants and animal species, including micro-organisms. They provide critical habitats for wildlife and ecosystem diversity, including wetlands for migratory species where they play a critical role in maintaining the integrity and sustainability of these regions. Unfortunately, these regions are under severe threat; the effect of the complex interplay of unsustainable land-use practices, harsh weather variability coupled with the effect of global climate change has put the flora of these regions in jeopardy causing inevitable loss of genetic diversity and reduction in the distribution of certain species in the wild. Untold number of arid and semi-arid zone species has become extinct in the last 100 years with limited research on their biological diversity, species composition, characteristics and endemism (Hassan, 2003). Indeed, biodiversity in the arid zone remains a largely unknown domain. According to Hassan, (2003), much of previous

1 | P a g e studies on these regions have been in the developed countries such as Australia, the central

Asian countries of the former Soviet Union and the United States with meagre contributions from only two developing countries: Kenya and Namibia. In this 21st century of biodiversity loss crisis and great difficulties in maintaining dry-lands, biodiversity studies on arid flora is therefore a need of the hour.

Sustaining and maintaining biodiversity in the arid and semi-arid region has been a complex intertwine battle: a fight against desertification; loss of plants and animals from the arid regions increases the vulnerability of the zones to drought and desertification. Hence, the knowledge on the biodiversity of arid and semi-arid regions including their genetic characteristics is an important tool in efforts to conserve biodiversity and combat drought and desertification.

Nigeria is one of the developing African countries located between latitudes 4’ 16’N and 13’

52’N; and between longitudes 2’ 49’E and 14’ 37’E, blessed with variable climatic conditions and physical features which have endowed her with a very rich biodiversity. By virtue of its geographical extent, it spans different climatic and ecological zones with a substantial part of its area extending into the Sudano-Sahelian belt, which, together with the neighbouring northern Guinea Savanna constitutes the dry lands of the country. Nigeria is one of the countries south of the Sahara, faced with advancing desert encroachment and geometric increase in biodiversity loss, with notable effects on the northern part of the country (Njidda and Haruna,

2010). Desertification phenomenon has been reported in the northern Nigeria since 1920s, but the impact has been more glaring since the famine of 1971 to 1973 in these parts of the country.

Currently, desertification affects fifteen northernmost states of the country (Jaiyeoba, 2002).

Although, the extent and severity of desertification in Nigeria has not been fully established neither the rate of progression has been properly documented. Nevertheless, some reports suggested the progression rate of desertification in Nigeria is about 0.6 km per year. Likewise,

Nwafor, (2006) reported that about 351,000 km2 of the country is already lost to desertification,

2 | P a g e accounting for almost one-fifth of the total Nigeria land area. In the same vein, the Food and

Agriculture Organization (FAO), World Meteorological Organization (WMO), and UNESCO reported that about 15% of Nigeria land is prone to desertification (Emodi, 2013). The visible sign of this phenomenon has been the gradual shift in vegetation from grasses, bushes and occasional trees, to grass and bushes; and in the final stage, expansive areas of desert-like sand.

It has been estimated that between 50 and 75% of the land mass of Bauchi, Borno, Gombe,

Jigawa, Kano, Katsina, Kebbi, Sokoto, Yobe and Zamfara States are being affected by varying degrees of desertification (Folaji, 2007). As illustrated in Figure 1.1 and Table 1, there are 15 desertification frontline States in Nigeria out of the total 36 States and the Federal Capital

Territory. These States accounts for about 63.83% of the total land area of Nigeria and about

62 million of Nigerians are either directly or indirectly affected by desertification problems

(Olagunju, 2015b).

Fig. 1.1: Map of Nigeria depicting desertification frontline states Source: (Olagunju, 2015b)

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Table 1: Desertification frontline states of Nigeria

State Geographical Land Population (2006) Rate of region Area % of Number Density Desertification (km2) Nigeria (km2) Sokoto North West 27,825 3.06 3,702,676 133 Severe Zamfara North West 37,931 4.17 3,278,873 86 Severe Kastina North West 23,287 2.59 5,801,584 246 Severe Jigawa North West 23,287 2.56 4361,002 7 Severe Kano North West 20,280 2.23 9,401,286 464 Moderate Kebbi North West 36,985 4.06 3,256,541 88 Severe Kaduna North West 42,481 4.67 6,113,503 144 Moderate Borno North East 72,609 7.98 4,171,104 57 Severe Yobe North East 46,609 5.12 2,321,339 50 Severe Bauchi North East 41,119 4.52 4,653,066 113 Moderate Gombe North East 17,100 1.88 2,365,040 138 Moderate Adamaw North East 38,700 4.26 3,178,950 82 Moderate a Taraba North East 56,282 6.19 2,294,800 41 Moderate Niger North Central 68,925 7.58 3,954,772 57 Moderate Plateau North Central 27,147 2.98 3,206,531 118 Moderate Total 580,841 63.83 62,061,067 107

Source: (Olagunju, 2015b)

However, several approaches and different policies such as Arid Zone Afforestation Project

(AZAP) in 1977, River Basin Development Authorities (RBDA) in 1987, Federal and State

Environmental Protection Agency (FEPA / SEPA), Great Green Wall Project, membership of the Desertification Convention in 1997 and tree planting project in 2005 amongst others have been put in place to reverse and combat desert encroachment in these regions (Medugu, 2009).

Yet the problem is rather aggravating because in-situ conservation in this area proves ineffective as a result of the notoriety of the herdsmen and various unsustainable activities 4 | P a g e apace. In view of the aforesaid, different quarters have called for new approaches and recommendations on how to tackle this menace, such as, the call for a paradigm shift in mitigation approaches by (Omijeh, 2008, Murtala, 2012, Nneji, 2013, Olagunju, 2015a amongst others). Hence, to prevent further degradation of arid and semi-arid lands and to sustain deserts biodiversity it is imperative to maintain the biological resources of this region.

It is necessary to understand, describe, document, and characterize the diversity of dry land species so as to help provide baseline information for effective and efficient conservation strategies. To achieve this, plant species of the bean family (Fabaceae), dominant in the arid zone of Nigeria, are used as exemplar species. This plant family includes some of the most important sources of vegetable protein and dietary fibre, fodder and fuel plants as well as green manures valuable in improving soil structure and fertility; they provide such an excellent scientific opportunity for devising mitigation strategies (Wojciechowski, 2003).

1.2 Economic Importance of Leguminosae

Of all the plants used by man, only the grasses are more important than legumes, few if any other plant families have as many species that are so economically important worldwide as the

Fabaceae; they are the most important family domesticated in the entire Dicotyledonae (Duke,

1982; Harborne, 1994; Pickersgill, 1996). Today, dry legume are used for food throughout the world. These are referred to as pulse, dry beans, legume seeds or legume grains.

Legume grains are nutritionally two or three times richer in protein than cereal grains (Singh and Jambunathan, 1991). Some members produce seeds such as soybeans, groundnuts and winged beans which are also sources of oil (Graham and Vance, 2003). They offer a variety of edible products in addition to seeds, many immature green pods are edible during two or three weeks before the fiber lignified and harden. At this stage, they are still green and succulent and can be used as a green vegetable (Singh and Singh, 1992). Although they have less protein than

5 | P a g e the mature , they are rich in vitamins and soluble carbohydrate (Grusak, 2002a). In addition, leaves of leguminous plants are eaten in some parts of the world, particularly in the tropics. Pterocarpus species are grown in Southeastern Nigeria mainly for their leaves (Arora et al., 1991). Other legume products such as pods from Senna angustifolia and related species are widely used for their laxative properties (Grusak, 2002b; Madar and Stark, 2002).

Rotenone, an insecticide and fish poison, is extracted from species of Derris, Lonchocarpus and Tephros (Dixon et al., 2002; Ndakidemi and Dakora, 2003). Many important processed foods like Mayonnaise and ice-cream contain gums from legumes such as Acacia senegal (gum

Arabic) or Cyamopsis tetragonoloba (guar). Legumes also provide huge source of income through exports, with an estimate of about two billion US dollars per annum in US alone

(Gowda et al., 2009). A key contribution of legumes to sustainable agriculture and the nitrogen cycle is their ability to fix atmospheric nitrogen, which saves an estimated US$10 billion annually in commercial fertilizer (Graham and Vance, 2003). Tropical legume trees play particularly important role as forage in arid regions (National Academy of Sciences, 1979). As abundant timber in humid areas (Doyle and Luckow, 2003), they play a secondary role in the ecological stability of the Sahelian arid zone especially in regard to its contribution to soil conservation, land degradation and desertification systems, afforestation, agroforestry species for soil conservation and regeneration of degraded lands (Beshai, 1984; Wojciechowski, 2003;

Githae et al., 2011; Traoré et al., 2012).

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1.3 Statement of problems

Quite a number of species are lost in the arid and semi-arid regions in the past few years with many species disappearing without being known and adequate conservation efforts provided.

World Bank (1992) projected that desertification and biodiversity loss would continue to be on the increase if nations fail to set up remedial actions to repair current damage hence the urgent need of the hour is to conserve the remnant resources of dry lands plants. This is crucial to worldwide efforts to combat desertification, to prevent further degradation of the fragile ecosystems and to sustain biodiversity in deserts,

Furthermore, the lack of adequate knowledge and basic information about the current conservation status of most habitats and species and key data usually frustrate management efforts or attempts put in place by policy makers. It is therefore expedient to understand, describe and document the genetic variation within and between populations as this will help to provide baseline information for effective and efficient conservation practices.

The exemplar plant species have been well studied but there are still controversies about the taxonomic position of some species within the family; whether the three sub-families should be elevated to the family rank as well as the placement of larger group clades in different genera and subfamilies. Hence, a phylogenetic analysis would be useful to elucidate on relationships among species.

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1.4 Aim and Objectives

This research aims at exploring and characterizing the diversity of arid legumes using morphological and molecular approaches.

The specific objectives are to:

1. give a systematic account of the diversity of family Fabaceae with emphasis on

collection and identification with a view to improving local knowledge on the

identification of species based on morphological characters;

2. explore the performance of three (3) cpDNA regions (rbcL, matK and trnL-F) and one

(1) nuclear region ITS to accurately identify species;

3. examine phylogenetic relationships among taxa, find out if they are consistent with

earlier classifications; and

4. red-listing of species according to IUCN categories and criteria.

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1.5 Significance of study

Dry land biodiversity is of tremendous global importance, it is central to the well-being and development of millions of people in developing countries. The loss of plants and animals in these regions limit development options and increases the vulnerability of the zones to drought and desertification. Therefore, this study will:

1. Provide information or detailed checklist of the legumes species present in the arid

region of Nigeria and their distribution to provide baseline information for effective

and efficient conservation practices.

2. Produce taxonomic keys for identification of arid land legumes based on exo-

morphological characters. The traditional bracketed key useful for taxonomists and an

electronic multi-access key useful for non-taxonomists including conservationists,

resource managers, herbalists, foresters, pharmacy students and teachers interested in

investigating plant properties.

3. Contribute information on key economic plant species to the consortium at Barcode of

Life (CBOL).

4. Elucidate species phylogenetic relationships among the Fabaceae, valuable information

in conservation assessments.

5. Fill the information gap on current conservation status of arid legumes.

6. Contribute to the Sustainable Development Goals and the United Nations Strategic

Goals for Biodiversity 2020.

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1.6 Research Questions

Results of this research aim to answer the following questions:

• How many genera/ species of legumes are represented in the arid and semi-arid region

of Nigeria?

• Can vegetative characters be built in an interactive multi-access key to aid the ease of

identification of arid legumes?

• Which gene region(s) is most effective in accurate identification of legumes?

• What relationship exists between species, are they monophyletic or polyphyletic?

• What is the conservation status of legumes in the semi-arid region of Nigeria?

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1.7 Operational definition of terms

Bootstrapping: a statistical method for estimating robustness of an estimator by sampling with replacement from the original sample. It allows assigning measures of accuracy defined in terms of bias, variance, confidence intervals, prediction error or some other such measure.

Chloroplast Deoxyribonucleic acid (cpDNA): is maternally inherited plastome containing genes that are involved in photosynthesis and protein-synthesis apparatus within the organelle.

Clade: a group of organisms consisting of a common ancestor and all its lineage descendants.

Consortium for the Barcode of Life (CBOL): an international initiative devoted to developing DNA barcoding as a global standard for the identification of biological species.

DELTA: DEscription Language for , a flexible method for encoding taxonomic descriptions for computer processing.

DNA Barcoding: a taxonomic method that uses a short genetic marker in an organism's DNA to identify it as belonging to a particular species.

EXON: parts of DNA that are converted into mature messenger RNA (mRNA).

Family: one of the 8 major taxonomic rank, consisting of one or more genera closely related to each other.

Genus: a collective taxonomic unit containing a number of similar related species.

Heuristic search: a computational method that optimizes a problem by iteratively trying to improve a candidate solution with regard to a given measure of quality.

Intergenic spacer: a region of non-coding DNA between genes.

INTRON: DNA region within a gene that is not translated into protein

ITS: refers to the spacer DNA situated between the small-subunit ribosomal RNA (rRNA) and large-subunit rRNA genes in the chromosome or the corresponding transcribed region in the polycistronic rRNA precursor transcript.

IUCN: is the world’s main authority on the conservation status of species.

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LUCID: software specifically designed for identification and diagnostic purposes. matK: The maturase K gene, formerly known as orfK located within the intron of the chloroplast gene trnK on the large single-copy section adjacent to the inverted repeat. The gene region is approximately 1500 base pairs bp.

Maximum Parsimony: an optimality criterion under which the phylogenetic tree, it usually minimizes the total number of character-state changes is to be preferred.

Monophyl: refers to a taxon that includes the most recent common ancestor and all of its descendants.

Paraphyl: refers to a taxon that includes the most recent common ancestor but not all of its descendants.

Parsimony: the parsimony principle chooses the simplest scientific explanation that fits the evidence i.e. the best hypothesis is the one that requires the fewest evolutionary changes.

Polyphyl: refers to a taxon composed of unrelated organisms that descended from more than one ancestor. rbcL: a single copy gene responsible to code the larger subunit of ribulose 1, 5 bisphosphate carboxylase/oxygenase RUBISCO or RuBPCase; its approximately 1430 bp.

SEVAG: 24 chloroform: 1 isoamyl-alcohol

TAXONDNA: is a collection of tools for manipulating datasets with multiple species.

Tribe: is a taxonomic rank above , but below family and subfamily. trnL-F: this gene region consists of trnL intron and the trnL-F intergenic spacer; region is located in the large single-copy region of the chloroplast genome. The intron has a conserved secondary structure and contains elements that are homologous across land plants while the spacer is highly variable in length and composition.

UPGMA: a simple agglomerative hierarchical clustering method attributed to Sokal and

Michener.

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1.7.1 Abbreviations and Acronyms

%: Percentage

µl: Microlitres

BI: Bayesian Inference

BP: Bootstrap percentage

BSA: Bovine Serum Albumin

CBD: Convention on Biological Diversity

CITES: Convention on International Trade in Endangered Species of wild fauna and flora

Cont’d: Continued

CTAB: Cetyl-Trimethyl-Ammonium-Bromide

DMSO: Dimethyl Sulfoxide

DNA: Deoxyribonucleic acid; the hereditary material in all living organisms g: Grammes hr: hours

ITS: Internal Transcribed Spacer

IUCN: International Union for Conservation of Nature kb: Kilobytes

L: Litres

M: Molar mins: Minutes ml: Millilitres mRNA: Messenger RNA

MSA: Multiple Species Alignment

Mya: Million years ago ng: Nanogram

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ºC: degree celcius

PAUP: Phylogenetic Analysis Using Parsimony

PCR: Polymerase Chain Reaction; an enzyme-catalyzed, in vitro copying process of specific DNA sequences that uses extremely small amounts of template DNA. rpm: rate per minutes

RNA: Ribonucleic Acid

PVP: Polyvinlpyrrolidone s.l.: Sensu lato (in the widest sense) s.s.: Sensu stricto (in the strict sense) s: seconds

SSC: Species Survival Commision

TAE: Trisbase Acetic acid EDTA

UPGMA: Unweighted Pair Group Method with Arithmetic mean

V: Volts

LUH: University of Lagos Herbarium, Lagos

RNG: University of Reading Herbarium, Reading

FHI: Forestry Herbarium Ibadan

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CHAPTER TWO LITERATURE REVIEW

2.1 The Family Fabaceae

The family Fabaceae is the third largest family of flowering plants after Orchidaceae and

Asteraceae with approximately 750 genera and 18,000 species (Polhill and Raven, 1981;

Humpreys, 1996). However, recent molecular classification recognized 36 tribes, 727 genera and 19,327 species (Lewis et al., 2005). Traditionally, the family is grouped into three sub- families (Caesalpinioideae, Mimosoideae and Papilionoideae) based on floral features. Most members of the sub-families Mimosoideae and Papilionoideae possess the nitrogen-fixing bacterial nodules (Rhizobium spp.), while only one quarter of the sub-family Caesalpinioideae nodulate (Allen, 1981; Giller and Wilson, 1991). Members of this family are cosmopolitan in distribution; they vary in habit from annual and perennial herbs to shrubs, trees, / and a few aquatics (the only known aquatic species is Neptunia oleracea) but there are no truly marine species. They also vary in size from some of the smallest plants of deserts and arctic/alpine regions to the tallest of rain forest trees (Lewis et al., 2005). They are conspicuous and often dominant in most of the vegetation types distributed throughout temperate and

Tropical regions of the world but noticeably poorly represented in mesic temperate habitats and among plant epiphytic communities (Rundel, 1989). They are particularly diverse in

Tropical forests and temperate lands with a great affinity for dry or arid climate. This great love and attraction for semi-arid to arid environments is not far-fetched from a high nitrogen-demanding metabolism, which is an adaptation to climatically variable or unpredictable habitats that allows an economical and opportunistic growth (McKey, 1994). The hallmark of legume biology, the fixation of atmospheric nitrogen via root-nodulating rhizobial bacteria, is just one of the several ways (in addition to arbuscular ,

15 | P a g e ectomycorrhizas, and uptake of inorganic nitrogen compounds) in which legumes obtain high levels of nitrogen to meet the demands of their metabolism (Sprent, 2001). The legumes constitute the largest number of plants in the arid and semi-arid zone where they play important role in the terrestrial nitrogen cycle regardless of whether they form root nodules or not (Sprent,

2001) by preventing soil erosion and maintaining its fertility (McKey, 1994; Sprent, 2001;

Freitas et al., 2010).

Leaves of most members of this family are predominantly alternate, pinnately compound as in

Caesaplinia, Delonix, Milletia and Peltohorum or bipinnate compound as in Acacia,

Dichrostachys. They could as well be simple as in Baphia or in some cases simple by suppression of leaflets reduced to one phyllode as in Acacia auriculiformis or reduced to a tendril as in . In some members, stipules are spirally arranged or sometimes large and leaf-like as in Pisum or developed into spines as in Prosopis, Robinia; some are unifoliate e.g.

Indigofera nummulariifolia, trifoliate as in Medicago, Trifolium but are rarely palmately compound. In some species, the leaves are able to close together at night (nyctinasty) as in some species of Mimosa that closes up whenever touched (Polhill and Raven, 1981).

Flowers are bisexual, usually irregular (zygomorphic) but regular (actinomorphic) in the

Mimosoideae, pentamerous, arranged singly or in , spikes, or heads. Perianth are biseriate, calyx gamosepalous and 5 lobed. Corolla are typically of five (rarely absent or reduced to single as in Swartzia, distinct or the two anterior ones basally connate, hypanthium sometimes present e.g. Arachis. Stamens are mostly 10 (sometimes numerous in

Mimosoideae) distinct or monoadelphous as in Sophora or diadelphous as in Zornia. Anthers are 2 –celled, dehiscing by longitudinal slits or infrequently by pores, sometimes with apical deciduous glands; pistil one with a superior ovary (hypogynous to perigynous), one locule, one carpel, placentation marginal along ventral sutures, ovules two to many in two alternating rows

16 | P a g e on a single placenta, ovule amphitropous, anatropous, or infrequently campylotropous, pendulous or ascending (Adelanwa, 2008).

The principal unifying feature of the family is the , the legume (Polhill, 1994). With a few exceptions, legumes are typically one-chambered pods (one locule), with parietal placentation along the adaxial suture, ovules 2 to many, in two alternating rows on a single placenta, typically dry and dehiscent along one or both sutures (legume), occasionally constricted into

1-seeded sections (loments) or indehiscent (samara, drupe, achene) (Watson and Dallwitz,

2007).

2.1.1 ECOLOGY OF THE LEGUMINOSAE

A remarkable feature of the Fabaceae is the high nitrogen metabolism that occurs in all members of this family (Allen and Allen, 1981; Giller and Wilson, 1991; Sprent and McKey,

1994; Sprent, 2009). This effect has made them evolve several mechanisms of efficiently scavenging organic and inorganic nitrogen from the soil. One of these is the formation of root nodules upon infection by rhizobia (McKey, 1994). Rhizobia is the collective name for bacteria that infect legume roots (rarely stem) and fixes atmospheric nitrogen, which then becomes available to the legume plant. This association occurs within a symbiotic relationship context, which benefits both the legume and rhizobium. Thus, many legume species play an important role in community dynamics where they are available for use in reforestation or re-vegetation projects worldwide (McKey, 1994). Their efficient ability to acquire nitrogen, phosphorus, and other essential nutrients, whether through nodulation or association with mycorrhizae

(symbiotic root fungi), allows them to be easily established on abused lands where soils are eroded, leached, or acidic (Sprent, 2001). They are mostly found to be pioneer species or as major constituents in secondary vegetation. This is a desired property because by killing the legume (naturally or intentionally), nitrogen is released for use by non-leguminous associates

(McKey, 1994; Sprent, 2001; Freitas et al., 2010).

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2.1.2 PHYTOCHEMISTRY OF THE LEGUMINOSAE

A great diversity of phytochemical compounds occurs among the Fabaceae; members alone produce about 28% of all known flavonoids and 95% of all isoflavonoid and aglycones

(Hegnauer and Grayer-Burkmeijer, 1993), with anthocyanin pigments widely distributed in their floral tissues. According to Hegnauer and Grayer-Burkmeijer, (1993) past studies have revealed presence of many flavones and flavonol glycosides obtained from both floral and leaf tissues e.g. Robinin obtained from the leaf of Robinia pseudoacacia. Flavonoids like chalcones and flavonones have also been chiefly found in woody members of the family where they are present in leaf, bark and heartwood although are absent from herbaceous legumes (Ingham,

1981). However, mainly among the Papilionoideae are the Isoflavonoids that is like a phytochemical signature of members of the subfamily (Dewick, 2002). In addition, within the

Leguminosae are some 850 compounds including 362 isoflavones (Dewick, 2002) which are usually obtained from root, wood, bark or seed rather than leaf or flower; a typical example is rotenoid, useful insecticidal or pesticidal agent (Harborne, 1993). Some of these phytochemical compounds (terpenoids, benzoquinones, neoflavonoids) are characteristically present in wood resins, heartwood of some species e.g. dalbergione is present in the heartwood of Dalbergia species (Harborne, 1993).

A variety of legume plants like Laburnum, Lupin are known to be poisonous to man or to farm animals, this has been traced to the presence of a nitrogen-containing secondary metabolites which may be cyanogenic glycosides, nitro compounds, non-protein amino acids or alkaloids

(Wolff, 1971; Grayer and Harborne, 1994). The most widely occurring cyanogens in legumes are linarin and lotaustralin recorded in species of Acacia, Lotus, Ornithopus, Phaseolus and

Trifolium. In addition, non-protein amino acids like cavanine have been isolated from

Canavalia ensiformis, albizzine from Albizzia, lathyrine from Lathyrus and mimosine from

Mimosa (Boutler and Parthier, 1982). Harmful effects of these amino acids include

18 | P a g e neurotoxicity in mammals, including man (Bell et al., 1978), poisonous and anti-feedant properties in insects (Rosenthal, 1982). Based on the research of (Olson, 1978), some seleno- cystathionine known to the family (Astragalus and Neptunia) has been the cause of livestock poisoning in North America. Alkaloids like quinolizidine or lupine have as well been recorded

(Southon and Buckingham, 1989; Kite et al., 2009), known to be responsible for both poisoning and teratogenic effects in cattle and sheep feeding on wild Lupin (Mears and Mabry, 1971;

Kinghorn and Smoleenski, 1981). Conversely, some of these compounds are of immense health benefits. Examples are anthraquinones and anthrones known for its purgative properties present in the pods, leaves and roots of Cassia, Chamaecrista and Senna spp. (Thaomson, 1971). Also, the non-protein amino acid L-dopa (L-3,4-dihydroxyphenylaline) from Mucuna pruriens has potential uses in the treatment of Parkinson's disease and has been tested against the malaria vector Anopheles gambiae. Polydroxyalkaloids have been isolated from Castanospermum and investigated for their use in the treatment of cancer, HIV-related conditions and diabetes

(Stevenson et al., 2010). As part of a review of 1,700 plant species used traditionally to treat diabetes, over 80 species from 47 genera are of the Leguminosae and identified to have anti- diabetic properties (Simmonds and Howes, 2006).

The importance of the phenolic constituents in the family has been extensively discussed as far back as 1962 in Bate-smith, (1962) where he noted that the Leguminosae alone among the families of the dicotyledons contain representatives of all classes of subsidiary phenolic compounds. The phytochemical richness of the family depicts a well-developed chemotaxonomy of these plants, an example is the leaf flavonoid pattern in the genus Battisia that has proven to be useful in the identification of natural hybrids between species, in a situation where morphological and cytological techniques fail to give answer (Bate-smith,

1962). In addition, the phytoalexin induction in the family has also enriched the understanding of taxonomic affinities in herbaceous legumes (Harbone et al., 1984).

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Overall, many bioactive constituents are known in legumes. However, future research using bioassay and some newly adopted techniques will undoubtedly reveal many more interesting structures some of which may become the plant drug of the future.

2.1.3 WHERE AND WHEN DID LEGUMES ORIGINATE?

As diverse as the Fabaceae family is today, it is believed that all of its members trace their origin to a single species that lived a long time ago in some specific place and adapted to a particular environment. Based on legume biogeographic studies, the time and place of origin of legumes remains something of a mystery. Legumes have not been among the families represented in the rich mid-Cretaceous (approximately 90 mya), its fossil record is not particularly rich until about 35 to 54 mya (Eocene or mid-Tertiary), when papilionoid and mimosoid legumes become abundant and diverse in both North American and European fossil florae (Crept and Taylor, 1985; Taylor, 1990; Herendeen et al., 1992). According to

Wojciechowski, 2003, based on molecular estimates, the major lineages had all diverged from one another by around 50 mya e.g. Lotus and Medicago lineages may have diverged from one another by around 40 mya (Wojciechowski, 2003; Schrire et al., 2005).

Until recently, there has been a belief that legumes originate from Africa in the late Cretaceous from where they migrated to and subsequently North America, leaving behind

“archaic” genera in Africa (Polhill and Raven, 1981). These were called a “western

Gondwanan” family (Fig. 2.1), and often were cited as an example of the biotic connection that existed between Africa and South America during the Cretaceous (65–145 mya), when these continents were in close proximity. Although, advances in the understanding of continental drift and the availability of more precise phylogenies for legumes have not supported the

Gondwanan hypothesis, and most recent biogeographic studies have concentrated on Eocene

(35–55 mya) or later events to explain legume distributions. However, in another biogeographic model, during the Eocene, a land bridge in the North Atlantic joined Africa,

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Europe, and North America in one continuous land mass (Tiffney, 1985) whose climate was much warmer and wetter than it is today. The Pacific Northwest of North America harbored

Tropical rainforests, whereas warm-temperate forest covered Wyoming similar in composition as seen in Southeast Asia of date. This mixed assemblage of plants, containing both temperate and Tropical elements, was termed the boreoTropical flora (Wolfe, 1975). This provides an alternative general explanation for the distribution of legumes, hypothesizing that continental breakup and progressively cooler climates since the Eocene have led to the current distribution patterns (Fig. 2.2). Under this model, any archaic African taxa are interpreted as relicts of the once widespread boreoTropical forest that have managed to survive the increased aridity and cooler temperatures that led to their extinction in the northern hemisphere (Lavin et al., 2000).

This boreoTropical hypothesis is also compatible with the finding that the Cercidieae is one of the first legume clades to have differentiated: The boreoTropical flora contains both Cercis and

Bauhinia, and Cercis (redbud) has persisted in the modern temperate forests of Europe, North

America, and East Asia. However, phylogenetic evidence (e.g. Lavin and Luckow, 1993) and a reinterpretation of Eocene fossils (Taylor, 1990) also indicate that many putatively “archaic”

South American taxa are in fact recent offshoots from northern hemisphere radiations; in contrast to the Gondwanan hypothesis, the direction of migration of legumes in the Americans is north to south rather than south to north. Finally, the few phylogenetic studies that have used molecular approaches to date legume radiations have found them to have Tertiary rather than late Cretaceous ages (Herendeen et al., 1992; Wojciechowski, 2003; Lewis et al., 2005). There are unquestionably “archaic” lineages of legumes in South America as well as in Africa, but whether they are relictual boreoTropical taxa pushed south by an increasingly inhospitable climate, Gondwanan elements that managed to surmount a large water barrier, or more recent examples of long-distance dispersal will probably never be known. What has become clear is

21 | P a g e that such distributions are the exception rather than the rule, and events during and after the

Tertiary provides the best general historical explanation for legume distributions.

Fig. 2.1: Gondwanan biogeographic hypothesis scenario for legumes, showing a phylogenetic tree superimposed on continental positions from 80 mya (Doyle and Lucknow, 2003).

Fig. 2.2: Boreotropics biogeographic hypothesis scenario for legumes, showing a phylogenetic tree superimposed on continental positions from the Eocene 50 mya (Doyle and Lucknow, 2003).

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2.1.4 EVOLUTION OF FABACEAE

The order Fabales closely related to a group of Rosid also contain nitrogen-fixing plants:

Rosales, Cucurbitales, and Fagales (Fig.2.3). Members of these orders do fix nitrogen, but use root-dwelling actinomycetes, typically Frankia, rather than Rhizobium and relatives used in legumes (Watson and Dallwitz, 2007). The order Fabales contains around 7.3% of eudicot species with its greatest diversity is contained mainly within the Fabaceae (Angiosperm

Phylogeny Group, 2009). This clade of Fabales also includes the Polygalaceae,

Surianaceae and Quillajaceae families (Fig. 2.3) which originated 94 to 89 mya, although its diversification started some 79 to 74 million years ago (Steele et al., 2000; Stevens, 2006). In fact, the Leguminosae have diversified during the early tertiary to become a ubiquitous part of the modern earth’s biota, along with many other families belonging to the flowering plants

(Herendeen et al., 1992 and Lewis et al., 2005). The first definitive legumes appear during the

Late Paleocene (ca. 56 Mya; Persson, 2001; Herendeen, 2001; Herendeen and Wing, 2001;

Wing et al., 2004) with representatives of all the three traditionally recognized subfamilies, the

Caesalpinioids, mimosoids, and Papilionoids (Polhill et al., 1981; Crept and Taylor, 1986) appearing from fossils soon afterwards, beginning around 50 to 55 Mya (e.g., Herendeen et al.,

1992). Therefore, the Fabaceae started their diversification approximately 60 million years ago and the most important clades separated some 50 million years ago (Bruneau et al., 2008a).

The age of the main Caesalpinioideae clades have been estimated 56 ± 34 million years, and the basal group of the Mimosoideae 44 ± 2.6 million years (Lavin et al., 2005; Bruneau et al.,

2008a). The division between Mimosoideae and is dated as occurring between 59 and 34 million years ago and the basal group of the Faboideae between 58.6 ± 0.2 million years ago (Crept and Herendeen, 1992; Wikström et al., 2001). Attempts to estimate the age of legumes and diversification in the family based on molecular sequence data have been published in recent years. Wikström et al., (2001) used a non-molecular clock based analysis

23 | P a g e of the three-gene data set (plastid atpB & rbcL, and nuclear 18S rDNA genes; Soltis et al.,

2000) with a minimum age of 84 mya for the split between Fagales and Cucurbitales as an internal calibration point, and estimated an age for Fabaceae of 74-79 Ma. Lavin et al., (2005) have also presented a comprehensive analysis of rates of molecular evolution and estimated ages for crown groups within the legume family. In other studies, tertiary macrofossils that showed distinctive combinations of apomorphic characters or features were used to constrain the minimum age of 12 specific internal nodes to estimate ages of a number of the clades identified in recent family-wide phylogenetic analyses of plastid matK (Wojciechowski et al.,

2004; Savolainen et al., 2000) and rbcL (Kajita et al., 2001) gene sequence data. Their findings indicate the age of the legume crown clade differs by only 1.0 to 2.5 Ma from the age of the stem clade and the oldest caesalpinioid, mimosoid, and papilionoid crown clades show approximately the same age range of 40 to 59 Ma. These findings are consistent with a rapid diversification of the family soon after its origin during the Late Paleocene. Remarkably, three large clades that include papilionoids traditionally considered derived (Polhill et al., 1981;

Polhill, 1994), the “dalbergioids” (Lavin et al., 2001), “Hologalegina” (Wojciechowski et al.,

2000), and “mirbelioids” (Crisp et al., 2000), all have ages estimates in the 50 Mya period or older. Molecular evidence also confirms the hypothesis that Caesalpinioideae includes the earliest diverging lineages among the legumes. This was also the prevailing theory prior to molecular studies, based on the group’s high diversity in the tropics, an extended fossil record, and the wide variation of floral and vegetative structures beyond the specializations in the other two subfamilies. The unique Rhizobium nitrogen-fixation symbiosis is under-developed in

Caesalpinioideae than in the other groups, which seems to have originated in this subfamily

(McKey, 1994; Sprent, 2001). What is becoming clearer, however, is that Caesalpinioideae legumes are more diverse than how previously thought they were to be, also, the other two subfamilies, Mimosoideae and Papilionoideae, radiated from particular lineages among the

24 | P a g e diverse Caesalpinioideae legumes. This strengthens the idea that legumes form a single family; however, the phylogenetic relationships within the family are more complex than the former simple division into three subfamilies (Herendeen et al., 1992; Soltis et al., 1995). As further molecular analyses are undertaken, a clearer picture will develop.

Fig. 2.3: Relationships of legumes and allies (Fabales, eurosid I clade) to other major clades of flowering plants (Doyle and Lucknow, 2003).

25 | P a g e

2.1.5 CLASSIFICATION OF LEGUMINOSAE

Most taxonomic systems including the APGIII systems placed the Fabaceae in the order

Fabales. The first formal classification of Bentham and Hooker (1876) reported 339 genera and

7,453 species, Polhill and Raven (1981) reported 650 genera distributed in 18,000 species, while Humpreys (1996) reported 750 genera and about 16,000 – 19,000 species. Hutchinson and Dalziel (1958) reported 165 genera and 769 species in West Africa out of which 135 genera and 494 species were reported in Nigeria, (Husaini, 1984; Adelanwa, 2008). The recent update of the tribal and generic classification of the family, having the benefit from more than 10 years of intensive molecular phylogenetic studies, recognizes 36 tribes, 727 genera and 19,327 species (Lewis et al., 2005). The family contains at least 4 genera of 500 or more species recorded in Acacia, Astragalus, Crotalaria, and Indigofera and at least 40 genera with 100 spp. or more. At the other extreme, nearly 500 genera are small, either being monospecific or containing up to 10 species (Lewis et al., 2005). Early classification of the family grouped members into three subfamilies, the Caesalpinioideae, Mimosoideae, and Papilionoideae

(although sometimes these have been ranked as separate families - Caesalpiniaceae,

Mimosaceae, and Papilionaceae), and considered most closely related to the Connaraceae and

Sapindaceae based on anatomical, morphological evidences and biogeographic distributions

(Polhill and Raven, 1981). The recognition of the three subfamilies is based mostly on flower features, including size, symmetry, aestivation of petals, (united or free), stamen number and heteromorphy, pollen (single or polyads), and also on the presence of a pleurogram, embryo radicle shape, leaf complexity, and presence of root nodules (Lewis et al., 2005).

Differences in these characteristics led to the view that the Mimosoideae and Papilionoideae are unique and distinct lineages in the family, which arose independently within a paraphyletic

"basal" caesalpinioid assemblage. The Dimorphandra group of tribe Caesalpinieae and papilionoid tribe Swartzieae were likely the transitional groups between them, respectively

26 | P a g e

(Polhill, 1994). However, the status of the Leguminosae representing a family of three subfamilies Mimosoideae, Caesalpinioideae and Papilionoideae or be treated as three different families Mimosaceae, Caesalpiniaceae and Fabaceae or Papilionaceae, remains a disputed issue. The Cronquist and Dahlgren systems elevated the three subfamilies to the familial level while the 20th and early century evidence from morphology and molecules support the legumes being one monophyletic family with the monophyletic Mimosoideae and Papilionoideae nested within the basal paraphyletic Caesalpinioideae. This view has been reinforced not only by the degree of interrelatedness of taxonomic groups within the legumes compared to that between legumes and its relatives, but also by recent molecular phylogenetic studies (Doyle et al., 2000;

Kajita et al.,2001; Wojciechowski, 2003; Wojciechowski et al., 2004). This shows strong support for a monophyletic family, closely related to Polygalaceae, Surianaceae, and

Quillajaceae, which together form the order Fabales (sensu Angiosperm Phylogeny Group,

2003). Within the confinement of this present study, the family Fabaceae is treated as one family comprising the three subfamilies:

Sub-family Caesalpinioideae: This sub-family comprises of about 180 genera and 3,000 species, mainly trees (Tamarindus indica) or shrubs (Parkinsonia spp.; Cajanus cajan) and rarely herbs (Cassia tora) of Tropical savanna and forest of South America, Tropical Africa and Asia. (William, 1983) Members are characterized with alternate, stipulate leaves with pulvinate base, could be compound unipinnate as in Cassia, Tamarindus, bipinnate as in

Delonix, Caesalpinia, there are rarely species with simple leaves. Inflorescence is mostly capitate but sometimes spicate or racemose; flowers are pedicellate, bracteate, zygomorphic, bisexual, pentamerous and hypognous. Their androecium comprises of 10 free stamens while gynoecium is monocarpellary with superior unilocular ovary, ovules arranged in marginal placentation; seeds are usually endospermic (Soni and Soni, 2010). Several species in this sub- family are well-known Tropical ornamentals such as the flamboyant tree (Delonix regia) and

27 | P a g e

Pride of Barbados (Caesalpinia pulcherrima). Alexandrian Senna () is a commercially grown medicinal plant, known for its purgative qualities. Hutchinson and Dalziel

(1958) reported 58 genera and 192 species from West Africa out of which 42 genera and 101 species occur in Nigeria. Giller and Wilson, (1991) divided the sub-family into five tribes, however recent classifications recognize four tribes namely: - Caesalpinieae, Cassieae,

Cercidieae and Detarieae. However, modern phylogenetics have shown their groupings are artificial; members are likely to be split into several sub-families upon more researches in the future (Wojciechowski et al., 2006).

Sub-family Mimosoideae: The sub-family contains 80 genera and 3,270 species distributed throughout, sub-Tropical and warm temperate regions of the world (Lewis et al., 2005).

Members are characterized by usually alternate, stipulate and bipinnately compound leaves. In the Acacias, stipules are usually modified into thorns while in the Australian Acacia, petioles are reduced to a leaf-like phyllode. Inflorescence is mostly capitate but sometimes spicate or racemose. Flowers are mostly insect pollinated, bracteate, sessile, complete, actinomorphic, hermaphroditic, mostly pentamerous and hypogynous. Calyx is usually 4-5, gamosepalous with valvate aestivation. Corolla comprise 4-5, gamopetalous (Acacia, Albizia) or polypetalous. Androecium usually between four - ∞, the number and cohesion of stamens show much variation and used in taxa delimitation. Gynoecium is monocarpellary, unilocular with superior ovary; ovules are in marginal placentation. Seeds are non-endospermic or with scanty endosperm (Soni and Soni, 2010). Many are tall canopy trees, while most are under storey trees or shrubs. Lianas are rare and very few are herbs. Examples of genera within this sub-family are Acacia and Mimosa. Several species are of high economic importance A. senegal for gum and several Acacia species are known for their timber (Acacia, Albizia) and gum exudates, A. sinuate for saponins while Mimosa pudica as ornamentals. Hutchinson and Dalziel, (1958) described 27 genera and 81 species from West Africa out of which Burkill, (1979) reported 22

28 | P a g e genera and 60 species found in Nigeria. Bentham, (1875) recognized five tribes while recent classifications divide the sub-family into four tribes namely: Acacieae, Ingeae, Mimoseae and

Mimozygantheae. However, modern phylogenetics have shown their groupings are artificial; several informal subgroups have been proposed but not yet formally assigned to the tribal level

(Lewis and Elias, 1981; Luckow et al., 2000, 2003; Hughes et al., 2003).

Sub-family Papilionoideae: is the largest of the three sub-families with about two-third of all the genera and species of the family (Bentham, 1865). It comprises of 476 genera and 13,860 species widely distributed from rain forest to the edges of dry and cold deserts. Leaves are usually alternate, simple as in Crotalaria, pinnately compound (Dalbergia, Abrus) or digitately compound (Trifolium). In some species like Vicia and Pisum, terminal leaves are modified to a leaflet. Leaves of several genera e.g. Calopogonium show sleeping movements. Inflorescence are usually racemose, flowers are zygomorphic polypetalous, papilionaceous, hermaphroditic, pentamerous. Calyx has five sepals united into a tube, corolla has five unequal and zygomorphic flower with the outer being the largest called standard, two lateral called wings while the two innermost are almost united called the keel. The corolla encloses the stamen and carpel. Androecium has 10 stamens mostly diadelphous (9+1). Gynoecium is monocarpellary with superior ovary; ovules are in marginal placentation. Seeds are non-endospermic or with scanty endosperm (Soni and Soni, 2010). Members are of the greatest economic importance among the Fabaceae, many species are pulses, vegetables, oils, fibres, fodder, timbers, dyes, medicine amongst others. Hutchinson and Dalziel (1958) described 80 genera and 496 species from West Africa out of which Hussaini (1984) reported 71 genera and 333 species found in

Nigeria. Bentham, (1865) and Hutchinson and Dalziel, (1958) recognized nine tribes:

Podalyrieae, Genistae, Trifolieae, Lotea, Hedysareae, Vicieae, , Dalbergieae,

Sophoreae. However, upon years of studies and surveys of cryptic features, chromosomes, anatomy, chemistry, pollen, evolutionary and diversification, Polhill, (1981) presented a

29 | P a g e supposed relationship among tribes and re-classified members into 32 tribes. However, modern phylogenetics recommend a clade-based classification as a superior alternative to the traditional tribal classification of Polhill, 1981 (Cardoso et al., 2012a, 2013; Wojciechowski,

2013 and LPWG, 2013a).

2.1.6 PHYLOGENY OF LEGUMINOSAE

Studies on the phylogeny of the family Fabaceae based on molecular data dates back as far as over 20 decades. It began with the analysis on higher level phylogenetic relationships based on the chloroplast rbcL gene (Doyle, 1995; Käss and Wink, 1995, 1996, 1997; Doyle et al., 1997).

Overall, the topology consistently supports a monophyletic Leguminosae, although not always strongly so, as suggested by bootstrap and/or jackknife analyses. The traditionally defined

(Polhill et al., 1981) subfamilies Mimosoideae and Papilionoideae have both been resolved as monophyletic, nested within a paraphyletic Caesalpinioideae (Klitgaard and Bruneau, 2003).

However, despite many years of studies, many issues in legume phylogeny remain unresolved.

This is particularly true for the relationships among the larger clades. According to

(Wojciechowski, 2003) there is the need for extensive studies on more variable genes and non- coding sequences, alone or in combination with morphological data.

The subfamily Caesalpinioideae is a paraphyletic group at the base of the Leguminosae and from which are derived the monophyletic subfamilies Mimosoideae and Papilionoideae. This subfamily is currently divided into four tribes: Cercideae, Detarieae, Cassieae and

Caesalpinieae. Of these, only the former two are supported as monophyletic in recent phylogenetic analyses of plastid sequence data which to date have sampled 166 genera

(Herendeen et al., 2003; Bruneau et al., 2001, 2008a; unpub. data from several researchers).

The tribal limits, informal generic groupings and the generic limits of certain large genera (e.g.,

Bauhinia L. s.l., Caesalpinia L. s.l.) as proposed by Lewis et al., (2005) are generally well supported and consistently resolved, whereas others are tentative arrangements that merit

30 | P a g e further study with thorough taxon sampling of the caesalpinoideae. For instance, relationships among the basal nodes of the legumes are not well supported (Fig. 2.4), with Cercideae,

Detarieae and Duparquetia Baill. alternatively resolved as the sister group to the remaining legumes depending on taxon sampling, locus sequenced and method of phylogenetic analysis

(Bruneau et al., 2008b; Bello et al., 2009, 2012). However, most recent phylogenetic evaluations of the family consider Cercideae to occupy the most basal clade position (e.g.,

Doyle et al., 2000; Kajita et al., 2001; Bruneau et al., 2001; Herendeen et al., 2003;

Wojciechowski, 2003; Wojciechowski et al., 2004; Bruneau et al., 2008a; Bello et al., 2009).

Although Cercideae and Detarieae have been individually strongly supported as monophyletic, but relatively few molecular or morphological characters support their wider relationships as seen in Herendeen et al., 2003. Albeit, the position of the monospecific and morphologically unique West African genus Duparquetia (Banks et al., 2003; Herendeen et al., 2003; Prenner and Klitgaard, 2008) still remains uncertain. Based on both rbcL and trnL analyses, the

Dimorphandra group of Caesalpinieae, a diverse assemblage of genera (many of which share characteristics with the Mimoseae) is paraphyletic with respect to the Mimosoideae (Fig. 2.4).

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Fig. 2.4: Phylogenetic relationships at the base of Leguminosae based on parsimony analyses of trnL intron data (Wojciechowski, 2003).

The subfamily Mimosoideae is the smallest of the subfamilies of legumes but probably the least understood from a phylogenetic perspective. While it has generally been accepted that the

Mimosoideae was monophyletic, and derived from caesalpinioid ancestors (Chappill, 1995;

Luckow et al., 2000), the traditional “boundary” between the two subfamilies, with mimosoids distinguished by valvate aestivation of the petals and usually sepals, is not as distinct as once

32 | P a g e believed (e.g., Elias, 1981). Early analyses of molecular data (Käss and Wink, 1996; Doyle et al., 1997) supported the monophyly of the subfamily, but these results were suspect because too few mimosoid taxa were included in these studies. More extensive sampling of both mimosoid and presumed closely related caesalpinioid taxa for trnL and trnK/matK sequences however, reveals there is no support for the monophyly of the Mimosoideae. With members of the Dimorphandra group of Caesalpinieae (Dimorphandra, Erythrophleum, Mora) nested within early diverging lineages of Mimoseae (Diniziya, Piptadeniastrum, Pentaclethra) or unresolved with respect to these and other caesalpinioid taxa (Luckow et al., 2000; Bruneau et al., 2001; Luckow et al., 2003). Several shared anatomical and morphological features such as bipinnate leaves, similar root nodules, and elongated spikes or paniculate of small bisexual flowers (Polhill et al., 1981; Chappill, 1995; Luckow et al., 2000) support a close relationship between the Dimorphandra group and mimosoids. In addition to the suspect monophyly of the subfamily, the monophyly and relationships of its constituent tribes have been problematic as well (Luckow et al., 2000, 2003). Within Mimosoideae, none of the traditionally recognised tribes, Acacieae, Ingeae, Mimoseae and Parkieae, are monophyletic (a fifth, Mimozygaantheae, is monotypic) based on analyses of trnL and trnK/matK data (Luckow et al., 2000, 2003). Indeed, these authors suggest their continued recognition may not be feasible. Mimoseae form a basal grade, with Parkieae, Ingeae and Acacieae nested within it.

Only Acacia s.s. is strongly supported as monophyletic (Luckow et al., 2003). Furthermore, all analyses agree that the other genus of Acacieae, the monotypic Faidherbia, is more closely related to Ingeae, a relationship that has been demonstrated by recent cladistic analyses of morphological data Chappill and Maslin, 1995 and Grimes, 1999 (Fig. 2.5).

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Fig. 2.5: Schematic consensus phylogeny of Mimosoideae based on Luckow et al., 2000, 2003; Simon et al., 2009; Bouchenak-Khelladi et al., 2010; Brown et al., 2011 and Miller et al., 2011. (Source: LPWG, 2013b)

The Papilionoideae, the largest and most widely distributed of the three traditionally recognised subfamilies of Leguminosae, is readily distinguished from the other subfamilies by vegetative, floral, and fruiting characters (Polhill, 1981; Chappill, 1995) including floral development

(e.g., Tucker, 1987, Tucker, 2003; Tucker and Douglas, 1994). While the subfamily has been the subject of intensive taxonomic research over the past few decades, higher-level phylogenetic relationships within Papilionoids have remained unclear. This may be due, in

34 | P a g e part, to a poor understanding of relationships among, and perhaps an over-emphasis on, the genera in the presumed basal tribes Sophoreae, Swartzieae, and to a lesser extent Dalbergieae

(Polhill, 1994; Pennington et al., 2001). Papilionoideae has been consistently resolved as monophyletic in analyses of rbcL data (Käss and Wink, 1995, 1996; Doyle et al., 1997, 2000;

Kajita et al., 2001; Lavin et al., 2003), but support for this conclusion has been low. Similarly, the overall topology of “Millettioids/Phaseoloids” (Kajita et al., 2001) that includes ,

Desmodieae subtribes Desmodiinae and Lespedezinae, Abreae, Phaseoleae, and Psoraleeae

(Wojciechowski et al., 1993; Lavin et al., 1998; Hu et al., 2000; Kajita et al., 2001); and

“Hologalegina” (“galegoid clade”, Doyle et al., 2000; Kajita et al., 2001). Likewise, the trnL data strongly support the monophyly of the Papilionoideae, as currently circumscribed (Polhill and Raven, 1981; Polhill, 1994) but clearly rejecting long-standing suggestions based on the rbcL strict consensus tree of Doyle et al., (2000) (Kajita et al., 2001). Here relationships among members of the subfamily is largely unresolved and weakly supported, although there is an emergent pattern involving the resolution of at least four major papilionoid clades (Lavin et al., 1990). Each of these clades has been the subject of recent investigations and further delimited and refined through extensive sampling and by the use of more informative molecular data. These include the “dalbergioids” (“aeschynomenoid group”, Doyle et al., 2000; Kajita et al., 2001), which comprises tribes Adesmieae, Aeschynomeneae, the Dalbergia group of

Dalbergieae, and subtribe Bryinae of Desmodieae (Lavin et al., 2001); the “genistoids”, tribes

Genisteae, Thermopsideae, Crotalarieae, Liparieae, Podalyrieae, and members of Sophoreae s.s. (Käss and Wink, 1997; Crisp et al., 2000). The phylogenetic position of a number of genera and small groups nested among the major clades of Papilionoids remain poorly resolved (Fig.

2.6). Many of these taxa include genera of Sophoreae, Swartzieae and Dalbergieae that are not members of the clades and they form groups whose relationships are not resolved within the

50-kb inversion clade (Fig. 2.6).

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Fig. 2.6: Schematic phylogeny of Papilionoideae compiled as a super tree based upon phylogenetic analyses of Lavin et al., 2001; Pennington et al., 2001; Crisp and Cook, 2003; Wojciechowski et al., 2004; Boatwright et al., 2008; Egan and Crandall, 2008; Simon et al., 2009. Dotted branches are weakly supported as measured by parsimony bootstrap or Bayesian posterior probabilities. (LPWG, 2013b).

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2.2 Legumes: Morphological Characterization and the identification of arid-land species

Plant identification is the bedrock of biology, and one of the primary objectives of systematics.

Its importance in plant conservation cannot be overemphasized as an unknown entity cannot be neither sustainably used nor preserved. Hence, correct identification of plants should be of prime importance to all users, resource managers, taxonomists and conservationists. However, from time immemorial, plants have been identified through either traditionally, based on distinct features or characteristics upon recognition, comparison with an already identified specimen, thirdly, by expert determination or by the use of keys or similar devices (Davies and

Heywood, 1967). In terms of reliability or accuracy, the best method of identification is expert determination. In principle, the expert will have prepared treatments (monographs, revisions, synopses) of the group in question, and it is probable that the more recent floras or manuals include the expert's concepts of taxa. Although of great reliability, this method presents problems by requiring the valuable time of experts and creating delays for identification.

Identification upon recognition is based on extensive, past experience of the identifier with the plant group in question. In some groups, this is virtually impossible. According to Morse,

(1971), comparing an unknown sample with a named specimen, photographs, illustrations or descriptions is a reliable method, but may be very time consuming or virtually impossible due to the lack of suitable materials for comparison. Its reliability is, of course, dependent on the accuracy and authenticity of the specimen, illustrations or descriptions used in the comparison.

The use of keys or similar devices (synopses, outlines, etc.) is by far the most widely used method and does not require the time, materials, or experience involved in comparison and recognition. However, traditional plant identification keys have been in a difficult and boring form to use, but with the advent of computers, electronic keys, which are user friendly, are becoming increasingly available. Pictures of whole plants and parts can now be attached to the

37 | P a g e keys to improve recognition of the plants. In addition, some electronic keys such as those using

Lucid can be deployed over the internet so that anyone with internet connection can have access to and make use of it.

Some members of the Fabaceae can sometimes be difficult to identify particularly due to the fact that their identification is based on a few reproductive characteristics as seen in the identification of Afzelia species (Donkpeng et al., 2008), as well are difficulties in identifying or separating some members of the genus Acacia (Steven and Subramanyam, 2009). In addition, since the identification keys for legumes are dependent on floral characters, the identification of sterile specimen is in most cases impossible. Even experienced curators sometimes misidentify or mix up species as was experienced during my field data collection.

2.2.1 Morphological Characters

Morphological characters of a plant are the external features that can be examined with the naked eye or with the aid of a magnifying instrument (Stuessy, 2009a). In the Fabaceae, many different types of morphological features are used including number of parts, shape, size, colour, texture and arrangement of vegetative and reproductive features (Lewis et al., 2005).

Reproductive characters often have the advantage of being more stable across environmental factors in many species of plants than vegetative characters (Stuessy, 2009a). In the

Leguminosae, there are usually slight variability in the floral features used in distinguishing species, an example is the counterpart West African species of Afzelia that are differentiated based on floral features and size and colour of seeds (Donkpeng et al., 2008) and number and attachments of stamen number in delimitating members of tribe Acacieae (LPWP, 2013a).

2.2.2 Taxonomic Keys

A key is an artificial arrangement or analytical device where by a choice is provided between two contradictory statements resulting in the acceptance of one and the rejection of the other.

A pair of contradictory statements in a key is called a couplet while each statement of a couplet

38 | P a g e is termed a lead. Keys do not only involve characters important to classification but rather characters that are easily observable and can be used by novices as well (Winston, 1999). Keys can be grouped into two: single access (dichotomous) and multiple access keys. Single access keys are often presented as bracketed or indented keys. In the bracketed key, the couplets are always next to each other in consecutive lines on the page, the end of each line in the key there is either a number or a name referring to the couplet. They are easier to follow for beginners

(Winston, 1999). On the other hand, in the indented keys, each of the couplet is indented a fixed distance from the left margin of the page, the “yes” alternatives are exhaustively dealt with first before the “no”. This makes them more difficult to use for novices. Single access keys can be printed or interactive using hypertext.

Multi-access keys include punch cards, tabular keys and computer based interactive keys

(Winston, 1999; Stace, 1991) of which computer-based keys are now the common kind. Their advantage over single access keys include the ability of the user to start from any point and progress in any format and not be limited when a character cannot be identified or categorized in an organism (Stace, 1991). However, in both single and multi-access keys descriptions and images are necessary to avoid misidentification.

Interactive keys can be computer programmes in which the user enters the characters of the specimen; the program eliminates taxa whose characters do not match those entered until one taxon is left (Watson and Dallwitz, 1992; Dallwitz et al., 2005). Their disadvantage is that the user may use easily observable characters but these may not necessarily contribute significantly to separating taxa. However, they have an advantage over traditional keys in the following ways (Dallwitz et al., 2007):

• The start, order and progress along characters are not restricted.

• Characters already entered could be corrected.

• Some level of error could still lead to correct identification.

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• Multiple states can be entered for some characters when in doubt.

• The program should be able to detect user and/or data entry errors.

• Updating the key is easily done by modifying the data matrix appropriately.

• Numeric characters do not have to be categorized.

2.3 Molecular Characterization

Molecular characterization is the process by which species are identified or differentiated employing molecular tools. Standard characterization and evaluation are carried out using different methods, including the use of descriptor lists of morphological characters, genetic descriptions based on specific DNA sequences, biochemical assays using isozymes or protein profiles or the application of molecular markers and the identification of particular sequences through diverse genomic approaches (Perera et al., 2000). However, genetic characterization clearly offers an enhanced power for detecting diversity (including genotypes and genes) that exceeds that of traditional methods. Likewise, genetic characterization with molecular technologies offers greater power of detection than do phenotypic methods (e.g. isozymes).

This is because molecular methods reveal differences in genotypes, that is, in the ultimate level of variation embodied by the DNA sequences of an individual and uninfluenced by environment. Conversely, differences revealed by phenotypic approaches are at the level of gene expression (proteins). Hence, the use of molecular biology techniques has made it possible to better characterize plant and construct relationship among and within different plant species.

Molecular characterization of species has been of huge advantage or concern to conservationists; it contributes to making informed decisions for conservation activities through identifying genes to adding value to genetic resources. It provides reliable information for assessing the amount of genetic diversity (Perera et al., 2000), the structure of diversity in samples and populations (Shim and Jorgensen, 2000; Figliuolo and Perrino, 2004), rates of

40 | P a g e genetic divergence among populations (Maestri, et al., 2002) and the distribution of diversity in populations found in different locations (Ferguson et al., 2003; Perera et al., 2000).

Molecular characterization also helps to determine the breeding behaviour of species, individual reproductive success and the existence of gene flow, that is, the movement of alleles within and between populations of the same or related species, and its consequences (Papa and

Gepts, 2003; Gepts et al., 2005). Molecular data usually improves or even allow the elucidation of phylogeny, and provide the basic knowledge for understanding taxonomy, domestication and evolution (Judd et al., 2002; Nwakanma et al., 2003). As a result, information from molecular markers or DNA sequences offers a good basis for better conservation approaches.

Recent advances in molecular biology, principally in the development of the polymerase chain reaction (PCR) for amplifying DNA, DNA sequencing and data analysis, have resulted in powerful techniques which can be used for the screening, characterization and evaluation of genetic resources. Many of these techniques have successfully been used to study the extent and distribution of variation in species gene pools and to answer typical evolutionary and taxonomic questions (Karp et al., 1996; Karp et al., 1997). An example is the report of

Coleman, (2000) where he investigated the taxonomic limit of Ulmus plotii. The study showed a distinction between the endemic species and the clone cultivar U. glabra and U. minor

(Culham, 2006). Different techniques employed in molecular characterization ranged from

DNA barcoding, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), microsatellites to single nucleotide polymorphisms (SNP) (Williams et al., 1990, 1993; Neuhaussen, 1992; Zietkiewicz et al., 1994; Vos et al., 1995) and phylogenetic studies to identify and infer evolutionary relationship between or among species

(Soltis et al., 1998).

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2.3.1 Molecular Characters

These are the molecules of organisms that can be isolated, characterized and analysed to establish genetic relationship between members of different taxonomic caregories. There are three different types of data mostly explored for molecular studies: DNA, RNA and proteins

(Mandelkern et al., 1981; Radman and Wagner, 1988).

DEOXYRIBONUCLEIC ACID (DNA)

The information dictating the structures of the enormous variety of cells found in all living organisms is encoded in and translated by the molecule known as DNA (Radman and Wagner,

1988). DNA is a nucleic acid; its chain is between 2.2 and 2.6 nm wide and one nucleotide unit is 0.33 nm long (Mandelkern et al., 1981). It is made up of long chain known as the nucleotides which consists of sugar and a phosphate group joined by ester bond (Fig. 2.7). It usually occurs as linear chromosomes in eukaryotes and circular chromosomes in prokaryotes. The set of chromosomes in a cell makes up its genome, the information carried by DNA is held in the sequence of pieces of DNA called genes (Cramer et al., 2000).

Transmission of genetic information in genes is achieved via complementary base pairing

(Watson, 1968). For example, in transcription, when a cell uses the information in a gene, the

DNA sequence is copied into a complementary RNA sequence through the attraction between the DNA and the correct RNA nucleotides. Usually, this RNA copy is then used to make a matching protein sequence in a process called translation which depends on the same interaction between RNA nucleotides. Alternatively, a cell may simply copy its genetic information in a process called DNA replication. Genomic DNA is located in the cell nucleus of eukaryotes, as well as small amounts in mitochondria and chloroplasts. In prokaryotes, the

DNA is held within an irregularly shaped body in the cytoplasm called the nucleoid. The genetic information in a genome is held within genes, and the complete set of this information in an organism is called its genotype (Cramer et al., 2000). A gene is a unit of heredity and is

42 | P a g e a region of DNA that influences a particular characteristic in an organism. Genes contain an open reading frame that can be transcribed, as well as regulatory sequences such as promoters and enhancers, which control the transcription of the open reading frame in many species

(Ptashne and Ganne, 1997). Some non-coding DNA sequences play structural roles in chromosomes. Telomeres and centromeres typically contain few genes, but are important for the function and stability of chromosomes. However a gene is a sequence of DNA that contains genetic information and can influence the phenotype of an organism. Within a gene, the sequence of bases along a DNA strand defines a messenger RNA sequence, which then defines one or more protein sequences. The relationship between the nucleotide sequences of genes and the amino-acid sequences of proteins is determined by the rules of translation, known collectively as the genetic code. The genetic code consists of three-letter 'words' called

''codons'' formed from a sequence of three nucleotides (e.g. ACT, CAG, and TTT). In transcription, the codons of a gene are copied into messenger RNA by RNA polymerase. This

RNA copy is then decoded by a ribosome that reads the RNA sequence by base-pairing the messenger RNA to transfer RNA, which carries amino acids. Since there are 4 bases in 3-letter combinations, there are 64 possible codons (4^3 combinations). These encode the twenty standard amino acids, giving most amino acids more than one possible codon. There are also three 'stop' or 'nonsense' codons signifying the end of the coding region; these are the TAA,

TGA and TAG codons (Agarwal et al., 1996). However, molecular data have advantages over morphological data in the following ways:

• Molecular entities are strictly heritable

• The description of molecular characters is unambiguous

• There is some regularity to the evolution of molecular traits

• Molecular data are amenable to quantitative treatment.

• Homology assessment is easier than with morphological traits

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• Molecular data are robust to evolutionary distance

• Molecular data are abundant

• Accumulation of molecular data is less time consuming

Fig. 2.7: DNA double helix structure Source: Watson and Crick (1953)

2.3.2 DNA Barcoding

DNA barcoding is one of the tools for characterization and identification of species using an internationally agreed protocol and regions of DNA to create a global database of living organisms (Hebert et al., 2003; Hebert and Gregory, 2005). The importance of plant barcoding is highlighted by the need for accurate species identification to both conserve and utilize plants.

In many parts of the world, this may be hindered by lack of taxonomic expertise (Chase and

May, 2009) as well as the difficulty in species identification from materials such as roots, seeds and pollen or at different life growth stage; or in the mixture of plants sampled from the air,

44 | P a g e soil or water (de Vere et al., 2012). The DNA barcoding expertise depend on using standard regions of DNA of an unknown species in comparison with a library of DNA sequences from taxonomically identified species (Hebert et al., 2004, Ratnasingham and Hebert, 2007). A close enough match ≤ 1% sequence divergence for animals (Hebert et al., 2004; Hajibabaei et al.,

2007; Ratnasinggham and Hebert, 2007) to any of the sequences in the library identifies the unknown species but this has proven to be difficult in plants. If the match to any of the stored sequences in the database to the unknown individual is not close enough, the sample cannot be identified and may need to be further investigated (DeSalle et al., 2005; Moritz and Cicero,

2004). A barcode will be ideal if the within species (intraspecific) variation does not overlap with the between species (interspecific) variation (Meyer and Paulay, 2005; Edwards et al.,

2008). Different genome regions evolve at different rates among groups (Shaw et al., 2005) and species diverge at different points in time. Intraspecific and interspecific divergences therefore differ from group to group (Meyer and Paulay, 2005). Finding a genome region of the right level of divergence (small variation within species but larger variation between species variation) to be used as a barcode in land plants has been difficult (Chase et al., 2007). Although in animals, the mitochondrial gene (cox1) appears to work well (Hebert et al., 2003) with notable exceptions as shown in Meier et al., (2006). Following the evaluation of several candidate markers, it did become evident that none was good enough as a single loci barcode region. Thus, the plant working group of the consortium for the barcoding of life recommended that regions of two plastid genes rbcL and matK be adopted as the standard plant DNA barcodes, with the recognition that supplementary markers may be required as research progresses (CBOL, 2009; Hollingsworth et al., 2011).

DNA barcoding of plants is already employed in a wide variety of applications such as being used for the verification of plant products ranging from medicinal plants (Asashina et al., 2010;

Chen et al., 2010) to kitchen species (De Mattia et al., 2011), berries (Jaakola et al., 2010),

45 | P a g e olive oil (Kumar, et al., 2011) and tea (Stoeckle et al., 2011). Ecological applications have included the identification of invasive species (Bleeker et al., 2008; Saunders, 2009; Van De

Wiel et al., 2009), characterization of below ground plant diversity using roots (Kesanakurti et al., 2011) and reconstruction of past vegetation and climate from plant remains in the soil

(Sonstebo et al., 2010). Genetic sequences obtained in the context of DNA barcoding have also been used to create phylogenetic trees for use in phylogenetic community ecology (Kress et al., 2009; Kress et al., 2010). The use of DNA barcoding as an identification tool is also dependent on the creation of high –quality reference databases of sequences (Hollingsworth et al., 2011). Essential to a database is that every DNA sequence should be associated with the plant specimen from which it came, indicating where it came from and by whom it was collected and identified. This is best done through the creation of a herbarium voucher alongside each DNA sample (Fig. 2.8), although sometimes for rare and threatened species a photograph might provide a substitute (de Vere et al., 2012). The laboratory procedures through which a sample is processed should also be recorded with the primers used, trace files and quality statistics for its DNA sequence all available to end users of the data (de Vere et al.,

2012). All data should be publicly available; Genbank provides a repository for DNA sequences but in addition, it is recommended to deposit data on to the Barcode of Life Data system (BOLD) (Ratnasingham and Hebert, 2007). BOLD provides a means of managing projects and trace file scans of herbarium specimen and photographs to be stored alongside

DNA sequences. (Ratnasingham and Hebert, 2007).

2.3.2.1 Barcode data analysis

The identification of animal species using barcodes has relied extensively on tree building approaches also called hierarchical clustering (DeSalle et al., 2005; Little and Stevenson,

2007). In plants, the analysis follows a similar trend as those used for animals and involves tree building using distance, parsimony or likelihood approaches Matz and Neilsen, 2005; Nielsen

46 | P a g e and Matz, 2006; Hajibabaei et al., 2007; Lahaye et al., 2008a; Lahaye et al., 2008b; CBOL plant Working group, 2009 and Gonzalez et al., 2009. Although, DeSalle et al., (2005), suggested the use of character based methods. However, the ongoing argument is that sometimes the basis for using phylogeny is unfounded, the method will fail to identify species correctly especially in paraphyletic species (Meyer and Paulay, 2005). Fifteen percent of the failure of barcode regions to classify plants correctly is attributed to paraphyly (Fazekas et al.,

2008, 2012).

Fig. 2.8: DNA Barcoding pipeline (BOLD homepage, 2013)

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Desalle et al., (2005) have also objected to the use of distance methods because cut-off points

(the proportion of within-species variation that is large enough to make an individual not belong to that species) have to be determined. Within species sequence, variation differs between different groups of organisms (Hebert et al., 2003) and therefore a cut-off would be difficult to find that will apply to all organisms (DeSalle et al., 2005; Little and Stevenson,

2007). Meyer and Paulay, (2005), have advocated for comprehensive sampling and the use of statistical procedures to fix a meaningful cut-off. In the case of the BOLD database, a 1%, cut- off is considered appropriate for animals based on comprehensive studies of Ratnasingham and

Hebert, 2007. To estimate the reliability of barcodes regions, several identification techniques have been employed by several authors which have always produced varied results (Meyer and

Paulay, 2005, Ross, et al., 2008 and Austerlitz et al., 2009) but it was however concluded that no single method has been found to be the best. The two most widely used techniques are tree based and sequence comparison. Based on the sequence comparison approach, the “best match” and “best close match” functions of TAXONDNA (Meier et al., 2006) can be used to assess the utility of different barcode regions for accurate species discrimination. The best match is a least stringent criterion; it finds the closest barcode match to each query sequence, if both sequences are from same species then identification is considered successful, if query sequence matches several sequences from different sequences identification is considered ambiguous whereas mismatch names and sequences are considered incorrect. Best close match defines set a threshold frequency of 95% of all intraspecific sequences before identification can be made (Meier et al., 2006). Query sequence with the smallest distance within the 95th percentile and conspecific is considered successful, sequences with the smallest distance within the 95th percentile but with a mixture of allospecific and conspecific sequences are considered ambiguous while sequences with the smallest distance within the 95th percentile and allospecific are incorrect.

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DNA barcoding has the potential to identify species rapidly (Cowan et al., 2006). This would complement the existing taxonomic systems and improve the study, use and conservation of living organisms (DeSalle et al., 2005). However, a species can only be declared unknown if all currently known species are included in the library. Most members of the Fabaceae of the

Nigerian arid flora are yet to be included to the database (to the best of my knowledge based on the information available me) and therefore would have to be added to the library.

2.3.3 Phylogeny Reconstruction

Phylogenetic studies using DNA is usually to characterize evolutionary relationships or histories among genomic group at different levels of taxonomic hierarchy: order, family, genus, species and population levels (Soltis et al., 1998; Avise, 2000; The Angiosperm Phylogeny

Group, 2009). It is also used for the reconstruction of evolutionary relationships among organisms living (extant) and dead (extinct). This technique has proven useful in many biodiversity conservation, taxonomic, medicine, ecological, forensics and biogeographical approaches (Fitch and Margoliash, 1967; Fitch, 1971). According to Hartman and Steel, (2006) and Purvis et al., (2005), it is of immense help in prioritizing conservation needs. Based on the report of Baker et al., (2007), a genetic evidence has proved the illegal trade in protected whales enroute Japan, the US and South Korea. This technique presents evolutionary relationships by a hierarchical tree-like structure based on shared characters or heritable traits that can be compared among organisms. Different data types based on morphology, behavioral traits, protein sequences or genetic sequences can be employed in phylogenetic studies (Stuessy,

2009b). Although morphological data were traditionally used (Stuessy, 2009b), molecular data have accumulated substantially and are now commonly used (Lemey et al., 2009). Molecular phylogenetics attempts to determine the rates and patterns of changes occurring in the sequences and to reconstruct the evolutionary history of genes and organisms (Fig. 2.9).

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Fig. 2.9: Phylogenetic workflow (Thompson, 2013) MSA= multiple sequence alignment

2.3.3.1 Methods for phylogeny reconstruction

The many and remarkably diverse methods employed in Phylogenetics are classified into three main categories based on their overall schema:

• Distance,

• Parsimony and

• Likelihood.

Distance methods: this uses the evolutionary distance between operational taxonomic units

(OTUs) to infer Phylogenetic history. The principle employed in this method is to first generate a distance matrix (i.e., a table of “evolutionary distances” between each pair of taxa) based on an algorithm used to generate a phylogenetic tree (Nei, 1972, 1991, 1996; Harrison and

Langdale, 2006). There are different algorithms used e.g.:

• Uncorrected (P): also referred to as the p-distance, is the uncorrected number of changes

between two sequences (Kluge and Parris, 1969).

• Jukes-Cantor (JC): assumes that the chance of any nucleotide changing into any other

anywhere in the sequence is equal to each other (Jukes and Cantor, 1969).

• Kimura 2-Parameter (K2P): this model as well tackles the problem with same approach

as the JC but assumes different rates for transitions and transversions (Kimura, 1980).

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• General Time-Reversible (GTR): this model relaxes the assumptions about the

correlation among rates of change from one nucleotide to another (Kidd and

Sgaramella-Zonta, 1971).

Parsimony also known as Maximum Parsimony (MP)

Parsimony methods are based on the Razor principle, which states that, lacking any other factors, the simplest explanation—the most parsimonious one—should be chosen. Application of this principle to evolutionary studies has led to the development and use of “parsimony” methods for phylogenetic inference (Fitch, 1975; Nei and Takezaki, 1994; Page and Holmes,

1998). These methods use only derived states of meristic characters to construct a tree, it infers the most parsimonious tree to be the one with the minimum number of substitutions (i.e., the one requiring the fewest evolutionary changes). It minimizes the number of character states that evolve on the tree or, in other words, finding the shortest tree or finding the tree that makes the fewest assumptions of homoplasy (Harrison and Langdale, 2006). However, this method has a problem of being prone to long branch attractions. If the sequences are evolving at very different rates, the probability of convergent substitutions is significant in long branches i.e.

The most parsimonious clustering can lead to false topology (Farris, 1983).

Maximum likelihood (ML)

This uses derived states of meristic characters or quantitative characters to construct a tree based on the probabilities of character states changing on the tree. The probability of change is estimated from the data based on the probability that a particular model of character change and the observed character states would give rise to a particular tree (Felsenstein, 1973a,

Felsenstein, 1973b). The tree with the highest probability, or likelihood, is the one favoured.

This is different from the MP methods due to its use of an explicit evolutionary model and allows variable substitution rates for each branch used to estimate reliability of tree. From a

51 | P a g e theoretical point of view, this is a best-justified method. Sequence simulation experiments have shown that ML works better than the others (Parsimony, NJ) in most cases but it is a very computer-intensive method, it is nearly always impossible to evaluate all possible trees because there are too many, a partial exploration of the space of possible trees is done (Thompson,

2013). An offshoot of this method is the Maximum Bayesian method which is gradually recently gaining popularity amongst phylogenetics. It seeks to calculate the actual probability of the hypothesis by attempting to assign a value to the prior odds term of the equation (Beerli and Felsenstein, 1999). A very powerful use of likelihood and Bayesian methods is that they allow for testing of a variety of evolutionary hypotheses within a statistical framework

(Thompson, 2013).

2.3.3.2 Assessing of the reliability of a tree

One approach to assessing how well a tree represents all of the data is to resample the data repeatedly and re-perform the phylogenetic analysis to see how often the same result is obtained from these resampled (and non-identical) datasets. Resampling by bootstrapping in which the characters (e.g., alignment columns) are resampled with replacement, or by jack- knifing in which the characters are resampled without replacement (Felsenstein, 1985a).

Frequently, 100 or 1000 of these new resampled datasets are generated and a phylogenetic tree is built from each of them. The new trees are then compared to determine in what fraction of the trees particular evolutionary groupings are found. It is very important to realize that these tests do not determine how accurate a tree is, just how well it reflects the underlying data. If the data are biased in some way (e.g., there has been significant convergent evolution), the result can be high bootstrap or jack-knife support for an incorrect tree. A bootstrap or jack- knife support value of ≥95% are widely considered to reflect correct relationships, although some authors have suggested that 70% may be a more realistic cut-off point (Nei and Kumar,

2000). This is likewise an alternative method of assessing the reliability of a tree using the

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Approximate Likelihood-Ratio Test. This test assumes two different types of hypothesis as seen in the Standard LRT where the null hypothesis infers branch has length of zero (0) and statistically calculates the difference of maximum log-likelihood values under alternative and null hypotheses. The second hypotheses, Approximate LRT assumes the null hypothesis to be the inferred branch is incorrect and statistically calculates the difference of likelihood values for best and second-best alternative arrangements around branch of interest. This test is implemented in PhyML (Thompson, 2013).

2.3.3.3 Gene Regions for Phylogeny Reconstruction

Certain gene regions are better suited for studying specific taxonomic levels than others (Shaw et al., 2005). Some regions have wider taxonomic range than others; this is because different genes evolve at different rates (Shaw et al., 2005). A fast-evolving region might be too saturated and slow evolving genes might be invariable to be useful for certain studies (Soltis et al., 1998). It is therefore helpful to use genes that have already proven useful based on previous studies as cited in literature. In addition, pseudo genes can result in the wrong tree if mixed with the functional gene region because of the differences in the rates of evolution. Since pseudo genes are non-coding, they are unlikely to be under the same selection pressures compared to functional genes and are more likely to accumulate mutations. It is therefore important to choose an appropriate gene region in other to detect a phylogenetic signal; the choice is dependent on the relationship among the taxonomic units being explored.

Relationships employed at higher level of taxonomic hierarchy (order or family) will require slowly evolving genes while relationships at lower level of taxonomic hierarchy (species, population) will require a fast-evolving gene (Soltis et al., 1998). In plants, the rbcL and matK genes have been used to explore wide range relationships from the species level up to the family level (Chase et al., 1993; Barracough et al., 1996; Hilu et al., 2003).

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2.4 CONSERVATION ASSESSMENTS

Conservation assessments are technical activities undertaken using a variety of methods to determine how likely a species will go extinct in the near future given current knowledge about population trends, range, and recent, current or projected threats (Akcakaya et al., 2006).

Results from these assessments serve as a spur to conservation action for threatened species. It could be done at different scales be it global, regional or at national levels (Butchart et al.,

2006). The most trusted global assessment programme is the IUCN Red List of Threatened

Species. The IUCN Red List uses a well-established, quantitative protocol that is universally applicable to all species (marine or terrestrial, plant or animal) (Hayward, 2011). Using this protocol, volunteer experts, including members of IUCN SSC Specialist Groups, those interested in conservation assessment of species work with IUCN staff and other experts to assess global populations (Hoffman et al., 2008). Conservation assessments are often undertaken at the scale of large geographic regions, regional or national levels. The purpose of such assessments is to determine whether species are at risk of regional extirpation due to the threats they face. Regional assessments make it easier to implement conservation actions in particular areas, which may not have been apparent at the global scale. Countries around the world also often undertake conservation assessments at the national scale in order to gauge their contribution to conservation and report on national biodiversity commitments such as the

Convention on Biological Diversity. Such assessments can help a country to determine whether their environmental stewardship is adequate for the species within their borders (Rodrigues et al., 2006). Although most countries follow the IUCN Categories and Criteria, others modify them based on their own needs, and some even take on their own assessments independent of

IUCN protocols. This research will be following the IUCN Categories and Criteria of 2014

(IUCN, 2014).

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2.4.1 IUCN Categories

IUCN Red List system consists of nine main categories (Fig. 2.10) which divided into two major groups; not evaluated and evaluated species. The evaluated species category is divided into two subgroups: data deficient species (DD) and adequate data species. The latter is then further divided into two main groups, which are; non-threatened [Least Concern (LC), Near

Threatened (NT)] and threatened [Vulnerable (VU, Endangered (EN) and CR and Extinct

(EX)].

Fig. 2.10: The International Union for Conservation Nature (IUCN) Red List Categories at the regional level (IUCN, 2014)

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2.4.2 IUCN Criteria

The most important categories; threatened, are based on five scientific criteria (A, B, C, D and

E) in which this criterion evaluate the risk of extinction of the species based on biological and ecological factors such as: A. declining population (past, present and/or projected); B.

Geographic range size and fragmentation, decline or fluctuations; C. Small population size and fragmentation, decline, or fluctuations; D. Very small population or very restricted distribution; and E. Quantitative analysis of extinction risk (e.g. Population Viability Analysis).

The five main criterion were then further divided into sub criteria and the listing of a particular species was justified more specifically with a set of quantitative thresholds, under a particular category. The species could be already Extinct (EX) or Extinct in the Wild (EW) if none of the thresholds was met. If it nearly meets the conditions for a threatened category it is Near

Threatened (NT) and if its current extinction risk is relatively low, it qualifies for Least Concern

(LC). However, if the available data are insufficient to list the species under any category, it qualifies as Data Deficient (DD). The evaluation of a certain species needs to consider all the available data against all five criteria. Although a species may not meet all five criteria to qualify as threatened, it may meet all the conditions for at least one criterion. The species of interest were evaluated against as much criteria as possible and the listing was interpreted with as many criteria permitted and applicable for a specific category (IUCN, 2014).

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Table 2: Summary of Criteria (A) in the International Union for Conservation of Nature (IUCN) criteria and categories

A. Population reduction. Declines measured over the longer of 10 years or 3 generations based on any of A1 to A4. Critically Endangered Endangered Vulnerable A1 ≥ 90% ≥ 70% ≥ 50% A2, A3 & A4 ≥ 80% ≥ 50% ≥ 30% A1. Population reduction observed, estimated, inferred, or suspected in the past where the causes of the reduction are clearly reversible AND understood AND have ceased, based on and specifying any of the following: - (a) Direct observation. - (b) An index of abundance appropriate to the taxon. - (c) A decline in area of occupancy (AOO), extent of occurrence (EOO) and/or habitat quality. - (d) Actual or potential levels of exploitation - (e) Effects of introduced taxa, hybridization, pathogens, pollutants, competitors or parasites.

A2. Population reduction observed, estimated, inferred, or suspected in the past where the causes of reduction may not have ceased OR may not be understood OR may not be reversible, based on (a) to (e) under Al.

A3. Population reduction projected or suspected to be met in the future (up to a maximum of 100 years) based on (b) to (e) under Al.

A4. An observed, estimated, inferred, projected or suspected population reduction (up to a maximum of 100 years) where the time period must include both the past and the future, and where the causes of reduction may not have ceased OR may not be understood OR may not be reversible, based on (a) to (e) under Al.

Source: IUCN, 2014

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Table 3: Summary of Criteria (B) in the International Union for Conservation of Nature (IUCN) criteria and categories

B. Geographic range in the form of either B1 (extent of occurrence) AND/OR B2 (area of occupancy) Critically Endangered Vulnerable Endangered B1. Extent of occurrence < 100 km² < 5,000 km² < 20,000 km² (EOO) B2. Area of occupancy < 10 km² < 500 km² < 2,000 km² (AOO) AND at least 2 of the following 3 conditions: (a), (b) and (c) - (a) Severely fragmented, = 1 ≤ 5 ≤ 10 OR Number of locations -(b) Continuing decline in any of: (i) extent of occurrence; (ii) area of occupancy; (iii) area, extent and/or quality of habitat; (iv) number of locations or subpopulations; (v) number of mature individuals. -(c) Extreme fluctuations in any of: (i) extent of occurrence; (ii) area of occupancy; (iii) number of locations or subpopulations; (iv) number of mature individuals.

Source: IUCN, 2014

Table 4: Summary of Criteria (C) in the International Union for Conservation of Nature (IUCN) criteria and categories

C. Small population size and decline Critically Endangered Vulnerable Endangered Number of mature < 250 < 2,500 < 10,000 individuals AND either C1 or C2: 20% in 5 years or 10% in 10 C1. An estimated continuing 25% in 3 years or 1 2 years or 3 decline of at least: generation generations generations C2. A continuing decline AND at least 1 of the following 3 conditions: -(ai) Number of mature individuals in each ≤ 50 ≤ 250 ≤ 1,000 subpopulation

-(aii) % individuals in one 90–100% 95–100% 100% subpopulation =

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-(b) Extreme fluctuations in the number of mature individuals.

Source: IUCN, 2014

Table 5: Summary of Criteria (D) in the International Union for Conservation of Nature (IUCN) criteria and categories

D. Very small or restricted population Critically Endangered Vulnerable Endangered D. Number of mature < 50 < 250 D1. < 1,000 individuals D2. Only applies to the VU category. Restricted area of occupancy or number of 10% in 10 locations with a plausible years or 3 future threat that could drive generations the taxon to CR or EX in a very short time.

Source: IUCN, 2014

Table 6: Summary of Criteria (E) in the International Union for Conservation of Nature (IUCN) criteria and categories

E. Quantitative Analysis Critically Endangered Vulnerable Endangered > 20% in 20 > 50% in 10 years or Number of mature years or 5 > 10% in 100 3 generations individuals generations years (100 years max.) (100 years max.)

Source: IUCN, 2014

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2.4.3 Components of a Red List assessment

Three main components are required for a red list assessment. Firstly, is to assign each species a Red List category based from the five main criteria as described above in 2.4.2. Secondly is to justify the assessment with supporting documentation on the geographical range of the species, population size and information to indicate population trends, description of the habitats and ecological requirements of the species, local uses and trades of the species, description of the threats affecting the species populations and habitats, documentation of the conservation and research actions currently in place and recommendation of some conservation actions needed in the future. Lastly is to present the map of species’ distribution (Hoffman et al., 2008).

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CHAPTER THREE METHODOLOGY

3.1 Sample Collection

Both herbarium and silica-gel dried samples were used in this study. Young fresh leaf tissues were collected randomly across the Sudano-sahelian region of Nigeria which spans across the arid and semi-arid regions of the country (Fig. 3.1). Leaf materials of Leguminosae were collected from field, botanical gardens and forest reserves from the study region. The plant parts, leaves, flowers, and stem were collected using secateurs and cutlass. The fresh specimens were pressed using a plant press, which consists of a wooden frame (for rigidity), blotting paper (to absorb moisture) and folded newspaper (to contain the plant material). The plant press was tightened using straps or twines. This is to extract moisture in the shortest period of time, while preserving the morphological integrity of the plant and to yield material that can be readily mounted on herbarium paper for long-term storage. The dried specimens were poisoned to prevent them from fungal and insect attack using a mixture containing 5 L of

Methylated spirit, 4-6 drops of phenol and 200 mL of mercury (II) chloride. Some of the specimens were poisoned by dipping the whole plants into a basin containing the poison and others were poisoned using a brush dipped in the mixture to brush both the adaxial and abaxial parts of the plant according to Bridson and Forman (1992). The poisoned specimens were then mounted on standard mounting sheet/cardboard (16.5x10.5 cm) using glue and kept in between newspapers for drying. The cardboard provides physical support that allows the specimens to be handled and stored with a minimum of damage. Identification and authentication of the species were done at the University of Lagos Herbarium (LUH) and Forest Herbarium Ibadan

(FHI). Voucher specimens were deposited at the University of Lagos Herbarium for future purposes (Ogundipe and Chase 2009).

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Samples for molecular studies were stored in a zip lock polythene bag containing silica gel and stored in the refrigerator at 4o C prior to use.

Five (5) different herbaria, (University of Lagos Herbarium, Lagos; Forestry Herbarium,

Ibadan; Ahmadu Bello University Herbarium, Zaria; University of Reading Herbarium,

Reading; Royal Botanic Gardens, Kew) were visited, representative samples were studied morphologically and collected in cases where permitted for molecular studies; 0.02g was collected and stored in a labelled brown envelope.

Photographs of samples were taken for those available as well as their Global Positioning

System (GPS) coordinates and Geo-referenced points were plotted on a Nigerian map using the ArcMapGIS software (Fig. 3.1).

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Fig. 3.1: Map depicting different ecological zones and sampling location in Nigeria Source: Modified from Balogun, 1999

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3.2 Morphological Characterization

Both qualitative and quantitative characteristics of each specimen was obtained and recorded.

Qualitative features such as leaf shape, leaf tip, leaf base, leaf texture, leaf margin, flower color was examined visually or with the aid of magnifying lens when necessary while quantitative features such as the length of leaf, leaflet, petiole were measured with a meter rule or thread.

Thirty-four (34) exo-morphological characters were scored and used to construct keys for

Nigerian arid legumes, a single access bracketed key and an electronic multi-access key based on the Lucid computer program. Keys were then validated by using them to identify specimens.

Electronic Multi-access Key

The electronic key was built based on the Lucid programme which can be deployed either on a CD or in the internet. The programme has two different suites (builder and player) for preparing and using the key respectively. The builder is used to develop a key where features, states and species names are entered, an empty lucid builder is shown below Fig. 3.2.

Features were entered by clicking on the features icon, typed in the space bar above and pressing the enter key on the keyboard, states are added by clicking on the toggle button which toggles between adding features and states before pressing the enter key. Names of understudied species are listed in entities box, this is done by clicking on the entities icon before typing name in the space bar above before pressing the enter button on the keyboard. After all the features, states and entities have been entered in, each character and states were scored against distinct species. One way of scoring quickly and efficiently is to use the spreadsheet scoring on the lower panel as seen in Fig. 3.3. Afterwards the lucid key is saved and ready to use.

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Fig. 3.2: Empty Lucid Builder v3.3

Fig. 3.3: Spreadsheet being scored

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3.3 Molecular Characterization

3.3.1 Source of Plant material

Specimens obtained from the field were used in this study complemented with an additional hundred and thirteen (113) replica legume samples obtained from the University of Reading

Herbarium. Outgroup was defined based on previous studies according to Wojciechowski,

2003; Wojciechowski et al., 2004 and Lavin et al., 2005. Sequences of members of the

Polygalaceae were downloaded from Genbank and included in the dataset.

3.3.2 DNA Isolation

Total genomic DNA was extracted following the modified 2X CTAB protocol of Doyle and

Doyle (1987). Approximately 0.0300g of silica-gel dried and 0.0180g of herbarium plant material were ground with 2 tungsten carbide beads in a 2 ml eppendorf tube using a bead beater at 30 Hz for 45 s. Upon completion, beads were removed and 800 µl of pre-warmed 2X

CTAB buffer was pipetted into the tubes and incubated for 1hr with occasional inversion at 5-

10 mins interval. Afterwards, they were centrifuged at 13000 rpm for 3 mins and the aqueous upper phase was carefully transferred into a clean 1.5 ml eppendorf tube, an equal volume of

SEVAG was then added, mixed well to obtain an emulsion and centrifuged at 13,000 rpm for

5 mins. The upper layer was again carefully transferred into another clean 1.5 ml eppendorf tube and the chloroform extraction phase was repeated while the aqueous phase was transferred into a clean 1.5 ml screw cap tube. Upon centrifugation, 0.08 volumes of cold 7.5 M ammonium acetate and 0.54 volumes (using the combined volume of aqueous phase and added AmAc) of cold propan-2-ol were added, mixed and kept in the freezer for one hour (one week in case of herbarium specimen) to precipitate the DNA. Afterwards, samples were centrifuged at 13000 rpm for 15 mins to form a pellet. Liquid was carefully poured out and 700 µl of cold 70% ethanol was then added to tubes, mixed and allowed to stand for a few minutes. The tubes were centrifuged, liquid was pipetted off and the ethanol phase was repeated to remove remnant

66 | P a g e contaminants. Dry pellets were dried in the centrivamp at 35 ºC and samples were resuspeneded in 100µl of TE buffer overnight. Resultant DNA samples were stored at -20 ºC for subsequent use.

3.3.3 Quantification of DNA Samples

This was done to ascertain the concentration and relative absorbance of the isolated DNA. This involved the use of a nanodrop to determine the absorbance of isolates at wavelengths of A230,

A260 and A280. To achieve this, 2µl of TBE buffer was used to blank the nanodrop after which

2µl of isolates was spotted on and readings were documented and repeated for all samples.

3.3.4 Gel Electrophoresis

This is a quality check to determine the integrity of the isolates. Electrophoresis of the isolated total genomic DNA products was done on 0.7% agarose gel, 0.63g agarose in 90 ml I XTAE Tris-

Acetate-EDTA (TAE) buffer. This was achieved by dissolving the 0.63g of agarose in 90ml

TAE buffer using a microwave and allowed to cool. Upon cooling, 1.5µl of ethidium bromide was added to the solution and swirled to mix; the agarose solution was then poured in a gel plate fixed with combs to create wells in the gel. When the gel had solidified, the combs in the gel were carefully removed and the gel plate was immersed into the gel electrophoresis tank containing 1X TAE buffer.

3.3.5 Loading of Samples and Running of Gel

Aliquots of 2µl of loading dye was spotted in each well of a loading rack for each 3µl of the extracted genomic DNA and mixed with a pipette. The mixture was taken into the well in the gel using a 1-10 µl range pipette. This was allowed to run at 100V for 60mins. The gel was then visualized under an ultraviolet transilluminator attached to a computer system and printer, photographed with gel documentation units (UVitec) and printed.

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3.3.6 Polymerase Chain Reaction (PCR)

This procedure was carried out to make more copies of targeted regions of DNA using a thermocycler at different temperature gradients with the help of the enzyme taq polymerase.

Prior to sample amplifications, a pilot study was conducted to screen primers and fourteen (14) different gene regions were understudied. Twenty-two (22) different primers were screened

(Table 7) and upon completion, four (4) (chloroplast rbcL, matK, trnL-F and nuclear ITS) regions (Table 8) were selected to be employed in the study based on their amplification and sequencing success rates.

Polymerase chain reaction (PCR) was performed in a 50µl reaction mixtures containing 25µl biomix, 1µl BSA, 2ul DMSO, 1.75µl of 10uM of each forward and reverse primer, 17.5µl of

Millipore H2O and 1µl of 30 – 50 ng template DNA. Amplifications were run on a Veriti® 96 well thermal cycler with profiles for each reaction as listed in Table 9. Upon completion of

PCR process, products were stored at -20ºC prior to further processes.

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Table 7: Gene regions and primers screened Gene region Primers (forward and reverse) trnGR trnG1F & trnG22R psbK-L psbL & psbK atpF-H atpF & atpH trnS-G trnSGCU & trnGUUC trnD-T trnD & trnT ndhJ-trnF ndhJ & trnFGAA trnH-psbA trnH & psbA petL-psbE petL & psbE rbcL rbcL1F & rbcL724R & 1460R rp132-trnL rp132F & trnLUAG rps16 rps16 & trnKUUU matK matK390F &1326R; matK_1R&3Fkim; matK_fabF&fabR trnL-F trnLc-f, tab c&d, tab e&f ITS ITS 3&4; 4&5; 17SE&26SE

Table 8: Gene regions and primer sequences employed in the study Region Primer sequence (5’ to 3’) References ITS ACGAATTCATGGTCCGGTGAAGTGTTCG/ Sun et al., 1994 ITS17SE & ITS26SE TAGAATTCCCCGGTTCGCTCGCCGTTAC

Kim KJ, unpublished data, matK CGTACAGTACTTTTGTGTTTACGAG/ Korea University, Seoul, 3F_KIM/1R_KIM AATATCCAAATACCAAATCC Korea

rbcL ATGTCACCACAAACAGAAAC/ Olmstead et al., 1992; Fay 1F/724R/1460R TCGCATGTACCTGCAGTAGC/ et al., 1997 TCCTTTTAGTAAAAGATTGGGCCGAG

trnL intron and spacer CGAAATCGGTAGACGCTACG/ Taberlet et al., 1991 trnLc-f GGGGATAGAGGGACTTGAAC

Table 9: Profiles of PCR reactions Initial denaturing Denaturation Annealing Extension Final extension No. of cycles Temp./time Temp./time Temp./time Temp./time Temp./time

ITS 970C/2:00 970C/1:00 550C/0:45 720C/0:45 720C/7:00 30

matK 940C/5:00 940C/0:40 480C/0:40 720C/0:40 720C/7:00 30

rbcL 960C/0:50 960C/50 530C/0:50 720C/2:00 720C/7:00 30

trnL-F 940C/2:00 940C/1:00 550C/1:00 720C/2:00 720C/10:00 30

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3.3.7 Gel Electrophoresis of PCR Product

The integrity of PCR products was checked on 1% agarose gel. This was achieved by dissolving

1g agarose in 100ml 1XTAE (TAE buffer) using a microwave and allowed to cool by passing tube across cold flowing tap. Upon cooling, 1.5µl of ethidium bromide was added to the solution and swirled to mix; the agarose solution was then poured in a gel plate fixed with combs to create wells in the gel. When the gel had solidified, the combs in the gel were carefully removed and the gel plate was immersed into the gel electrophoresis tank containing 1XTris-borate-

EDTA (TAE) buffer. Samples were loaded into well and electrophoresis process was ran as described in 3.3.5.

3.3.8 Cleaning and quantification of PCR product

This was achieved using the Qiagen plant DNAeasy mini kit. Successfully amplified PCR products were transferred into column silica membrane tubes labelled accordingly, 5 times volume of solution B was transferred into same tube and centrifuged at 13,000 rpm for 1min. thereafter, 750ml of Buffer E was transferred into tube and centrifuged at 13,000 rpm for 1min, again centrifugation at 13,000 rpm for 1min was done to allow remnant buffer to be dislodged.

Column silica membrane tubes were inserted in freshly Eppendorf tubes and labelled accordingly and 50ul of elution buffer was added to the membrane tube. It was ensured buffer was released in the middle of the membrane. Tubes were centrifuged at 13,000rpm for 1min after which membrane tubes were discarded while purified DNA has been eluted into the

Eppendorf tube and kept in the freezer at -20ºC for subsequent use. The spectrophotometric analysis of product was checked on a nanodrop as described in 3.3.3., but this time blanking was done with elution buffer. Products were diluted to the required sequencing requirements,

70 | P a g e lids of tubes sealed with a parafilm and samples were sent to the Source Bioscience Cambridge laboratory for bidirectional sequencing using the same primer used in PCR.

3.3.9 Sequence Contiging, Editing and Alignment

Raw sequence data were assembled and edited using SeqMan™ II, one of the programmes of the Lasergene®software package (DNASTAR, Inc.) where forward and reverse sequences were contiged and manually edited should be the case of ambiguities. Consensus file was saved and sequences were verified by a Blastn search on Genbank. The edited sequences were aligned using the multiple alignment Clustal W algorithm (Thompson et al., 1994) as implemented in

MegAlign™ (Lasergene®, DNASTAR, Inc.), with further visual adjustment. This was done so as so form a hypothesis about homology (Lemey et al., 2009) so that sequences with the same ancestral states are put in the same column in all the taxa.

3.4 Data Analysis

3.4.1 DNA Barcoding analysis

Firstly, aligned sequences were trimmed to be of equal lengths to allow comparison within each dataset for each region singly and combined; a total of 10 datasets were used (4 single regions, 5 double regions and 1 combined of all the four regions). The four regions were first concatenated to form a single data (rbcL+matK+trnL-F+ITS), two regions concatenations were also done (rbcL+matk; matk+ITS; rbcL+trnL-F; matK+trnL-F; rbcL+ITS). The different gene concatenations and single region analysis were performed on data by partitioning the characters and running the analysis on the appropriate partition. The variability of each of the barcode region understudied was assessed based on the number of variable and parsimony informative sites, using MEGA 5. All genetic distances were calculated using the Kimura two parameters

(K2P) distance method as implemented in MEGA 5.0 (Tamura et al., 2011). Intra and inter specific genetic distances was calculated for species that are represented with more than one individual. Firstly, to characterize the inter-specific divergence, three parameters were used

71 | P a g e according to Chen et al., 2010; Meyer and Paulay, 2005, Meier et al., 2008: (i) average inter- specific distance (K2P distance) between all species in each genus with at least two species;

(ii) average theta prime (θ′), where theta prime is the mean pairwise distance within each genus with more than one species, thus eliminating biases associated with different numbers of species among genera; and (iii) smallest inter-specific distance, i.e., the minimum inter-specific distance within each genus with at least two species. Likewise, intraspecific distance was characterized by three parameters: (i) average intra-specific difference (K2P distance), that between all samples collected within each species with more than one individual; (ii) theta (θ), where theta is the mean pairwise distance within each species with at least two representatives;

θ eliminates biases associated with unequal sampling among a species; and (iii) average coalescent depth, which is the maximum intra-specific distance within each species with at least two individuals (Chen et al., 2010; Meyer and Paulay, 2005, Meier et al., 2008).

Furthermore, Wilcoxon signed rank tests were performed as described in Lahaye, 2008a; Kress and Erickson, 2007.

To assess the barcoding gap, the distribution of intra-specific versus interspecific variability was assessed. Minimum interspecific distance was plotted against the maximum intraspecific distance as recommended by CBOL.

To estimate the reliability of barcodes regions, several identification techniques have been employed by several authors which has always produced varied results (Meyer and Paulay,

2005; Ross et al., 2008; Austerlitz et al., 2009) but it was however concluded that no single method has been found to be the best. The two most widely used techniques (tree based and sequence comparison) were tested to estimate the resolving power of the regions. The “best match” and “best close match” functions of TAXONDNA (Meier et al., 2006) was used to assess the utility of DNA barcoding for accurate species discrimination. These sequence comparison approach was performed on all the 10 barcode data using uncorrected pairwise

72 | P a g e distance and a minimum 300bp sequence overlap. For the tree based analysis, a Neighbour

Joining and UPGMA tree was estimated with Kimura’s 2-parameter (K2P) (Kimura, 1980) nucleotide evolution model using PAUP (Swofford, 2002), bootstrap replicate of 1,000 was used to access the node support. Maximum parsimony was also conducted in PAUP with 100 random sequence addition and other default parameters, node support was obtained from heuristic searches of 1000 bootstrap replicate. Maximum Bayesian inference was conducted in

Mr Bayes v3.1.2, MrModel test was run to specify the best fit model of evolution for each region with the Akaike Information criterion, the GTR+I+G model was chosen for the each of the four regions in the combined data. Two independent runs with four Markov chains (one cold and three heated) run for 10,000,000 generations and each tree sampled every 1000th generation were performed. The first 2,500,000 samples trees were discarded as ‘burn in’ while the remaining trees were used to build a 50% majority rule consensus tree with posterior probability for nodes.

3.4.2 Phylogenetic analysis

Three data matrices were used for the analysis, all chloroplast regions (rbcL+matk+trnL-F), nuclear region (ITS) and the combined matrix of chloroplast and nuclear genes

(rbcL+matk+trnL-F+ITS). Each of the three matrices was analysed separately using Parsimony and Bayesian methods.

Parsimony analysis was performed using PAUP 4b10 (Swofford, 2002) with nucleotide substitutions equally weighted and unordered. Heuristic search was used with tree-bisection- and-reconnection and random sequence addition. Bootstrap analysis was also performed to test the robustness of each clade with random addition of sequences 1000 replicates.

A Bayesian analysis was carried out by first determining the optimal substitution model using

MrModeltest v2.3 (Nylander, 2004) and the Akaike information criterion. The model selected for the chloroplast region was the general reversible model with a proportion of invariable sites

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(GTR + I) while general reversible model with a gamma shape and a proportion of invariable sites (GTR + I +L) was specified for the nuclear region. Four discrete states were used for the gamma substitution. The data were therefore partitioned into two for the Bayesian analysis and the correct substitution model as specified by MrModel test was specified for each partition.

The partitions were unlinked so that each parameter could be specified separately. Analysis was run for 25,000,000 generations with sampling every 30,000 generations. Metropolis coupling with four chains, one cold and three heated were used with the two independent runs running simultaneously. The runs, however stopped (split standard deviation set at 0.01) after

7705000 generations and 7705 sampled trees in each run. Approximately 24.7% (6,000,000) trees were used as burnin in summarizing the parameters and tree. The robustness of each clade was ascertained based on the posterior probability as an inference on the validity of the tree.

3.5 Conservation Assessment

Species were evaluated according to IUCN criteria and categories of 2014 with subsidiary documentations and distribution for each species. The guidelines of the IUCN Red List

Categories and Criteria were followed without deviation or modification. For national assessments, risk of extinction of such endemic species were assessed following the threshold values which listed under each of the criterion provided as provided in Tables 2 -6. In this study, criterion A or E were not used for any of the species assessed as the first two requirements were not met: the generation length and the population reduction rate in the past, present or future due to the lack of quantitative data and population trend rates. Criterion B was the most used due to data availability: distribution range points (collected from herbarium labels, databases and localities visited) and number of location of the species. Criteria C and D were used for some species with very restricted distribution, small population size and number of mature individuals and the percentage of mature individuals in each subpopulation.

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CHAPTER FOUR RESULTS 4.1 Sample collection

Extensive field surveys in the Sudano-sahelian region of Nigeria yielded a total of three hundred and twenty-four (324) samples of a hundred and eighteen (118) species distributed in twenty-two, thirty-eight and fifty-eight species of subfamilies Mimosoideae, Caesalpinioideae and Papilionoideae respectively as seen in Plates 1-3. Herbarium specimens were deposited at the University of Lagos Herbarium (LUH). Appendix I shows the voucher numbers of specimens deposited in the herbarium.

A B

C D

Plate 1 (A-D): Photographs of some members of the Fabaceae (A=Pods of Tamarindus indica; B=Leaves and inflorescence of Senna italica; C=Leaves of Daniellia oliveri; D= Flowers and foliage of Chamaecrista mimosoides)

Magnification x1.5

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E F

G H

I J

Plate 2 (E-J): Photographs of some members of the Fabaceae (E=Pods of Bauhinia rufescens; F=Foliage of Burkea africana; G=Foliage and leaves of Detarium macrocarpum; H= Pods of Piliostigma reticulatum; I=Pods and leaves of of Piliostigma thonningii; J=Pods and leaves of Isoberlina doka)

Magnification x1.5

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K L

M N

O P

Plate 3 (K-P): Photographs of some members of the Fabaceae (K=Pod and leaves of Isoberlina tomentosa; L=Flowers, pod and leaves of Senna sieberiana; M=Inflorescence and foliage of Leucaena leucocephala; N=Pods of Mucuna pruriens; O=Leaves of Acacia senegal; P=Inflorescence and leaves of Desmodium velutinum) Magnification x1.5

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4.2 Morphological Characterization

Members of Fabaceae occur mostly as trees, shrubs, climbers or herbs; members of the sub- family Mimosoideae are mostly trees, deciduous (Albizia lebbeck) or evergreen tree (Acacia auriculiformis) or small trees or shrubs as in Acacia ataxacantha. However, members are rarely herbs. Among the Caesalpinioideae, members were mostly trees and shrubs with a few herbaceous or sub-shrub species (Chamaecrista nigricans). The Papilionoideae were mostly herbaceous plant with straggling, creeping or climbing habit and shrubs. However, a few tree species were recorded e.g. Dalbergia sissoo. Types of leaves recorded among species ranged from a few unifoliate or simple leaves to mostly compound leaves that are pinnately, bipinnately, imparipinnately, paripinnately or trifoliate. Most mimosoids are bipinnately compound although imparipinnate compound leaf type was encountered in Pithecellobium dulce. Likewise, Acacia auriculiformis has its leaves reduced to a falcate phyllode which were arranged singly on the branch. Among the Caesalpinioideae, leaf type ranged from pinnately compound, bipinnate compound to paripinnate compound; no imparipinnate leaf type was recorded. However, members of the tribe Cercidieae are characterized with simple bilobed leaves. Papilionoids are the most variable in terms of leaf type, members are mostly adorned with trifoliate leaves, while others are either simple, pinnately or imparipinnate compound.

Leaves were arranged mostly alternate with a few opposite while leaflets are usually opposite within the Mimosoideae. The Caesalpinioideae leaves were mostly alternate with a few opposite. Leaves of the Papilionoideae are mostly arranged alternate, rarely opposite; among the species with trifoliate leaves, some are arranged pinnate-trifoliate. Flowers are usually arranged either singly as a globose head or in group with colours ranging from an array of cream to pale pink, pale yellow, bright yellow, golden yellow or reddish yellow; bicolored flowers of yellow and pink was observed in Dichrostachys cinerea. However, some species have white, lilac, purple, blue, purple, green or orange flowers. Inflorescence is usually in form

78 | P a g e of a , panicle, spikes or corymb in some species. Distinct flower types differentiate members of the three sub-families. Flowers of Mimosoideae are usually actinomorphic with showy numerous free stamens, Caesalpinioideae and Papilionoideae has zygomorphic flowers which were also distinguishable, Papilionoids possess distinct papillionaceous flowers in form of a butterfly. The type of fruit is however uniform among members which is in form of a pod but with varying morphological features. Some were oblong, linear, cylindrical, stout, moniliform, falcate, spirals or twisted in different forms and shapes (Plates 1-3).

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4.2.1 Systematic Descriptions

Family: FABACEAE Lindl. (Leguminosae Jussieu, nom. cons.)

Fabaceae Lindl. (1836), nom. cons. Intr. Nat. Syst. Bot. 148. (1836).

Type Genus: Faba Mill.

Sub Family I: Mimosoideae DC.

Mimosoideae DC. Prodr. Sys. Nat. Reg. Veg. 2:424. (1825).

Type Genus: Mimosa L. F.T.A. 2: 335.

Description:

Trees or shrubs; leaves bipinnate and stipulate, stipule may be modified into spines, often falling early; inflorescence cymose head or head; flowers actinomorphic, hermaphrodite, small, tetra or pentamerous; calyx and corolla valvate; petals connate below, stamens number varies from 4 (Mimosa) to many (Acacia, Albizzia); separate or the filaments all united toward base, exceeding the corolla, carpel one and fruit legume.

Tribe I: Acacieae Dumort.

Acacieae Dumort. Anal. Fam.Pl. 40. (1829).

Type: Acacia Mill. F.T.A. 2: 337; Trans. Linn. Soc. 30: 444 (1875).

1. Genus: Acacia Mill.

Acacia Mill.

Type Species: Acacia nilotica (L.) Delile Fl. Aegypt. Illustr. 79. (1813).

Description:

Trees or shrubs with free staminal or shortly united at the base filaments; green

vertically oriented phyllodes, broadened leaf petioles, yellow flowers in rounded head.

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1. Acacia ataxacantha DC. F.T.A. 2: 343

Syn.: Acacia eriadenia Benth.

Acacia lugardiae N.E. Br

Description:

A scandent shrub or small tree, 2-10m tall armed with claw shaped prickles on stem

and branch. Leaves pinnate compound, opposite, with about 24 – 36 pairs of leaflets.

Leaflets are (0.5-0.8) X 0.1cm, sessile, opposite, elliptic with an obtuse or sub-acute

tip. Leaf base cuneate, flat upper and lower midrib, greenish brown under surface with

a papery texture. Leaves are sometimes reduced to phyllode. Flower colour cream to

yellow and inflorescence arranged in spikes or globose head. Pod linear to oblong with

pointed tip.

Habitat: Within forest rregions as well as dry Savanna vegetation

Distribution: West Tropical Africa

Flowering period: December

Specimens examined: Oshingboye 001; Ogundipe & others LUH 4159; LUH 4174;

Onochie FHI 40209; Barret RNG 423

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a b

Plate 4: Acacia ataxacantha, arrow showing (a) claw shaped prickles and pod with pointed tip (b) foliage and inflorescence

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2. Acacia auriculiformis A. Cunn. ex Benth.

Syn: Acacia auriculaeformis Benth.

Acacia moniliformis Griseb.

Racosperma auriculiforme (Benth.) Pedley

Description:

An evergreen tree 15-25m tall, leaves are reduced to phyllode which are falcate and

alternate. Phyllodes are sessile about 17-21 per branch, 0.8-1.9cm wide and 8 – 20cm

long, acute tip and attenuate base, glabrous, flat midrib on both upper and lower surface,

greenish grey under-colour and leathery textured. The inflorescence spike or globose

head, flowers are golden yellow in colour. Pods are twisted in irregular spirals.

Habitat: Moister Savanna regions

Distribution: West Tropical Africa, Papau New Guinea

Flowering period: April - August

Specimens examined: Oshingboye 031; Ogundipe & others LUH 4171; LUH 4159;

LUH 4170

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Plate 5: Acacia auriculiformis leaves reduced to falcate phyllode

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3. Faidherbia albida Del. A. Chev. Rev. Bot. Appliq. 14: 876 (1934).

Syn: Acacia gyrocarpa Hochst.

Acacia leucocephala Benth.

Acacia albida Del.

Description:

One of the largest Acacia trees in Africa up to about 30 m high, leaves are bipinnate

compound, opposite, petiole 1mm. There are about 10-24 pairs of leaflets, opposite, 0.6-

0.9cm long and 0.1-0.2cm wide. Leaflets are oblong with an obtuse or sub-acute tip, obtuse

or round base, greenish grey, flat upper and lower surface, entire margin and papery texture.

Presence of stipules and spines adorned on branches. Inflorescence is panicles with a cream

to white or at times yellow flower. Pods are thick and twisted.

Habitat: Savanna

Distribution: Sudano-Sahelian region of Sub-Saharan Africa, Sudan, Angola

Flowering period: June

Specimens examined: Oshingboye 015; Ogundipe & others LUH 4159; LUH 4172;

Onwudinjoh FHI 24016

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Plate 6: Foliage of Faidherbia albida; arrow show inflorescence

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4. Acacia sieberiana DC. - F.T.A. 2: 847

Syn: Acacia abyssinica sensu auct.

Acacia sieberana DC.

Acacia vermoesenii De Wild.

Description:

A deciduous tree between 12-25m high, armed with spines in pairs at the nodes. Leaves are

pinnate compound, alternate with about 27 – 38 pairs of leaflets, petiole 1mm. Leaflets

oblong, (0.3-0.4) X 0.1cm, sessile, opposite, elliptic with an obtuse or rounded tip, leaf base

obtuse with flat upper and lower midrib, margin entire, green under surface with a papery

texture. Flower colour is cream to white sometimes yellow and inflorescence arranged in a

globose head. Pod linear and slightly falcate.

Habitat: Deciduous woodlands and wooded grasslands

Distribution: West Tropical Africa

Flowering period: April - May

Specimens examined: Oshingboye 017; Ogundipe & others LUH 4150; LUH 5211; Keay

FHI 16251

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Plate 7: Foliage of Acacia sieberiana, arrow showing long pointed thorns

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5. Acacia senegal (L.) Willd. Sp. Pl. 4: 1077 (1806).

Syn: Acacia circummarginata Chiov.

Acacia volkii Suess.

Mimosa senegal L.

Description:

A small deciduous tree or shrub 5 – 7m tall armed with thorns placed just below the

nodes usually in threes. Leaves are pinnate compound, opposite with about 7 – 19 pairs

of leaflets. Leaflets are (1.0-1.9) X (0.1-0.2) cm, sessile, opposite, elliptic or oblong

with an obtuse tip. Leaf base cuneate with flat upper and lower midrib, leaf margin

entire, thickened, greenish grey under surface with a leathery texture. Stipules minute

or absent. Flower colour cream to white and inflorescence arranged in spikes. Pod linear

to oblong with an acute tip.

Habitat: Semi-desert regions of Sub-Saharan Africa

Distribution: Sudano-Sahelian Sub-Saharan Africa

Flowering period: April, July

Specimens examined: Oshingboye 017; Ogundipe & others LUH 4173; LUH 4175;

Barret RNG 386; Keay FHI 16145; Gachathi RNG 92

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Plate 8: Acacia senegal pod, arrow showing paired spines

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6. Acacia nilotica (L.) Delile Fl. Aegypt. Illustr. 79 (1813).

Syn: Acacia arabica (Lam.) Willd.

Mimosa arabica Lam.

Mimosa nilotica L.

Description:

A deciduous tree 25m high armed with thorns in axillary pairs, which is usually lacking

in mature tree. Leaves are bipinnate, alternate with about 9-16 pairs of leaflets. Leaflets

are alternate, greenish grey under surface, 0.8-1.5cm long, 0.1-0.3cm wide, sessile,

elliptical, obtuse or acute tip and obliquely cuneate at the base. Both upper and lower

midrib is flat, margin entire and papery in texture. Inflorescence is a globose head while

flowers are golden yellow in colour, pods are spirals twisted in a distinct neck-lace like.

Habitat: Savanna

Distribution: Dry regions of Sub-Saharan Africa extending throughout the drier parts

of North Africa into Eygpt.

Flowering period: December – January

Specimens examined: Oshingboye 019; Ogundipe & others LUH 4177; Keay FHI

6682; Jackson FHI 14935; Barret RNG 183; Monier RNG 2402; Parker RNG 4685

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Plate 9: Acacia nilotica flowers

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7. Acacia dudgeoni Craib ex Holl. Kew Bull., Add. Ser. 9: 291 (1911)

Syn: Acacia samoryana A. Chev.

Senegalia dudgeoni (Craib ex Holland) Kyal. & Boatwr.

Description:

A small tree or shrub 9 m high armed with prickles 0.3 – 0.6 m and curved inwards.

Leaves are bipinnate, alternate with about 7-10 pairs of leaflets. Leaflets are alternate,

greenish grey under surface, 0.8-1.5cm long, 0.1-0.3cm wide, sessile, elliptical, obtuse

or acute tip and obliquely cuneate at the base. Both upper and lower midrib is flat,

margin entire and papery in texture. Inflorescence is a globose head while flowers are

white, pods are oblong and flattened.

Habitat: Savanna

Distribution: Sudanian and Guinean ecozones

Flowering period: January – February

Specimens examined: Oshingboye 020; Ogundipe & others LUH 4159, LUH 4176;

LUH 5188; Rosevear FHI 26604; Keay FHI 16001

Tribe II: Igneae Benth. & Hook. f.

Igneae Benth. & Hook. f. Gen. Pl. 1: 437. (1865).

Type: Inga Mill. Gard. Dict. Abr. 2. (1754).

2. Genus: Albizia Durazz.

Albizia Durazz. Mag. Tosc. 3(4): 11–14. 1772.

Type Species: Albizia julibrissin Durazz.

Description:

Small trees or shrubs with pinnate or bipinnate compound leaves. Flowers are usually

small with more than 10 stamens in bundles, which are joined at the base stamens and

much longer than the petals. The stamens are usually showy.

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8. Albizia lebbeck Benth. F.T.A. 2: 358

Syn: Acacia lebbeck (L.) Willd.

Albizia latifolia B. Boivin

Inga borbonica Hassk.

Description:

A deciduous tree attaining 30m in height. Leaves pinnate compound, opposite with

about 2 – 11 pairs of leaflets, petiole 1mm. Leaflets are obliquely oblong, (4.1-4.8) X

(2.0-2.7) cm, opposite, obtuse tip, leaf base is obliquely round with a flat upper and

raised lower midrib. Greenish yellow under surface, entire and thickened leaf margin

with a leathery texture. Flowers are white, inflorescence arranged in a capitate head

while pods are flat to oblong.

Habitat: Moister Savanna regions

Distribution: native of Tropical Asia, now widespread in the tropics.

Flowering period: November

Specimens examined: Oshingboye 021; Odewo FHI 96195; Daramola FHI 63867;

Silva RNG 59954; Bisby RNG FB1288; Hatschbach RNG 47340

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Plate 10: Foliage of Albizia lebbeck, arrow showing pod

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9. Albizia zygia (DC.) Macbr. Contrib. Gray Herb. 59: 8 (1919).

Description:

A deciduous tree 9-30m in height. Leaves are pinnate compound, opposite with about

4 – 7 pairs of leaflets that are broadening towards the apex, petiole 10-12mm and

stipulate. Leaflets are obovate to obliquely rhombic, (2.1-5.0) X (1.0-2.2) cm, opposite,

with an acute or obtuse tip, leaf base is obliquely round to cuneate with a flat upper and

raised lower midrib, green to greenish grey under surface, entire and thickened leaf

margin with a leathery texture. Flowers are white or pink, inflorescence arranged in a

capitate head while pods are flat to oblong.

Habitat: Regrowth and fringing forest

Distribution: West Tropical Africa

Flowering period: February - March

Specimens examined: Oshingboye 139; Ogundipe & others LUH 4392; Soladoye FHI

10134: Daramola FHI 63154

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Plate 11: Foliage of Albizia zygia showing pinnate obliquely rhombic leaflets

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3. Genus: Pithecellobium Mart.

Pithecellobium Mart. Flora 20: 114–117. (1837).

Description:

Trees or shrubs with united stamens in a tube, leaves bipinnate and coil or ear

shaped fruit pods.

10. Pithecellobium dulce (Roxb.) Benth.

Syn: Acacia obliquifolia M. Martens & Galeotti

Albizia dulcis (Roxb.) F. Muell.

Inga dulcis (Roxb.) Willd.

Inga javanica DC.

Description:

A small to medium-sized semi-evergreen tree, 15-20m high, armed with spines at the

base of each leaf. Old leaves are shed almost simultaneously as new leaves grows, this

gives the tree an evergreen appearance. Leaves are pinnate compound, opposite with

about 2-3 pairs of leaflets, petiole 8-17mm and stipulate. Leaflets are oblong or

obliquely elliptic, (2.3-2.7) X (1.0-1.4) cm, opposite with an obtuse tip. Leaf base

cuneate, flat upper and raised lower midrib, greenish grey under surface, entire and not

thickened leaf margin with a papery texture. Flowers are white to green, inflorescence

is panicles while pods are spirals.

Habitat: Fringing forest

Distribution: West Tropical Africa and America

Flowering period: February - March

Specimens examined: Oshingboye 025; Lekan FHI 105251; Mallam FHI 31773; Parker

RNG 4687

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Plate 12: Pithecellobium dulce leaves, arrow showing spiral pods

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4. Genus: Prosopis L.

Prosopis L. F.T.A. 2: 331; Benth. in Trans. Linn. Soc. 30: 376 (1875).

Type Species: Prosopis spicigera L.

Description:

Spiny trees or shrubs, leaves bipinnate with small flowers in axillary cylindrical spikes,

calyx slightly pubescent; petals glabrous; and 10 free stamens.

11. Prosopis africana Taubert. E. & P. Pflanzenfam. 3, 3: 119 (1892).

Description:

A big tree reaching up to 20m high. Leaves are pinnate compound, alternate with about

8-14 pairs of leaflets, petiole 6-7mm. Leaflets oblong or lanceolate (1.5-1.6) X (0.5-

1.0) cm, opposite, acute tip, leaf base cuneate with a flat upper and slightly raised lower

midrib, greenish brown under surface, entire and thickened leaf margin with a leathery

texture. Flowers are greenish white to yellow, inflorescence is a spike while pods are

stout and cylindrical.

Habitat: Savanna

Distribution: West Tropical Africa, Tropical America

Flowering period: April - May

Specimens examined: Oshingboye 029; Ogundipe & others LUH 4194; Keay FHI

16179

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Plate 13: Prosopis africana pinnate leaves

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Tribe III: Mimoseae Bronn.

Mimoseae Bronn. Integr. Syst. Class. (1981).

Type: Mimosa L. F.T.A. 2: 335

5. Genus: Dichrostachys (DC.) Wight & Arn.

Dichrostachys (DC.) Wight & Arn. Prodr. Fl. Ind. Orient. 1:271. (1834).

Description:

Shrubs or small trees with spines terminating short lateral branchlet, leaves

bipinnate. Inflorescence are axillary spikes, composed of bisexual

flowers, yellow, upper part broader, neuter, mauve or pink or sometimes white.

Calyx short 5-toothed, petals 5 and 10 stamens.

12. Dichrostachys cinerea Wight et Arn. Ann. Bot. Rom. 13: 409 (1915).

Syn: Cailliea glomerata (Forssk.) J.F. Macbr.

Dichrostachys glomerata (Forssk.) Chiov.

Dichrostachys platycarpa Welw.

Mimosa cinerea L.

Mimosa glomerata Forssk.

Description:

A semi deciduous to deciduous small tree or thorny shrub up to 7m tall, thorns are

alternate and almost at right angles. Leaves are pinnate compound opposite, petiole 5-

7mm with about 14 – 25 pairs of leaflets. Leaflets are (0.3-0.5) X 0.1cm, sessile,

opposite, elliptic or oblong with an obtuse tip. Leaf base is round with a flat upper and

lower surface midrib, leaf margin entire and not thickened, greenish brown on under

surface with a papery texture. Flowers are bicolored with pink upper and yellow lower,

inflorescence arranged in spikes. Pod are spirals and tightly twisted.

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Habitat: Savanna and distrurbed scrublands

Distribution: West Tropical Africa, parts of South Africa

Flowering period: April

Specimens examined: Oshingboye 030; Ogundipe & others LUH 4139; Teniife RNG

SFS02 Barret RNG 22116

Plate 14: Dichrostachys cinerea leaves, arrow showing tightly twisted pod

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6. Genus: Entada Adans.

Entada Adans. Fam. Pl. 2: 318, 554. (1763).

Description:

Trees or shrubs, leaves bipinnate. Inflorescences of spikes, axillary or supra-

axillary, solitary or clustered. Flowers bisexual or unisexual. Calyx gamosepalous,

with 5 teeth, petals 5 and 10 stamens. Pod with valves splitting transversely into 1-

seeded segments, the segments leaving a persistent empty frame.

13. Entada abyssinica Steudel ex A. Rich - F.T.A. 2: 327; Aubrév. l.c. 292.

Syn: Entadopsis abyssinica (A. Rich.) G.C.C. Gilbert & Boutique

Description:

A small to medium sized deciduous tree 3-15m tall. Leaves are pinnate compound,

opposite, petiole 10-18mm with about 8 – 25 pairs of leaflets. Leaflets are (0.9-1.3) X

0.3cm, opposite, oblong or linear with an obtuse to slightly mucronate tip, leaf base is

obtuse to round to cuneate with a flat upper and lower midrib, leaf margin entire and

slightly thickened, green or greenish yellow on under surface with a leathery texture.

Flowers are cream to white and inflorescence arranged in raceme. Pod are straight and

flat.

Habitat: Savanna

Distribution: West Tropical Africa

Flowering period: March - June

Specimens examined: Oshingboye 033; Ogundipe & others LUH 5179; Keay FHI

28340; Lungu RNG 12; RNG 8

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Plate 15: Entada abyssinica pinnate leaves

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14. Entada africana Guill & Perr. F.T.A. 326

Syn: Entada sudanica Schweinf

Entadopsis sudanica (Schweinf.) G.C.C. Gilbert & Boutique

Description:

A small to medium sized deciduous tree 3-15m tall. Leaves are pinnate compound,

opposite, petiole 10-18mm with about 8 – 25 pairs of leaflets. Leaflets are (0.9-1.3) X

0.3cm, opposite, oblong or linear with an obtuse to slightly mucronate tip, leaf base is

obtuse to round to cuneate with a flat upper and lower midrib, leaf margin entire and

slightly thickened, green or greenish yellow on under surface with a leathery texture.

Flowers are cream to white and inflorescence arranged in raceme. Pod are straight and

flat.

Habitat: Savanna

Distribution: West Tropical Africa

Flowering period: April - May

Specimens examined: Oshingboye 034; Ogundipe & others LUH 4146; Keay FHI

16123; Olorunfemi FHI 24413

7. Genus: Leucaena Benth.

Leucaena Benth. J. Bot. 4: 416–417. (1842).

Type Species: Leucaena glauca Benth. Nom. 2: 94 (1872).

15. Leucaena leucocephala (Lam.) de Wit F.T.A. 2: 337; Benth. l.c. 443

Syn: Acacia frondosa Willd.

Acacia glauca (L.) Willd.

Acacia leucocephala (Lam.) Link

Leucaena glauca (Willd.) Benth.

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Description:

Leucaena leucocephala is a small highly branched shrub or a small evergreen tree high up to 10m. Leaves are bipinnate compound, opposite with about 18–29 pairs of leaflets, petiole 1-2mm, stipules present. Leaflets are (0.5-0.8) X 0.2cm, sessile, opposite, elliptic or oblong with an acute tip, leaf base cuneate with a flat upper and lower surface midrib. Leaf margin entire and not thickened, greenish brown under surface with a papery texture. Flower colour is cream to white or yellow with inflorescence arranged in globose head. Pod linear and flat.

Habitat: Road-side

Distribution: Native of America; now common in the warmer parts

Flowering period: April, October - November

Specimens examined: Oshingboye 035; Ogundipe & others LUH 4332; LUH 1709;

Ibhanesebhor FHI 31483; Ekwuno FHI 92681; Parker RNG 2678; Hansen RNG 1970;

Bisby RNG 1199; Dremman RNG 10; Larimer RNG 28

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Plate 16: Leucaena leucocephala bipinnate leaves, arrow showing globose inflorescence

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8. Genus: Mimosa L.

Mimosa L. F.T.A. 2: 335.

Description:

Herbs or shrubs; stems armed with prickles. Leaves bipinnate and sensitive.

Inflorescences of ovoid or subspherical heads which are axillary or arranged in

clusters. Flowers bisexual or male. Calyx very small. Corolla gamopetalous.

Pods flat, bristly or prickly and splitting transversely into 1-seeded segments.

16. Mimosa pigra L. - Cent. Pl. 1: 13 (1755).

Syn: Mimosa pellita Humb. & Bonpl. ex Willd.

Description:

Mimosa pigra is a branched prickly shrub up to 6m high, armed with prickles on both

stem and leaves. Leaves are bipinnate compound, alternate, with about 13-27 pairs of

leaflets, petiole 5-7mm. Leaflets are alternate, with rusty under surface, 0.3-0.5cm long,

0.1cm wide, sessile, linear or oblong-linear, acuminate tip and truncate base. Both upper

and lower midrib is flat, margin entire and not thickened with a papery texture.

Inflorescence is a globose head while flowers are pink in colour. Pods are flat and

oblong-clustered.

Habitat: Sudanian freshwater swamp and aquatic vegetation

Distribution: Tropical Africa

Flowering period: June - September

Specimens examined: Oshingboye 035; Ogundipe & others LUH 4158; Witt and

Daramola FHI 78387: Daramola FHI 92125: Latilo FHI 61412: Olorunfemi and

Onijamowo FHI 67552; Parker RNG 1212; Sulaimon RNG SFS12; Kaison RNG 19;

RNG 98

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Plate 17: Mimosa pigra bipinnate leaves, arrow showing oblong clustered pod

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17. Mimosa pudica L. - F.T.A. 2: 336

Syn: Mimosa hispidula Kunth

Description:

Mimosa puduca is a low much branched perennial shrub or herb 2m in height, armed

with prickles on both stem and leaves. Leaves are bipinnate compound, alternate, with

about 13-27 pairs of leaflets, petiole 5-7mm. Leaflets are alternate with rusty under

surface, 0.3-0.5cm long, 0.1cm wide, sessile, linear or oblong-linear, acuminate tip and

truncate base. Both upper and lower midrib is flat, margin entire and not thickened with

a papery texture. Inflorescence is a globose head while flowers are pink or lilac in

colour, pods are flat and oblong-clustered.

Habitat: Open waste ground

Distribution: Tropical America and Africa

Flowering period: May - August

Specimens examined: Oshingboye 036; Ogundipe & others 4161; Witt FHI 27917;

Cheung RNG 16; Prker RNG 7142; Webb RNG 67; Bisby RNG FB1298

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Plate 18: Mimosa pudica bipinnate leaves

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9. Genus: Parkia R.Br.

Parkia R.Br. F.T.A. 2: 323.

18. Parkia biglobosa (Jacq.) R. Br. ex G. Don f - F.T.A. 2: 324

Syn: Mimosa biglobosa Jacq.

Description:

A deciduous tree 7-20m in height. Leaves are pinnate compound, alternate with about

20 – 32 pairs of leaflets, petiole 5mm and non-stipulate. Leaflets are oblong, (1.8-2.1)

X (0.5-0.6) cm, opposite with an obtuse tip, leaf base round with flat upper and raised

lower midrib, greenish brown under surface, leaf margin entire and thickened with a

leathery texture. Flowers are orange or red, inflorescence is raceme while pods are

linear to oblong.

Habitat: Savanna

Distribution: West Tropical Africa

Flowering period: March and June

Specimens examined: Oshingboye 037; Ogundipe & others LUH 4191; Keay FHI

16205; Pilz FHI 99265

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Plate 19: Parkia biglobosa pinnate oblong leaves

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Sub Family II: Caesalpinioideae DC.

Caesalpinioideae DC. Prodr. Sys. Nat. Reg. Veg. 2:473. (1825).

Type Genus: Caesalpinia L. Sp. Pl. 1: 380. (1753).

Description:

Trees or shrubs, less often climbers or herbs. Leaves are usually pinnate, rarely simple

or bipinnate. Inflorescences usually spikes or panicles of racemes, rarely of spikes or

capitate. Flowers are zygomorphic, sepals usually free; petals imbricate in bud, usually

with the dorsal within and overlapped by the lateral, free or sometimes connate below,

usually 5, sometimes reduced to 1 or 0. Stamens usually 10 or fewer (rarely numerous).

Pods various and seeds generally without areoles.

Tribe I: Cercidieae Bronn.

Cercidieae Bronn. Form. Pl. Legumin. 131. (1822).

Type: Cercis L. Sp. Pl. 1: 374. (1753).

10. Genus: Bauhinia L.

Bauhinia L. Sp. Pl. 1: 374 (1753).

Type Species: Bauhinia divaricata L. Sp. Pl. 1: 374. (1753).

Description:

Shrubs or small trees, sometimes scrambling or climbing. Leaves alternate, simple, and

conspicuously 2-lobed. Flowers usually large and showy, bisexual, in short racemes or

solitary. Pods oblong-linear, woody, dehiscent with flat seeds.

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19. Bauhinia monandra Kurz.

Syn: Bauhinia kappleri Sagot

Bauhinia porosa Baill.

Bauhinia punctiflora Baker

Description:

A small fast-growing evergreen tree or shrub 10.5m high. Leaves are orbicular, bilobed

split up to one third or half its length with obtuse or rounded lobe and arranged in

alternate. Leaf length ranged between 5.4-12.5cm long and 6.6-12.1cm wide, petiole

23-53mm, leaf base sub-cordate to round. Greenish grey under surface, flat upper and

slightly raised lower midrib, margin bilobed and thickened with a leathery texture.

Inflorescence is a corymb with pale pink flowers, pods are flat and slightly inflated.

Habitat: Grasslands and riparian areas

Distribution: West Tropical Africa

Flowering period: October – January

Specimens examined: Oshingboye 038; Witt FHI 66874; Mbadugba and Molokwu

FHI 94990; Gbile and Daramola FHI 73641

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Plate 20: Bauhinia monandra obbicular bilobed leaves, arrowing showing flowers

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20. Bauhinia purpurea L. - Sp. Pl. 1: 375. (1753).

Syn: Bauhinia castrata

Description:

A medium-sized deciduous tree or shrub 10-12m high. Leaves are orbicular, deeply

bilobed with obtuse lobes and alternate. Leaf length ranged between 5.5-8.0cm long

and 7.5-9.1cm wide, petiole 16-32mm, leaf base cordate to round. Greenish grey under

surface, flat upper and raised lower midrib, margin bilobed and thickened with a

leathery texture. Inflorescence is axillary panicles with lavender, purple or blue flowers,

pods are flat.

Habitat: Mountane forest, savanna, scrublands and dry deciduous foretss

Distribution: Tropical and sub-Tropical climates

Flowering period: September - November

Specimens examined: Oshingboye 039; Ogundipe & others LU 4139; LUH 6646

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Plate 21: Bauhinia purpurea orbicular bilobed leaves, arrow showing flowers

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21. Bauhinia rufescens Lam. F.T.A. 2: 289

Syn: Adenolobus rufescens (Lam.) A. Schmitz

Bauhinia adansoniana Guill. & Perr.

Piliostigma rufescens (Lam.) Benth.

Description:

A shrub 3m high. Leaves are small, orbicular and deeply bilobed up to the base with

obtuse lobes and arranged in alternate. Leaf length ranged between 1.5-2.1cm long and

1.9-2.3cm wide, petiole 18-30mm, leaf base cordate to round. Greenish grey under

surface, flat upper and lower midrib, margin bilobed and not thickened with a papery

texture. Inflorescence in racemes with greenish yellow to white flowers, pods are

narrow and twisted.

Habitat: Drier savannna

Distribution: Sudano-sahelian ecozones

Flowering period: December - January

Specimens examined: Oshingboye 040; Ogundipe & others LUH 5124; Edith FHI

106137

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Plate 22: Bauhinia rufescens orbicular bilobed leaves, arrow showing narrow, twisted pods

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22. Bauhinia tomentosa L. - Sp. Pl. 1: 375. (1753).

Syn: Bauhinia pubescens DC.

Description:

A scrambling shrub up to 4m. Leaves are orbicular, bilobed up to half of leaf blade with

oval to elliptic lobes and alternate. Leaf length ranged between 4.2-5.7cm long and 3.5-

6.5cm wide, petiole 20-35mm, leaf base truncate or slightly cordate. Greenish yellow

under surface, flat upper and lower midrib, margin bilobed and not thickened with a

papery texture. Inflorescence in racemes with yellow flowers, pods are narrow and

slender.

Habitat: Woodland

Distribution: Tropical Africa and Asia

Flowering period: December - March

Specimens examined: Oshingboye 041; Ogundipe& others 5167; Gbile FHI 2716;

Gbile FHI 224454

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Plate 23: Bauhinia tomentosa orbicular bilobed leaves

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23. Bauhinia vahlii Wight and Arn.

Syn: Bauhinia racemosa Vahl

Description:

A climber up to 20m in full length. Leaves are orbicular, bilobed with a broad cut and

obtuse lobes arranged in alternate. Leaf length ranged between 11.4-12.0cm long and

10.3-15.1cm wide, petiole 35-40mm, leaf base cordate. Greenish grey under surface,

flat upper and raised lower midrib, margin bilobed and thickened with a leathery

texture. Inflorescence in corymiform cyme with cream to white flowers, pods are flat.

Habitat: Climax monsoon-deciduous forests

Distribution: Eastern Asia

Flowering period: April - June

Specimens examined: Oshingboye 042; Ogundipe & others LUH 4392

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Plate 24: Bauhinia vahlii orbicular bilobed leaves, arrow showing flowers

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11. Genus: Piliostigma Hochst.

Piliostigma Hochst. Flora 29: 598. (1846)

Type Speices: Piliostigma reticulatum (DC.) Hochst. Flora 29: 599. (1846).

Description:

Trees without tendrils, leaves simple, and conspicuously 2-lobed with rust-coloured

indumentum beneath. Flowers medium to small, unisexual in racemes or

panicles. Pods indehiscent, linear to oblong, woody, and many-seeded.

24. Piliostigma reticulatum (DC.) Hochst. Hook. Ic. Pl. sub. t. 3460. (1846).

Syn: Bauhinia reticulatum

Bauhinia reticulata DC.

Bauhinia glabra A. Chev.

Bauhinia glauca A. Chev.

Description:

An evergreen shrub or small tree 9m. Leaves are alternate, obcordate, bilobed with

obtuse lobes. Leaf length ranged between 7.0-8.1cm long and 8.1-9.0cm wide, petiole

28-35mm, leaf base round. Greenish grey under surface, flat upper and slightly raised

lower midrib, margin bilobed and thickened with a leathery texture. Inflorescence in

axillary raceme with white with pink stripes flowers, pods are hard, long and straight.

Habitat: drier savanna regions

Distribution: Sudano-sahelian region, Cameroons, Ubangi-Shari and Sudan

Flowering period: December

Specimens examined: Oshingboye 043; Ogundipe & others LUH 4262; Keay FHI

22010; Binuyo FHI 43514

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Plate 25: Piliostigma reticulatum obcordate bilobed leaves

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25. Piliostigma thonningii (Schum.) Milne-Redh. Hook. Ic. Pl. 35: 3460 (1947).

Syn: Bauhinia thonningii Schum.

Description:

A deciduous small tree or shrub 6m. Leaves are alternate, obcordate, and bilobed, leaf

length ranged between 9.6-11.2cm long and 9.0-12.0cm wide, petiole 35-40mm, leaf

base cordate and tip emarginate. Greenish brown under surface, flat upper and raised

lower midrib, margin bilobed and thickened with a leathery texture. Inflorescence in

axillary raceme with white flowers, pods are oblong, short and pedicellate.

Habitat: Savanna ecozone

Distribution: Tropical Africa from Abyssinia to Transvaal.

Flowering period: August - October

Specimens examined: Oshingboye 044; Ogundipe & others LUH 5181; LUH 4216;

LUH 4388; Latilo FHI 46144

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Plate 26: Piliostigma thonningii obcordate bilobed leaves, arrow showing pod

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Tribe II: Detarieae. DC.

Detarieae. DC. Cand. Aug. Pyr. (1825).

Type: Detarium Juss. F.T.A. 2: 312.

12. Genus: Afzelia Sm.

Afzelia Sm. F.T.A. 2: 301; Léonard in Reinwardtia 1: 61–66 (1950).

Description:

Trees with paripinnate leaves; leaflets subequal, mostly in 4−6 pairs, with distinct

petiolules; flowers with enlarged red dorsal petal; pod thick, woody; seeds black cupped

at base by the red or orange aril, and embedded in white pithy endosperm.

26. Smith ex Pers. F.T.A. 2: 302

Syn: Pahudia africana Pers.

Description:

Afzelia africana is a large deciduous tree, 10-18m high. Leaves are paripinnate

compound, alternate, 6-30cm in length with about 4-11 pairs of leaflets, petiole 8mm.

Leaflets opposite, green to greenish yellow under surface, 6.3-9.6cm long 3.4-5.0cm

wide, elliptic or ovate, obtuse tip and cuneate base. Flat upper and raised lower midrib,

margin entire and thickened with a leathery texture. Inflorescence is a paniculiform

cyme with whitish to yellow flowers, pods are flat.

Habitat: Savanna and fringing forest

Distribution: West Tropical Africa, Ubangi-Shari, French Cameroons, A.-E. Sudan,

Uganda, Belgian Congo and Tanganyika

Flowering period: March - April

Specimens examined: Oshingboye 045; Ogundipe & others LUH 4308; Osaji and

Okafor FHI 060348; Gbile FHI 80072

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Plate 27: Afzelia africana paripinnate leaves

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13. Genus: Cynometra L.

Cynometra L. Sp. Pl. 1: 382. (1753).

Type Speices: Cynometra cauliflora L. Scient. Surv. Porto Rico 363 (1926).

27. Cynometra megalophylla Harms Engl. Bot. Jahrb. 26: 262 (1899).

Description:

A medium sized tree up to 12m. Leaves are alternate, 10.1-16.7cm long, paripinnate

compound with about 3-4 pairs of leaflets per pinna. Leaflets obovate (5.7-7.5) X (2.2-

3.2) cm, opposite with an acuminate tip, leaf base oblique. Greenish grey under surface,

flat upper and lower surface midrib, margin entire and thickened with a leathery texture.

Inflorescence is raceme with off white to cream flowers, pods are corky.

Habitat: Riverine and forest

Distribution: West Tropical Africa

Flowering period: May and June

Specimens examined: Oshingboye 047; Ogundipe & others 4187; Ejiofor and Latilo

FHI 32031

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a b

Plate 28: Cynometra megalophylla (a) showing paripinnate leaves (b) leaves, arrow showing corky pod

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14. Genus: Daniellia Benn.

Daniellia Benn. F.T.A. 2: 299.

Type Species: Daniellia thurifera Benn.

28. Daniellia oliveri (Rolfe) Hutch. & Dalziel. F.W.T.A., ed. 1, 1: 341 (1928).

Syn: Paradaniellia oliveri Rolfe

Description:

A deciduous tree up to 25m. Leaves are alternate, up to 32cm long, paripinnate

compound with about 6-9 pairs of leaflets per pinna, petiole 39-45mm, stipules present.

Leaflets oblong to ovate (12.3-15.3) X (5.8-7.5) cm, opposite, acuminate tip, and

oblique leaf base. Greenish grey under surface, flat upper and raised lower midrib,

margin entire and thickened with a leathery texture. Inflorescence is raceme with off-

white to cream flowers, pods are samara-like.

Habitat: Savanna

Distribution: Sudano-sahelian region, French Cameroons, Ubangi-Shari, and Angola

Flowering period: December

Specimens examined: Oshingboye 048; Ogundipe & others LUH 5195; LUH 2784:

Daramola FHI 44886: Pilz FHI 99257: Olorunfemi FHI 88321: Emwiogbn FHI 61776

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Plate 29: Daniellia oliveri paripinnate leaves

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15. Genus: Detarium Juss

Detarium Juss. F.T.A. 2: 312.

29. Detarium macrocapum Harms Engl. Bot. Jahrb. 30: 78 & fig. (1901).

Description:

A medium sized to large deciduous tree up to 50m. Leaves are alternate, 16.1-20.5cm

long, paripinnate compound with about 3-4 pairs of leaflets per pinna, petiole 20-25mm.

Leaflets ovate (9.0-12.4) X (3.2-4.5) cm, alternate, acute tip, and oblique leaf base.

Brown to dark brown under surface, channelled upper and raised lower layer midrib,

margin entire and thickened with a leathery texture. Inflorescence is panicle with off-

white to cream flowers, pods are ovoid.

Habitat: Drtier parts of Savanna

Distribution: West Tropical Africa

Flowering period: December - January

Specimens examined: Oshingboye 049; Ogundipe & others LUH 4140; LUH 4246;

LUH 4308; LUH 4390; Daramola FHI 81205

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Plate 30: Detarium macrocarpum paripinnate leaves

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16. Genus: Craib & Stapf ex Holland

Isoberlinia Craib & Stapf ex Holland Bull. Misc. Inform. Ser. 9: 266. (1911).

30. Isoberlinia tomentosa (Harms) Craib & Stapf Kew Bull. Add. Ser. 9: 267 (1911).

Syn: dalzielii (Craib & Stapf) Baker f.

Description:

A medium sized tree up to 18m. Leaves are alternate, 20.1-29.5cm long, paripinnate

compound with about 3-4 pairs of leaflets per pinna, petiole 30-38mm. Leaflets are

ovate to elliptical (9.3-15.0) X (4.5-6.2) cm, opposite, acute or short acuminate tip, and

rounded to oblique leaf base. Brown under surface, channelled upper and raised lower

midrib, margin slightly undulate and thickened with a leathery texture. Inflorescence is

an axillary panicle with white to pink flowers, pods are flat and elongated.

Habitat: Savanna

Distribution: Sudano-sahelian ecoregions, French Cameroons, Ubangi-Shari.

Flowering period: April

Specimens examined: Oshingboye 050; Chapman FHI 56078: Macgregror FHI 1449:

Latilo FHI 34435: Ejiofor FHI 19841

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Plate 31: Isoberlina tomentosa paripinnate leaves

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31. Isoberlinia doka Craib & Stapf Kew Bull. Add. Ser. 9: 267 (1911).

Syn: Berlinia doka (Craib & Stapf) Bak. f.

Berlinia kerstingii Harms.

Description:

A medium sized tree up to 18 – 20 m. Leaves are alternate, 20.1-29.5cm long,

paripinnate compound with about 3-4 pairs of leaflets per pinna, petiole 60-65mm.

Leaflets are ovate to elliptical (6.3-20.0) X (3.5-9.5) cm, opposite, obtuse or short

acuminate tip, and rounded to slightly round leaf base. Brown under surface,

channelled upper and raised lower midrib, margin slightly undulate and thickened with

a leathery texture. Inflorescence is an axillary panicle with white flowers, and pods

oblong.

Habitat: Savanna woodland

Distribution: Sudano-sahelian ecoregions, French Cameroons, Ubangi-Shari.

Flowering period: April - May

Specimens examined: Oshingboye 051; Ogundipe & others LUH 1326A; Chapman

FHI 56078: Macgregror FHI 1449: Latilo FHI 34435: Ejiofor FHI 19841

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17. Genus: Tamarindus L.

Tamarindus L. F.T.A. 2: 307.

Type Species: Tamarindus indica L.

32. Tamarindus indica L. F.T.A. 2: 308

Description:

An evergreen tree up to 30 m high. Leaves are alternate, 20.1-29.5cm long, paripinnate

compound with about 10-18 pairs of leaflets per pinna, petiole 30-38mm. Leaflets are

narrowly oblong (10.8-25.1) X (3.5-9.8) cm, opposite, round tip, and rounded to oblique

leaf base. Brown under surface, channelled upper and raised lower midrib, margin

entire and slightly thickened with a leathery texture. Inflorescence is spike with pale

yellow to pink flowers, pods are subcylindrical and elongated.

Habitat: Savanna

Distribution: Sudano-sahelian ecoregions, French Cameroons, Ubangi-Shari.

Flowering period: January, June

Specimens examined: Oshingboye 052; Ogundipe & others LUH 4303; LUH 4134;

LUH 4370; Latilo and Odewo FHI 69403

Tribe III: Caesalpinieae Rchb.

Caesalpinieae Rchb. Fl. Germ. Excurs. 2(2): 544. (1832).

Type: Caesalpinia L. Sp. Pl. 1: 380. (1753).

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18. Genus: Burkea Hook.

Burkea Hook. F.T.A. 2: 319.

Description:

Unarmed trees with rusty-tomentose young . Leaves bipinnate with alternate

leaflets. Inflorescence of small flowers in an elongated, pendulous, catkin-like spike,

usually aggregated into a panicle. Pod indehiscent, elliptic, flattened, and coriaceous.

33. Burkea africana Hook. f. F.T.A. 2: 320

Description:

Burkea africana is a medium sized tree up to 20m high. Leaves are bipinnate

compound, opposite with about 9-15 pairs of leaflets per pinna, petiole 13-18mm,

presence of minute stipules. Leaflets are elliptic or ovate (3.0-4.8) X (1.8-2.2) cm,

alternate with an obtuse or emarginate tip. Leaf base is round or asymmetrical with a

flat midrib upper and lower surface, greenish brown under surface, entire and thickened

leaf margin with a leathery texture. Flowers are cream to white, inflorescence is an

elongated spike while pods are flat.

Habitat: Woodland and wooded grassland

Distribution: Tropical Affica

Flowering period: December - March

Specimens examined: Oshingboye 053; Ogundipe & others LUH 4483; Keay FHI

19261

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Plate 32: Burkea africana bipinnaate leaves

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19. Genus: Caesalpinia L.

Caesalpinia L. F.T.A. 2: 262.

Description:

Trees or climbing or sprawling shrubs, unarmed or bearing prickles. Leaves bipinnate

with opposite leaflets. Inflorescence of medium to large flowers arranged in a raceme

or panicle. Pod not winged.

34. Caesalpinia pulcherrima (L.) Sw. F.W.T.A 1: 481

Syn: Poinciana pulcherrima Linn.

Poinciana bijuga Lour.

Poinciana elata Lour.

Description:

Caesalpinia pulcherrima is an evergreen low-branching shrub up to 20m high. Leaves

are bipinnate compound, opposite with about 6-18 pairs of leaflets per pinna, petiole

32-54mm. Leaflets are oblong or ovate (1.5-1.8) X (0.7-1.1) cm, opposite with an

obtuse tip. Leaf base is round or asymmetrical, flat upper and lower midrib, greenish

yellow under surface, entire and not-thickened leaf margin and papery texture. Flowers

are large yellow, orange or red, inflorescence is a corymb while pods are flat.

Habitat: Dry wastelands and roadside

Distribution: introduced to the tropics and subtropics

Flowering period: throughout the year

Specimens examined: Oshingboye 054; Felix FHI 107134; Silva RNG 51664

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Plate 33: Caesalpinia pulcherrima bipinnate leaves, arrow showing flowers

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20. Genus: Delonix Raf.

Delonix Raf. Fl. Tellur. 2: 92 (1837).

Type Species: Delonix regia (Bojer ex Hook.) Raf.

Description:

Unarmed trees with imparipinnate or bipinnate leaves. Inflorescences of all species

arranged in axillary raceme. Pods are large, linear- oblong, somewhat flattened, woody

and pendulous, containing many ellipsoidal, pale brown, finely motley seeds in

individual chamber.

35. Delonix regia (Bojer ex Hook.) Raf. F.T.A 2: 266.

Syn: Delonix regia Hook. var. flavida Stehle

Delonix regia Hook. var. genuina Stehle

Poinciana regia Hook.

Description:

A medium sized deciduous tree up to 15m. Leaves are opposite, up to 28.9-35.1cm

long, bipinnate compound with about 19-28 pairs of leaflets per pinna, petiole 20-

35mm, stipules present. Leaflets are oblong to ovate (4.3-7.3) X (0.2-0.3) cm, opposite,

obtuse tip, and cuneate leaf base. Greenish yellow under surface, flat upper and lower

midrib, margin entire and not thickened with a papery texture. Inflorescence is corymb

with red flowers, pods are flattened.

Habitat: dry deciduous forest but now widely cultivated

Distribution: native of Madagascar but now grown as ornamental in the tropics

Flowering period: February - June

Specimens examined: Oshingboye 055; Daramola FHI 060752: Witt FHI 64396

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Plate 34: Delonix regia bipinnate leaves

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Tribe III: Cassieae Bronn.

Cassieae Bronn. Form. Pl. Legumin. 78: 127. (1822).

Type: Cassia L. F.T.A. 2: 268; Benth. in Trans. Linn. Soc. 27: 503–591 (1871).

21. Genus: Cassia L.

Cassia L. F.T.A. 2: 268; Benth. in Trans. Linn. Soc. 27: 503–591 (1871).

Type Species: Cassia fistula Taxon 37: 975 (1988).

Description:

Unarmed trees or shrubs with paripinnate leaves. Flowers in many-flowered racemes;

bracteoles 2 at base of pedicels. Pod long, cylindric or elongated.

36. Cassia arereh Del. F.T.A. 2: 270.

Description:

A small to medium sized tree 10m high. Leaves are alternate, 16.2-26.2cm long,

bipinnate compound with about 5-8 pairs of leaflets per pinna, petiole 20-33mm.

Leaflets are elliptic (4.2-5.2) X (2.6-2.8) cm, opposite with an acute tip, leaf base

cuneate. Greenish yellow under surface, channeled upper and raised lower midrib,

margin entire and thickened with a leathery texture. Inflorescence is corymb with bright

yellow flowers, pods are narrow and longitudinal.

Habitat: Savanna

Distribution: Sudano-sahelian regions

Flowering period: February - April

Specimens examined: Oshingboye 056; Kennedy FHI 8052

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Plate 35: Cassia arereh bipinnate leaves

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37. Cassia mannii Oliv. F.T.A. 2: 272 (1871).

Description:

A tree up to 25m high. Leaves are alternate, 16.2-26.2cm long, bipinnate compound

with about 5-12 pairs of leaflets per pinna, petiole 20-33mm. Leaflets are ovate to

elliptic or oblong to elliptic (3.5-8.2) X (2.2-4.8) cm, opposite with an acute to obtuse

or obtusely acuminate tip, leaf base cuneate. Greenish yellow under surface, flat upper

and lower midrib, margin entire and thickened with a leathery texture. Inflorescence is

raceme with pink or white flowers, pods are cylindrical.

Habitat: Savanna

Distribution: West Tropical Africa

Flowering period: November - December

Specimens examined: Oshingboye 057; Ogundipe & others 5131;

38. Cassia sieberiana DC. F.T.A. 2: 270.

Syn: Cassia kotschyana Oliv. F.T.A. 2: 271 (1871).

Description:

A deciduous shrub or small tree up to 12m high. Leaves are alternate, paripinnate

compound with about 5-14 pairs of leaflets per pinna, petiole 20-33mm, stipules

present. Leaflets are elliptic to ovate (3.5-10.1) X (2.0-4.8) cm, opposite with an acute

to round tip, leaf base cuneate. Greenish yellow under surface, flat upper and lower

midrib, margin entire and thickened with a leathery texture. Inflorescence is axillary

raceme with bright yellow flowers, pods are cylindrical.

Habitat: Savanna

Distribution: Sudano-sahelian regions

Flowering period: February - May

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Specimens examined: Oshingboye 058; Ogundipe & others LUH 4248; LUH 1323A;

Obaseki FHI 27911

39. Cassia italica (Mill.) Lam. ex F. W. Andr. Fl. Pl. A.-E. Sud. 2: 117. (1952).

Syn: Cassia italica (Mill.) Spreng.

Senna italica Mill.

Description:

A small deciduous perennial herb 6m high. Leaves are alternate, pinnate compound

with about 4-6 pairs of leaflets per pinna, petiole 3-9mm. Leaflets are elliptic (4.2-5.2)

X (2.6-2.8) cm, opposite with an acute tip, leaf base cuneate. Greenish yellow under

surface, flat upper and lower midrib, margin entire and not thickened with a papery

texture. Inflorescence is axillary raceme with yellow or orange flowers, pods are oblong

or ellipsoidal.

Habitat: Savanna

Distribution: Sudano-sahelian regions extends to Nprth West

Flowering period: April -July

Specimens examined: Oshingboye 059; Ogundipe & others LUH 5202; LUH 5202;

Keay FHI 15667; Davies RNG 911; Dwerry RNG 13111; Fitzgeralg RNG 29; Barret

RNG 126

40. Cassia fistula Linn. Sp. Pl. 1: 378 (1753).

Description:

A small to medium sized tree 14m high. Leaves are alternate, bipinnate compound with

about 3-8 pairs of leaflets per pinna, petiole 8-13mm. Leaflets are elliptic to ovate (6.2-

20.2) X (3.6-8.8) cm, opposite with an acute tip, leaf base cuneate. Greenish yellow

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under surface, flat upper and lower midrib, margin undulate and thickened with a

leathery texture. Inflorescence is axillary raceme with bright yellow flowers, pods are

broad.

Habitat: Dry deciduous forest, grasslands, cultivated in gardens

Distribution: West Tropical Africa

Flowering period: November - December

Specimens examined: Oshingboye 060; Ogundipe & others LUH 5212

41. Cassia absus F.T.A. 2: 279.

Syn: Chamaecrista absus (L.) H.S. Irwin & Barneby

Description:

A viscid herb or weak shrub, 0.5-0.8m high. Leaves are alternate, bipinnate compound

with about 2 pairs of leaflets per pinna. Leaflets are elliptic (1.5-3.0) X (1.0-1.5) cm,

opposite with an acute tip, leaf base oblique. Greenish yellow under surface, flat upper

and lower midrib, margin entire and not thickened with a papery texture. Inflorescence

is axillary raceme with small red or yellow flowers, pods are oblong.

Habitat: Wastelands

Distribution: West Tropical Africa, Asia and Australia

Flowering period: August - September

Specimens examined: Oshingboye 061; Ogundipe & others LUH 5146; Keay FHI

22997; Latilo FHI 17961: Onochie FHI 43796: Ibhanesebhor and Osanyinlusi FHI

100890

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22. Genus: Senna Mill.

Senna Mill. Gard. Dict. Abr. 3: 28 (1754).

Type Species: Senna alexandrina Mill. Gard. Dict. Abr. 8:1 (1768).

Description:

Unarmed trees, shrubs, subshrubs or herbs with paripinnate leaves. Flowers in racemes

with pedicels without bracteoles. Pods shorter than in Cassia, or flattened, indehiscent

or tardily dehiscent, valves not twisting.

42. Senna tora (L.) Roxb. F.T.A. 2: 275.

Syn: Cassia obtusifolia L.

Cassia tora Linn

Description:

An erect shrub 2.5m high. Leaves are opposite, 3.9-4.2cm long, pinnate compound with

about 3-4 pairs of leaflets per pinna, petiole 12-18mm, stipules present. Leaflets are

obovate (1.8-2.5) X (0.8-1.5) cm, opposite with an obtuse, or bluntly oval tip, leaf base

oblique. Greenish yellow under surface, flat upper and lower midrib, margin entire and

not thickened with a papery texture. Inflorescence is raceme with bright yellow flowers,

pods are narrow and longitudinal.

Habitat: Disturbed sites, waste areas, roadsides

Distribution: widespread in tropics

Flowering period: November

Specimens examined: Oshingboye 063; Ogundipe & others LUH 4667; Chueng RNG

159; Hussaini RNG 1958

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Plate 36: Senna tora pinnate leaves, arrow showing pod

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43. Senna alata (L.) Roxb. F.T.A. 2: 275

Syn: Cassia alata Linn.

Description:

A large spreading shrub 2-3m. Leaves are alternate, up to 52cm long, pinnate compound

with about 9-13 pairs of leaflets per pinna, petiole 15-18mm. Leaflets are oblong or

elliptic (5.8-8.7) X (2.8-4.2) cm, opposite with an obtuse or retuse tip, leaf base round.

Greenish yellow under surface, flat upper and raised lower midrib, margin entire and

not-thickened with a papery texture. Inflorescence is axillary raceme with golden

yellow flowers, pods are elongated and winged.

Habitat: Dry forest regions

Distribution: widespred in the tropics

Flowering period: October

Specimens examined: Oshingboye 064; Gbile FHI 2756; FHI 94313; Odewo,

Ibhanesebhor and Raji FHI 88147; Emwiogbon/Osanyilusi FHI 1248; Gbile FHI

27052

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Plate 37: Senna alata pinnate leaves

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44. Senna hirsita (L.) H.S. Irwin & Barneby Sp. Pl. 1: 378 (1753).

Syn: Cassia hirsuta Linn.

Description:

An erect herb or a perennial shrub 0.5-3m. Leaves are alternate, up to 18.6cm long,

pinnate compound with about 3-5 pairs of leaflets per pinna. Leaflets are ovate or

elliptic (3.4-5.8) X (1.7-2.2) cm, opposite, acuminate tip, and round leaf base. Greenish

yellow under surface, flat upper and raised lower midrib, margin entire and not-

thickened with a papery texture. Inflorescence is axillary raceme with yellow or range

flowers, pods are slightly falcate.

Habitat: Wasteland

Distribution: Tropics and subtropic region

Flowering period: November

Specimens examined: Oshingboye 065; Latilo FHI 27300; Odewo FHI 102005;

Olorunfemi FHI 186686; Emwiogbon/Osanyilusi FHI 86991; Ariwaodo/Adesina FHI

97148; Tamajong FHI 16761

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Plate 38: Senna hirsuta pinnate leaves, arrow showing pod

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45. Senna obtusifolia L. F.W. T.A 2nd ed.1 (2): 455

Syn: Cassia obtusifolia L.

Description:

A short-lived shrub 2.5m. Leaves are alternate, up to 8.6cm long, pinnate compound

with about 3-4 pairs of leaflets per pinna, petiole 34-37mm. Leaflets are obovate (2.5-

3.0) X (1.5-1.9) cm, opposite, obtuse tip, and oblique leaf base. Greenish yellow under

surface, flat upper and raised lower midrib, margin entire and not-thickened with a

papery texture. Inflorescence is axillary raceme with yellow flowers, pods are falcate.

Habitat: Wasteland

Distribution: Tropics and sub-tropics

Flowering period: July - November

Specimens examined: Oshingboye 066; Ogundipe & others LUH 4138; LUH 4305;

LUH 4373; Olorunfemi FHI 43709: Eimunjeze and Ekwuno FHI 68068: Oyayomi,

Fagbemi; Daramola and others FHI 78423: LUH 3964; Gonzales RNG 29; Terry

RNG 1898; Parker RNG 2163; RNG 1394; RNG 2763; RNG 2726

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Plate 39: Senna obtusifolia pinnate leaves, arrow showing pod

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46. Senna occidentalis (L.) Link F.T.A. 2: 274.

Syn: Cassia occidentalis L.

Description:

A short-lived shrub 2m. Leaves are alternate, up to 14.6cm long, pinnate compound

with about 3-5 pairs of leaflets per pinna, petiole 55-61mm. Leaflets are lanceolate (4.5-

6.4) X (2.0-2.9) cm, opposite, acuminate tip, and round leaf base. Dark brown under

surface, flat upper and raised lower surface midrib, margin entire and not-thickened

with a papery texture. Inflorescence is diachasium with yellow flowers, pods are

slightly falcate.

Habitat: Wasteland

Distribution: Tropics and subtropic region

Flowering period: April – June and September - November

Specimens examined: Oshingboye 067; Latilo FHI 65644: Shodeinde FHI 40739:

Ekwuno FHI 88959: Parker RNG 4535; Terry RNG 3667; Parker RNG C2309; Terry

RNG 5008; Barret RNG 531

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Plate 40: Senna occidentalis pinnate leaves, arrows showing flower and pods

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47. Senna siamea (Lam.) H.S. Irwin & Barneby Encycl. Méth. Bot. 1: 648 (1783).

Syn: Cassia siamea Lam.

Description:

A medium-sized tree 20m. Leaves are alternate, up to 25.6cm long, pinnate compound

with about 9-11 pairs of leaflets per pinna, petiole 15-20mm. Leaflets are oblong (4.8-

5.4) X (1.9-2.2) cm, opposite, with a rounded or slightly retuse tip, and round leaf base.

Greenish grey under surface, flat upper and raised lower surface midrib, margin entire

and thickened with a leathery texture. Inflorescence is axillary raceme with yellow

flowers, pods are slightly falcate.

Habitat: Wasteland

Distribution: Tropics and subtropic region

Flowering period: October - November

Specimens examined: Oshingboye 068; Ogundipe & others 5168; Odewo FHI

102270: Morton FHI 15987: Odewo FHI 8326: LUH, 2320; Terry RNG 3667; RNG

5008

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Plate 41: Senna siamea pinnate leaves

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48. Senna singueana (Del.) Lock Cent. Pl. Afr. 28 (1826).

Syn: Cassia singueana Del.

Description:

A shrub or small tree 15m. Leaves are alternate, up to 19.2cm long, paripinnate

compound with about 5-9 pairs of leaflets per pinna, petiole 55-61mm. Leaflets are

elliptic or obovate (3.2-4.3) X (1.7-2.0) cm, opposite, with a round or retuse tip and leaf

base oblique. Greenish brown under surface, channelled upper and raised lower surface

midrib, margin entire and thickened with a papery texture. Inflorescence is axillary

raceme with yellow flowers, pods are linear.

Habitat: Wasteland

Distribution: Tropics and subtropic region

Flowering period: December - January

Specimens examined: Oshingboye 069; Oshingboye & others LUH 5131; Hacker RNG

231; Parker RNG 2309; RNG 7488

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Plate 42: Senna singueana paripinnate leaves, arrow showing pod

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23. Genus: Chamaecrista Monech.

Chamaecrista Monech Methodus 272 (1974)

Type Species: Chamaecrista nictitans (L.) Moench.

Description:

Herbs with paripinnate leaves, often with ± cup-shaped sessile or stalked glands on

petiole and/or rhachis. Flowers in one to many flowered racemes; pedicels with 2

bracteoles near or above the middle. Pods flattened, elastically dehiscent, with twisting

valves.

49. Chamaecrista nigricans (Vahl.) Greene F.T.A. 2: 280.

Syn: Cassia nigricans Vahl.

Description:

An erect herb or undershrub 1.5m high. Leaves alternate, 2.8-5.0cm long, paripinnate

compound with about 5-14 pairs of leaflets per pinna, petiole 8-10mm, stipules present.

Leaflets are oblong (0.8-1.2) X (0.1-0.3) cm, opposite, obtuse, round or mucronate tip,

and round leaf base. Greenish yellow under surface, flat upper and lower midrib,

margin entire and not thickened with a papery texture. Inflorescence is raceme with

bright yellow flowers, pods are flattened.

Habitat: Wooded Savanna, grassland, waste places, roadsides, disturbed soil

Distribution: Tropical Africa, Arabia and India

Flowering period: February, October

Specimens examined: Oshingboye 070; Ogundipe & others LUH 4369; Olorunfemi

FHI 24358

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Plate 43: Chamaecrista nigricans paripinnate leaves

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50. Chamaecrista mimosoides (L.) Greene Sp. Pl. 1: 379 (1753); F.T.A. 2: 280

Syn: Cassia mimosoides Linn

Description:

An erect short-lived perennial herb 1.7m high. Leaves are alternate, 10cm long,

bipinnate compound with about 20-30 pairs of leaflets per pinna, stipules present.

Leaflets are elliptic or linear (0.2-0.4) X (0.1-0.15) cm, opposite, acute or mucronate

tip, leaf base obliquely truncate. Dark brown under surface, flat upper and lower

surface midrib, margin entire and not thickened with a papery texture. Inflorescence is

raceme with bright yellow flowers, pods are flattened.

Habitat: mostly on sandy soil

Distribution: Tropical Africa and Asia

Flowering period: February

Specimens examined: Oshingboye 071; Ogundipe & others LUH 5201; Keay FHI

22563; Terry RNG 113; RNG 3945; Riches RNG 370; Terry RNG 5; Parker RNG 204;

Iwens RNG N100; Hacker RNG 236; RNG 52

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Plate 44: Chamaecrista mimosoides bipinnate leaves

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51. Chamaecrista rotundifolia (Pers.) Greene Trans. Linn. Soc. 27: 570. (1805).

Syn: Cassia rotundifolia Pers.

Description:

A short-lived herb 1m. Leaves are alternate, bifoliate, obovate (1.8-1.9) X (1.0-1.2) cm,

petiole 4mm with a round tip and oblique base. Greenish yellow under surface, flat

upper and raised lower layer midrib, margin entire and not-thickened with a papery

texture. Inflorescence is axillary raceme with yellow flowers, pods are linear.

Habitat: Savanna

Distribution: Tropical Africa and Asia

Flowering period: February, August

Specimens examined: Oshingboye 072; Ekwuno FHI 91985: Witt FHI 27889:

Emwiogbon FHI 33146: Emwiogbon FHI 87394: Ekwuno and others FHI 95991;

Ajulo FHI 102269

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Plate 45: Chamaecrista rotundifolia bifoliate leaves

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24. Genus: L.

Dialium L. F.T.A. 2: 282; Bull. Soc. Roy. Bot. Belg. 84: 29–45 (1951).

Type Species: Dialium indum L.

Description:

Unarmed trees with imparipinnate or sometimes 3-foliolate leaves. Inflorescence

panicle, flowers brown outside, greenish-white inside, petals minute and

inconspicuous. Pod sessile, indehiscent, slightly flattened, and densely pubescent.

52. Dialium guineense Willd – F. T. A. 2: 283

Description:

An erect herb or undershrub 1.5m high. Leaves alternate, 2.8-5.0cm long, paripinnate

compound with about 5-14 pairs of leaflets per pinna, petiole 8-10mm, stipules present.

Leaflets are oblong (0.8-1.2) X (0.1-0.3) cm, opposite, obtuse, round or mucronate tip,

and round leaf base. Greenish yellow under surface, flat upper and lower midrib,

margin entire and not thickened with a papery texture. Inflorescence is raceme with

bright yellow flowers, pods are flattened.

Habitat: Forest and also persistent in Savanna vegetation

Distribution: West Tropical Africa, Principe ans S. Tomé

Flowering period: April – May and September - November

Specimens examined: Oshingboye 072; Emwiogbon FHI 65999: Fagbemi and

Osanyinlusi FHI 82811: Odewo and others FHI 88148

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Sub Family III: Faboideae Rudd (Papilioideae nom. cons,)

Faboideae Rudd Rhodora 70: 496 (1968).

Type Genus: Faba Mill. Gard. Dict. Abr. 4:1 (1754).

Description:

Trees, shrubs, climbers or, most often, herbs. Leaves alternate, most often pinnate or

pinnately trifoliolate, less often pinnate-trifoliate, occasionally unifoliolate, simple or

digitate. Pulvinus always present at the base of each leaflet, and usually present at base

of petiole. Inflorescences of racemes or panicles, less often umbellate, capitate or

spicate. Flowers zygomorphic, papilionaceous, with a large, often erect, adaxial petal

(standard), two lateral petals (wings) parallel with each other, and two lower petals,

which are often connate by their lower margins to form a keel. Sepals united at

base. Petals imbricate in bud. Stamens usually 10, rarely fewer, either all fused

(monadelphous) or 1 free and 9 fused (diadelphous) or rarely all free. Pods

various. Seeds without areoles.

Tribe I: Abreae Hutch.

Abreae Bronn. Form. Pl. Legumin. 131. (1964).

Type: Abrus Adans. F.T.A. 2: 174.

25. Genus: Abrus Adans.

Abrus Adans. F.T.A. 2: 174.

Type species: Abrus precatotious Linn.

Description:

Woody subshrubs or lianes. Leaves paripinnate with numerous leaflets. Inflorescences

axillary or terminal; bracts and bracteoles present. Pod linear or oblong.

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53. Abrus precatorious (L.) W.F. Wight - F.T.A. 2: 175.

Description:

A high climbing, twining, perennial herb up to 3.3m when fully stretched. Leaves

alternate, pinnate compound with 7-16 pairs of leaflets per pinnae, stipules present and

petiole 3-4mm. Leaflets are opposite, oblong, (1.1-1.7) X (0.4-1) cm, tip obtuse and

base round. Dry leaf under colour greenish yellow, flat upper and lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is a compound

cyme with white or pink or red flower, pods are oblong and flat.

Habitat: wastelands and low forests

Distribution: Tropics and sub-tropics

Flowering period: April – August

Specimens examined: Oshingboye 073; Ogundipe & others LUH 4493; Jones FHI

16649: Olorunfemi FHI 57085: Onochie FHI 35922: Emwiogbon FHI 87127; Terry

RNG 3575

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Plate 46: Abrus precatorious pinnate leaves, arrow showing flowers and pod

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Tribe II: Phaseoleae Bronn ex DC.

Phaseoleae Bronn ex DC. Prodr. 2: 381 (1825).

Type: Phaseolus L. F.T.A. 2: 191.

26. Genus: Adenodolichos Harms

Adenodolichos Harms. Engl. Bot. Jahrb. 33: 179 (1902)

Description:

Herbs, subshrubs or shrubs. Leaves trifoliolate; leaflets covered with gland dots

beneath. Inflorescences axillary or terminal. Ovary 2-ovuled, Pods oblong-falcate,

flattened, much narrowed towards the base.

54. Adenodolichos paniculatus (Hua) Hutch. F.W.T.A., ed. 1, 1: 411 (1928).

Syn: Dolichos paniculatus Hua

Adenodolichos macrothyrsus Harms

Description:

A perennial woody shrub or subshrub up to 4.5m. Leaves opposite with pinnate-

trifoliate leaflets, and petiole 45-50mm. Leaflets ovate, (10.8-12.0) X (4.7-7.1) cm, tip

obtuse or retuse and base rounded-oblique. Dry leaf under colour dark brown, flat upper

and lower midrib, leaf margin entire and not-thickened with a papery texture.

Inflorescence is an axillary raceme with distinct papillionaceous flower bearing a

pinkish keel and a yellowish standard, pods are flat.

Habitat: Savanna vegetation

Distribution: West Tropical Africa, French Cameroons and Sudan

Flowering period: December

Specimens examined: Oshingboye 074; Ogundipe & others LUH 4195; LUH 4786

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Plate 47: Adenodolichos paniculatus pinnate-trifoliate leaves

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27. Genus: Cajanus DC.

Cajanus DC. F.T.A. 2: 215.

Type Species: Cajanus cajan (L.) Huth.

Description:

Shrub, pinnately trifoliolate leaves, covered with small yellow glands beneath.

Inflorescence a terminal panicle with many numerous short axillary racemes. Pod

grooved between the seeds, somewhat inflated, with a long apical bristle.

55. Cajanus cajan (L.) Millsp. F.T.A.2:216

Syn: Cajanus cajan (L.) Huth

Cajanus indicus Spreng.

Description:

An erect annual shrub up to 2m. Leaves alternate with pinnate-trifoliate leaflets, stipules

present and petiole 30-47mm. Leaflets oblong or lanceolate, (7.3-10.0) X (2.0-2.8) cm,

tip aristate or acuminate and cuneate base. Dry leaf under colour greenish yellow, flat

upper and lower midrib, leaf margin entire and not-thickened with a papery texture.

Inflorescence is an axillary raceme with yellow or cream coloured flowers, pods are

lenticular to ovoid.

Habitat: dryer regions of the tropics and subtropics and temperate regions, cultivated

Distribution: cultivated throughout the tropics and subtropics

Flowering period: variable, within 90 and 240 days after planting

Specimens examined: Oshingboye 075; Felix and others FHI 106993; Daggash FHI

35025: Kzog FHI 91432

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Plate 48: Cajanus cajan pinnate-trifoliate leaves, arrow showing flowers

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28. Genus: Eriosema (DC) Desv.

Eriosema (DC) Desv. F.T.A. 2: 223.

Type Species: Eriosema rufum (Kunth) G. Don.

Description:

Shrub, pinnately trifoliolate leaves, covered with small yellow glands beneath.

Inflorescence a terminal panicle with many numerous short axillary racemes. Pod

grooved between the seeds, somewhat inflated, with a long apical bristle.

56. Eriosema psoraloides (Lam) G. Don F.W.T.A. 1:403

Syn: Crotalaria psoraloides Lam.

Eriosema cajanoides (Guill. & Perr.) Hook. f.

Eriosema proschii Briq.

Rhynchosia psoraleoides (Lam.) DC.

Description:

A much-branched perennial shrub up to 2.5m. Leaves alternate with pinnate-trifoliate

leaflets, stipules present and petiole 30-47mm. Leaflets elliptic to oblong or obovate,

(2.3-90.0) X (2.0-5.8) cm, tip acute or obtuse and cuneate base. Dry leaf under colour

greenish yellow, flat upper and lower midrib, leaf margin entire and not-thickened with

a papery texture. Inflorescence is an axillary raceme with golden yellow coloured

flowers, pods are ovate to almost circular.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa extending to South Africa

Flowering period: November - December

Specimens examined: Oshingboye 076; Daramola FHI 99018: Jos, Emwiogbon FHI

87137

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29. Genus: Calopogonium Desv.

Calopogonium Desv. Ann. Sci. Nat., sér. 1, 9: 423 (1826).

Type Species: Calopogonium mucunoides Desv.

57. Calopogonium mucunoides Desv. Amshoff in Fl. Suriname 2: 196 (1826).

Description:

A creeping perennial herb up to 5m. Leaves alternate with pinnate-trifoliate leaflets,

stipules present and petiole 42-125mm. Leaflets ovate to rhomboid-ovate, (2.8-5.8) X

(3.9-6.6) cm, tip obtuse and cuneate to round at the base. Dry leaf under colour greenish

yellow, flat upper and lower midrib, leaf margin entire and not-thickened with a papery

texture. Inflorescence is a raceme with purple or blue flower colour, pods are linear to

oblong.

Habitat: Wastelands

Distribution: Tropical Africa and America

Flowering period: December

Specimens examined: Oshingboye 077; Ogundipe & others LUH 4186; Ekwuno and

others FHI95678: LUH 5524

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Plate 49: Calopogonium mucunoides pinnate-trifoliate leaves, arrow showing flowers

183 | P a g e

30. Genus: Glycine Linn.

Glycine Linn. F.T.A. 2: 178.

Type Species: Glycine javanica Linn. F.T.A, 2:178

58. Glycine max (L.) Merr. Desv. Amshoff in Fl. Suriname 2: 196 (1826).

Syn: Dolichos sofa L.

Dolichos soja L.

Glycine angustifolia Miq.

Glycine gracilis Skvortsov

Description:

An erect annual herb up to 1.5m. Leaves opposite with trifoliate leaflets, stipules

present and petiole 42-125mm. Leaflets ovate, (2.8-5.8) X (3.9-6.6) cm, tip obtuse and

cuneate to round at the base. Dry leaf under colour greenish yellow, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is a

raceme with white to purple pink flower colour, pods are linear to slightly curved and

flattened.

Habitat: Cultivated

Distribution: Esat Asia, Tropical Africa and America

Flowering period: July - September

Specimens examined: Oshingboye 078; Ogundipe & others LUH 1304A

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31. Genus: Dolichos Linn.

Dolichos Linn. F.T.A. 2: 209.

Type Species: Dolichos trilobatus L. Mant. Pl. 1: 101 (1767)

Description:

Herbs, shrubs or suffrutices, sometimes climbing. Leaves pinnately or rarely digitately

trifoliolate, often appearing after the flowers. Inflorescences axillary. Pod flattened

laterally.

59. Dolichos stenophyllus Harms in Engl. Bot. Jahrb. 26: 314 (1899).

Syn: Macrotyloma stenophyllum (Harms) Verdc

Description:

A slender annual or climbing perennial herb up to 0.9m. Leaves alternate with trifoliate

leaflets, stipules present and petiole 7-15mm. Leaflets ovate to oblong, narrowly oblong

to lanceolate, (2.8-7.2) X (1.9-2.6) cm, tip acute and cuneate base. Dry leaf under colour

greenish brown, flat upper and lower midrib, leaf margin entire and not-thickened with

a papery texture. Inflorescence is an axillary raceme with yellow flower colour, pods

are linear.

Habitat: Savanna

Distribution: Sudanian wooded grassland

Flowering period: October

Specimens examined: Oshingboye 079; Ogundipe & others LUH 5162; Jones FHI

5892

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32. Genus: Phaseolus Linn.

Phaseolus Linn. F.T.A. 2: 191.

Type Species: Phaseolus vulgaris Linn.

60. Phaseolus vulgaris Linn. Sp. Pl. 2: 273 (1753).

Syn: Phaseolus compessus DC.

Phaseolus esculentus Salisb.

Phaseolus nanus L. & Jusl.

Description:

An annual herb up to 3m. Leaves alternate with pinnate-trifoliate leaflets, stipules

present and petiole 12-45mm. Leaflets ovate, (15.8-20.8) X (9.9-15.0) cm, tip obtuse

and cuneate to round at the base. Dry leaf under colour greenish yellow, flat upper and

lower midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence

is a raceme with pale purple or red-purple flower colour, pods are linear.

Habitat: Widely cultivated

Distribution: New world, Central Mexico

Flowering period: September - October

Specimens examined: Oshingboye 080; Swarbrick FHI 53940: Emwiogbon FHI

55859

186 | P a g e

33. Genus: Vigna Savi

Vigna Savi F.T.A. 2: 194.

Type Species: Vigna villosa Savi

Description:

Climbing, prostrate or erect herbs or subshrubs. Leaves pinnately (rarely digitately)

trifoliolate or unifoliolate. Inflorescences axillary. Pods linear, usually terete and not

flattened.

61. Vigna subterranea (L.) Verdc. F.T.A. 2: 207

Syn: Glycine subterranea L.

Voandzeia subterranea (L.) Thouars

Description:

An annual creeping herb 0.1-0.3m. Leaves alternate with pinnate-trifoliate leaflets,

petiole 50-180mm. leaflets elliptic to ovate, (4.8-5.2) X (1.1-2.3) cm, tip emarginate

and cuneate base. Dry leaf under colour greenish brown, flat upper and raised lower

midrib, leaf margin entire and slightly thickened with a leathery texture. Inflorescence

is an axillary raceme with yellow flower colour, pods are round.

Habitat: Savanna

Distribution: Sudanian woodland and cultivated

Flowering period: July

Specimens examined: Oshingboye 081; Ogundipe & others LUH 5194

187 | P a g e

Plate 50: Vigna subterranea pinnate-trifoliate leaves

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62. Vigna ambacensis Welw. ex Bak. F.T.A. 2: 201 (1871)

Syn: Vigna heterophylla A. Rich

Description:

An annual climbing herb 0.9-7m. Leaves alternate with pinnate-trifoliate leaflets,

petiole 40-60mm and stipules present. Leaflets oblong to ovate-lanceolate, (4.5-15) X

(1.1-5.3) cm, tip narrowly rounded to acuminate or apiculate and round or subcordate

base. Dry leaf under colour greenish brown, flat upper and lower midrib, leaf margin

entire and slightly thickened with a leathery texture. Inflorescence is an axillary raceme

with pink to mauve or violet flower colour, pods are linear to cylindrical.

Habitat: Savanna

Distribution: Grasslands and Guinea-Congolia/Sudania regional transition zone

Flowering period: November - December

Specimens examined: Oshingboye 085; Ogundipe & others LUH 5195; Keay FHI

28145

63. Vigna gracilis (Guill. & Perr.) Hook. f. — F.T.A. 2: 205

Syn: Dolichos gracilis Guill. & Perr.

Vigna afzelii Bak.

Vigna parvifolia Planch. ex Bak

Vigna occidentalis Bak. f.

Description:

A slender twinning herb 0.1-0.3m. Leaves alternate with pinnate-trifoliate leaflets,

petiole 14-28mm. leaflets broad-ovate to ovate-rhomboidal, (1.4-2.9) X (0.3-1.1) cm,

tip obtuse to acute and cuneate base. Dry leaf under colour greenish brown, flat upper

and lower midrib, leaf margin entire and slightly thickened with a leathery texture.

189 | P a g e

Inflorescence is an axillary raceme with pink or blue turning yellow flower colour, pods

are linear.

Habitat: Savanna

Distribution: Sudanian grasslands, Sudanian regional transition zone

Flowering period: November

Specimens examined: Oshingboye, 089; Barret RNG 5184

64. Vigna racemosa (G. Don) Hutch. & Dalziel F.W.T.A., ed. 1, 1: 409 (1928)

Syn: Clitoria racemosa G. Don

Vigna donii Baker

Vigna strophiolata Piper

Description:

A perennial climbing herb up to 7m. Leaves alternate with pinnate-trifoliate leaflets,

petiole 1.5-11cm, stipules present. Leaflets oblong, lanceolate to ovate, (3.0-13.0) X

(1.0-8.0) cm, tip acute or acuminate and round or truncate base. Dry leaf under colour

greenish brown, flat upper and lower midrib, leaf margin entire and slightly thickened

with a leathery texture. Inflorescence is an axillary raceme with blue flower colour,

pods are linear to cylindrical and curved.

Habitat: Savanna

Distribution: Grasslands

Flowering period: November

Specimens examined: Oshingboye 089; Ogundipe & others LUH 5134

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65. Vigna unguiculata (Linn.) Walp. Rep. 1: 779 (1842);

Syn: Dolichos unguiculatus Linn. (1753).

Dolichos biflorus Linn.

Dolichos sinensis Linn.

Vigna sinensis (Linn.) Savi ex Hassk

Description:

An annual herb up to 4m high. Leaves alternate with pinnate-trifoliate leaflets, petiole

5-25cm. leaflets elliptic to ovate, (4.8-5.2) X (1.1-2.3) cm, tip acute and hastate to

oblique base. Dry leaf under colour greenish brown, flat upper and lower midrib, leaf

margin entire and slightly thickened with a leathery texture. Inflorescence is an axillary

raceme with pink to purple flower colour, pods are cylindrical.

Habitat: Savanna and cultivated

Distribution: Sudanian woodland, thickets and cultivated

Flowering period: October - November

Specimens examined: Oshingboye 092; Olorunfemi and others FHI 96469: Daramola

FHI 18127

34. Genus: Mucuna Adans.

Mucuna Adans. F.T.A. 2: 184.

Type Species: Mucuna urens (L.) DC.

Description:

Climbers or lianes. Leaves pinnately trifoliolate. Inflorescences axillary. Pods covered

with irritant hairs, usually dehiscent.

191 | P a g e

66. Mucuna pruriens (L.) DC. F.T.A. 2: 187

Syn: Dolichos pruriens Linn

An annual climbing herb 2-3m. Leaves alternate with trifoliate leaflets, stipules minute

and petiole 20-26mm. Leaflets rhombic to ovate, (12.8-16.4) X (7.5-8.5) cm, tip acute,

lateral leaflets with asymmetrical rhombic base while terminal ones are cuneate. Dry

leaf under colour greenish grey, flat upper and raised lower midrib, leaf margin entire

and thickened with a leathery texture. Inflorescence is an axillary raceme with cream

or purple flower colour, pods are oblong to elliptic.

Habitat: Savanna

Distribution: Sudanian wooded grassland and tropics

Flowering period: December - January

Specimens examined: Oshingboye 092; Ogundipe & others LUH 4472; Latilo

FHI47778: Daramola FHI34264

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Plate 51: Mucuna pruriens trifoliate leaves, arrow showing pod

193 | P a g e

35. Genus: Erythrina L.

Erythrina L. F.T.A. 2: 181

Type Species: Erythrina corallodendron L.

Description:

Deciduous trees or shrubs; bark usually corky and often armed with prickles borne on

woody bosses. Leaves pinnately trifoliolate, often prickly. Inflorescences usually

terminal, bearing flowers in fascicles. Pod constricted between the seeds, and dehiscent.

67. Erythrina senegalensis DC. F.T.A. 2: 181

Description:

A small tree up to 7m armed with prickles on bark. Leaves alternate with pinnate-

trifoliate leaflets, petiole 49-60mm. leaflets oblong to ovate, (6.8-11.2) X (3.6-6.3) cm,

tip obtuse and obliquely round base. Dry leaf under colour greenish yellow, flat upper

and raised lower midrib, leaf margin entire and thickened with a leathery texture.

Inflorescence is an axillary raceme with bright red flower colour, pods are twisted.

Habitat: Savanna vegetation

Distribution: Sudanian woodland

Flowering period: November

Specimens examined: Oshingboye 095; LUH 6275; Daramola FHI 10471:

Olorunfemi and others FHI 88526: Daley FHI 32259

194 | P a g e

Plate 52: Erythrina senegalensis leaves, arrow showing inflorescence

195 | P a g e

68. Erythrina sigmoidea Hua Bull. Mus. Hist. Nat., sér. 1, 3: 327 (1897)

Syn: Erythrina dybowskii Hua

Erythrina eriotricha Harms

Description:

A small tree 3-6m armed with prickles on the stem. Leaves opposite with pinnate-

trifoliate leaflets, petiole 72-96mm. leaflets ovate, broader than long (6.8-7.7) X (7.3-

10.6) cm, tip round, emarginate or mucronate and broadly cuneate base. Dry leaf under

colour greenish grey, flat upper and lower midrib, leaf margin entire and thickened with

a leathery texture. Inflorescence is an axillary raceme with red flower colour, pods are

moniliform.

Habitat: Savanna

Distribution: Sudanian woodland, French Cameroons and Chad

Flowering period: April

Specimens examined: Oshingboye 097; Hepper FHI 53146: Ariwaodo FHI100724:

Ogbogu and Odewo FHI 106182

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Plate 53: Erythrina sigmoidea pinnate-trifoliate leaves

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Tribe III: Desmodieae (Benth). Hutch.

Desmodieae (Benth). Hutch. 1964

Type: Desmodium Desv. F.T.A. 2: 159

36. Genus: Alysicarpus Neck.

Alysicarpus Neck. F.T.A. 2: 169

Type Species: Alysicarpus bupleurifolius (L.) DC.

Description:

Herbs, stipules free or joined. Leaves unifoliolate, rarely pinnately trifoliolate.

Inflorescences axillary, terminal, falsely racemose or rarely paniculate. Pod

transversely jointed, subcylindric or somewhat flattened, and constricted.

69. Alysicarpus glumaceus (Vahl.) DC. Prod. 2: 353 (1825)

Syn: Alysicarpus violaceus (Forssk.) Schindl.

Description:

An erect annual shrubby legume up to 1.5m. Leaves alternate, unifoliate, stipules

present and petiole 10-25mm; oblong or elliptic, (1.6-3.2) X (0.7-1.7) cm, tip acute and

cordate base. Dry leaf under colour greenish yellow, flat upper and lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with red or yellow flower colour, pods are oblong and with constrictions.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye 101; Ogundipe & others 5158; Hacker RNG 96;

RNG 1355; RNG 174

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Plate 54: Alysicarpus glumaceus leaves

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70. Alysicarpus rugosus (Willd.) DC. Prod. 2: 353 (1825); F.T.A. 2: 171

Syn: Hedysarum rugosum Willd.

Alysicarpus violaceum Schindl.

Description:

An erect annual legume up to 2m. Leaves alternate with pinnate-trifoliate leaflets,

stipules present and petiole 30-35mm. Leaflets oblong or lanceolate, (8.9-10.8) X (0.2-

0.3) cm, tip acuminate and cuneate base. Dry leaf under colour greenish grey, flat upper

and lower midrib, leaf margin entire and thickened with a leathery texture.

Inflorescence is an axillary raceme with red or yellow flower colour, pods are oblong

and with constrictions.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa

Flowering period: April, October

Specimens examined: Oshingboye 104; Ogundipe & others LUH 4384; Onochie FHI

8132; Keay FHI 5467; Barret RNG 348; Fortune RNG 2331; Iwens RNG 126; Hacker

RNG 2; Parker RNG 2; Parker RNF E107; RNG 465; RNG 3842

200 | P a g e

Plate 55: Alysicarpus rugosus leaves

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71. Alysicarpus ovalifolius (Schumach) J. Léonard Bull. Jard. Bot. Brux. 24: 88 (1954).

Syn: Hedysarum rugosum Willd.

Alysicarpus violaceum Schindl.

Description:

An erect annual legume up to 6m. Leaves simple, alternate, stipules present and petiole

4-12mm. Leaves oblong or suborbicular, (1.0-2.5) X (0.5-1.0) cm, tip acute or round

and cordate base. Dry leaf under colour greenish grey, flat upper and lower midrib, leaf

margin entire and thickened with a leathery texture. Inflorescence is an axillary raceme

with red flower colour, pods are oblong.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa

Flowering period: September

Specimens examined: Oshingboye 107; Terry RNG 3133; Davies RNG 398; Terry

RNG 3228; Webb RNG 103

72. Alysicarpus vaginalis (Linn.) DC. F.T.A. 2: 170

Syn: Alysicarpus nummularifolius (L.) DC.

Alysicarpus nummularifolius (L.) DC. var. angustatus Ohwi

Hedysarum cylindricum Poir.

Hedysarum vaginale L

Description:

An erect or spreading perennial legume up to 6m. Leaves simple, alternate, stipules

present and petiole 10mm. Leaves ovate, (0.5-6.5) X (0.3-2.5) cm, tip obtuse and

cuneate base. Dry leaf under colour greenish grey, flat upper and lower midrib, leaf

202 | P a g e

margin entire and thickened with a leathery texture. Inflorescence is an axillary raceme

with reddish yellow or pale purple flower colour, pods are oval to oblong.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa

Flowering period: August

Specimens examined: Oshingboye 109; Nolile FHI 26380: Leeuw FHI 18003: Ohaeri

FHI 72884: Macaulay FHI 62070: Latilo FHI 62696; Lee RNG 2771; Bowen RNG

3806; Kaison RNG 101; RNG 31; Nam-Matra& Kaison RNG 9

37. Genus: Desmodium Desv.

Desmodium Desv. F.T.A. 2: 159

Type Species: Desmodium scorpiurus (Sw.) Poir.

Description:

Shrubs or herbs, leaves unifoliolate or pinnately trifoliolate. Inflorescences axillary or

terminal, falsely racemose or paniculate. Pod transversely jointed, flattened with (1-)2-

many 1-seeded articles, often with hooked hairs.

73. Desmodium gangeticum (L.) DC. Prod. 2: 327 (1825); F.T.A. 2: 161

Syn: Aeschynomene gangetica Poir.

Aeschynomene maculata (L.) Poir

Description:

A sub-erect undershrub 0.6-1.2m. Leaves alternate, unifoliate, ovate to lanceolate,

petiole 10-22mm, (4.6-15.4) X (2.9-4.6) cm, tip acute or slightly acuminate and round

base; stipules present. Dry leaf under colour greenish brown, flat upper and lower

203 | P a g e midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an axillary raceme with cream, white or pink flower colour, pods are sub-falcate.

Habitat: Sudanian wooded grassland

Distribution: Tropics and sub-tropics

Flowering period: March - April

Specimens examined: Oshingboye 111; Latilo and Fagbemi FHI66274: Jones FHI

13715; FHI 27906

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Plate 56: Desmodium gangeticum leaves

205 | P a g e

74. Desmodium scorpiurus (Sw.) Desv. Journ. de Bot. 1: 122 (1813).

Syn: Desmodium akoense Hayata

Desmodium arenarium Kunth

Desmodium parviflorum M.

Desmodium virgatum Desv.

Description:

A straggling herb 0.6m. Leaves alternate with pinnate-trifoliate leaflets, petiole 10-

18mm; stipules present. Leaflets lanceolate to elliptic, (1.1-2.2) X (0.5-1.0) cm, tip

acute and cuneate base. Dry leaf under colour greenish grey, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an

axillary raceme with white, blue, pink or purple flower colour, pods are linear.

Habitat: wasteland

Distribution: Tropical America and Africa

Specimens examined: Oshingboye 113; Gbile FHI57595: Macaulay FHI 62066:

Ariwaodo and Adesina FHI97348

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Plate 57: Desmodium scorpiurus leaves

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75. Desmodium tortuosum (Sw.) DC. Prod. 2: 332 (1825)

Syn: Hedysarum tortuosum Sw.

Desmodium ospriostreblum Steud. ex Chiov.

Desmodium abyssinicum (Hochst. ex A. Rich.) Hutch. & Dalz

Description:

An erect short-lived herb 0.6-3m. Leaves alternate with pinnate-trifoliate leaflets,

petiole 20-44mm; stipules present. Leaflets lanceolate to elliptic, (4.0-6.3) X (1.9-2.5)

cm, tip acute and cuneate base. Dry leaf under colour greenish grey, flat upper and

lower midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence

is a panicle with purple flower colour, pods are indented in chain-like form.

Habitat: Grassland, wasteland

Distribution: Tropics and subtropics

Flowering period: October

Specimens examined: Oshingboye 114; Magaji FHI 17982: Fagbemi FHI89828:

Arasi FHI 95195

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Plate 58: Desmodium tortuosum leaves (pinnate-trifoliate leaflets)

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76. Desmodium velutinum (Willd.) DC. F.T.A. 2:162.

Syn: Desmodium lasiocarpum (P. Beauv.) DC.

Desmodium latifolium (Roxb.) DC.

Hedysarum deltoides Poir.

Description:

An erect shrub or subshrub up to 3m. Leaves alternate, unifoliate, rarely trifoliate,

petiole 14-20mm; stipules present, ovate, (4.0-20.2) X (4.6-5.6) cm, tip acute and

bluntly cuneate base. Dry leaf under colour greenish grey, channelled upper and raised

lower midrib, leaf margin serrated and not-thickened with a papery texture.

Inflorescence is an axillary raceme or panicle with violet, red or blue flower, pods are

narrowly oblong.

Habitat: Grassland, wasteland

Distribution: Tropics and subtropics

Flowering period: September - November

Specimens examined: Oshingboye 115; Ogundipe & others LUH 5196; LUH 4363;

Olorunfemi and others FHI96340: Olorunfemi and Oguntayo FHI91898: Ondo,

Onijamowo and others FHI 79997: Latilo and others FHI 67247

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Plate 59: Desmodium velutinum leaves

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Tribe IV: Crotalarieae Hutch

Crotalarieae Hutch (1964).

Type: Crotalaria L. F.T. A. 2:7

38. Genus: Crotalaria L.

Crotalaria L. F.T. A. 2:7

Description:

Herbs, or shrubs, leaves simple, unifoliolate or digitately 3(-7)-foliolate, and usually

petiolate. Flowers arranged in axillary racemes or heads. Pod usually inflated, 1 to

many seeded.

77. Crotalaria comosa Baker F.T.A. 2: 34 (1871).

Description:

An erect annual herb 0.6-0.9m. Leaves alternate with trifoliate leaflets, stipules absent

and petiole 12-16mm. Leaflets oblong to oblanceolate, (4.8-5.0) X (1.8-2.6) cm, tip

obtuse or sub-cuspidate and cuneate to round base. Dry leaf under colour greenish

yellow, flat upper and lower midrib, leaf margin entire and thickened with a leathery

texture. Inflorescence is an axillary raceme with bright yellow flower colour, pods are

linear to oblong.

Habitat: Savanna and cultivated land

Distribution: Tropical Africa, Belgian Congo and Angola

Flowering period: December

Specimens examined: Oshingboye 118; Olorunfemi FHI 24361

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Plate 60: Crotalaria comosa trifoliate leaves

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78. Crotalaria hyssopifolia Klotzsch F.T.A. 2: 24

Syn: Crotalaria pseudotenuirama Torre

Description:

An erect annual herb 0.5-0.6m. Leaves alternate with trifoliate leaflets, stipules absent

and petiole 5-8mm. Leaflets oblanceolate, (1.8-3.0) X (0.3-0.6) cm, tip obtuse and

cuneate to round base. Dry leaf under colour greenish yellow, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an

axillary raceme with bright yellow flower colour, pods are linear.

Habitat: Sudanian wooded grassland

Distribution: Tropical Africa

Flowering period: October - December

Specimens examined: Oshingboye 119; Cole FHI8779; Keay FHI 25510; Darter

FHI42312; Gbile and Odewo FHI102444

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Plate 61: Crotalaria hyssopifolia trifoliate leaves

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79. Crotalaria lachnosema Stapf Kew Bull. 1910: 329

Description:

A woody undershrub 0.6-1.8m. Leaves alternate with trifoliate leaflets, stipules present

and petiole 5 -10mm. Leaflets oblanceolate, (2.8-5.4) X (1.3-2.4) cm, tip cuspidate and

cuneate base. Dry leaf under colour greenish yellow, flat upper and raised lower midrib,

leaf margin entire and thickened with a leathery texture. Inflorescence is an axillary

raceme with yellow flower colour, pods are linear.

Habitat: Sudanian woodland, roadside, wasteland

Distribution: West Tropical Africa, Sudan

Flowering period: June - July

Specimens examined: Oshingboye 121; Odewo FHI 97375: Ekwuno FHI 77099;

Sofoluwe FHI 38185; Ekwuno FHI 70883; FHI 77767; Odewo FHI 97375: Ekwuno

FHI 77099

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Plate 62: Crotalaria lachnosema trifoliate leaves

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80. Crotalaria macrocalyx Benth. F.T.A. 2: 20

Description:

An erect herb 0.9m. Leaves alternate with trifoliate leaflets, stipules present and petiole

26-45mm. Leaflets oblanceolate to elliptic, (2.7-5.7) X (0.2-1.2) cm, tip obtuse and

cuneate base. Dry leaf under colour greenish grey, flat upper and lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with bright yellow flower colour, pods are round to oblong.

Habitat: Sudanian woodland and grasslands

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye, 124; Keay FHI 5464; Olorunfemi FHI 24352;

Daggash FHI 22004

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Plate 63: Crotalaria macrocalyx trifoliate leaves

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81. Crotalaria pallida Aiton F.T.A. 2: 40

Syn: Crotalaria falcata DC.

Crotalaria mucronata Desv.

Crotalaria pisiformis Guill. & Perr.

Description:

An erect shrub 1.5m. Leaves alternate with trifoliate leaflets, stipules present and

petiole 50-62mm. Leaflets elliptic to lanceolate, (6.0-8.1) X (1.6-1.8) cm, tip acute and

cuneate base. Dry leaf under colour greenish yellow, flat upper and lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with yellow flower colour, pods are cylindrical.

Habitat: Sudanian woodland, grasslands and wastelands

Distribution: Tropics and subtropics

Flowering period: December - January

Specimens examined: Oshingboye 125; Keay FHI 5451; Jones FHI 769

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Plate 64: Crotalaria pallida trifoliate leaves

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82. Crotalaria naragutensis Hutch. Kew Bull. 1921: 363

Syn: Crotalaria lynesii Baker f. & Martin

Description:

An erect branched undershrub 0.9-1.8m. Leaves alternate with trifoliate leaflets,

stipules tiny or absent and petiole 22-75mm. Leaflets elliptic, (2.8-7.2) X (0.9-1.8) cm,

tip acute and cuneate base. Dry leaf under colour greenish yellow, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an

axillary raceme with yellow flower colour, pods are linear.

Habitat: Open savana, grassland

Distribution: West Tropical Africa, Chad and Sudan

Flowering period: November

Specimens examined: Oshingboye 127; Ogundipe & others LUH 1314A

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a b

Plate 65: Crotalaria naragutensis (a) trifoliate leaves (b) arrow showing inflorescence

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83. Crotalaria retusa L. F.T.A. 2: 13

Syn: Crotalaria cuneifolia (Forssk.) Schrank

Description:

An erect annual woody herb 1.3m. Leaves alternate, simple, oblanceolate to spathulate,

stipules present, petiole 2-3mm, (4.4-6.7) X (2.1-2.6) cm, tip round and cuneate base.

Dry leaf under colour greenish grey, flat upper and lower midrib, leaf margin entire and

not-thickened with a papery texture. Inflorescence is an axillary raceme with bright

yellow flower, pods are oblong to cylindrical.

Habitat: Guinea-Congolian anthropic landscapes; roadside

Distribution: Tropics and subtropics,

Flowering period: September

Specimens examined: Oshingboye 129; Ogundipe & others LUH 5207; Ekwuno and

others FHI 95669: Gbile and Daramola FHI 73639: Daramola FHI 54676: Daramola

FHI 78550: Ariwaodo FHI 73220

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Plate 66: Crotalaria retusa leaves

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84. Crotalaria arenaria Benth. — F.T.A. 2: 11

Description:

An erect annual or perennial undershrub 1.3m. Leaves alternate, simple, oblong,

stipules present, sessile, (4.4-6.7) X (2.1-2.6) cm, tip round and cuneate to obtuse base.

Dry leaf under colour greenish grey, flat upper and lower midrib, leaf margin entire and

not-thickened with a papery texture. Inflorescence is an axillary raceme with pale

yellow flower colour, pods are oblong and sessile.

Habitat: Sahel regional transition zone; grassland; semi-desert

Distribution: Sudano-sahelian sub-sahara Africa

Flowering period: July

Specimens examined: Oshingboye 129; Ogundipe & others LUH 4221

85. Crotalaria senegalensis (Pers.) Bacle ex DC- F.T.A, 2:31

Syn: Crotalaria shamvaensis sensu Torre

Crotalaria uncinella Lam. var. senegalensis Pers.

Description:

An erect annual herb 1.3m. Leaves alternate with trifoliate leaflets, elliptic-oblong to

lanceolate, stipules present, petiole 1.5-5cm, (2.0-6.5) X (0.6-2.5) cm, tip round and

cuneate base. Dry leaf under colour greenish grey, flat upper and lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with bright yellow flower colour, pods are oblong to ellipsoid.

Habitat: Dry fields; roadside

Distribution: Tropics and subtropics, Belgian Congo, Sudan

Flowering period: September - December

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Specimens examined: Oshingboye 130; Ekwuno and Fagbemi FHI94052: Soladoye

and others FHI83756: Latilo FHI62698: Jackson FHI 77548

86. Crotalaria falcata Vahl. F.T.A, 2: 40

Syn: Crotalaria pallida var. obovata (G. Don) Polhill

Crotalaria obovata G. Don

Description:

An erect annual woody herb 1.3m. Leaves alternate, with pinnate-trifoliate leaflets,

oblanceolate to spathulate, stipules present, sessile, (2.54 - 7.7) X (1.27-2.0) cm, tip

mucronate and cuneate base. Dry leaf under colour greenish grey, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an

axillary raceme with yellow flower tinged with red, pods are oblong and sessile.

Habitat: Sudanian grassland, often in coastal sands

Distribution: Tropics and subtropics,

Flowering period: September - November

Specimens examined: Oshingboye 131; Oyayomi and Osanyinlusi FHI 82378:

Adebusuyi FHI58692: Jones FHI 18842

Tribe V: Dalbergieae Bronn ex DC.

Dalbergieae Bronn ex DC Prodr. 2: 415 (1825).

Type: Dalbergia Linn. f. F.T.A. 2: 231.

39. Genus: Dalbergia Linn. f.

Dalbergia Linn. f. F.T.A. 2: 231.

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Description:

Climbers, shrubs or small trees. Leaves imparipinnate. Inflorescences paniculate. Fruit

indehiscent, elliptic-oblong to oblong, usually flattened.

87. Dalbergia sissoo (Roxb.) Kuntze Fl. Ind., ed. Carey, 3: 223 (1832).

Syn: Amerimnon sissoo (Roxb.) Kuntze

Description:

A medium to large sized deciduous tree up to 30m. Leaves alternate, imparipinnate

compound with 3-7 leaflets, stipules absent and petiole 18-25mm. Leaflets broad ovate,

(4.5-7.8) X (3.2-5.6) cm, tip acuminate and round base. Dry leaf under colour greenish

grey, flat upper and lower midrib, leaf margin entire and thickened with a leathery

texture. Inflorescence is an axillary raceme with cream or pale yellow flower colour,

pods are linear and flat.

Habitat: Eroded and gullied areas, cultivated

Distribution: Africa, India, Australia and Southern USA

Flowering period: May

Specimens examined: Oshingboye 133; Ogundipe & others 4265; Ujo FHI 21935:

Kennedy FHI8513: Odewo and Adedeji FHI 96902: Brown FHI 57327: Odewo and

Adedeji FHI 96908

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a b

Plate 67: (a) Dalbergia sisso imparipinnate leaves (b) arrow showing inflorescence

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40. Genus: Pterocarpus Jacq.

Pterocarpus Jacq. F.T.A. 2: 237

Type Speceis: Pterocarpus officinalis Jacq.

Description:

Trees, leaves imparipinnate; leaflets alternate to subopposite. Flowers in racemes

or panicles. Fruit flattened, indehiscent with a variously thickened, ± central, seed-

bearing part and a surrounding wing.

88. Pterocarpus erinaceus Poir. F.T.A. 2: 239

Description:

A deciduous tree 15-25m. Leaves alternate, imparipinnate compound with about 7-15

leaflets, stipules minute and petiole 28-35mm. Leaflets alternate, ovate to elliptic, (6.0-

8.6) X (2.5-3.1) cm, tip acute or acuminate and cuneate base. Dry leaf under colour

greenish grey, flat upper and raised lower midrib, leaf margin sinuate and thickened

with a leathery texture. Inflorescence is an axillary panicle with golden yellow flower

colour, pods are circular and flattened.

Habitat: Sudanian woodland

Distribution: Tropical Africa

Flowering period: February - May

Specimens examined: Oshingboye 135; Olorunfemi and others FHI 88320: Eyoh FHI

54534: Onochie FHI 7635: Daramola FHI 104872

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Plate 68: Pterocarpus erinaceus imparipinnate leaves

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41. Genus: Stylosanthes Sw.

Stylosanthes Sw. F.T.A. 2: 155.

Type Speceis: Stylosanthes procumbens Sw.

Description:

Perennial herbs or subshrubs. Leaves pinnately trifoliolate. Inflorescences dense,

axillary or terminal with persistent primary and secondary bracts. Fruit oblong,

flattened, with 1-2 articles, the upper article narrowed into a distinct hooked beak.

89. Stylosanthes erecta P. Beauv. F.T.A. 2: 156

Description:

An annual herb or subshrub 0.1-1.5m. Leaves alternate with trifoliate leaflets, stipules

present; petiole 3-5mm. Leaflets elliptic, (1.2-2.8) X (0.3-0.7) cm, tip acute and obtuse

base. Dry leaf under colour greenish grey, flat upper and raised lower midrib, leaf

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with orange or yellow flower colour, pods are oblong.

Habitat: Savanna woodland, grassland

Distribution: West and Central Tropical Africa

Flowering period: November - June

Specimens examined: Oshingboye 137; Wmwiogbon FHI 87401: Okafor FHI 58754:

Gbile and others FHI 65334: Witt and others FHI 64211: Odewo and Arasi FHI

90989

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Plate 69: Stylosanthes erecta trifoliate leaves

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42. Genus: Aeschynomene Linn.

Aeschynomene Linn. F.T.A. 2: 145

Type Speceis: Aeschynomene aspera L.

Description:

Annual or perennial herbs, subshrubs or shrubs. Leaflets subopposite to alternate,

asymmetric at base. Inflorescences axillary, less often terminal. Pod linear or

elliptic.

90. Aeschynomene indica Linn. F.T.A. 2: 147

Syn: Aeschynomene cachemiriana Cambess.

Aeschynomene glaberrima Poiret

Aeschynomene hispida Willd

Aeschynomene macropoda DC.

Description:

An annual or perennial herb or subshrub 0.3-2.5m. Leaves pinnate and alternate,

stipules present; petiole 3-5mm. Leaflets elliptic to oblong, (0.3-1.8) X (0.1-0.3) cm,

tip acute and cuneate base. Dry leaf under colour greenish grey, flat upper and lower

midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an

axillary raceme with reddish or purple-streaked yellow flower colour, pods are linear

and slightly curved.

Habitat: Sudanian grasslands, sudanian fresh water swamp and acquatic vegetation

Distribution: Africa, North America, Asia

Flowering period: November - June

Specimens examined: Oshingboye 140; Ogundipe & others LUH 4711

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91. Aeschynomene lateritia Harms in Engl. Bot. Jahrb. 26: 292 (1899)

Description:

An annual herb 0.1-8.9m. Leaves pinnate and alternate, stipules present; petiole 3-5mm.

Leaflets elliptic, (0.2-1.8) X (0.1-0.3) cm, tip acute and slightly oblique base. Dry leaf

under colour greenish grey, flat upper and lower midrib, leaf margin entire and not-

thickened with a papery texture. Inflorescence is an axillary raceme with very small

yellow flower colour, pods are linear.

Habitat: Savanna woodland, Sudanian bushland, grassland and thicket

Distribution: West and Central Tropical Africa

Flowering period: September

Specimens examined: Oshingboye 142; Ogundipe & others LUH 5200; Olorunfemi

FHI 24375

43. Genus: Zornia J. F. Gmel.

Zornia J. F. Gmel. F. T. A. 2:158

Type Speceis: Zornia bracteata J.F. Gmel.

Description:

Erect or prostrate, annual or perennial herbs. Leaves digitately 2-or 4-foliolate.

Inflorescence spicate, terminal or axillary. Pods sessile.

92. Zornia latifolia Sm. Rees Cycl. 39, No. 4 (1819)

Syn: Zornia gracilis DC.

Description:

A perrenial herb 0.2-3.5m. Leaves bifoliate, stipules present; petiole 3-5mm. Leaflets

lanceolate to oblong, (1.2-2.8) X (0.3-0.7) cm, tip acute and cuneate base. Dry leaf

235 | P a g e

under colour greenish grey, flat upper and flat lower midrib, leaf margin entire and not-

thickened with a papery texture. Inflorescence is a terminal spike with yellow flower

colour, pods are two to eight jointed and shortly beaked.

Habitat: Sudanian anthropic landscapes, wastelands, Savanna

Distribution: West Tropical Africa; Central and Noth America

Flowering period: October - December

Specimens examined: Oshingboye 143; Keay FHI 28063; FHI 25526

44. Genus: Arachis Linn.

Arachis Linn. F.T.A. 2: 157

Type Speceis: Arachis hypogaea L.

Description:

Annual or perennial herbs. Leaves paripinnate, 4-foliolate. Inflorescences axillary

dense sessile 2-7-flowered spikes. Pod 1-6-seeded, maturing underground,

constricted between the seeds, but not jointed, indehiscent; the walls thick and

reticulate.

93. Arachis hypogaea Linn. F. T. A. 2: 158

Syn: Arachis hypogaea L. subsp. oleifera A.Chev.

Arachis nambyquarae Hoehne

Description:

An annual herb or subshrub 0.1-0.5m. Leaves alternate with trifoliate leaflets, stipules

present; petiole 3-5mm. Leaflets pblong to ovate, (3.2-5.8) X (1.3-2.7) cm, tip acute

and obtuse base. Dry leaf under colour greenish green, flat upper and flat lower midrib,

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leaf margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with yellow flower colour, pods are contracted between the seeds.

Habitat: Cultivated

Distribution: South and , cultivated

Flowering period: September

Specimens examined: Oshingboye 145; Ogundipe & others LUH 1317A; Olorunfemi

FHI 24375

Tribe VI: Robinieae Hutch.

Robinieae Hutch. (1964).

Type: Robina Cothen. Disp. 21. (1790).

45. Genus: Gliricidia Kunth

Gliricidia Kunth Nov.Gen. Sp. 6:393 (1824).

Type Species: Gliricidia sepium (Jacq.) Kunth ex Walp.

94. Gliricidia sepium (Jacq.) Kunth ex Walp.

Syn: Galedupa pungam Blanco

Gliricidia maculata (Kunth) Walp.

Lonchocarpus sepium (Jacq.) DC.

Description:

A small to medium large deciduous tree 10-12m. Leaves opposite, imparipinnate

compound with 5-19 leaflets, stipules present and petiole 17-28mm. Leaflets opposite,

elliptic to ovate, (4.1-6.7) X (2.1-2.6) cm, tip acuminate and cuneate base. Dry leaf

under colour rusty, flat upper and lower midrib, leaf margin entire and not-thickened

with a papery texture. Inflorescence is an axillary raceme with pink to lilac flower

colour, pods are flat and elongated.

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Habitat: cultivated

Distribution: Tropical America

Flowering period: November - January

Specimens examined: Oshingboye 146; Ohaeri FHI 78914: Goddy FHI 41717: Witt

FHI 64393

Plate 70: Gliricidia sepium leaves, arrow showing pod

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Tribe VII: Indigofereae Benth.

Indigofereae Benth. (1859)

Type: Indigofera Linn. F.T.A. 2: 65

46. Genus: Indigofera Linn. F.T.A. 2: 65

Description:

Herbs or shrubs. Hairs usually medifixed. Leaves usually imparipinnate. Flowers

usually in axillary racemes. Corolla usually red. Fruit usually dehiscent.

95. Indigofera arrecta Hochst. ex A. Rich. Tent. Fl. Abyss. 1: 184 (1847); F.T.A. 2: 97

Syn: Indigofera arecta A. Rich.

Indigofera madagascariensis Colla

Indigofera umbonata Baker

Description:

An erect woody shrub up to 3m. Leaves alternate, imparipinnate compound with 7-21

leaflets, stipules present and petiole 11-12mm. Leaflets opposite, elliptic to oblong,

(1.4-2.4) X (0.4-0.9) cm, tip obtuse or round and cuneate base. Dry leaf under colour

greenish grey, flat upper and lower midrib, leaf margin entire and not-thickened with a

papery texture. Inflorescence is an axillary raceme with pink or red flower colour, pods

are linear.

Habitat: Sudanian bushland and thicket

Distribution: Tropical Africa, Middle East

Flowering period: October

Specimens examined: Oshingboye 149; Odewo and Binuyo FHI 100953: Olorunfemi

and others FHI94291: Latilo FHI 62818: Clayton FHI 39905: Witt and others FHI

77612: Okafor FHI 59083: Parker RNG 4666; Fortune RNG 2220

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Plate 71: Indigofera arrecta imparipinnate leaves, arrow showing flowers

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96. Indigofera hirsuta L. Sp. Pl. 2: 1062 (1753); F.T.A. 2: 88

Syn: Anila hirsuta (L.) Kuntze

Indigofera astragalina DC.

Indigofera ferruginea Schum. & Thonn.

Indigofera fusca G. Don

Indigofera hirta Bojer

Indigofera indica Mill.

Description:

An erect annual herb 1.5m. Leaves alternate, imparipinnate compound with 5-9 leaflets,

stipules present and petiole 1mm. Leaflets opposite, elliptic to oblong, 0.4 X 0.25 cm,

tip obtuse or sub-acute and cuneate base. Dry leaf under colour greenish grey, flat upper

and lower midrib, leaf margin entire and slightly thickened with a leathery texture.

Inflorescence is an axillary raceme with pink or brick-red flower colour, pods are linear.

Habitat Sudania regional transition zone, wasteland

Distribution: Tropical Africa, India, South America

Flowering period: September

Specimens examined: Oshingboye 150; Ogundipe & others 4237; LUH 1302A;

Okeke and others FHI7260: Lowe FHI27512: Jackson FHI 15846: Latilo FHI63549:

Magaji FHI17973: Jackson FHI54370: Daramola FHI 60291; Hacker RNG 184

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Plate 72: Indigofera hirsuta imparipinnate leaves, arrow showing inflorescence

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97. Indigofera nummulariifolia (L.) Livera ex Alston Trim. Fl.Ceyton 6, suppl .72 (1931).

Syn: Hedysarum nummularifolium Linn.

Indigofera echinata Willd.

Description:

An annual branched herb 0.6-0.7m. Leaves alternate, simple, obovate to orbicular-

obovate, stipules present, petiole 1-2mm, (2.0-2.7) X (1.3-1.9) cm, tip obtuse and

cuneate base. Dry leaf under colour greenish yellow, flat upper and raised lower midrib,

leaf margin entire and thickened with a leathery texture. Inflorescence is an axillary

raceme with red or pink flower colour, pods are linear.

Habitat: wasteland,

Distribution: Tropical Africa, Asia, India

Flowering period: August

Specimens examined: Oshingboye 152; Ogundipe & others LUH 4260; LUH 5210;

Keay FHI 12733; 28010; Ujor FHI 34119

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Plate 73: Indigofera nummulariifolia leaves

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98. Indigofera spicata Forssk. l.c. 138 (1775).

Syn: Jacq.

Description:

A perennial creeping herb up to 0.5m. Leaves alternate, imparipinnate compound with

9-13 leaflets, stipules present and petiole 4-5mm. Leaflets opposite, elliptic to obovate,

(0.9-5.0) X (0.4-0.6) cm, tip obtuse or mucronate and cuneate base. Dry leaf under

colour greenish grey, flat upper and lower midrib, leaf margin entire and thickened with

a leathery texture. Inflorescence is an axillary raceme with pink or orange-red flower

colour, pods are descending and shortly pointed.

Habitat: Sudanian anthropic landscapes, wasteland, bushland

Distribution: Tropical Africa, Middle East, Central America

Flowering period: May - July

Specimens examined: Oshingboye 154; Ogundipe & others 4703; Tamajong FHI

16793

245 | P a g e

Plate 74: Indigofera spicata imparipinnate leaves

246 | P a g e

99. Indigofera suffruticosa Mill. Gard. Dict., ed. 8: 2 (1768)

Syn: Indigofera angolensis D. Dietr.

Indigofera anil L.

Description:

A perennial herb or woody shrub up to 2m. Leaves alternate, imparipinnate compound

with about 13 leaflets, stipules minute and petiole 10-15mm. Leaflets opposite, obovate

to elliptic, (2.6-4.0) X (1.1-1.6) cm, tip mucronate and cuneate base. Dry leaf under

colour greenish grey, flat upper and lower midrib, leaf margin entire and thickened with

a leathery texture. Inflorescence is an axillary raceme with pink or red flower colour,

pods are sickled shaped.

Habitat: cultivated

Distribution: America, introduced in Tropical Africa

Flowering period: April

Specimens examined: Oshingboye 155; Ogundipe & others LUH 4631; Jones &

Onochie FHI 17244

247 | P a g e

Plate 75: Indigofera suffruticosa imparipinnate leaves

248 | P a g e

100. Indigofera conferta Gillelt l.c. 579 (1956)

Syn: Indigofera grisea Bak.

Indigofera bracteolate DC.

Description:

A perennial branching herb up to 1m. Leaves alternate, imparipinnate compound with

about 13 leaflets, stipules minute and petiole 10-15mm. Leaflets opposite, obovate to

ovate, (2.6-3.0) X (1.1-1.6) cm, tip acute and cuneate base. Dry leaf under colour

greenish grey, flat upper and lower midrib, leaf margin entire and thickened with a

leathery texture. Inflorescence is an axillary raceme with pink or red flower colour,

pods are slightly sickled shaped.

Habitat: wasteland

Distribution: Tropical Africa

Flowering period: April

Specimens examined: Oshingboye 156; Ogundipe & others LUH 4631; Ejiofor FHI

29389

101. Indigofera dendroides Jacq. Ic. Pl. Bar. 3, t. 571 (1788–9); F.T.A. 2: 100

Syn: Indigofera dalabaca A. Chev.

Indigofera kengeleensis De Wild.

Indigofera sesbaniifolia A. Chev.

Description:

A branched annual herb up to 1.0m. Leaves alternate, imparipinnate compound with

numerous leaflets. Leaflets opposite, obovate to elliptic, (1.6-3.3) X (0.3-1.0) cm, tip

mucronate and cuneate base. Dry leaf under colour greenish grey, flat upper and lower

249 | P a g e midrib, leaf margin entire and not thickened with a papery texture. Inflorescence is an axillary raceme with pink or red flower colour, pods are sickled shaped.

Habitat: wasteland

Distribution: Tropical Africa

Flowering period: August - September

Specimens examined: Oshingboye 167; Ogundipe & others LUH 4675; Olorunfemi

FHI 24368; Iwens RNG 101

102. Indigofera nigritana Hook. f. Fl. Nigrit. 294 (1849); F.T.A. 2: 78

Description:

An erect perennial herb up to 1.5m. Leaves alternate, imparipinnate compound with about 5-7 pairs of leaflets, stipules minute and linear, petiole 10-15mm. Leaflets opposite, oblanceolate to elliptic, (2.6-4.0) X (1.1-1.6) cm, tip round and cuneate base.

Dry leaf under colour greenish grey, flat upper and lower midrib, leaf margin entire and not-thickened with a papery texture. Inflorescence is an axillary raceme with red flower colour, pods are oblong.

Habitat: wasteland

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye 169; Ogundipe & others LUH 4469

103. Indigofera macrocalyx Guill. & Perr. Fl. Seneg. 1: 175, 46 (1832); F.T.A. 2: 71

Description:

A perennial herb up to 1m. Leaves alternate, imparipinnate compound with about 3-6 pair of leaflets, stipules minute and petiole 1-6mm. Leaflets opposite, obovate to elliptic, (2.6-4.0) X (1.1-1.6) cm, tip mucronate or round and cuneate base. Dry leaf

250 | P a g e under colour greenish grey, flat upper and lower midrib, leaf margin entire and thickened with a leathery texture. Inflorescence is an axillary raceme with pink or red flower colour, pods are roundish pointed.

Habitat: wasteland

Distribution: Tropical Africa

Flowering period: October - November

Specimens examined: Oshingboye 171; Ogundipe & others LUH 4629; Iwens RNG

17

104. Indigofera paniculata Vahl ex Pers. Syn. 2: 325 (1807)

Syn: Indigofera procera Schum. & Thonn

Description:

An erect much branched annual herb up to 1.4m. Leaves alternate, simple, stipules minute and petiole 1-2mm, linear to lanceolate, (2.6-5.0) X (0.1-0.6) cm, tip acute and cuneate base. Dry leaf under colour greenish grey, flat upper and lower midrib, leaf margin entire and thickened with a leathery texture. Inflorescence is an axillary raceme with pink or red flower colour, pods are linear.

Habitat: wasteland

Distribution: Tropical Africa, Angola

Flowering period: October

Specimens examined: Oshingboye 172; Ogundipe & others LUH 4675; Iwens RNG

N104

251 | P a g e

105. Indigofera pulchra Willd. – F.T.A 2:76

Syn: Indigofera dupuisii Mich.

Description:

An erect annual herb up to 1m. Leaves usually dimorphic, alternate, imparipinnate

compound with about 13 leaflets, stipules minute and petiole 3-6mm. Leaflets opposite,

oblanceolate to elliptic, (0.26-0.6) X (0.2-0.6) cm, tip mucronate and cuneate base. Dry

leaf under colour greenish grey, flat upper and lower midrib, leaf margin entire and

non-thickened with a papery texture. Inflorescence is an axillary raceme with red flower

colour, pods are oblong.

Habitat: Sudanian woodland,

Distribution: Tropical Africa

Flowering period: May - July

Specimens examined: Oshingboye 176; Emwiogbon and others FHI87044:

Olorunfemi FHI 86835: Soladoye and others FHI 84702: Daramola and Emwiogbon

FHI 72397

Tribe VIII: Swartzieae DC.

Swartzieae DC. Prodr. 2: 422 (1825).

Type: Swartzia Schreb. F.T.A. 2: 256.

47. Genus: Bobgunnia J.H. Kirkbr. & Wiersema

Bobgunnia J.H. Kirkbr. & Wiersema Brittonia 49(1): 1. (1997)

Description:

Trees or shrubs. Leaves imparipinnate; leaflets alternate without translucent dots or

streaks. Flowers in lateral racemes. Pod indehiscent, cylindric or beaded, woody.

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106. Bobgunnia madagascariensis Desv. F.T.A. 2: 257

Syn: Swartzia madagascariensis Desv.

Description:

A small deciduous tree 10-15m. Leaves alternate, imparipinnate compound with about

7-15 leaflets, stipules present and petiole 35-46mm. Leaflets alternate, ovate to elliptic,

(3.9-5.0) X (2.0-2.6) cm, tip round or slightly retuse and cuneate base. Dry leaf under

colour greenish yellow, channelled upper and raised lower midrib, leaf margin entire

and thickened with a leathery texture. Inflorescence is an axillary raceme with white

flower colour, pods are cylindrical.

Habitat: Sudanian wooded grassland

Distribution: West Tropical Africa, Mascarene Islands

Flowering period: March - May

Specimens examined: Oshingboye 178; Ohaeri FHI 78740: Eyoh FHI 54537:

Oyayomi and others FHI 79910: Olorunfemi FHI 55761: Ogbe and Jaiyesimi FHI

15891: Latilo FHI 27430:

253 | P a g e

Plate 76: Bobgunnia madagascariensis imparipinnate leaves

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Tribe IX: Millettieae Miq.

Millettieae Miq. (1855).

Type: Millettia Wight & Arn. Prodr. 1:263-264 (1834).

48. Genus: Tephrosia Pers.

Tephrosia Pers. F.T.A. 2: 104.

Description:

Herbs, subshrubs or shrubs. Leaves trifoliolate; leaflets covered with gland dots

beneath. Inflorescences axillary or terminal. Ovary 2-ovuled, Pods oblong-falcate,

flattened, much narrowed towards the base.

107. Tephrosia elegans (Pers.) Schum. F.T.A. 2: 118

Description:

An annual herb 0.9-1.2m. Leaves alternate and pinnate compound, stipules present,

petiole 2-4mm. Leaflets (2) opposite, elliptic, oblong or oblanceolate (2.5-5.5) X (0.5-

1.6) cm, tip obtuse or narrowly round and cuneate base. Greenish brown dry leaf under

colour, flat upper and raised lower midrib, leaf margin entire and thickened with a

leathery texture. Inflorescence is an axillary raceme with apricot orange flower colour,

pods are linear to oblong.

Habitat: sudanian grassland

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye 180; Jones & Keay FHI 4874 Onochie FHI 8162;

Latilo FHI 23534

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Plate 77: Tephrosia elegans leaves, arrow showing stipule

256 | P a g e

108. Tephrosia bracteolata Guill. & Perr. F.T.A. 2: 116

Description:

An annual herb 2-3m. Leaves are alternate, up to 31.2cm long, pinnate compound with

about 10-16 pairs of leaflets per pinna, petiole 8mm; stipules present. Leaflets elliptic

(5.4-7.8) X (0.4-0.6) cm, opposite, with an emarginate tip, leaf base cuneate. Greenish

grey under surface, flat upper and lower midrib, margin entire and slightly thickened

with a leathery texture. Inflorescence is pseudo-raceme, pale pink flowers, pods are flat

and linear.

Habitat: Sudaian grassland

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye 182; Geerling FHI 38391: Odewo and Oni FHI

79313: Emwiogbon and Osanyinlusi FHI 87022

257 | P a g e

Plate 78: Tephrosia bracteolata pinnate leaves

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109. Tephrosia linearis (Willd.) & Pers. F.T.A. 2: 120

Syn: Galega linearis Willd.

Tephrosia discolor E. Mey.

Tephrosia rutenbergiana Vatke

Tephrosia seminuda Baker

Description:

An erect annual herb up to 1.5m. Leaves are alternate, up to 17.6cm long, pinnate

compound with about 6-13 pairs of leaflets per pinna, petiole 1-2mm; stipules present.

Leaflets are linear-oblong to elliptic (3.2-4.3) X (0.3-0.4) cm, opposite, with a narrowly

round tip, leaf base cuneate. Greenish grey under surface, flat upper and lower midrib,

margin entire and not-thickened with a papery texture. Inflorescence is pseudo-raceme,

orange, yellow, red or pink flowers, pods are flat and linear.

Habitat: Sudanian grassland, wasteland

Distribution: Tropical Africa

Flowering period: September

Specimens examined: Oshingboye 183; Ogundipe & others LUH 4310; LUH 4684;

Keay FHI 22274

259 | P a g e

Plate 79: Tephrosia linearis pinnate leaves

260 | P a g e

110. Tephrosia platycarpa Guill. & Perr. F.T.A. 2: 109; Bak. f. l.c. 187.

Description:

An annual herb 0.3-0.4m. Leaves are alternate, up to 18.5cm long, pinnate compound

with about 5-6 pairs of leaflets per pinna, petiole 27-35mm; stipules present. Leaflets

are elliptic (6.2-7.0) X (0.2-0.3) cm, opposite, with an acute tip, and cuneate leaf base.

Greenish grey under surface, flat upper and lower midrib, margin entire and not-

thickened with a papery texture. Inflorescence is a raceme, corolla not seen, pods are

linear to oblong.

Habitat: Sudanian grassland, wasteland

Distribution: Tropical Africa

Flowering period: August - September

Specimens examined: Oshingboye 185; Oguntayo and others FHI 81256: Latilo and

Fagbemi FHI 67251: Latilo FHI 62513: Charter FHI 38752: Okafor and Daramola

FHI 72415

261 | P a g e

Plate 80: Tephrosia platycarpa pinnate leaves

262 | P a g e

111. Tephrosia pedicellata Bak – F.T.A. 2: 117

Description:

A perennial herb 0.3-0.4m. Leaves are alternate, pinnate compound with about 5-6 pairs

of leaflets per pinna, petiole 27-35mm; stipules present. Leaflets are oblanceolate (2.5-

3.5) X (0.2-0.3) cm, opposite, with an acute or round tip, and cuneate leaf base.

Greenish grey under surface, flat upper and lower midrib, margin entire and not-

thickened with a papery texture. Inflorescence is a raceme, with pink flower colour.

Pods are linear to oblong.

Habitat: Sudanian grassland, wasteland

Distribution: Tropical Africa

Flowering period: September - October

Specimens examined: Oshingboye 186; Latilo FHI 46904: Afolayan FHI 25947:

Ibhanesebhor and others FHI100969: Olorunfemi FHI 48101: De Leeuw FHI 18005:

Daramola FHI 45542: Rojer FHI 48461

49. Genus: Lonchocarpus H. B. & K.

Lonchocarpus H.B. & K. F.T.A. 2:241

Description:

Mostly trees, less often shrubs, leaves imparipinnate. Inflorescences axillary or

terminal. Ovary 2-ovuled, pods clustered, oblong, flattened, much narrowed towards

the base.

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112. Lonchocarpus sericeus (Poir.) Kunth ex DC. F.T.A. 2: 241.

Syn: Derris sericea (Poir.) Ducke

Lonchocarpus cruentus Lundell

Lonchocarpus formosanus DC.

Robinia sericea Poir.

Description:

An evergreen shrub or small tree up to 15m. Leaves are alternate, up to 28.5cm long,

paripinnate compound with about 4-6 pairs of leaflets per pinna, petiole 10-17mm;

stipules present. Leaflets are elliptic to ovate (9.1-14.0) X (3.2-6.8) cm, opposite, with

terminal leaflets bigger than lateral ones, tip shortly acuminate, leaf base broadly

cuneate or round. Greenish grey under surface, flat upper and slightly raised lower

midrib, margin sinuate and not-thickened with a papery texture. Inflorescence is an

axillary raceme with lilac flower colour; pods are flat and clustered.

Habitat: fringing forest, sudania transition forest

Distribution: Tropical Africa,

Flowering period: May - August

Specimens examined: Oshingboye 187; Felix FHI 106534: Howard FHI 56787:

Ariwaodo FHI 94767: Onochie FHI 32035

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Plate 81: Lonchocarpus sericeus paripinnate leaves

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113. Lonchocarpus cyanescens (Schumach. & Thonn.) Benth. F.T.A. 2: 243

Syn: Philenoptera cyanescens (Schum. & Thonn.) Roberty

Robinia cyanescens Schum. & Thonn.

Description:

A deciduous scandent shrub or medium-sized tree up to 20m tall. Leaves are alternate,

up to 19.2cm long, paripinnate compound with about 4-6 pairs of leaflets per pinna,

petiole 8-18mm; stipules present. Leaflets are obovate (5.8-9.0) X (2.5-3.9) cm,

opposite, with terminal leaflets bigger than lateral ones, tip shortly acuminate, leaf base

cuneate. Greenish grey under surface, flat upper and raised lower midrib, margin

sinuate and thickened with a leathery texture. Inflorescence is an axillary raceme with

lilac or blue flower colour; pods are oblong and pointed.

Habitat: Sudanian forest, bushland and thicket, cultivation

Distribution: Tropical Africa

Flowering period: April – May

Specimens examined: Oshingboye 188; Jones FHI 484

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Plate 82: Lonchocarpus cyanescens leaves

267 | P a g e

50. Genus: Millettia Wight & Arn.

Millettia Wight & Arn. F.T.A. 2: 126; Dunn J. Linn. Soc. 41: 123 (1912)

Description:

Trees or shrubs, sometimes scandent. Leaves imparipinnate. Flowers arranged in

terminal racemes or panicles. Pod flattened, coriaceous, 2-valved but often tardily

dehiscent.

114. Millettia bateri (Benth.) Dunn J. Bot. 49: 221 (1911)

Description:

A large , rarely a shrub or small tree3m tall. Leaves are alternate, imparipinnate

compound with about 2-7 pairs of leaflets per pinna, petiole 20-66m; stipules present.

Leaflets are elliptical to oblong (5.8-9.0) X (2.5-3.9) cm, opposite, tip obtusely

acuminate, leaf base cuneate. Greenish grey under surface, flat upper and lower midrib,

margin entire and not thickened with a papery texture. Inflorescence is an axillary

raceme with pale pink or red colour; pods are linear and flat.

Habitat: Sudania regional transition zone; forest

Distribution: Tropical Africa

Flowering period: March – April

Specimens examined: Oshingboye 189; Ogundipe & others LUH 4680; Onochie FHI

15542

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51. Genus: Ostryoderris Dunn.

Ostryoderris Dunn. Kew. Bull. 1911:363

Description:

Deciduous tree; leaves imparipinnate; leaflets varying from opposite, subopposite to

alternate. Fruit flattened, indehiscent, and linear-oblong.

115. Ostryoderris stuhlmannii (Taub.) Dunn ex Harms Engl. Pflanzenw. Afr. 3, 1:

644 (1915)

Syn: Xeroderris stuhlmannii (Taub.) Mendonca & Sousa

Ostryoderris chevalieri Dunn.

Description:

A deciduous tree up to 20m tall. Leaves are alternate, imparipinnate compound with

about 4-9 pairs of leaflets per pinna, petiole 8-18mm. Leaflets are oblong to ovate (5.8-

12.8) X (3.5-5.2) cm, opposite, tip obtuse, leaf base asymmetric. Greenish grey under

surface, flat upper and raised lower midrib, margin entire and thickened with a leathery

texture. Inflorescence is an axillary raceme with white to greenish white flower colour;

pods are oblong and flat.

Habitat: deciduous woodland, grassland and Savanna

Distribution: West Tropical Africa

Flowering period: February

Specimens examined: Oshingboye 190; Ogundipe & others LUH 4602; Jaiyesimi FHI

15825; Keay FHI 16172; Ogua FHI 7817

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Tribe X: Sesbanieae Hutch.

Sesbanieae Hutch. (1964).

Type: Sesbania Scop. F.T.A. 2: 133

52. Genus: Sesbania Scop.

Sesbania Scop F.T.A. 2: 133

Description:

Herbs, shrubs, or small soft woody prickly trees, leaves pinnate; leaflets numerous,

linear, oblong, mucronate and deciduous. Flowers in axillary racemes.

116. Sesbania bispinosa (Jacq.) W. F. Wight Bur. Pl. Indust. Bull. 137; 15 (1909).

Syn: Aeschynomene aculeata Schreb.

Aeschynomene bispinosa Jacq.

Aeschynomene spinulosa Roxb.

Sesbania aculeata (Willd.) Pers.

Description:

An erect annual shrub up to 4m tall. Leaves are alternate, up to 35.2cm long, pinnate

compound with about 15-22 pairs of leaflets per pinna, petiole 8-18mm; stipules

present. Leaflets are oblond to oblong linear (1.2-2.5) X (0.3) cm, opposite, tip obtuse,

emarginate or apiculate, leaf base obtuse. Greenish grey under surface, flat upper and

raised lower midrib, margin sinuate and thickened with a leathery texture. Inflorescence

is an axillary raceme with yellow and purple-spotted flower colour; pods are slightly

curved.

Habitat: Muddy swamp, road side

Distribution: Tropics and sub-tropics

Flowering period: July - September

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Specimens examined: Oshingboye 192; Lowe FHI 43094: Latilo FHI 63529: Ekwuno

and Fagbemi FHI 93931: Daramola FHI 98990

117. Sesbania dalzielii Phill. & Hutch. l.c. 49 (1921).

Syn: Sesbania sudanica subsp. occidentalis J. B. Gillett

Description:

A slender shrub up to 2m high. Leaves are alternate, up to 20cm long, imparipinnate

compound with about 40 pairs of leaflets per pinna, petiole 2-5mm; stipules present.

Leaflets are linear to oblong (0.8-2.5) X (0.2-0.4) cm, tip round or acutely mucronate,

leaf base obliquely cuneate. Greenish grey under surface, flat upper and lower midrib,

margin entire and not-thickened with a papery texture. Inflorescence is an axillary

raceme with lilac or blue flower colour; pods are torulose and pointed.

Habitat: Muddy swamp, road side

Distribution: Tropics and sub-tropics

Flowering period: October

Specimens examined: Oshingboye 193; Ogundipe & others LUH 5128

Tribe XI: Sophoreae Spreng. ex DC.

Sophoreae Spreng. ex DC. Prodr. 2: 94 (1825).

Type: Sophora L. Sp. Pl. 1: 373 (1753).

53. Genus: Pericopsis Thwaites

Pericopsis Thwaites Enum. Pl. Zeyl. 413 (1864).

Description:

Trees or shrubs. Leaves imparipinnate; leaflets alternate or subopposite. Inflorescence

a terminal panicle. Fruit flat, winged along upper or both margins, indehiscent.

271 | P a g e

118. Pericopsis laxiflora (Benth.) Meeuwen Notizbl. 21, 2: 64 & fig. (1911)

Syn: Afrormosia laxiflora (Benth. ex Bak.) Harms.

Description:

A deciduous scandent shrub or medium-sized tree up to 14m tall. Leaves are alternate,

imparipinnate compound with about 9-13 pairs of leaflets per pinna, petiole 8-18mm;

stipules present. Leaflets are narrowly lanceolate to broadly ovate (5.8-9.0) X (2.5-3.9)

cm, tip shortly acuminate to acute, leaf base cuneate. Greenish grey under surface, flat

upper and raised lower midrib, margin entire and thickened with a leathery texture.

Inflorescence is terminal panicle with greenish white or cream flower colour; pods are

oblong.

Habitat: Savanna woodland and fringing forest

Distribution: Tropical Africa

Flowering period: May - June

Specimens examined: Oshingboye 195; Ogundipe & others LUH 4219; Eimunjeze and

Oguntayo FHI 71316: Gbile FHI 18519: Eimunjeze and Adebusuyi FHI 69984: Latilo

FHI 58705: Macaulay FHI 59913

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4.2.2 Keys

4.2.2.1 Bracketed Keys to Sub-Families:

1a. Flower actinomorphic; petals valvate in bud, often united at the base………………

………...……………………………………………………...……………... Mimosoideae

1b. Flower zygomorphic; petals imbricate in bud, free or some of them united;

………………………………….………………………………...... ………. 2

2a. Adaxial petal over-lapped by adjacent lateral petals; sepals generally free; leaves

bipinnate or pinnate, rarely simple or unifoliate………………………... Caesalpinioideae

2b. Adaxial petal not over-lapped but outside the adjacent lateral petals; sepals united at

the base; leaves simple, pinnate, imparipinnate, trifoliate or pinnate-

trifoliate…...………...... Papilionoideae

Sub-Family: Mimosoideae

Members are trees or shrubs, very rarely simple pinnate leaves, often with large glands on the rachis. Flowers are hermaphroditic, small, spicate, racemose or capitate, actinomorphic, usually pentamerous. Calyx tubular, valvate or very rarely imbricate, 5 – lobed or toothed.

Petals are valvate, free or cornate into short tube. Stamens equal in number to the sepals or more numerous, free or monadelphous; anthers small, 2-celled, opening lengthwise, often with a deciduous gland at the apex. Ovary superior, of one carpel, fruit mostly dehiscent and seeds with scanty or no endosperm.

Keys to Genera: 1a. Flowers bicolour with upper pink and lower yellow...... Dichrostachys

1b. Flowers not bicolor ……………………………………………………….………… 2

2a. Habit shrub ………………………………………………………………………….. 3

2b. Habit tree …………………………………………………………………………….. 4

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3a. Shrub between 2- 10m, leaf elliptic with obtuse tip ………………..……... Leucaena

3b. Shrub or herb not more than 6m, leaf oblong, elliptic or lanceolate …….…… Mimosa

4a. Mostly trees between 5 – 20m, leaves reduced to phyllodes, armed with thorns and prickles, flower colours yellow to golden yellow …………………...... …... Acacia

4b. Low branching tree, not more than 12m, leaf oblong, elliptic or lanceolate ..…... Entada

5a. Tree between 15 – 20m, leaf obliquely elliptic with obtuse tip ………. Pithecellobium

5b. Tree at least 20m, lanceolate leaf with acuminate tip and attenuate base ……. Prosopis

6a. Decidious tree up to 20m, leaf oblong or obliquely rhombic, obtuse tip, round to oblique base, flower colour off-white ……………………………………………….………... Albizia

6b. Decidious tree 35m, leaf oblong, obtuse tip, round base, flower colour orange or red …

…………………………………………………………………………………………. Parkia

Sub-Family: Caesapinioideae

Members are Tropical or subTropical trees and shrubs, root nodules uncommon. Leaves bipinnate or paripinnate, rarely simple, 1-foliolate or 2-foliolate, sometimes with specialized glands, but very rarely with a tendril. Flowers often bilaterally symmetrical, rarely radially symmetrical, with five petals which are not differentiated into standard, wings and keel. Sepals generally free; petals imbricate in bud, the median petal overlapped by the lateral 4; stamens are visible externally, (1-)10(-50). Seeds often without a pleurogram, but if pleurogram present then closed.

Keys to Genera: 1a. Simple leaves …………………………………………………………………….... 2

1b. Compound leaves …………………………………………………………….…….. 3

2a. Leaves orbiculate, bilobed margin, with pale pink, or purple flower colour …..Bauhinia

2b. Leaves orbcordate, entire margin, with white flower colours…………….... Piliostigma

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3a. Leaves paripinnate compound ……………….…………………………………….... 4

3b. Leaves bipinnate compound, obtuse tip, oblique to round base, entire margin, papery texture, red flower colour ……………………………………………………………..….... 8

4a. Leaves elliptic or ovate ………………………………………………………………. 5

4b. Leaves oblong or ovate ………………………………………………………………. 6

5a. Obtuse tip, cuneate base, margin entire, whitish to yellowish flower colour …. Afzelia

5b. Acute or short acuminate tip, slightly oblique base, undulated margin, white or pale pink flower colour …………………………………………………………………. Isoberlinia

6a. Round tip, assymetric round base, entire margin, stipules present, yellow or pink flower colour ……………………….…………………………………………………… Tamarindus

6b. Acuminate tip, oblique base, entire margin, stipules present, white to cream flower colour, samara pod …………………………………………………………….…... Daniellia

7a. Ovate leaves, acute tip, oblique base, entire margin, stipules absent, cream to white flower colour ……….……………………………………………………..………… Detarium

7b. Obovate leaves, acuminate tip, oblique base, entire margin, stipules absent, cream to white flower colour, corky pod …………………………………………..………… Cynometra

8a. Leaves armed with spines or prickles …………….…………………….… Caesalpinia

8b. Leaves not armed with spines or prickles …..……………………..….……... Delonix

9a. Habit tree, elliptic to ovate leaves …………………………………………………... 10

9b. Habit shrub or herb ………………………………………………………………….. 11

10a. Obtuse or emarginate tip, oblique to round base, entire margin, minute stipules present, cream to white flower colour …………………………………………….……………. Burkea

10b. Acute tip, round or cuneate base, entire margin, stipules absent, bright yellow flower colour ………………………...……………………………………………………….... Cassia

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11a. Habit herb, sessile leaf attachment, oblong, elliptic or obovate leaves, round or mucronate tip, round or obliquely truncate base, yellow to bright yellow flower.. Chamaecrista

11b. Habit shrub, petiolate, oblong, obovate, elliptic, lanceolate leaves, obtuse, retuse, or acuminate tip, round or oblique base, yellow to orangey-yellow flower colour ……… Senna

Sub-Family: Papilionoideae

This is the largest of the three sub-families with about two – thirds of all the genera and species in the family. Members are mostly herbs and shrubs with a few tree species, they are mostly easily recognized by their papilionaceous (butterfly-like) flowers. The flower is irregular

(zygomorphic) and is made up of five petals, free or adnate to the staminal tube, posterior petal

(standard) outside in bud, the two lateral (wings) intermediate, the two lower inside and usual cohering by short lobed margin (keel), stamens usually 9 + 1, the one vexillary, opposite the standard.

Keys to Genera: 1a. Flowers bicolour with pink keel and yellow standard……………...... Adenodolichos

1b. Flowers monocolour ranging from purple to yellow, red, and pink...... 2

2a. Flowers streaked with red or purple ……………...... Aeschyonomene

2b. Flowers not streaked …………………………………………………………………. 3

3a. Leaf trifoliate compound ………………..…………………………………………... 4

3b. Leaf pinnate, bifoliate or imparipinnate compound …...……………………………. 13

4a. Leaflets pinnate-trifoliate…………………………………………………………..… 5

4b. Leaflets not pinnate-trifoliate …...…………………………………………………... 10

5a. Habit herb ……………………………………………………………………………. 6

5b. Habit shrub, sub-shrub or small tree ………………………………………………… 9

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6a. Erect herb, leaves oblong to lanceolate, acuminate tip, cuneate base, red flower colour

…………………………………………………………………………..……… Alysicarpus

6b. Creeping herb, leaves ovate to rhomboid ovate, obtuse tip, round to cuneate base, blue or purple flower colour ………………….…………………………………... Calopogonium

7b. Straggling herb, leaves ovate to elliptic, acute tip, cuneate bae, blue, purple, violet, white or pink flower colours …………………………………….………………... Desmodium

7a. Creeping herb, leaves ovate to elliptic, emarginate tip, cuneate base, yellow flower colour …………………………………………………………..………………………. Vigna

8a. Shrub and sub-shrub ……………………………………………………………… 9

8b. Small tree, leaves oblong to ovate, obtuse or round tip, emarginate or mucronate tip, broadly cunueate or obliquely round base, presence of prickles on bark and stem, red to bright red flower colour ……………………………………………….…………………... Erythrina

9a. Sub-shrub, leaves oblong to lanceolate, aristate or apiculate tip, cuneate base, yellow to cream flower colour ……………………………..………………………….……... Cajanus

9b. Shrub, leaves elliptic to oblong obovate, obtuse tip, cuneate base, golden yellow flowers ……………………………………………………………………………… Eriosema

10a. Habit branched herb, leaves ovate to oblong, round, emarginate or mucronate tip, cuneate or round base, yellow flower colour ………………………………………... Arachis

10b. Habit erect herb, leaves oblanceolate, elliptic, spathulate or oblong, obtuse, cuspidate or acute tip, cuneate base, yellow, bright yellow or pale yellow flower colour ……... Crotalaria

11a. Habit creeping herb, leaves ovate or rhomboid, acute tip, base of terminal leaves are cuneate while lateral ones are asymmetric rhombic, purple or cream flower colour…. Mucuna

11b. Habit rect herb, leaves elliptic, acute tip, obtuse base, orange or yellow flower colour

………………………………………………………………………………….... Stylosanthes

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12a. Erect herb, leaves ovate, tip obtuse, cuneate base, purple or pink to white flower colour

………………………………………………………………………………………… Glycine

12b. Climbing herb, leaves oblong to lanceolate, tip acute, cuneate base, yellow flower colour ………………………………………………………………………………. Dolichos

13a. Leaves bifoliate or pinnate compound …………………………………………….. 14

13b. Leaves imparipinnate compound ………………………………………………...... 17

14a. leaves bifoliate, leaves lanceolate to oblong, acute tip, cuneate base, yellow flower colour …………………………………………………………………………………... Zornia

14b. Leaves pinnate compound and habit herb …….…………………………………….. 15

15a. Leaves elliptic, flowers yellow with purple spots, pod slightly curved …….... Sesbania

15b. Leaves oblong or oblanceolate, flowers unspotted, pod flat to oblong ………….…. 16

16a. Obtuse leaf tip, round base, white, pink or red flower colour ………….…...…… Abrus

16b. Acute, emarginate or narrowly round tip, cuneate base, orange, yellow, red or pink flower colour ………………………………………………………………………. Tephrosia

17a. Habit tree, deciduous or evergreen ………………….………….…………………... 18

17b. Habit shrub or liana .…...………………………………………………...... ……... 21

18a. Leaves broad ovate, acuminate tip, round base, cream or pale yellow flower.. Dalbergia

18b. Leaves ovate or elliptic, acute or acuminate tip, cuneate base, golden yellow flower colour ……………………………..…………………………………………….... Pterocarpus

19a. Leaves oblong to ovate, tip obtuse, obliquely cuneate base, white to greenish white flower colour …………………………………………………………………..… Ostryoderris

19b. Leaves elliptic or ovate ……………………………………………………………... 20

20a. Acuminate tip, cuneate base, pink or lilac flower colour ……………………. Gliricidia

20b. Round or slightly retuse tip, cuneate base, white flower …………………..… Swartzia

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21a. Shrub, leaves obovate, ovate or elliptic, shortly acuminate tip, cuneate or round base, terminal leaflets bigger than the lateral ones, lilac or blue flower colour ……. Lonchocarpus

22b. Scandent shrub, leaves elliptic, oblong or ovate, acute, obtuse or mucronate tip, cuneate base, brick red, rose or pink flower colour …………………………………...... … Indigofera

23a. Liana, leaves elliptic to oblong, tip obtusely acuminate, base cuneate, purple or red flower colour …………………………………………………………………………. Millettia

23b. Scandent shrub, leaves narrowly lanceolate to broad ovate, tip acute, base cuneate, cream to greenish white flower colour …………………………………………...…. Pericopsis

4.2.2.2. Electronic multi-access key

The saved key was opened and used for identification in the lucid player suite (Fig. 4.1). The features and states are on the top right panel while the entities in the top right panel. As features and states are selected on the left top panel, they fall to the lower left panel and entities are split such that only entities matching the criterion only remains in the top right panel while others which do not meet the criterion are moved to the lower right panel. As seen in Fig. 4.2 when state elliptic of leaf shape was selected, only four species were left in the upper right panel, furthermore, upon the selection of round leaf base Entada africana was selected (Fig. 4.3). In another example in Fig. 4.4 Acacia auriculiformis is distinct and was identified in one step by clicking on falcate leaf shape.

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Fig. 4.1: Lucid player when no feature was selected

Fig. 4.2: Lucid player when elliptic was selected, only four entities were left in the upper right panel

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Fig. 4.3: Lucid player when round leaf base was further selected, E. africana was identified in two steps

Fig. 4.4: Lucid player when falcate leaf shape was selected, A. auriculiformis was identified just in one step

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4.2.3 UPGMA Relationship based on morphological data

The recorded features in a pair-wise analysis showed similarity among species based on

morphological characters; Morphological characters recorded in details are recorded in

Appendix II. Likewise, information on the scoring of data as used in the DELTA software

is detailed in Appendix III - V. Relationship among species based on morphological

characters using an unweighted pair-wise analysis are shown in Figs. 4.5 – 4.7.

Acacieae

Mimosieae

Igneae

Acacieae

Fig. 4.5: Unweighted pair group method of arithmetic averages (UPGMA) dendogram of Mimosoideae species based on morphological characters

Key: ASE=Acacia senegal; AAT=Acacia ataxacantha; ANI=Acacia nilotica; AAL=Acacia albida; ASI=Acacia sieberiana; MPU=Mimosa pudica; MPI=Mimosa pigra; LLE= Leucaena leucocephala; DCI= Dichrostachys cinerea; PBG=Parkia biglobosa; PBC=Parkia bicolor; EAF=Entada africana; EAB=Entada abyssinica; AZY=Albizia zygia; ALE=Albizia lebbeck; PAF=Prosopis africana; PDU=Pithecellobium dulce; AAU=Acacia auriculiformis

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Cassieae

Caesalpinieae

Cassieae

Detarieae

Cassieae

Resin producing Detarieae Dialiinae Dimorphandra group

Cercidieae

Fig. 4.6: Unweighted pair group method of arithmetic averages (UPGMA) dendogram of Caesalpinioideae species based on morphological characters

Key: CRO=Chamaecrista rotundifolia; SOB=Senna obtusifolia; STO=Senna tora; SHI=Senna hirsuta; SAL=Senna alata; CMI=Chamaecrista mimosoides; CNI=Chamaecrista nigricans; DRE=Delonix regia; CPU=Caesalpinia pulcherrima; SOC=Senna occidentalis; TIN=Tamarindus indica; ITO=Isoberlina tomentosa; IDO=Isoberlina doka; DMA=Detarium macrocarpum; DOL=Danielii oliveri; CSI=Cassia sieberiana; SSI=Senna siamea; SSN=Senna singuena; CAR=Cassia arereh; CME=Cynometra megalophylla; AFZ=Afzelia africana; DGU=Dialium guineensis; BAF=Burkea africana; BTO=Bauhinia tomentosa; BRU=Bauhinia rufescens; BVA=Bauhinia vahlii; BMO=Bauhinia monandra; BPU=Bauhinia purpurea; PTH=Piliostigma thonningii; PRE=Piliostigma reticulatum

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Swartzieae Desmodieae Millettieae Desmodieae Dalbergieae Millettieae Dalbergieae Desmodieae Phaseoleae

Desmodieae

Crotalarieae Dalbergieae

Phaseoleae Crotalarieae

Milletieae

Indigofereae

Milletieae Dalbergieae Sesbanieae Indigofereae Abreae

Phaseoleae

Robinieae Phaseoleae

Fig. 4.7: Unweighted pair group method of arithmetic averages (UPGMA) dendogram of Papilionoideae species based on morphological characters

Key: SMA=Swartzia madagascariensis; DVE=Desmodium velutinum; LSE=Lonchocarpus sericeus; DGA=Desmodium gangeticum; PER=Pterocarpus erinaceus; LCY=Lonchocarpus cyanescens; DSI=Dalbergia sissoo; DSC=Desmodium scorpiurus; CMU=Calopogonium mucunoides; DTO=Desmodium tortuosum; ARU=Alysicarpus rugosus; AGL=Alysicarpus glumaceus; CLA=Crotalaria lachnosema; AHY=Arachis hypogaea; CNG=Crotalaria naragutensis; CPA=Crotalaria pallida; CCA=Cajanus cajan; VSU=Vigna

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4.3 Molecular Characterization

4.3.1 DNA Isolation

Total genomic DNA was successfully isolated from 437 samples in 119 species. In summary,

a 98.09% success was recorded, isolates are of high molecular weight and purity level DNA

both from silica dried samples and some herbarium samples. Isolates were deposited at the

DNA bank of the University of Lagos, Nigeria and University of Reading, England. The list of

samples and their resultant DNA bank number are shown on Appendix VI.

Qualitative analysis using agarose gel electrophoresis revealed mostly high molecular weight

DNA as shown in Plate 83. Likewise, quantitative analysis using spectrophotometry revealed

DNA concentration ranged between 10.1-1780.4ng/µl; the purity ratio at absorbance ratio of

260 against 280 revealed isolates of high purity level with ratio ranging between 1.75 and 2.01.

1Kb hyper- Isolated DNA ladder 10,000bp

a. b.

1Kb hyper- Isolated DNA ladder 10,000bp

c. d. Plate 83(a-d): Electropherogram of extracted DNA

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4.3.2 DNA Amplification

Results of targeted DNA regions amplified with specific primers revealed products with

distinct bands (Plate 84). Mimosoideae yielded 91% successful amplifications while

Caesalpinioideae samples yielded 75.23% success and Papilionoideae samples yielded 77.44%

success rates of amplification.

1Kb hyper-ladder PCR product 1500bp PCR product -ve 800bp Control (-ve) a. b.

1Kb hyper-ladder

PCR product PCR product 1000bp 800bp -ve Control (-ve) c. d.

Plate 84(a-d): Electropherogram of PCR product; a=ITS; b=rbcL; c=matK; d=trnL-F

4.3.3 DNA Barcoding Analysis

Determination of Genetic Divergence Using Six Parameters

According to Gao et al., 2010, a favorable barcode should exhibit a high inter-specific

divergence so as to be able to distinguish different species. Results from the six parameters

used to characterize inter- versus intraspecific variations revealed ITS exhibited significantly

the highest inter-specific discrimination higher than matK and trnL while rbcL was the least.

Likewise, intra-specific variations were similar, ITS exhibited the largest, while rbcL again

revealed the smallest variations Table 10.

Furthermore, Wilcoxon signed rank tests confirmed that ITS and matK exhibited the highest

inter-specific divergence between congeneric species, whereas rbcL exhibited the

lowest, Table 11. Similarly, rbcL exhibited the lowest variation between conspecific

individuals, whereas ITS and trnL-F showed the highest based on Wilcoxon signed rank tests

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(Table 12). These indicated that the nuclear ITS region could be proposed as the most suitable

DNA barcodes to distinguish the species of Nigerian arid land Fabaceae.

Table 10. Analysis of inter-specific divergence between congeneric species and intra-specific variation of candidate barcodes

Marker ITS matK RbcL trnL-F All inter-specific distance 0.126192±0.047004 0.017475±0.010025 0.013615±0.006273 0.025907±0.018742

Theta prime 0.080250±0.030837 0.010667±0.006022 0.008500±0.003416 0.009600±0.005177

Minimum inter-specific distance 0.309500±0.384869 0.015000±0.015086 0.009750±0.011026 0.020400±0.025861

All intra-specific distance 0.143570±0.136360 0.00500±0.012005 0.000667±0.001496 0.001488±0.002145

Theta 0.014357±0.136360 0.000444±0.011991 0.000667±0.001496 0.001294±0.001863

Coalescent depth 0.022000±0.022464 0.000833±0.021761 0.001333±0.003155 0.002000±0.003123

Table 11. Wilcoxon signed rank tests for inter-specific divergence

W+ W- Relative Ranks, n, P value Result

ITS MatK W+ = 114, W- = 6, n = 15, P ≤ 0.002 ITS>matK

ITS RbcL W+ = 90, W- = 1, n = 13, P ≤ 0.00188 ITS>rbcL

ITS trnL-F W+ = 114, W- = 6, n = 15, P ≤ 0.00214 ITS>trnL

matK rbcL W+ = 117, W- = 6, n = 15, P ≤ 0.0012 matK>rbcL

matK trnL-F W+ = 27, W- = 126, n = 17, P ≤ 0.01928 matK>trnL

rbcL trnL-F W+ = 0, W- = 120, n = 15, P ≤ 0.00064 rbcL

Table 12. Wilcoxon signed rank tests for intra-specific divergence

W+ W- Relative Ranks, n, P value Result

ITS matK W+ = 99.5, W- = 5.5, n = 14, P ≤ 0.00318 ITS>matK

ITS rbcL W+ = 91, W- = 0, n = 13, P ≤ 0.00148 ITS>rbcL

ITS trnL-F W+ = 108.5, W- = 11.5, n = 15, P ≤ 0.00596 ITS>trnL

matK rbcL W+ = 14, W- = 14, n = 18, P ≤ 1.00 matK=rbcL

matK trnL-F W+ = 0, W- = 45, n = 20, P ≤ 0.005 trnL>matK

rbcL trnL-F W+ = 11, W- = 44, n = 10, P ≤ 0.09295 rbcL

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Assessment of Barcoding Gap

To assess the barcoding gap that exists between species, the level of divergence between and within species was calculated by pairwise analysis of sequences based on all the tested barcode regions. For an accurate analysis of sequence identification, the minimum interspecific divergence should be lower than the maximum intraspecific distance exhibiting a separate, non-overlapping distributions between intra- and inter-specific samples. From the analysis, ITS region exhibited the highest barcoding gap (Fig. 4.8) while rbcL region was the least with no barcoding gap at 0.0% distance (Fig. 4.13). The combination of ITS with both rbcL and matK increased the interspecific divergence by 1.0% (Figs. 4.9 & 4.13) while a combination of all the four regions as well depicted a barcoding gap of 1.0% (Fig. 4.17).

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70

60

50

40

30 proportion(%) 20 INTRASPEC INTERSPEC 10

0

pairwise distance (%)

Fig. 4.8: Distribution of inter-specific divergence between congeneric species and intra- specific variation for ITS region showing 2% barcoding gap

60

50

40

30

proportion(%) 20 INTRASPEC INTERSPEC 10

0

pairwise distance (%)

Fig. 4.9: Distribution of inter-specific divergence between congeneric species and intra- specific variation for ITS+matK region showing 1% barcoding gap

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160 140 120 100 80 60

proportion(%) INTRASPEC 40 INTERSPEC 20 0

pairwise distance (%)

Fig. 4.10: Distribution of inter-specific divergence between congeneric species and intra- specific variation for matK region showing no barcoding gap

120

100

80

60

proportion(%) 40 INTRASPEC INTERSPEC 20

0

pairwise distance (%)

Fig. 4.11: Distribution of inter-specific divergence between congeneric species and intra- specific variation for matK+trnL-F region showing no barcoding gap

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80

70

60

50

40

30 proportion(%) INTRASPEC 20 INTERSPEC 10

0

pairwise distance (%)

Fig. 4.12: Distribution of inter-specific divergence between congeneric species and intra- specific variation for matK+rbcL region showing no barcoding gap

160

140

120

100

80

60 proportion(%) INTRASPEC 40 INTERSPEC 20

0

pairwise distance (%)

Fig. 4.13: Distribution of inter-specific divergence between congeneric species and intra- specific variation for rbcL region showing no barcoding gap

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60

50

40

30

20 proportion(%) INTRASPEC 10 INTERSPEC

0

pairwise distance (%)

Fig. 4.14: Distribution of inter-specific divergence between congeneric species and intra- specific variation for rbcL+ITS region showing 1% barcoding gap

80 70 60 50 40 30 proportion(%) INTRASPEC 20 INTERSPEC 10 0

pairwise distance (%)

Fig. 4.15: Distribution of inter-specific divergence between congeneric species and intra- specific variation for rbcL+trnL-F region showing no barcoding gap

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160

140

120

100

80

proportion(%) 60 INTRASPEC

40 INTERSPEC

20

0

pairwise distance (%)

Fig. 4.16: Distribution of inter-specific divergence between congeneric species and intra- specific variation for trnL-F region showing no barcoding gap

70

60

50

40

30 proportion(%) 20 INTRASPEC INTERSPEC 10

0

pairwise distance (%)

Fig. 4.17: Distribution of inter-specific divergence between congeneric species and intra- specific variation of COMBINED (rbcL + ITS + matK + trnL-F) dataset showing 1% barcoding gap

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Evaluation of Species Authentication Capability of Barcodes

Results of the best match and best close match of sequences performed on TAXONDNA are

shown on Table 13. There was a little bit of difference between results of best match and best

close match. The best match is a least stringent criterion; it finds the closest barcode match to

each query sequence, if both sequences are from same species then identification is considered

successful, if query sequence matches several sequences from different sequences

identification is considered ambiguous whereas mismatch names and sequences are considered

incorrect. Best close match defines set a threshold frequency of 95% of all intraspecific

sequences before identification can be made (Meier et al., 2006). Query sequence with the

smallest distance within the 95th percentile and conspecific are considered successful,

sequences with the smallest distance within the 95th percentile but with a mixture of allospecific

and conspecific sequences are considered ambiguous while sequences with the smallest

distance within the 95th percentile and allospecific are incorrect.

Table 13: Identification success of the 10 barcodes based on the best match and best close match program in TAXONDNA

Best match (%) Best close match (%) Barcodes N of sequences Correct Ambiguous Incorrect Correct Ambiguous Incorrect No match & species ITS 201 (86) 100 0 0 99.62 0 0 1.37 matK 262 (99) 97.46 1.26 1.26 94.93 0 1.26 3.79 rbcL 318 (104) 87.01 11.68 1.29 87.01 11.68 1.20 0 trnL-F 263 (98) 98.63 1.36 0 98.63 1.36 0 0 rbcL + matK 194 (84) 98.18 0 1.81 98.18 0 1.81 0 rbcL + ITS 194 (84) 99.72 0 0.27 81.81 0 5.45 12.72 rbcL + trnL-F 194 (84) 92.72 1.81 5.45 92.72 1.81 5.45 0 matK + trnL-F 194 (84) 98.18 0 1.81 98.18 0 1.81 0 matK + ITS 194 (84) 99.36 0 0.63 92.9 0 1.81 7.27 rbcL + ITS + matK 194 (84) 96.54 1.81 1.63 92.72 1.81 1.81 3.63 + trnL-F

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Tree Based Analysis

The tree based analysis based on three different approaches of Neighbour Joining (NJ),

UPGMA and Bayesian inference (BI) showed clustering of conspecifics at mostly >70% bootstrap support (NJ & UPGMA) and >0.65 posterior probability for BI. For the NJ analysis, most species were grouped at varying bootstrap support values while other single regions were between 80 – 90% support at varying values and points. The combined regions (rbcL + matK

+ITS +trnL-F) revealed a well-defined species cluster at both high and low bootstrap value

(Fig. 4.20). With the UPGMA analysis, ITS showed clustering of species at mostly 100% support value, matK had species clustered at support value range 70-99% while rbcL had an average support of 87.8% species clustering. Combination of all the four regions in the analysis gave a support of different percentages for all species cluster (Fig. 4.18).

The Bayesian analysis of all the combined regions (rbcL + matK +ITS +trnL-F) showed the most robust resolution of species cluster, species were clustered at distinct node at a high posterior probability of within 0.60 – 1.0 as shown in Fig. 4.19.

In all the tree based analysis, all species clustered with their conspecifics at the same points and clustered with their tribe members though the clustering orientation and support values differing due to the difference in analysis and evolution specified.

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Fig. 4.18: UPGMA analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data

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Fig. 4.18: UPGMA analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.18: UPGMA analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.18: UPGMA analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.19: Bayesian analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data

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Fig. 4.19: Bayesian analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.19: Bayesian analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.20: Neigbour joining analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data

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Fig. 4.20: Neigbour joining analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.20: Neigbour joining analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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Fig. 4.20: Neigbour joining analysis of combined regions (rbcL + matK +ITS +trnL-F) sequence data (cont’d)

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4.3.4 Phylogenetic Analysis

Two different phylogenetic analysis were carried out on three data matrices which revealed varying levels and clustering among the different species, however, analysis resolves a fairly conformity to each other in topology,

For the parsimony analysis, ITS had a total of 1130 characters out of which 324 are constant,

116 are variable and parsimony uninformative while 690 are parsimony informative. The tree

(Fig.4.21) had a total length of 7580, with a retention index of 0.7248 and a rescaled consistency index of 0.1718. The homoplasy index was 0.7629 (0.7248 excluding uninformative characters). In the combined chloroplast regions dataset, there are a total of 3946 characters out of which 2055 are constant, 394 are variable and parsimony uninformative while

1497 are parsimony informative. The tree (Fig.4.23) had a total length of 6025, with a retention index of 0.8529 and a rescaled consistency index of 0.4053. The homoplasy index was 0.5248

(0.4053 excluding uninformative characters). The combination matrix of all the understudied regions has a total of 5076 characters out of which 2379 are constant, 510 are variable and parsimony uninformative while 2187 are parsimony informative. The tree (Fig.4.25) had a total length of 13984, with a retention index of 0.7807 and a rescaled consistency index of 0.2601.

The homoplasy index was 0.6668 (0.6952 excluding uninformative characters).

The Bayesian analysis showed the best resolution of species clustering at distinct node with a quite high posterior probability of within 0.70-1.0. Although the trees from the parsimony analysis are somewhat different in topology from the Bayesian analysis.

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4.4 Conservation Assessment of species

Conservation assessment based on the IUCN 2014 red-list categories and criteria are as summarized in Table 14.

Table 14: Conservation status of species S/N Species IUCN status S/N Species IUCN status 1. Pterocarpus erinaceus CR and CITES. II 10. Mimosa pudica LC 2. Prosopis africana EN 11. Bauhinia purpurea LC 3. Afzelia africana EN 12. Acacia auriculiformis LC 4. Erythrina senegalensis VU 13. Cassia mimosoides NE 5. Daniellia oliveri VU 14. Abrus precatorious NE 6. Parkia biglobosa VU 15. Acacia nilotica NE 7. Detarium macrocarpum LC 16. Burkea africana NE 8. Dichrostachys cinerea LC 17. Isoberlinia doka NE 9. Dalbergia sisso LC 18. Senna tora + 100 more NE

Key: EN: Endangered; VU: Vulnerable; CR=Critically Endangered; LC=Least Concerned; NE= Not Evaluated; CITES. II= CITES appendix II.

Pterocarpus erinaceus Poir

Common names: Senegal rosewood, African rosewood, black camwood, African teak,

African Kino

CR- Critically Endangered, A1cd+A2+A3; B2b (i,ii,iii,iv); C2(ai); D1

Pterocarpus erinaceus is an endemic savanna tree species up to 16m high, and up to 2m in

girth. Bole slightly buttressed in large trees. Bark dark and rough, flaking off in thick pieces 7-

10cm; slashes brown with fine red lines, slowly oozing drops of blood-red gum. Branches are

smooth and grey. Young branchelets densely and shortly hairy. Flowers golden yellow, turning

pale, fragrant; in copious racemes and panicles at ends of .

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Species occurs in the woodland savanna of Nigeria: Nupe, Zungeru, Zamfara, Bauchi, Plateau,

Taraba, Oyo (Agboola, 1979, Hutchinson and Dalziel, 1979, Burkill, 1995). It grows abundantly and flowers from February to May in the north and south-western parts of Nigeria.

Pterocarpus erinaceus is under severe numerous threats especially of over-exploitation for its timber, habitat fragmentation, deforestation, random cutting, fodder, medicine, fires, human interference, multiple local utilizations. The population size is drastically decreasing with

>90% of mature individuals lost from the wild. Therefore P. erinaceus is assigned CR.

Prosopis africana Taubert

Common names: Ironwood

EN- Endangered, A1cd+A3; B2b (iii,iv); C2(ai)

Prosopis africana is an endemic Savanna tree species up to 12-18m high and 1m in girth, branching very low down and up to crooked limbs forming an irregular open crown, but with clean bole in forest condition. Bark black, strongly fissured, flaking off in ragged patches leaving light brown scars; slash reddish darkening to red-brown with pale drooping foliage and very hard wood. Flower-spikes yellowish, fragrant; pods stout, roughly cylindrical, with loose rattling seeds.

Prosopis africana is more abundant in northern parts of Nigeria: Nupe, Zamfara, Kaduna,

Katagum, Yola (Agboola, 1979, Hutchinson and Dalziel, 1972, Burkill, 1995). It grows abundantly and flowers from April to May.

Prosopis africana is a multi-purpose species of great socio-economic importance, threatened with extinction from its natural habitat in the Guinea Savanna of Nigeria due to overexploitation. The fruit of the tree is used as feed for animals, while the seeds are fermented to make ukpehe, a highly proteinaceous condiment. The tree is not cultivated. The products from the hard wood, such as some wooden farm implements, kitchen utensils, and planks for

316 | P a g e construction, are extensively traded. The tree is a good source of firewood and charcoal; the secondary roots are used for medicinal purposes. Currently, >70% of P. africana have disappeared from extensive parts of its range due to over-exploitation. Hence, P. africana is assigned EN.

Afzelia africana Smith ex Pers.

Common names: Africanoak, African mahogany, counter wood, Mahogany bean tree

EN- Endangered, A1cd+A3; B2b (iii,iv); C2(ai)

Afzelia africana is an endemic humid and dry forest, especially in the forest-savanna borders or semi-decSavanna tree species up to 15m high with large leaves, with 3 – 10 pairs of leaflets, all of same the size. Flowers very fragrant, green, except for the petal which is white with reddish markings, broad at the top and narrowed at the base into a claw, fruits and seeds black, shining with orange aril.

Afzelia africana is found in fringing and drier parts of the forest in Nupe, Zaria, Lokoja,

Katagum (Agboola, 1979, Hutchinson and Dalziel, 1972, Burkill, 1995). It grows abundantly and flowers from Feb to May.

Afzelia africana is threatened with extinction from its natural habitat majorly due to overexploitation of its timber for the international market and other economic usages. The flour from seeds is used as a substitute for wheat flour in biscuits and doughnuts. Leaves, fruits and seeds are browsed by wildlife. Afzelia seed (containing 31% fat) may be a source of seed oil for domestic and industrial use. The tree is a good source of firewood, charcoal, and construction purposes. The leaves, bark and roots are used for medicinal purposes. Currently,

>70% of A. africana have disappeared from extensive parts of its range due to over- exploitation. Hence, A. africana is assigned EN.

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Erythrina senegalensis A.DC

Common names: Coral tree, Coral flower

VU- Vulnerable, A1d+A3; B2c (iv); C2b

Erythrina senegalensis is a small tree, up to 5m high, but sometimes attaining a height of 15m armed with stout prickles slightly recurved from a woody base; conspicuous with scarlet flowers, usually when leafless; standard petal folded flat. It usually occurs in dry open savanna woodlands, burned savanna, plateau with fine gravel, degraded regrowths, coastal savanna, bank of streams and roadsides by grassland.

Erythrina senegalensis is grown abundantly and flowers from Sept to Oct in the Savanna regions of northern Nigeria as a hedge plant Borgu, Kontagora, Abeokuta, Zabolo, Jos, Ondo,

Kwara, Niger, Mambilla, Zaria, Adamawa, Zaria, Bailey, Plateau, Niger, Oyo (Agboola, 1979,

Hutchinson and Dalziel, 1972, Burkill, 1995, Adelanwa, 2008).

Erythrina senegalensis has a large number of traditional medicinal uses in West Africa, the bark and root is used to cure several diseases: amenorrhoea, malaria, jaundice, infections, abortion, wound, and body pain (chest pain, back pain, abdominal pain etc). The wood is used for making knife handles, seeds are made into necklaces and used as game counters.

This species is threatened with various activities hindering its regeneration such as debarking, increased exploitation, habitat fragmentation, deforestation, random cutting, human interference, the population size is decreasing with a small number of healthy mature individuals, over 50% of Erythrina senegalensis has been debarked. Hence, E. senegalernsis is assigned VU.

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Daniellia oliveri (Rolfe) Hutch. & Dalz.

Common names: African copaiba balsam, West African copal, wood oil tree

VU- Vulnerable, A1d+A3; B2c (iv); C2b

Daniellia oliveri is large fire resistant Savanna tree, up to 35m high, with copious rather flat panicles of white scented flowers and smooth pale flat 1- seeded fruits of horny consistence. It flowers between November and December in northern Nigeria and found in abundance along the coastal areas of southern Nigeria Nupe, Kontagora, Lokoja, Bauchi, Plateau, Lagos, Ishan,

Onitsha (Agboola, 1979, Hutchinson and Dalziel, 1972, Burkill, 1995, Adelanwa, 2008).

Daniellia oliveri is a multipurpose tree species, important both as timber and forest enrichment tree. Its wood is used as pole, fuel wood, charcoal, food, shade and shelter (Bojang, 2000).

Daniellia oliveri is important in agroforestry systems, soil and water conservation and of high medicinal value being potent in treating gastrointestinal ailments (Ahmadu et al., 2004), as antiaborifacients in pregnancy, pain-killer, skin mucosae and as sedative (Burkill, 1985), for the cure of rheumatism/ pains (MacDonald and Olorunfemi, 2000) and active as antimicrobial agent (Ahmadu et al., 2004).

This species is threatened with various activities apace hindering its regeneration such as debarking, increased exploitation, habitat fragmentation, deforestation, random cutting, human interference. The population size is decreasing with over 50% lost in the wild. Hence, D. oliveri is assssigned VU.

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Parkia biglobosa (Jacq.) R Br. ex Don

Common names: African locust bean, nitta tree, monkey cutlass, West African locust bean

VU- Vulnerable, A1d+A3; B2c (iv); C2b

Parkia biglobosa is a tree up to 18m high. The bole is short and crooked with twisted spreading branches forming a wide crown. It is readily recognised by its red pendulous flowers with individual flowers stalkless, each with narrow spoon-shaped bracteole. It is scattered all over the country and flowers between March and June in northern Nigeria (Agboola, 1979,

Hutchinson and Dalziel, 1972, Burkill, 1995, Adelanwa, 2008).

Parkia biglobosa is a multipurpose tree species, its seeds are consmed, used as condinent. The pods are used as sponges and strings, dyes, and for fishing, and also for preparing insecticide powder (Don, 2007). The bark is used as a mouthwash, vapour inhalant for toothache, or for ear complaints. It is macerated in baths for leprosy and used for bronchitis, pneumonia, skin infections, sores, ulcers, and washes for fever, malaria, diarrhoea, and sterility. Roots are used in a lotion for sore eyes (Ntui et al., 2012).

However, despite these economic values, Parkia biglobosa population in the wild is declining at an alarming rate, and there are no conservation measures. One reason for this decline is uncontrolled bush burning through farming, which is a common feature in Nigeria during the dry season. Bush burning destroys the young seedlings and reduces the population density.

Another factor is excessive grazing by cattle and other domestic stock. These animals not only eat up the young and tender seedlings, but may also physically destroy them during movement within the vegetation. A more important factor is the fact that only a small percentage of the seeds germinate in the field, a lot more are dormant (Etejere et al., 1982). Over 50% of Parkia biglobosa are debarked with seeds usually harvested. Hence, P. biglobosa is assssigned VU.

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CHAPTER FIVE DISCUSSION

Conservation of the genetic resources of endemic desert plants is crucial to worldwide efforts to combat desertification and to prevent further degradation of the fragile ecosystems in arid and semi-arid regions. Among the first steps towards protecting and benefiting from these resources are sampling, accurate identification and studying biological specimen. In this study, extensive exploration of eleven (11) states in Northern Nigeria yielded a total of three hundred and twenty-four (324) samples distributed in 118 species and 53 genera; 22, 38 and 58 species are members of the Mimosoideae, Caesalpinioideae and Papilionoideae respectively. This somewhat contradicts Adelanwa, 2008 who recorded 56 genera in 106 species; the disparity in species samples might be as a result of larger sample size (more states explored) and edge effect as some of the samples might have been cleared over time.

Morphological features recorded among members supports Orwa et al., 2008 on the habit and life forms of Fabaceae. Features recoded are relatively distinct to each subfamily. The distinctive floral feature is the primary feature used to separate members into three different subfamilies as reported from the 18th century (Bentham, 1842, 1875; Cronquist, 1981). This is because some available exo-morphological features are overlapping among members of the family. It was observed many of the quantitative features were not useful in delimitating species which is also due to overlapping of species features even among tribes. However, some qualitative exo-morphological features were distinct and useful to delimit or differentiate among species as observed among the members. These characters were used in developing keys, a bracketed and an electronic multi-access key. Testing of the keys revealed many species can be identified using these keys. In the bracketed key, it was based mostly on qualitative features with 6, 11 and 23 couplets involved in the Mimosoideae, Caesalpinioideae and

Papilionoideae respectively. One of the challenges of using the bracketed key was that the order

321 | P a g e and progression along characters and numbering must be duly followed for correct identification to be made unlike in the electronic multi-access where any random or most easily observed character could be picked among the available options in the key; this corroborates

Dallwitz et al., 2007. Also, identification was easier and interactive using the electronic key because pictures or images of each species are provided against each entity, this allows for an easy comparison between specimen and images.

Furthermore, results of pair-wise analysis based on unweighted algorithm depicted various relationships and clustering of species taxonomically. The Mimosoideae tree in Fig. 4.5 supports, Lewis et al., 2005 on the relationships among Mimosoideae based on molecular data.

Members were clustered into three tribes with species within the tribe clustering in the same clade. It also supports the polyphyl of tribe Acacieae (Wojciechowski, 2003; Lewis et al., 2005) as another Acacia species Acacia auriculiformis clustered at the base of the tree. This is due to the reduction of its leaves to a falcate phyllode which is obviously different from other Acacia species studied. Based on molecular data, Faidherbia albida is more closely related to members of the Igneae than the Acacieae but results based on the exo-morphological approach shows

Faidherbia as more closely related to members of the Acacieae. However, some misidentifications and nomenclatural change was encountered among some members of the

Mimosoideae such as:

• Acacia albida has been moved to the genus Faidherbia now accepted as Faidherbia

albida.

Also, samples M033 and M034 were misidentified as Neptunia oleracea but results from molecular analysis revealed they were Aeschynomene spp., (members of the

Papilionoideae). However, these samples were excluded from the analysis as their correct identification is yet to be confirmed.

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Relationships among the Caesalpinioideae also depicts clustering of distinct species within its clade or tribe members. The tree separated members of the Detarieae into two different clades, the Detarieae and the resin producing Detarieae clade (Fig.4.8). Results also somewhat supports the phylogeny of Caesalpinioideae according to Tucker and Douglas, 1994; Chaphill,

1995; Lewis and Schirire, 2003 as members of the tribe Cercidieae clustered as the basal-most group of the sub-family. Results depicted a polyphyletic Detarieae while molecular data presents a monophyletic Detarieae; the tree revealed polyphyletic Cassieae. However, some synonyms and nomenclatural changes were observed among members of the Cassieae; the genus Cassia has been a waste basket taxon for a long time used to classify tribe members that did not fit in other genera (Roskov et al., 2009). Over the years many taxa have been transferred to more appropriate genera (Senna and Chamaecrista), such as:

• Cassia mimosoides is synonymous to Chamaecrista mimosoides but now accepted as

Chamaecrista mimosoides.

• Cassia hirsuta is synonymous to Senna hirsuta now accepted as Senna hirsuta.

• Cassia italica is synonymous to Senna italica now accepted as Senna italica.

• Cassia nigricans is synonymous to Chamaecrista nigricans but now accepted as

Chamaecrista nigricans.

• Cassia obtusifolia is synonymous to Senna obtusifolia but now accepted as Senna

obtusifolia.

• Cassia occidentalis is synonymous to Senna occidentalis but now accepted as Senna

occidentalis.

• Cassia singueana is synonymous to Senna singueana now accepted as Senna

singueana.

• Senna alata is an accepted name and synonymous to Cassia alata.

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• Senna rotundifolia is synonymous to Chamaecrista rotundifolia and now accepted as

Chamaecrista rotundifolia.

The Papilionoideae UPGMA tree based on morphological data was not able to thoroughly resolve relationships among Papilionoideae when in comparison with the current phylogeny of

Papilionoideae; members of the Dalbergieae, Phaseoleae and Milletieae were clustered in different clades (Fig. 4.9). Although current molecular studies revealed polyphyly among members, results from molecular data inferred monophyly in the above tribes. Albeit, within some clades, species-cluster of all of its tribe members was observed within the Crotalarieae and Indigofereae clades. This none uniform species-cluster might be due to the fact that members of Papilionoideae are the most variable in terms of morphological features (Orwa et al., 2008). Also, some synonyms and nomenclatural changes were encountered in the following:

• Crotalaria falcata is synonymous to Crotalaria mucronata with their accepted name

as Crotalaria pallida.

• Afromosia laxiflora has been transferred to the genus Pericopsis now accepted as

Pericopsis laxiflora.

• Swartzia madasgascariensis has been transferred to genus Bobgunnia now accepted as

Bobgunnia madgascariensis.

• Lonchocarpus cyanescens has been transferred to genus Philenoptera now accepted as

Philenoptera cyanescens.

• Vigna ambacensis is being renamed and accepted as Vigna heterophylla.

Also, a case of misidentification of taxa was recorded with samples (P75, P76, P147, P148) being species of Lonchocarpus; L. sericeus and L. cyanescens. Results of molecular analysis

324 | P a g e revealed the samples are members of the family Sapindaceae (see plate 81 and 82); however, the correct identification is yet to be confirmed.

For the molecular characterization, the manipulation of DNA in the laboratory, the basis of molecular biology, depends upon a solid understanding of how the properties of DNA affect the way it is handled and analyzed. The quality of the genomic DNA when tested on 0.8% agarose showed high molecular weight bands of 10,000 bp following electrophoresis. Majority of the samples had distinct bands showing the presence of DNA, except for a few with weak or smeared bands, Plate 83(a-d). The smeared bands indicated degraded DNA, isolates of low molecular weight which were mostly from herbarium specimen. This is also a confirmation that 0.8% agarose gel concentration is suitable for an effective resolution of DNA samples under consideration as reported by Oboh et al., 2009 and Ogunkanmi et al., 2008. This shows different DNA of different sizes; the brightness and intensity of a particular DNA signifies the difference among the samples. There is a relationship between the size of a DNA and its migration distance on the gel. There is variation in the movement of DNA on the gel this indicates samples are of different molecular weight. Low molecular weight DNA migrates faster while high molecular weight DNA migrates slower, thus DNA are of different molecular weights. The variation in the intensity of these samples is due to the differences in the concentration of DNA extracted from each sample. The purity of DNA was determined using a comparison of the optical density values of the DNA at various wavelengths. For pure DNA, the observed 260/280 nm ratio will be near 1.8. Elevated ratios usually indicate the presence of RNA, which can be tested by running the sample, on an agarose gel. A260/280 ratios below

1.8 often signal the presence of a contaminating protein or phenol (Clark and Christopher,

2000). Hence, isolated DNA were of good quality and purity with A260/280 ratio from 1.75 –

2.01.

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One of the criteria in establishing a suitable barcode marker is successful amplification and sequencing of regions from samples. In this study, all the four regions examined were quite successful in PCR reactions. Amplifications revealed clear and distinct bands, the ITS region had the highest success rate of 92.71% while rbcL and matK were 90.63% and trnL-F region occurred as the least successful. However, samples which failed to amplify were re-amplified with their subsequent internal primers for shorter fragments amplification which were later contiged and concatenated to produce its longer fragment. Failures were mostly due to degradation of isolated DNA predominantly from herbarium specimen. However, some isolates didn’t just amplify. In terms of sequencing, trnL-F region was the least successful while rbcL and matK the highest. This supports Kress et al., 2005; Newmaster et al., 2008 on successful amplification and sequencing of the matK and rbcL and further confirm why these two regions were selected as the ideal plant barcode region for all land plants (CBOL, 2009).

ITS region has demostrated to be successful and fairly easy to work with based on the reports of Gao et al., 2010 and Yao et al.. 2010 but in this study, some difficulties were encountered with sequencing and aligning ITS raw sequence data. Conversely, due to its high rates of evolution, ITS region recorded the highest variable characters and parsimony informative sites compared to other understudied regions supporting Chen et al., 2010.

To determine the utility and efficiency of each barcode loci, both tree building and distance methods were explored. In this study both methods have been used for comparison. Studies on the genetic divergence of region revealed the existence of high genetic distance among species based on the ITS region. Wilcoxon signed rank tests further confirmed that ITS exhibited the highest intra and inter-specific divergence between congeneric species, whereas rbcL exhibited the lowest. This is due to the high variability and very wide genetic distance which was significant enough to distinguish between inter and intraspecific genetic divergence which corroborates the findng of Sun et al., 2015 on the successful use of ITS to identify members of

326 | P a g e economically important species of Brassicaceae. Based on the distribution of intra and interspecific divergence analysis, ITS region again exhibited the highest barcoding gap of 2.00,

ITS+matK, ITS+rbcL, and ITS+matK+rbcL+trnL-F gap of 1.00 while other regions revealed no barcoding gap. This implies ITS region can either be used singly or in combination with either matK or rbcL as a potential plant barcode marker. In comparison with all other tested chloroplast regions, ITS performed more accurately than others. This further reinforces previous reports that cpDNA regions cannot establish genetic delimitations between closely related species. This is probably attributable to the maternal inheritance of cpDNA in most angiosperms (Petit et al., 2005) and suggests possible reason for the low discriminatory power of plastid plant barcodes (Sun et al., 2015). Conversely, results contradict Shinwari et al., 2014 which recommends rbcL as an efficient identification region for subfamily Mimosoideae

(Family: Fabaceae). Based on all the examined parameters, rbcL region exhibited the least in performance except in amplification and sequencing.

Based on TAXONDNA analysis, ITS region again revealed a 100% best match but a 99.62% best close match which was due to species matching with their conspecifics but outside the specified 3% set threshold value. trnL-F gave a second value of 98.63% resolving next to ITS for both best match and best close match; matK as a single region gave a match of 97.46% and

94.93% best match and best close match respectively while rbcL was the least success rate of

87.01% when used singly. However, based on previous findings, the plant barcoding community recognized the need for more than one barcode region (Chase et al., 2005, 2007;

Kress et al., 2005, Cowan et al., 2006; Newmaster et al., 2006; Kress and Erickson, 2007,

Taberlet et al., 2007; Lahaye et al., 2008a) due to the failure and challenge in finding a suitable single barcode locus for all land plants as was the case in for animal species. Although the choice of barcoding regions for all land plants is yet to be made but some region combinations

(ITS & rbcL; ITS & psbA-trnH; matK, atpF-atpH & pshA-trnH amongst others) have been

327 | P a g e suggested by Chase et al., 2005, 2007; Newmaster et al., 2006. During the third International

Barcoding of Life Conference in Mexico City, the CBOL plant working group (2009) proposed the use of the combination of rbcL & matK as a core plant barcode and suggested trnH-psbA and ITS should be investigated as complementary markers (Li et al., 2011). In this study, out of all the fifteen barcode regions screened, only five regions were successfully amplified and readily sequenced. However, the multi-tiered loci barcode approach improved the resolving power of the chloroplast regions. The combination of rbcL+matK increased the resolving power of both regions to 98.18% slightly above the 95% cut-off value and further strengthen their utility as a barcode region thus supporting reports of (Kress and Erickson, 2007; Chen et al., 2010 amongst others).

For the tree based analysis, the Bayesian method resulted in higher identification rates compared to the distance methods. Within the distance methods, the K2P neigbour joining approach resulted to higher identification rates when compared to the UPGMA analysis. Tree based analysis yielded congruent results by clustering of conspecifics to a distinct node although at varying support values and orientation due to the different approaches and model used. Overall, tree based analysis presented a higher identification rates than using

TAXONDNA. Tree building approaches have given higher rates of correct identification in some other studies as seen in Meier et al., 2006 and Gonzalez et al., 2009. In principle, tree methods do not depend on the existence of a barcode gap i.e. the non-overlap of intra and inter specific divergence (Wimers and Fiedler, 2007), it assumes monophyly in the species (Desalle et al., 2005).

However, according to CBOL, (2009) an ideal DNA barcode should be universal, reliable, cost-effective and show good discriminatory power. Despite the highest list species-level identification efficiency (100%) of ITS, in this study, none of the four DNA barcode candidates met all these criteria. In terms of overall performance, the ITS appears to be more promising

328 | P a g e than other tested regions. The ITS region has been repeatedly suggested as a barcode locus for all land plants (Kress et al., 2005; Chen et al., 2010) due to its wide genetic divergence but have not been enlisted as an ideal plant barcode due to some primary concerns. However, from this studY, amplification and direct sequencing of ITS region was quite successful in all our sampled species and no fungal contamination was detected. Overall, DNA barcoding has proven to be useful in identification of Nigerian arid-land legumes. Based on this study, the nuclear ITS region is the most useful region within our broad samples. ITS region can be potentially used as a standard DNA locus; results supports the proposal of ITS as an ideal barcode or to be used as a complementary locus to the other approved plant barcode regions.

Taking into account the paramount ecological and economic importance of legumes, the evolution and systematics of the family have been topics of long-standing interest to a large community of academia and researchers. The need to elucidate on the phylogenetic relationships of this important family is essential for understanding the evolutionary history, origin and diversification of members. Albeit, attempts to unravel the phylogeny of this family dates back over two decades and began with higher level relationships based on rbcL sequence data (Doyle, 1995; Doyle et al., 1997; Kass and Wink, 1995, 1996,

1997). However, despite the many years on intense studies and research, some higher level and infrafamiliar relationships within some groups have remained unclear and unresolved (Lewis and Schrire, 2003, Lewis et al., 2005). Based on previous researches, the overall topology of the family has been consistently supported as monophyletic although with low bootstrap or jackknife support values. The result presented in this work is probably the first record of phylogenetic study on Nigerian arid-land Fabaceae based on the current information available to the author. In the study, three data matrices were assessed: single nuclear gene (ITS); combinations of all chloroplast regions (matK, rbcL and trnL-F) and the combinations of all regions comprising chloroplast and nuclear regions (ITS, matK, rbcL, trnL-F). Results of the

329 | P a g e three matrices analyzed depicts two different topologies, the nuclear ITS region gave a different tree topology from the combination of all the four regions and combined chloroplast regions

(fig. 4.21 – 4.26). The phylogenetic analysis supports the monophyly of the family Fabaceae as supported by both molecular and morphological data (Wojciechowski, 2003; Lewis et al.,

2005). In all recent molecular studies, the subfamilies Mimosoideae and Papilionoideae have both been resolved as monophyletic nested within a paraphyletic Caesalpinioideae (Kass and

Wink, 1996; Doyle et al., 1997, 2000; Kajita et al., 2001). The sub-family Caesalpinioideae was said to comprise the basal element in the Fabaceae phylogenies with the relationships among major group not well supported. In many molecular data findings to date, the tribe

Cercidieae has consistently been resolved and strongly supported as the basal-most clade in the family (Wojciechowski, 2003). However, in this present study, the parsimonious analysis based on ITS sequence data did not clearly resolve relationships among species as the members of the Caesalpinioid transitional species were clustered within the monophyletic clade (1; Fig.

4.21) among members of the Mimosoideae at 25.13% BP in a polyphyletic relationship. The other two clades consist of members of the Caesalpinioideae and Papilionoideae with some members of Caesalpinioideae forming a grade within Papilionoideae clade but at a low support value of 19.47% BP. Although this supports the previous submissions of Caesalpinioideae as the basal elements in the tree (Lewis et al., 2005) but the Cercidieae were not the most basal clade. This is the probably the first submissions in the use of ITS sequence data in elucidating on the relationships among legumes, based on the information available to the author. Previous studies using ITS data were on some members or group among the Papilionoideae (Sanderson and Wojciechowski, 1996; Allan and Porter, 2000). The ITS Bayesian tree is congruent with the parsimony tree except that species resolution are clearer, species clustered into different sub-families as seen in the three different clades. Although members of the Caesalpinioideae occurred as the basal entity on the tree with a high support value of 0.99 BI but members of the

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Cercidieae did not cluster as the most basal clade on tree. The both trees based on ITS data depict a monophyletic Fabaceae with Papilionoideae nested within a paraphyletic

Caesalpinioideae, this is in contrast to previous reports such as Bruneau et al., 2000, 2001;

Kajita et al., 2001; Wojciechowski, 2003 amongst others.

Furthermore, in the study, the parsimony analysis of combined chloroplast regions revealed a monophyletic family tree with a monophyletic Papilionoideae and Mimosoideae nested within a paraphyletic Caesalpinioideae; however, the most basal clade consists of members of the tribe

Detarieae instead of Cercidieae as the most basal clade of the family. Although, this supports findings of Bruneau et al., 2008b; Bello, 2009, 2012, based on the trnL intron data; in his study, the basal nodes of the legume family tree are not well separated consisting of Cercidieae and

Detarieae with both resolved as sister group to other remaining legumes. In contrast, results from the Bayesian analysis supports the current phylogeny of Fabaceae where members of the

Cercidieae clustered as the most basal entities of the tree.

Analysis of the combined data comprising the four understudied regions corroborates earlier reports on the legume super tree. Based on the Bayesian analysis result, it inferred similar relationships to the combined chloroplast regions but members of the Cercidieae were not the most basal entities on the tree. Although clustering of some tribe members were highly resolved while some clade remains not well resolved. Overall, results of the phylogenetic analysis supports the polyphyly of tribe Acacieie in the Mimosoideae, Cassieae among the

Caesalpinioideae and the basal group of the Papilionoideae.

However, in comparing both methods employed, the maximum likelihood inferred a more robust phylogeny with high support values than the maximum parsimony analysis. This could be due to the influence of distinct region’s model of evolution specified prior in the analysis

(Thompson, 2013).

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The phylogenetic analysis results of this study resolve the status of the three subfamilies within the family, the three separate clades representing each subfamily and cluster of different tribes and their respective members. The subfamily Mimosoideae resolved three tribes with a polyphyletic Acacieae and genus Faidherbia more closely related to the tribe Ignae than the

Acacieae at 41.9% bp and 1 BP. Likewise, polyphly existed among members of the three tribes within Mimosoideae. Four tribes were recognized in the Caesalpinioideae, which corroborates previous studies and reiterated the monophyletic basal entities of tribes Cercidieae and

Detarieae while members of the other two tribes are polyphyletic. Eleven tribes were recognized within the Papilionoideae, while Dalbergieae occurred as the most basal entity and closely related to the Swartzieae (Crisp et al., 2009; Cardoso et al., 2012a&b). Results inferred a monophyletic Crotalarieae and Indigofereae closely related to the milletoid and Phaseoleae clades. Although previous studies reported Phaseoleae and Milletieae are polyphyletic, only a comprehensive revision at the tribal level would help elucidate (Crisp et al., 2009). The present study revealed a polyphyletic Phaseoleae and a monophyletic Milletieae, this is probably because out of the thirty papilionoid tribes only eleven are represented in our study and many unresolved genera are of temperate origin and not inclusive in this investigation. In order to explicitly unravel the phylogeny of Fabaceae, more species over a larger geographical area should be sampled and a full-species revision undertaken. However, according to

Wojciechowski et al., 2006, it is too early or premature to recognize any newly identified groups and proposed relationships among species due to the ongoing studies to further clarify, resolve and re-construct the phylogeny of Fabaceae. Hence, results of this phylogenetic study is a great contribution to the ongoing project.

The IUCN red-listing of species is the most comprehensive and objective global approach for evaluating the conservation status of plant and animal species with the goal to provide information and analyses on the status, trends and threats to species in order to inform and

332 | P a g e catalyze action for biodiversity conservation. Species were assessed using the IUCN red-list categories and criteria. Of all the species, Pterocarpus. erinaceus was assigned the IUCN status of CR. The major threat confronting this species is incessant logging, it is one of the most sought after species for timber. Due to its recent exploitation and decline in the wild,

Pterocarpus erinaceus is now enlisted in CITES Appendix II (CITES, 2015). Afzelia africana and Prosopis africana were assigned EN. The timber of A. africana is one of the best sold in open markets and mostly looped tree by herdsmen (Sinsin et al., 2004). Frequent complete lopping of trees results in rapid drain of stored reserves leading to reduction in growth of tree girth and production of leafy biomass (Bhat, et al., 1995). The multipurpose plant Prosopis africana is exploited for its iron wood, high quality fuel wood and seeds used as condiment for local puddings. Currently, these species are disappearing from extensive parts of their range due to over-exploitation (Adenle, 2012). The species in the VU category are still quite many in the wild but highly demanded and confronted with various threats hindering their regeneration.

Parkia biglobosa is currently exploited for its fruit; seeds are consumed and not allowed to regenerate (Teklehaimanot, 2004). Many Erythrina senegalensis trees are debarked and exploited for its Ethno-pharmacological properties (Adiaratou et al., 2008). If remedial actions are not planned, these species will be confronted with rapid declination in the wild and thus extinction. Of all the evaluated species, only Afzelia africana was previously assessed and assigned to the VU category hence, results from this study would be submitted and updated to the IUCN redlist database.

The overall global biodiversity goal is for mankind to live in harmony, preserve and continue to sustain the earth’s natural endowment. Throughout the years, governments or nations are encouraged to develop, implement and communicate the results of strategies for implementing the Strategic Plan for Biodiversity and encourage stakeholders at different levels to play a role

333 | P a g e in biodiversity preservation. This study has contributed directly to the United Nations Srategic goals for Biodiversity 2011 - 2020, specifically Achi bidoversity strategic (Padma, 2012):

• Goal A, target 1 (By 2020, at the latest, people are aware of the values of biodiversity

and the steps they can take to conserve and use it sustainably).

• Goal B, target 5 (By 2020, the rate of loss of all natural habitats, including forests, is

at least halved and where feasible brought close to zero, and degradation and

fragmentation is significantly reduced).

• Goal C, target 12 (By 2020 the extinction of known threatened species has been

prevented and their conservation status, particularly of those most in decline, has been

improved and sustained); 13 (By 2020, the genetic diversity of cultivated plants and

farmed and domesticated animals and of wild relatives, including other socio-

economically as well as culturally valuable species, is maintained, and strategies have

been developed and implemented for minimizing genetic erosion and safeguarding

their genetic diversity).

• Strategic goal D, target 15 (By 2020, ecosystem resilience and the contribution of

biodiversity to carbon stocks has been enhanced, through conservation and restoration,

including restoration of at least 15 per cent of degraded ecosystems, thereby

contributing to climate change mitigation and adaptation and to combating

desertification.

334 | P a g e

CHAPTER SIX 6.1 Summary of Findings Table 15: Summary of findings based upon the specific objectives of this study

S/N Objectives Summary of Findings

1. To give a systematic account One hundred and eighteen species of Fabaceae distributed

of the diversity of family in 118 species and 53 genera were systematically

Fabaceae with emphasis on documented from 324 samples collected in the course of

collection and identification. this study.

An updated checklist of legume species as well as

photographs and an electronic multi access key are

presented for easy identification of taxa by locals

especially where reproductive structures are not available.

2. To explore the performance of In comparisons of inter-specific genetic distances among

three cpDNA regions (rbcL, congeneric species using four candidate barcodes, the ITS

matK and trnL-F) and one region exhibited the highest inter-specific divergence

nuclear region ITS to while rbcL region provided the lowest.

accurately identify species In the multi-tiered loci barcode approach, ITS increased

the performance of the chloroplast regions.

ITS showed superiority to discriminate species more

accurately than the CBOL accepted rbcL+matK.

Results of this study proved ITS as a potential barcode

locus for Nigerian Arid-land Fabaceae.

335 | P a g e

3. To examine phylogenetic Phylogenetic analysis based on Maximum Parsimony and

relationships among taxa, find Bayesian Inference analysis present monophyly of the

out if they are consistent with family Fabaceae.

earlier classifications Topology inferred from the nuclear ITS region revealed a

monophyletic Mimosoideae and paraphyletic

Caesalpinioideae nested within a monophyletic

Papilionoideae while members of Cercidieae were not the

basal-most clade.

Results employing chloroplast rbcL, matK, and trnL-F

regions revealed a monophyletic Papilionoideae and

paraphyletic Caesalpinioideae nested within a

monophyletic Mimosoideae with members of the

Cercidieae occurring as the basal-most entities.

4. To red-list species according to Assessment of the redlisting of species revealed one, two,

the IUCN categories and three and six species assigned in Critically Endangered,

criteria. Endangered, Vulnerable and Least Concerned categories

of threats respectively while 89% of species are yet to be

evaluated.

336 | P a g e

6.2 Contributions to Knowledge

This research contributes to the knowledge of legume systematics in the following ways:

1. This thesis achieved an ex-situ conservation of arid legumes in Nigeria. A total of 440

genomic DNA has been deposited at the DNA bank of the University of Lagos

(www.dnabank.ng.org) and University of Reading; 334 DNA sequences deposited to

the NCBI Genbank and 173 barcode sequences submitted to the Barcode of Life

Database (BOLD).

2. The thesis presented an electronic multi-access key for identification of arid-land

legumes for the first time and an updated checklist and conservation status of the

legumes present in the arid and semi-arid region of Nigeria.

3. The thesis inferred phylogenetic relationship within Fabaceae employing a highly

variable gene (ITS) for the first time.

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Appendices

APPENDIX I: Voucher and Genbank number of species

S/N SPECIES Voucher Number ITS rbcL matK trnL-F

1 Acacia ataxacantha LUH 4176 KX057821 KX119246 KX119338 KX268135 2 Acacia auriculiformis LUH 4159 - KX119247 KX119339 KX268136 3 Faidherbia albida LUH 4173 KX057872 KX119296 KX119382 KX268187 4 Acacia sieberiana LUH 4150 KX057825 KX119251 KX119343 KX268140 5 Acacia senegal LUH 4175 KX057824 KX119250 KX119342 KX268139 6 Acacia nilotica LUH 4177 KX057823 KX119249 KX119341 KX268138 7 Acacia dudgeoni LUH 4176 KX057822 KX119248 KX119344 KX268141 8 Albizia lebbeck LUH 4172 KX057828 KX119256 KX119347 KX268144 9 Albizia zygia LUH 4174 KX057829 KX119258 KX119349 KX268146 10 Pithecellobium dulce LUH 5188 KX057895 KX119321 KX119403 KX268206 11 Prosopis africana LUH 4194 KX057896 - KX119404 - 12 Dichrostachys cinerea LUH 5188 KX057868 KX119293 KX119379 KX268182 13 Entada abyssinica LUH 5179 KX057869 - - KX268183 14 Entada africana LUH 4146 - - KX119380 KX268184 15 Leucaena leucocephala LUH 4332 KX057887 KX119311 KX119394 KX268196 16 Mimosa pigra LUH 4158 KX057888 KX119312 KX119395 KX268197 17 Mimosa pudica LUH 4170 KX057889 KX119313 KX119396 KX268198 18 Parkia biglobosa LUH 4191 - KX119316 KX119400 KX268201 19 Bauhinia monandra LUH a154 KX057835 KX119264 - KX268152 20 Bauhinia purpurea LUH 4388 KX057836 KX119265 - KX268153 21 Bauhinia rufescens LUH 5124 KX057837 KX119266 - KX268154 22 Bauhinia tomentosa LUH a161 KX057838 KX119268 - KX268155 23 Bauhinia vahlii LUH 4390 - KX119267 - KX268137 24 Piliostigma reticulatum LUH 4262 KX057894 KX119319 - KX268204 25 Piliostigma thonningii LUH 5181 - KX119320 - KX268205 26 Afzelia africana LUH 4308 - KX119255 - KX268143 27 Cynometra megalophylla LUH 4187 - KX119285 - KX268175 28 Daniellia oliveri LUH 5195 KX057861 KX119287 KX119373 KX268177 29 Detarium macrocapum LUH 4246 KX057867 KX119292 - KX268181 30 Isoberlinia tomentosa LUH 4308 KX057885 KX119309 KX119392 KX268194 31 Isoberlinia doka LUH 1326A KX057884 KX119308 KX119393 KX268193 32 Tamarindus indica LUH 4303 - - KX119340 - 33 Burkea africana LUH 4483 KX057840 KX119270 KX119356 KX268157 34 Caesalpinia pulcherrima LUH 4134 KX057841 KX119271 KX119357 KX268158 35 Delonix regia LUH 5209 KX057862 KX119288 KX119374 KX268178 36 Cassia arereh LUH 4373 KX057844 KX119274 KX119360 KX268162 37 Cassia mannii LUH 5201 KX057845 KX119275 - KX268161 38 Cassia sieberiana LUH 1323A KX057846 KX119276 KX119410 KX268163

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39 Cassia italica LUH 5202 KX057899 KX119324 KX119407 KX268210 40 Cassia fistula LUH 5212 - - - - 41 Cassia absus LUH 5146 - - - - 42 Senna tora LUH 5131 - KX119328 - KX268215 43 Senna alata LUH 6546 KX057897 KX119322 KX119405 KX268208 44 Senna hirsita LUH 6363 KX057898 KX119323 KX119406 KX268191 45 Senna obtusifolia LUH 4138 KX057900 KX119325 KX119408 KX268211 46 Senna occidentalis LUH 4305 KX057901 KX119326 KX119409 KX268212 47 Senna siamea LUH 5168 - - - KX268213 48 Senna singueana LUH 4134 KX057902 KX119327 KX119411 KX268214 49 Chamaecrista nigricans LUH 4369 KX057848 - KX119362 KX268165 50 Chamaecrista mimosoides LUH 4248 KX057847 KX119277 KX119361 KX268164 51 Chamaecrista rotundifolia LUH 5470 KX057849 KX119278 KX119363 KX268166 52 Dialium guineense LUH 4140 - - - - 53 Abrus precatorious LUH 4493 - KX119245 KX119337 KX268134 54 Adenodolichos paniculatus LUH 4171 KX057827 KX119253 KX119345 - 55 Cajanus cajan LUH a162 KX057842 KX119272 KX119358 KX268159 56 Eriosema psoraloides LUH 5149 - - - - 57 Calopogonium mucunoides LUH 5224 KX057843 KX119273 KX119359 KX268160 58 Glycine max LUH 1304A KX057874 KX119298 KX119384 KX268189 59 Dolichos stenophyllus LUH 5162 - - - - 60 Phaseolus vulgaris LUH a251 KX057893 KX119318 KX119402 KX268203 61 Vigna subterranea LUH 5132 KX057912 KX119335 - - 62 Vigna ambacensis LUH 5194 KX057909 - KX119417 KX268222 63 Vigna gracilis LUH 5184 KX057907 KX119336 KX119416 KX268221 64 Vigna racemosa LUH 5134 KX057910 KX119334 KX119418 KX268223 65 Vigna unguiculata LUH 5182 KX057911 - KX119419 KX268224 66 Mucuna pruriens LUH 4472 KX057890 KX119314 KX119397 KX268199 67 Erythrina senegalensis LUH 5159 KX057870 KX119294 KX119381 KX268185 68 Erythrina sigmoidea LUH a213 KX057871 KX119295 - KX268186 69 Alysicarpus glumaceus LUH 4186 KX057830 KX119259 KX119350 KX268147 70 Alysicarpus rugosus LUH 4384 KX057832 KX119261 KX119352 KX268149 71 Alysicarpus ovalifolius LUH 5174 KX057831 KX119260 KX119351 KX268148 72 Alysicarpus vaginalis LUH 5149 KX057833 KX119262 KX119353 KX268150 73 Desmodium gangeticum LUH 5141 KX057863 KX119289 KX119375 KX268179 74 Desmodium scorpiurus LUH 1311A KX057864 - KX119376 - 75 Desmodium tortuosum LUH 1309A KX057865 KX119290 KX119377 - 76 Desmodium velutinum LUH 5196 KX057866 KX119291 KX119378 KX268180 77 Crotalaria comosa LUH 4193 KX057851 - KX119364 KX268168 78 Crotalaria hyssopifolia LUH 4185 KX057853 KX119281 KX119365 - 79 Crotalaria lachnosema LUH a184 KX057854 KX119282 KX119366 KX268170 80 Crotalaria macrocalyx LUH 4156 KX057855 KX119283 KX119367 81 Crotalaria pallida LUH 5176 KX057857 KX119284 KX119368 KX268169 82 Crotalaria naragutensis LUH 1314A - - - KX268172

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83 Crotalaria retusa LUH 5207 KX057858 - KX119369 - 84 Crotalaria arenaria LUH 5145 KX057850 KX119279 - 85 Crotalaria senegalensis LUH 4392 - - - KX268173 86 Crotalaria falcata LUH 4672 KX057852 KX119280 - KX268171 87 Dalbergia sissoo LUH 4265 KX057860 KX119286 KX119372 KX268176 88 Pterocarpus erinaceus LUH 1312A - - - - 89 Stylosanthes erecta LUH 4313 KX057903 - KX119413 -

90 Aeschynomene indica LUH 4711 - - - -

91 Aeschynomene lateritia LUH 5200 - - - - 92 Zornia latifolia LUH 1308A - - - - 93 Arachis hypogaea LUH 1317A KX057834 KX119263 KX119354 KX268151 94 Gliricidia sepium LUH a234 KX057873 KX119297 KX119383 KX268188 95 Indigofera arrecta LUH 5210 KX057875 KX119299 KX119385 KX268190 96 Indigofera hirsuta LUH 4237 KX057878 KX119302 KX119387 KX268191 97 Indigofera nummulariifolia LUH 4260 - - - - 98 Indigofera spicata LUH 1302A KX057882 KX119306 KX119390 - 99 Indigofera suffruticosa LUH 4673 KX057883 KX119307 KX119391 - 100 Indigofera conferta LUH 4676 KX057876 KX119300 KX119386 - 101 Indigofera dendroides LUH 4674 - KX119301 - - 102 Indigofera nigritana LUH 5167 KX057880 KX119304 KX119388 KX268192 103 Indigofera macrocalyx LUH 4629 KX057879 KX119303 - - 104 Indigofera paniculata LUH 4675 KX057881 KX119305 KX119389 - 105 Indigofera pulchra LUH 1307A - - - - Bobgunnia KX119269 KX119355 106 madagascariensis LUH a258 KX057839 KX268156 107 Tephrosia elegans LUH 4182 - KX119333 - KX268220 108 Tephrosia bracteolata LUH 4684 KX057905 KX119330 KX119414 KX268217 109 Tephrosia linearis LUH 4310 KX057906 KX119331 KX119415 KX268218 110 Tephrosia platycarpa LUH 4667 - - - - 111 Tephrosia pedicellata LUH 4469 - KX119332 - KX268219 112 Lonchocarpus sericeus LUH 4195 - - - - 113 Lonchocarpus cyanescens LUH 4139 - - - - 114 Milletia bateri LUH 4680 - - - - 115 Ostryoderris stuhlmannii LUH 4602 - - - - 116 Sesbania bispinosa LUH 5128 KX057904 KX119329 KX119412 KX268216 117 Sesbania dalzielii LUH 5128 - - - - 118 Pericopsis laxiflora LUH 4219 KX057892 KX119317 KX119401 KX268202

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APPENDIX II: Character and states used in the key

#1. Average length of leaf/ /

#2. Type of leaf present/ 1. simple/ 2. bilobate/ 3. pinnately/ 4. bipinnately/ 5. imparipinnately/ 6. paripinnately/ 7. unifoliate/ 8. bifoliate/ 9. trifoliate/

#3. Number of leaflet per branch/ /

#4. Leaf attachment/ 1. leaf attached to the stem without a petiole / 2. leaf attached to the stem with a petiole / 3. leaf entirely sorrounding the stem /

#5. Length of leaflet/ /

#6. Leaflet width/ /

#7. Petiolule length/ /

#8. Petiole length/ /

#9. Leaf blade with degree of curvature/ 1. no curvature, parallel / 2. widest curvature at the middle / 3. widest curvature above middle / 4. widest curvature below middle / 5. widest curvature both above and below middle / 6. near circle in all outlins / 7. none/

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#10. Leaf blade narrow or wide with positioning of curvature/ 1. narrow and widest above middle / 2. narrow and widest below middle / 3. wide and widest below middle / 4. wide and widest above middle / 5. none/

#11. Leaf modification: leaf reduced to phyllodes/ 1. present/ 2. absent/

#12. Leaf modification: leaf reduced to sickled phyllodes/ 1. present with sickled blade / 2. absent/

#13. Leaf blade heart-shaped with positioning of curvature/ 1. widest at the base / 2. widest above the middle / 3. none/

#14. Leaf shape with relation to base symmetry/ 1. symmetrical / 2. asymmetrical /

#15. Leaf bilobed from the apex/ 1. presence/ 2. absent/

#16. Leaf with lateral asymmetrical while terminal symmetrical/ 1. present/ 2. absent/

#17. Leaf with terminal leaflet bigger than lateral leaflets/ 1. present/ 2. absent/

#18. Leaf tip/ 1. rounded/ 2. mucronate/ 3. obtuse/ 4. acute/ 5. acuminate/ 6. emarginate/ 7. aristate/ 8. cuspidate/

#19. Leaf Base/ 1. truncate/ 2. rounded/

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3. obtuse/ 4. cuneate/ 5. attenuate/ 6. cordate/

#20. Midrib Upper Surface/ 1. channelled/ 2. raised/ 3. flat/

#21. Midrib Lower Surface/ 1. channelled/ 2. slightly raised/ 3. raised/ 4. flat/

#22. Leaf margin thickening/ 1. thickened/ 2. slightly thickened/ 3. not thickened/

#23. Leaf margin/ 1. serrated/ 2. entire/ 3. bilobed/ 4. sinuate/

#24. Leaf texture/ 1. leathery/ 2. papery/

#25. Dry leaf under surface: green/ 1. green/ 2. greenish yellow/ 3. greenish brown/ 4. greenish grey/ 5. none/

#26. Dry leaf under surface: brown/ 1. brown/ 2. dark brown/ 3. rusty/ 4. none/

#27. Leaf arrangement/ 1. alternate/ 2. opposite/

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#28. Leaflet arrangement/ 1. alternate/ 2. opposite/ 3. trifoliate/ 4. pinnate-trifoliate/ 5. none/

#29. Accessory organs/ 1. present/ 2. absent/

#30. Types of accessory organs on leaf branch/ 1. prickles/ 2. stipule/ 3. spines/ 4. none/

#31. Presence of accessory organs on stem/ 1. prickles/ 2. spines/ 3. none/

#32. Visibleisible corolla/ 1. present/ 2. absent/

#33. Bicolored flowers/ 1. present/ 2. absent/

#34. Flower colour: yellow in relation to intensity/ 1. cream/ 2. yellow/ 3. bright yellow/ 4. greenish yellow/ 5. reddish yellow/ 6. golden yellow/ 7. none/

#35. Flower colour: red in relation to intensity/ 1. orange-red/ 2. bluish red/ 3. red/ 4. bright red/ 5. brick red/ 6. none/

#36. pure line flower colour/ 1. white/ 2. pink/

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3. lilac/ 4. purple/ 5. blue/ 6. orange/ 7. violet/ 8. green/ 9. none/

#37. constrictions in pods/ 1. present/ 2. absent/

#38. Pod twisting shape/ 1. irregular spirals/ 2. necklace-like/ 3. tightly-twisted/ 4. chain-like/ 5. moniliform/ 6. none/

#39. orientation of pod shape/ 1. flat/ 2. linear/ 3. clustered/ 4. spiral or twisted/ 5. falcate/ 6. ovoid/ 7. stout/ 8. lenticular/ 9. samara/

#40. flat Pod shape/ 1. oblong/ 2. round/ 3. cylindrical/ 4. round/ 5. moniliform/ 6. none/

#41. Tree height/ /

#42. Habit/ 1. herb/ 2. shrub/ 3. tree/ 4. climber/

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#43. habit of herb/ 1. erect 2. branched/ 3. perennial/ 4. creeping/ 5. climbing/ 6. straggling/ 7. none/

#44. habit of shrub/ 1. erect/ 2. scandent/ 3. branched/ 4. woody/ 5. thorny/ 6. prickly/ 7. none/

#45. Habit of tree/ 1. deciduous tree/ 2. evergreen tree/ 3. none/

#46. Inflorescence/ 1. axillary panicle/ 2. axillary raceme/ 3. raceme/ 4. pseudoraceme/ 5. spikes/ 6. corymb/ 7. dischasium/ 8. globose head/ 9. capitate head/ 10. panicles/

#47. flower type/ 1. actinomorphic/ 2. zygomorphic/ 3. papillionaceous/

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APPENDIX III: Character descriptions scored for each species

# AAT / 1,0 2,3 3,48-68 4,1 5,0.5-0.8 6,0.1 7,0 8,0 9,2 10,5 11,1 12,2 13,3 14,1 15,2 16,2 17,2 18,3/4 19,4 20,3 21,3 22,3 23,2 24,2 25,3 26,4 27,2 28,2 29,1 30,1 31,3 32,1 33,2 34,1 35,6 36,9 37,2 38,6 39,2 40,1 41,2-10 42,2/3 43,7 44,2 45,3 46,5/8 47,1

# AAU / 1,0 2,1 3,17-21 4,1 5,0 6,0.8-1.9 7,0 8,0 9,6 10,5 11,1 12,1 13,3 14,2 15,2 16,2 17,2 18,4 19,5 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,6 35,6 36,9 37,1 38,1 39,4 40,6 41,15-30 42,3 43,7 44,7 45,2 46,5/8 47,1

# AAL / 1,0 2,3 3,20-48 4,1 5,0.6-0.9 6,0.1-0.2 7,0 8,0.1 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3/4 19,2/3 20,3 21,3 22,3 23,2 24,2 25,4 26,4 27,2 28,2 29,1 30,2/3 31,3 32,1 33,2 34,1/2 35,6 36,1 37,1 38,3 39,4 40,6 41,30 42,3 43,7 44,7 45,1 46,10 47,1

# ASI / 1,0 2,3 3,54-72 4,1 5,0.3-0.4 6,0.1 7,0 8,0.1 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/3 19,3 20,3 21,3 22,3 23,2 24,2 25,1 26,4 27,1 28,2 29,1 30,3 31,3 32,1 33,2 34,1/2 35,6 36,1 37,2 38,6 39,5 40,6 41,12-25 42,3 43,7 44,7 45,1 46,8 47,1

# ASE / 1,0 2,3 3,9-17.0 4,1 5,1-1.9 6,0.1-0.2 7,0 8,0 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,2 28,2 29,1 30,1/2 31,3 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,2 40,1 41,7 42,2/3 43,7 44,7 45,3 46,5 47,1

# ANI / 1,0 2,3 3,18-28 4,1 5,0.8-1.5 6,0.1-0.3 7,0 8,0 9,1 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3/4 19,4 20,3 21,3 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,1 30,3 31,3 32,1 33,2 34,6 35,6 36,9 37,1 38,4 39,3 40,6 41,25 42,3 43,7 44,7 45,1 46,8 47,1

# ALE / 1,0 2,3 3,0 4,2 5,4.1-4.8 6,2-2.7 7,0 8,0.1 9,2 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,2 20,3 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,1 40,1 41,20-30 42,3 43,7 44,7 45,1 46,9 47,1

# AZY / 1,0 2,3 3,8.0-12 4,2 5,2.1-5 6,1.0-2.2 7,0 8,1.0-1.2 9,3/5 10,4 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3/4 19,2/4 20,3 21,3 22,1 23,2 24,1 25,1/4 26,4 27,2 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,2 37,2 38,6 39,1 40,1 41,9-30 42,3 43,7 44,7 45,1 46,9 47,1

# DCI / 1,0 2,3 3,28-48 4,2 5,0.3-0.5 6,0.1 7,0 8,0.5-0.7 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,2 20,3 21,3 22,3 23,2 24,2 25,3 26,4 27,1 28,2 29,1 30,3 31,3 32,1 33,1 34,2 35,6 36,2 37,1 38,3 39,4 40,6 41,7 42,2 43,7 44,5 45,3 46,5 47,1

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# EAB / 1,0 2,3 3,16-36 4,2 5,0.9-1.3 6,0.3-0.35 7,0 8,1.0-1.8 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,2/3 19,2/3/4 20,3 21,3 22,2 23,2 24,1 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,1 40,6 41,17 42,3 43,7 44,7 45,3 46,3 47,1

# EAF / 1,0 2,3 3,14-22 4,1 5,2-2.9 6,0.6-0.7 7,0 8,0 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1 19,2 20,3 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,1/5 35,6 36,1 37,2 38,6 39,5 40,6 41,10 42,3 43,7 44,7 45,3 46,3/5 47,1

# LLE / 1,0 2,3 3,36-56 4,2 5,0.5-0.8 6,0.2 7,0 8,0.1-0.2 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,3 22,3 23,2 24,2 25,3 26,4 27,1 28,2 29,1 30,2 31,1 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,1 40,6 41,2-10 42,2/3 43,7 44,7 45,3 46,8 47,1

# MPI / 1,0 2,3 3,26-40 4,2 5,0.3-0.5 6,0.75-0.1 7,0 8,0.5-0.7 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,1 20,3 21,3 22,3 23,2 24,2 25,5 26,3 27,1 28,2 29,1 30,1 31,1 32,1 33,2 34,7 35,6 36,2 37,2 38,6 39,1/3 40,1 41,6 42,2 43,7 44,4 45,3 46,8 47,1

# MPU / 1,0 2,3 3,26-46 4,2 5,0.8-0.9 6,0.1 7,0 8,1.6-2.0 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,3 22,3 23,2 24,2 25,5 26,1/2 27,1 28,2 29,1 30,1 31,3 32,1 33,2 34,7 35,6 36,2/3 37,2 38,6 39,1/3 40,1 41,1.5 42,3 43,7 44,3/6 45,3 46,8 47,1

# PBC / 1,0 2,3 3,32-52 4,2 5,1.5-1.9 6,0.3-0.4 7,0 8,0.2-0.4 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/3 19,2 20,3 21,3 22,1 23,2 24,1 25,5 26,2 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,1/2 36,6 37,2 38,6 39,2 40,6 41,35 42,3 43,7 44,7 45,3 46,3 47,1

# PBG / 1,0 2,3 3,40-56 4,2 5,1.8-2.1 6,0.5-0.6 7,0 8,0.5 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,2 20,3 21,3 22,1 23,2 24,1 25,3 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,3 36,6 37,2 38,6 39,1/2 40,1 41,7-20 42,3 43,7 44,7 45,1 46,3 47,1

# PDU / 1,0 2,6 3,4 4,2 5,2.3-2.7 6,1.0-1.4 7,0 8,0.8-1.7 9,1/2 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,4 20,3 21,3 22,3 23,2 24,2 25,4 26,4 27,2 28,2 29,1 30,2/3 31,3 32,1 33,2 34,7 35,6 36,1/8 37,1 38,1 39,4 40,6 41,15-20 42,3 43,7 44,7 45,3 46,10 47,1

# PAF / 1,0 2,3 3,16-26 4,2 5,1.5-1.6 6,0.5-1.0 7,0 8,0.6-0.7 9,1 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,3 22,2 23,2 24,1 25,3 26,4 27,2 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1/8 37,2 38,6 39,7 40,3 41,20 42,3 43,7 44,7 45,3 46,5 47,1

# AFZ / 1,16-30 2,4 3,6-18 4,1 5,6.3-9.6 6,3.4-5 7,0 8,0.5-1 9,2/4 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,1 40,6 41,10-18 42,3 43,7 44,7 45,1 46,1 47,2

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# BMO / 1,5.4-12.5 2,1 3,0 4,1 5,0 6,6.6-12.1 7,0 8,2.3-5.3 9,6 10,5 11,2 12,2 13,3 14,1 15,1 16,2 17,2 18,1/3 19,2/6 20,3 21,2 22,1 23,3 24,1 25,4 26,4 27,1 28,5 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,2 37,2 38,6 39,1 40,6 41,10.5 42,3 43,7 44,7 45,3 46,6 47,2

# BPU / 1,5.5-8 2,1 3,0 4,1 5,0 6,7.5-9.1 7,0 8,1.6-3.2 9,6 10,5 11,2 12,2 13,3 14,1 15,1 16,2 17,2 18,3 19,2/6 20,3 21,3 22,1 23,3 24,1 25,4 26,4 27,1 28,5 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,3/4/5 37,2 38,6 39,1 40,6 41,10-12 42,3 43,7 44,7 45,3 46,1 47,2

# BRU / 1,4-12.3 2,2 3,5-15 4,2 5,1-2.1 6,1.9-2.3 7,0.7-1.5 8,1.8-3 9,6 10,5 11,2 12,2 13,3 14,1 15,1 16,2 17,2 18,1/3 19,2/6 20,3 21,4 22,3 23,3 24,2 25,4 26,4 27,1 28,1 29,2 30,4 31,3 32,1 33,2 34,4 35,6 36,1 37,1 38,1 39,4 40,6 41,3 42,2/3 43,7 44,1 45,3 46,3 47,2

# BTO / 1,4.2-5.7 2,1 3,0 4,1 5,0 6,3.5-6.5 7,0 8,2-3.5 9,6 10,5 11,2 12,2 13,3 14,1 15,1 16,2 17,2 18,3/4 19,1/6 20,3 21,4 22,3 23,3 24,2 25,2 26,4 27,1 28,3 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,2 40,6 41,4 42,2/3 43,7 44,1 45,3 46,3 47,2

# BVA / 1,11.4-12 2,1 3,0 4,1 5,0 6,10.3-15.1 7,0 8,3.5-4 9,6 10,5 11,2 12,2 13,3 14,1 15,1 16,2 17,2 18,3 19,6 20,3 21,3 22,1 23,3 24,1 25,4 26,4 27,1 28,3 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,1 40,6 41,20 42,3 43,7 44,7 45,3 46,6 47,2

# BAF / 1,14-22.6 2,4 3,9-15 4,2 5,3-4.8 6,1.8-2.2 7,0.1-0.3 8,1.3-1.8 9,2/4 10,4 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3/6 19,2 20,3 21,4 22,1 23,2 24,1 25,3 26,4 27,2 28,1 29,1 30,2 31,3 32,1 33,2 34,1 35,6 36,9 37,2 38,6 39,1 40,6 41,20 42,3 43,7 44,7 45,3 46,5 47,2

# CPU / 1,18.2-19.8 2,4 3,12-36 4,2 5,1.5-1.8 6,0.7-1.1 7,0.1 8,3.2-5.4 9,1/4 10,4 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,2 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,2 28,2 29,1 30,1/2 31,3 32,1 33,2 34,2 35,3 36,9 37,2 38,6 39,1 40,6 41,3 42,3 43,7 44,1 45,2 46,6 47,2

# CAR / 1,16.2-26.2 2,4 3,10-18 4,2 5,4.2-5.2 6,2.6-2.8 7,0.3-0.6 8,2-3.3 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,1 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,2 40,6 41,10 42,3 43,7 44,7 45,3 46,6 47,2

# CNI / 1,2.8-5 2,4 3,10-28 4,1 5,0.8-1 6,0.1-0.3 7,0 8,0.8-1 9,1 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,1/2/3 19,2 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,1 40,6 41,1.5 42,1/2 43,7 44,7 45,3 46,3 47,2

# STO / 1,3.9-4.2 2,4 3,6 4,2 5,1.8-2.5 6,0.8-1.5 7,0.1-0.2 8,1.2-1.8 9,3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,2 28,2 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,2 40,6 41,1.5-2.5 42,1/2 43,1 44,7 45,3 46,3 47,2

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# CMI / 1,10 2,4 3,40-60 4,1 5,0.2-0.4 6,0.1-0.10 7,0 8,0.1-0.2 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,2/4 19,1 20,3 21,4 22,3 23,2 24,2 25,5 26,2 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,1 40,6 41,1.7 42,1 43,1 44,7 45,3 46,3 47,2

# CME / 1,10.1-16.7 2,6 3,6 4,1 5,5.7-7.5 6,2.2-3.2 7,0 8,0.5-0.8 9,3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,1 35,6 36,9 37,2 38,6 39,8 40,1 41,12 42,3 43,7 44,7 45,3 46,10 47,2

# DOL / 1,32 2,6 3,6-14 4,2 5,12.3-15.3 6,5.8-7.5 7,1.1-1.2 8,3.9-4.5 9,1/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,4 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,9 40,6 41,9-25 42,3 43,7 44,7 45,1 46,10 47,2

# DRE / 1,28.9-35.1 2,4 3,36-52 4,1 5,4.2-7.3 6,0.2-0.3 7,0 8,7-10 9,1/4 10,3 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,2 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,3 36,9 37,2 38,6 39,1/2 40,6 41,15 42,3 43,7 44,7 45,3 46,6 47,2

# DMA / 1,16.1-20.5 2,6 3,8 4,2 5,9-12.4 6,3.2-4.5 7,0.3-0.5 8,2-2.5 9,4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,1 21,3 22,1 23,2 24,1 25,5 26,1/2 27,1 28,1 29,2 30,4 31,3 32,1 33,2 34,1 35,6 36,1 37,2 38,6 39,6 40,6 41,50 42,3 43,7 44,7 45,3 46,10 47,2

# DGU / 1,10-13.1 2,5 3,5 4,2 5,4.2-7.3 6,3.3-6.2 7,0.2-0.3 8,1.1-1.2 9,2/3 10,4 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,2 20,3 21,4 22,1 23,2 24,1 25,3 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,1 40,6 41,30 42,3 43,7 44,7 45,3 46,10 47,2

# IDO / 1,25.2-29.2 2,6 3,6-8 4,2 5,9-12 6,4.4-5.8 7,0.8-1.2 8,2.8-4 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4/5 19,4 20,3 21,2 22,1 23,2 24,1 25,5 26,1 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,2 40,1 41,10-20 42,3 43,7 44,7 45,3 46,1 47,2

# ITO / 1,20-29 2,6 3,6-8 4,2 5,9.3-15 6,4.5-6.2 7,1.2-1.8 8,3-3.8 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4/5 19,2 20,1 21,3 22,1 23,2 24,1 25,5 26,1 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1/2 37,2 38,6 39,1/2 40,6 41,18 42,3 43,7 44,7 45,3 46,1 47,2

# PRE / 1,7-8.1 2,1 3,0 4,1 5,0 6,8.1-9 7,0 8,2.8-3.5 9,7 10,5 11,2 12,2 13,2 14,2 15,1 16,2 17,2 18,3 19,2 20,3 21,2 22,1 23,3 24,1 25,4 26,4 27,1 28,5 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1/2 37,2 38,6 39,2 40,6 41,9 42,3 43,7 44,7 45,3 46,2 47,2

# PTH / 1,9.6-11 2,1 3,0 4,1 5,0 6,9-12 7,0 8,3.5-4 9,7 10,5 11,2 12,2 13,2 14,2 15,1 16,2 17,2 18,6 19,6 20,3 21,3 22,1 23,3 24,1 25,3 26,4 27,1 28,5 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,2 40,1 41,6 42,2/3 43,7 44,1 45,3 46,2 47,2

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# SAL / 1,44.5-52.4 2,3 3,18-26 4,2 5,5.8-8.7 6,2.8-4.2 7,0.1-0.2 8,1.5-1.8 9,1/2 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3/6 19,2 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,6 35,6 36,9 37,2 38,6 39,9 40,6 41,2-3 42,2 43,7 44,1 45,3 46,2 47,2

# SHI / 1,17.5-18.6 2,3 3,6-10 4,2 5,3.4-5.8 6,1.7-2.2 7,0.1-0.2 8,3-6 9,1/2 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,5 19,2 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,6 37,2 38,6 39,5 40,6 41,0.5-3 42,2 43,7 44,1 45,3 46,2 47,2

# SOB / 1,7.9-8.6 2,3 3,4-6 4,2 5,2.5-3 6,1.5-1.9 7,0.1-0.2 8,3.4-3.7 9,3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,5 40,6 41,2.5 42,2 43,7 44,1 45,3 46,2 47,2

# SOC / 1,9.1-24.8 2,3 3,6-10 4,2 5,4.5-6.4 6,2-2.7 7,0.2-0.3 8,5.5-6.1 9,7 10,1 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,5 19,2 20,3 21,4 22,3 23,2 24,2 25,5 26,2 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,5 40,6 41,2 42,2 43,7 44,1 45,3 46,7 47,2

# CRO / 1,2.3-2.6 2,8 3,2 4,1 5,1.8-1.9 6,1-1.2 7,0 8,0.3-0.5 9,3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,1/2 40,6 41,1 42,1 43,1 44,7 45,3 46,2 47,2

# SSI / 1,23.5-25.5 2,3 3,18-22 4,2 5,4.8-5.5 6,1.9-2.2 7,0.2-0.3 8,1-3 9,1 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,1/2 19,2 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,5 40,6 41,15-20 42,3 43,7 44,7 45,3 46,2 47,2

# CSI / 1,17.5-22.5 2,6 3,6-12 4,2 5,2.6-5.2 6,1.6-2.9 7,0.5-0.6 8,2-3 9,2/4 10,3 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,1/4 19,2 20,3 21,3 22,1 23,2 24,1 25,3 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,1 40,3 41,15-20 42,2/3 43,7 44,7 45,3 46,2 47,2

# SSN / 1,12.8-19.2 2,6 3,10-18 4,2 5,3.2-4.3 6,1.7-2 7,0.3 8,2-4 9,2/3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/2 19,4 20,1 21,3 22,1 23,2 24,1 25,3 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,1/2 40,6 41,15 42,2/3 43,7 44,1 45,3 46,2 47,2

# TIN / 1,10-11 2,6 3,22-28 4,1 5,1.4-2.2 6,0.6-0.8 7,0 8,0.7-0.8 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1 19,2 20,3 21,4 22,3 23,2 24,2 25,5 26,1 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,2 37,2 38,6 39,1 40,3 41,15-25 42,3 43,7 44,7 45,2 46,5 47,2

# APR / 1,5-13 2,3 3,30 4,2 5,1.1-1.7 6,0.4-1.1 7,0.1 8,0.3-0.4 9,1 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,2 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,1/2 37,2 38,6 39,1 40,1 41,3.3 42,4 43,7 44,5 45,3 46,1 47,3

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# APA / 1,18.2-19.6 2,9 3,3 4,2 5,10.8-12 6,4.7-7.1 7,0.5-2.8 8,4.5-5 9,4 10,3 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,2 20,3 21,4 22,3 23,2 24,2 25,5 26,2 27,2 28,4 29,2 30,4 31,3 32,1 33,1 34,2 35,6 36,2 37,2 38,6 39,1 40,6 41,1.5-4 42,2 43,4 44,1 45,3 46,2 47,3

# AGL / 1,8.6-13.1 2,7 3,7 4,2 5,1.6-3.2 6,0.7-1.7 7,0.3-0.4 8,1-2.5 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,6 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,1 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,9 37,1 38,1 39,4 40,6 41,1 42,1 43,1 44,5 45,3 46,2 47,3

# ARU / 1,10-11 2,9 3,3 4,2 5,8.9-10.8 6,0.2-0.3 7,0.5-0.6 8,3-3.5 9,1 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,9 37,1 38,1 39,4 40,6 41,0.3-2 42,1 43,1 44,5 45,3 46,2 47,3

# AHY / 1,4.4-4.6 2,9 3,6 4,2 5,1.8-3.7 6,1.1-1.4 7,0.1-0.2 8,1.8.2.2 9,1/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/2/6 19,2/4 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,9 37,1 38,1 39,6/8 40,6 41,4 42,1 43,2 44,5 45,3 46,2 47,3

# CCA / 1,13-16.4 2,9 3,3 4,2 5,7.3-10 6,2-2.8 7,0.2-1.9 8,3-4.7 9,1 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5/7 19,4 20,3 21,4 22,3 23,2 24,1 25,2 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,1/2 35,6 36,9 37,2 38,6 39,8 40,1 41,4 42,2 43,7 44,1 45,3 46,2 47,3

# CMU / 1,14-18.3 2,9 3,3 4,2 5,2.8-5.8 6,3.9-6.6 7,0.3-0.7 8,4.2-13 9,4/5 10,3 11,2 12,2 13,3 14,1/2 15,2 16,1 17,2 18,3 19,2/4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,4/5 37,2 38,6 39,2 40,1 41,3-5 42,1 43,4 44,5 45,3 46,3 47,3

# CCO / 1,3.5-9 2,9 3,3 4,1 5,4-8.5 6,1.8-2.6 7,0 8,1.2-1.6 9,1 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3/8 19,4 20,3 21,4 22,1 23,2 24,1 25,2 26,4 27,1 28,3 29,2 30,4 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,2 40,6 41,0.6-0.9 42,1 43,1 44,5 45,3 46,3 47,3

# CHY / 1,2.3-3.8 2,9 3,3 4,1 5,1.8-3 6,0.3-0.6 7,0 8,0.5-0.8 9,7 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,3 29,2 30,4 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,2 40,6 41,0.5-0.6 42,1 43,1 44,5 45,3 46,2 47,3

# CLA / 1,3.5-5.3 2,9 3,3 4,2 5,2.8-5.4 6,1.3-2.4 7,0.1-0.2 8,0.5-1 9,7 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,8 19,4 20,3 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,2 40,6 41,0.6-1.8 42,2 43,7 44,4 45,3 46,2 47,3

# CMA / 1,7-8.6 2,9 3,3 4,1 5,2.7-5.7 6,0.2-1.2 7,0 8,2.6-4.1 9,2 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,6 40,1/4 41,0.9 42,1 43,1 44,5 45,3 46,2 47,3

391 | P a g e

# CPA / 1,12.3-14.8 2,9 3,3 4,2 5,6-8.1 6,1.6-1.8 7,0.1 8,6-6.2 9,2 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,2 40,1 41,1.5 42,1 43,7 44,5 45,3 46,2 47,3

# CNG / 1,7-10.2 2,9 3,3 4,1 5,2.8-7.2 6,0.9-1.8 7,0 8,2.2-7.5 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,3 23,2 24,2 25,2 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,2 40,6 41,0.9-1.8 42,2 43,7 44,3 45,3 46,3 47,3

# CRE / 1,4.4-6.7 2,1 3,16 4,2 5,U 6,2.1-2.6 7,0 8,0.2-0.3 9,7 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,5 29,1 30,2 31,3 32,1 33,2 34,3 35,6 36,9 37,2 38,6 39,2 40,1/3 41,1.3 42,1 43,1 44,5 45,3 46,2 47,3

# DSI / 1,5.9-16.9 2,5 3,7 4,2 5,4.5-7.8 6,3.2-5.6 7,0.3-2 8,1.8-2.5 9,4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,2 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,1 29,2 30,4 31,3 32,1 33,2 34,1/2 35,6 36,9 37,2 38,6 39,1/2 40,6 41,30 42,3 43,7 44,5 45,1 46,1 47,3

# DGA / 1,4.6-11.4 2,9 3,1 4,1 5,6.2-15.5 6,2.9-4.6 7,0 8,1-2.2 9,4 10,2/3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4/5 19,2 20,3 21,4 22,3 23,2 24,2 25,3 26,4 27,1 28,1 29,1 30,2 31,3 32,1 33,2 34,1 35,6 36,1/2 37,2 38,6 39,5 40,6 41,0.6-1.2 42,2 43,7 44,1 45,3 46,2 47,3

# DSC / 1,2.9-4.1 2,9 3,3 4,2 5,1.1-2.2 6,0.5-1 7,0.2-4 8,1-1.8 9,2 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,1/2/4/5 37,2 38,6 39,2 40,6 41,0.6 42,1 43,6 44,5 45,3 46,3 47,3

# DTO / 1,9.8-12.3 2,9 3,3 4,2 5,4-6.3 6,1.9-2.5 7,0.4-1.3 8,2-4.4 9,2 10,2 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,4 37,1 38,4 39,4 40,6 41,0.6-3 42,1 43,1 44,5 45,3 46,1 47,3

# DVE / 1,9.2-10.4 2,7 3,11 4,2 5,4-20 6,4.6-5.6 7,2-3 8,1.4-2 9,4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,1 21,3 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,5/7 37,3 38,6 39,2 40,1 41,0.5-3 42,2 43,7 44,1 45,3 46,1/2 47,3

# ESE / 1,14.7-16.3 2,9 3,3 4,2 5,6.8-11.2 6,3.6-6.3 7,0.2-2.8 8,4.9-6 9,1/4 10,3 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,3 19,2 20,3 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,4 29,1 30,1 31,3 32,1 33,2 34,7 35,4 36,9 37,1 38,6 39,4 40,6 41,7 42,3 43,7 44,5 45,3 46,1 47,3

# ESI / 1,17.5-22 2,9 3,3 4,2 5,6.8-7.7 6,7.3-10.6 7,0.2-4.8 8,7.2-9.6 9,4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/2/6 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,2 28,4 29,1 30,1 31,3 32,1 33,2 34,7 35,3 36,9 37,1 38,5 39,4 40,5 41,3-6 42,3 43,7 44,5 45,3 46,3 47,3

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# GSE / 1,14.5-21 2,5 3,13 4,2 5,4.1-6.7 6,2.1-2.6 7,0.3-0.4 8,1.7-2.8 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,5 19,4 20,3 21,4 22,3 23,2 24,2 25,5 26,3 27,2 28,2 29,2 30,4 31,3 32,1 33,2 34,7 35,6 36,2/3 37,2 38,6 39,1 40,6 41,10-12 42,2/3 43,7 44,1 45,3 46,3 47,3

# IAR / 1,5.4-9.1 2,5 3,18 4,2 5,1.4-2.4 6,0.4-0.9 7,0.1-0.2 8,1.1-1.2 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/3 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,2 37,2 38,6 39,2 40,6 41,3 42,2 43,7 44,1 45,3 46,2 47,3

# IHI / 1,1.2-2.3 2,5 3,7 4,1 5,0.4 6,0.25 7,0 8,0.1 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3/4 19,4 20,3 21,4 22,2 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,5 36,2 37,2 38,6 39,2 40,6 41,1.5 42,1 43,1 44,5 45,3 46,3 47,3

# IMA / 1,2.8-3.7 2,5 3,10 4,2 5,0.9-1.8 6,0.4-0.6 7,0.1 8,0.2-0.3 9,1/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,2 37,2 38,6 39,6/8 40,2 41,1 42,1 43,1 44,5 45,3 46,2 47,3

# INU / 1,2-2.7 2,1 3,11 4,1 5,U 6,1.3-1.9 7,0 8,0.1-0.2 9,3/6 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3 19,4 20,3 21,4 22,1 23,2 24,1 25,2 26,4 27,1 28,5 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,2 37,2 38,6 39,1 40,6 41,0.6-0.7 42,1 43,2 44,5 45,3 46,2 47,3

# ISP / 1,4-7.8 2,3 3,13 4,1 5,0.9-5 6,0.4-0.6 7,0.1-0.2 8,0.4-0.5 9,2/3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,2/3 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,1 36,2 37,2 38,6 39,2 40,6 41,0.5 42,1 43,1 44,5 45,3 46,3 47,3

# ISU / 1,8.2-9 2,5 3,11 4,2 5,2.6-4 6,1.1-1.5 7,0.1-0.2 8,1-1.5 9,2/3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,2 19,4 20,3 21,4 22,1 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,3 36,2 37,2 38,6 39,5 40,6 41,2 42,1/2 43,1 44,1 45,3 46,2 47,3

# LCY / 1,11.2-19.2 2,5 3,12 4,2 5,5.8-9 6,2.5-3.9 7,0.3-0.4 8,0.8-1.8 9,3 10,4 11,2 12,2 13,3 14,1 15,2 16,2 17,1 18,4/5 19,4 20,3 21,3 22,1 23,4 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,3/5 37,2 38,6 39,8 40,1 41,15-20 42,2 43,7 44,2 45,1 46,1 47,3

# LSE / 1,18.5-28.5 2,5 3,10 4,2 5,9.1-14 6,3.8-6.8 7,0.3-0.5 8,1-1.7 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,1 18,4/5 19,2/4 20,3 21,2 22,3 23,4 24,2 25,2 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,3 37,2 38,6 39,1/3 40,6 41,12-15 42,2/3 43,7 44,1 45,2 46,2 47,3

# MPR / 1,18-18.1 2,9 3,3 4,2 5,12-16.4 6,7.5-8.5 7,0.3-0.4 8,2-2.6 9,4/5 10,3 11,2 12,2 13,3 14,1/2 15,2 16,1 17,2 18,4 19,4 20,3 21,3 22,1 23,2 24,1 25,4 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,4 37,2 38,6 39,2 40,1 41,2-3 42,1 43,7 44,5 45,3 46,2 47,3

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# PER / 1,20-22.4 2,5 3,13 4,2 5,6-8.6 6,2.5-3.1 7,0.5-0.6 8,2.8-3.5 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4/5 19,4 20,3 21,3 22,1 23,4 24,1 25,4 26,4 27,1 28,1 29,1 30,2 31,3 32,1 33,2 34,6 35,6 36,9 37,2 38,6 39,1 40,4 41,15-25 42,3 43,7 44,5 45,1 46,1 47,3

# SBI / 1,9.5-29.5 2,3 3,46 4,2 5,1.4-2.6 6,0.3 7,0.1 8,0.9-1.2 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,3/6 19,3 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,4 37,2 38,6 39,5 40,6 41,8 42,1/2 43,1 44,1 45,3 46,3 47,3

# SER / 1,1.8-3.8 2,9 3,3 4,2 5,1.2-2.8 6,0.3-0.7 7,0.1 8,0.3-0.5 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,3 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,3 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,6 37,2 38,6 39,2 40,1 41,0.1-1.5 42,1/2 43,1 44,1 45,3 46,2 47,3

# SMA / 1,15.8-23.9 2,5 3,11 4,2 5,3.9-5 6,2.2.6 7,0.2-0.4 8,3.5-4.6 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/3 19,4 20,1 21,3 22,1 23,2 24,1 25,2 26,4 27,1 28,1 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,1 37,2 38,6 39,2 40,3 41,10-15 42,2/3 43,7 44,1 45,3 46,2 47,3

# TEL / 1,3.8-6.1 2,8 3,4 4,1 5,2.5-5.5 6,0.5-1.5 7,0 8,0.2-0.4 9,1/2 10,1 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1/3 19,4 20,3 21,3 22,1 23,2 24,1 25,3 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,6 37,2 38,6 39,2 40,1 41,0.9-1.2 42,1 43,7 44,5 45,3 46,2 47,3

# TBR / 1,20.7-31.2 2,3 3,32 4,2 5,5.4-7.8 6,0.4-0.6 7,0.2 8,0.8 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,6 19,4 20,3 21,4 22,2 23,2 24,1 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,1 33,2 34,7 35,6 36,2 37,2 38,6 39,1/2 40,1 41,2-3 42,1 43,1 44,5 45,3 46,4 47,3

# TLI / 1,14.8-17 2,3 3,16 4,2 5,3.2-4.3 6,0.3-0.4 7,0.6-1 8,0.1-0.2 9,1/2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,1 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,2 30,4 31,3 32,1 33,2 34,2 35,3 36,2/6 37,2 38,6 39,1/2 40,6 41,1.3-1.5 42,1 43,1 44,5 45,3 46,4 47,3

# TPL / 1,10.7-18.5 2,3 3,12 4,1 5,6-7 6,0.2-0.3 7,0 8,2.7-3.5 9,2 10,5 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,4 19,4 20,3 21,4 22,3 23,2 24,2 25,4 26,4 27,1 28,2 29,1 30,2 31,3 32,2 33,2 34,7 35,6 36,9 37,2 38,6 39,1 40,6 41,0.3-0.4 42,1 43,1 44,5 45,3 46,3 47,3

# VSU / 1,13.6-15.1 2,9 3,3 4,2 5,4.8-5.2 6,1.1-2.2 7,0.1-0.9 8,5-30 9,2/4 10,3 11,2 12,2 13,3 14,1 15,2 16,2 17,2 18,6 19,4 20,3 21,4 22,2 23,2 24,1 25,3 26,4 27,1 28,4 29,1 30,2 31,3 32,1 33,2 34,2 35,6 36,9 37,2 38,6 39,6 40,4 41,0.1-0.3 42,1 43,4 44,5 45,3 46,3 47,3

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APPENDIX IV: Items List used in DELTA

*NUMBER OF CHARACTERS 47 *MAXIMUM NUMBER OF STATES 10 *MAXIMUM NUMBER OF ITEMS 88

*CHARACTER TYPES 1,RN 2,OM 3,RN 4,OM 5-8,RN 9-40,OM 41,RN 42-47,OM

*NUMBER OF STATES 2,9 4,3 9,7 10,5 11,2 12,2 13,3 14,2 15,2 16,2 17,2 18,8 19,6 20,3 21,4 22,3 23,4 24,2 25,5 26,4 27,2 28,5 29,2 30,4 31,3 32,2 33,2 34,7 35,6 36,9 37,2 38,8 39,9 40, 6 42,4 43,7 44,7 45,3 46,10 47,3

*DEPENDENT CHARACTERS 2,1:5'

APPENDIX V: Tokey List used in DELTA

*COMMENT: HEADING legume species. Data converted #TIME #DATE.

*LISTING FILE tokey.lst

*INPUT FILE specs

*TRANSLATE INTO KEY FORMAT

*CHARACTER RELIABILITIES 1,8 2,8, 3,8 4,8 5,8 6,9 7,5 8,5 9,9 10,9 11,9 12,9 13,9 14,9 15,9 16,9 17,9 18,9 19,9 20,6 21,6 22,6 23,7 24,7 25,7 26,7 27,7 28,7 29,7 30,7 31,7 32,6 33,6 34,6 35,6 36,6 37,6 38,6 39,6 40,6 41,7 42,8 43,8 44,8 45,8 46,6 47,9

*COMMENT: KEY STATES 1,~0-0.1/0.1-5/5-9.1/9.1-20/20-36/36-43/43-55~ 3,~0-0.1/0.1-5/5-15/15-30/30-40/40-60~ 5,~0-0.1/0.1-3/3-5.5/5.5-16/16-20~ 6,~0.1-0.5/0.5-1.0/1.0-2.5/2.5-8.0/8.0-10/10-16~ 7,~0-0.1/0.1-0.5/0.5-0.8/0.8-1.9~ 8,~0-0.1/0.1-0.5/0.5-2/2-3.5/3.5-5.5/5.5-6.5/6.5-7/7-10/10-30~ 41,~0.1-0.3/0.3-0.5/0.5-1/1-8/8-20/20-35/30-35/35-50~

*KEY OUTPUT FILE kchars *INPUT FILE chars

*KEY OUTPUT FILE kitems *INPUT FILE items

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APPENDIX VI: DNA Bank number and Spectrophotometric readings

Sample Name DNA bank number Purity (A260/280) Conc. (ng/µl) Acacia albida M005 2.00 927.2 Acacia albida M006 1.86 583.4 Acacia albida M043 2.01 346.9 Acacia albida M033 2.01 387.7 Acacia ataxacantha M001 1.94 840.6 Acacia ataxacantha M002 1.97 1780.4 Acacia ataxacantha M045 2.00 424.3 Acacia ataxacantha M046 1.98 693.2 Acacia ataxacantha H089 2.01 179.5 Acacia auriculiformis M003 2.00 474 Acacia auriculiformis M004 2.00 995.7 Acacia auriculiformis M047 2.00 39.2 Acacia auriculiformis M048 1.98 125.7 Acacia ehrenbergiana M009 1.97 540.3 Acacia ehrenbergiana M010 2.01 330.9 Acacia nilotica M011 1.98 882.2 Acacia nilotica M012 1.98 1197.7 Acacia nilotica M049 1.80 699.3 Acacia nilotica M050 1.91 754 Acacia nilotica H086 2.01 164.2 Acacia nilotica H087 1.95 341.2 Acacia senegal M051 1.84 131.2 Acacia senegal M052 1.88 224 Acacia senegal H088 1.93 329.4 Acacia senegal H090 1.77 160.4 Acacia sieberiana M007 1.92 578.7 Acacia sieberiana M008 1.77 531.3 Acacia sieberiana M053 1.95 1045.5 Acacia sieberiana M054 2.01 449.9 Acacia villosa M055 2.00 312.1 Albizia lebbeck M013 1.98 239.6 Albizia lebbeck M014 1.97 177 Albizia lebbeck H091 1.98 10.1 Albizia lebbeck H092 1.81 6.9 Albizia lebbeck H093 1.78 19.9 Albizia lebbeck H096 2.01 271.5 Albizia zygia M015 2.01 391.6 Albizia zygia M016 1.88 226.5 Dichrostachys cinerea M017 1.94 283.9 Dichrostachys cinerea M018 2.00 251.1 Dichrostachys cinerea M031 1.98 404.4 Dichrostachys cinerea M032 1.98 334.7 Dichrostachys cinerea M056 2.00 172.6

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Dichrostachys cinerea M057 1.96 177.3 Dichrostachys cinerea H094 1.87 306.7 Dichrostachys cinerea H095 1.95 311.7 Entada abyssinica M019 1.75 194.1 Entada abyssinica M020 1.83 346.1 Entada abyssinica H097 2.01 261.3 Entada abyssinica H098 2.00 356.1 Entada africana M021 2.00 149.9 Entada africana M022 2.01 180.8 Entada africana M059 2.00 305.6 Entada africana M060 1.85 109.8 Leucaena esculenta M023 2.00 294 Leucaena esculenta M024 2.00 280.5 Leucaena esculenta M025 2.00 251.5 Leucaena esculenta M061 1.88 764 Leucaena esculenta M062 1.89 389.4 Leucaena leucocephala M026 1.89 271.2 Leucaena leucocephala M027 1.82 251.2 Leucaena leucocephala M028 2.00 184.3 Leucaena leucocephala H099 1.84 552.5 Leucaena leucocephala H100 1.94 467 Leucaena leucocephala H101 1.91 191.7 Leucaena leucocephala H102 1.99 675.3 Leucaena leucocephala H103 1.99 349.7 Mimosa pigra M029 2.01 234.3 Mimosa pigra M030 1.95 348.1 Mimosa pigra H108 2.01 319.5 Mimosa pigra H109 2.00 204.6 Mimosa pigra H110 1.87 157.6 Mimosa pigra H111 1.84 207.9 Mimosa pudica H104 1.85 380.8 Mimosa pudica H105 1.98 139.5 Mimosa pudica H106 1.77 61.8 Mimosa pudica H107 1.96 168 Neptunia oleracea M033 1.98 133.4 Neptunia oleracea M034 1.98 212.7 Neptunia oleracea H112 2.16 66.8 Parkia bicolor M035 1.83 225.4 Parkia bicolor M036 1.88 248.6 Parkia biglobosa M037 1.94 150.5 Parkia biglobosa M038 1.98 166.2 Parkia biglobosa M063 1.96 101.7 Parkia biglobosa M064 1.97 106.6 Pithecellobium dulce M039 1.83 209.3 Pithecellobium dulce M040 1.85 218.3 Pithecellobium dulce M065 1.86 386.2

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Pithecellobium dulce M066 1.85 468.1 Pithecellobium dulce H113 1.96 239.4 Prosopis africana M041 1.87 189.8 Prosopis africana M042 1.97 270.3 Prosopis africana M058 1.92 85.8 Afzelia africana C025 1.85 405.2 Bauhinia monandra C001 1.99 620.8 Bauhinia monandra C002 1.89 341.8 Bauhinia purpurea C009 1.88 271.4 Bauhinia purpurea C010 1.84 239.5 Bauhinia rufescens C003 1.99 312 Bauhinia rufescens C004 1.88 474 Bauhinia rufescens C005 1.98 362.4 Bauhinia rufescens C075 1.98 428.2 Bauhinia tomentosa C006 1.93 381.4 Bauhinia tomentosa C007 1.98 109.1 Bauhinia tomentosa C008 1.89 83.1 Bauhinia tomentosa C073 1.82 662.5 Bauhinia tomentosa C074 1.42 665.7 Bauhinia vahlii C011 1.86 248.9 Bauhinia vahlii C012 1.81 290 Burkea africana C013 1.95 142.9 Burkea africana C014 1.94 119.7 Burkea africana C076 1.97 279.9 Burkea africana C077 1.94 263.5 Caesalpinia pulcherrima C026 1.85 177.8 Caesalpinia pulcherrima C027 2.01 175.9 Caesalpinia pulcherrima C087 1.96 275.3 Cassia arereh C015 1.87 249.6 Cassia arereh C016 1.88 238.4 Cassia arereh C078 1.97 216.2 Cassia arereh C079 1.87 176.1 Cassia hirsuta C084 1.99 442.7 Cassia hirsuta C085 1.91 373.5 Cassia hirsuta C086 1.93 614.2 Cassia italica H065 1.83 131.2 Cassia italica H066 2.01 232.6 Cassia italica H067 1.84 200.8 Cassia italica H068 2.00 670.6 Cassia manii C019 1.94 186.3 Cassia manii C020 1.84 197.6 Cassia mimosoides C017 1.88 232.6 Cassia mimosoides C018 2.01 162.3 Cassia mimosoides C080 1.97 341.8 Cassia mimosoides C081 1.94 178 Cassia mimosoides H061 1.94 44

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Cassia mimosoides H062 2.01 44.6 Cassia nigricans C021 1.93 179.8 Cassia nigricans C022 1.99 146.7 Cassia nigricans C082 1.88 377.7 Cassia nigricans C083 1.87 243.1 Cassia obtusifolia H069 1.99 304.9 Cassia obtusifolia H070 1.81 128.6 Cassia obtusifolia H071 2.01 403.7 Cassia obtusifolia H072 1.95 220.3 Cassia fistula H073 2.01 150.4 Cassia fistula H074 1.73 277.1 Cassia fistula H075 1.74 240 Cassia absus H048 1.92 200.1 Cassia absus H049 1.85 171.3 Cassia absus H050 1.88 113.5 Cassia singueana H051 2.00 44.1 Cassia singueana H064 2.01 405.5 Cassia tora C023 1.90 863 Cassia tora C024 1.91 417.4 Cassia tora H059 1.96 399 Cassia tora H060 1.96 142.5 Cassia sieberiana C028 1.94 200.6 Cassia sieberiana C029 1.90 626.6 Chaemacrista rotundifolia C030 1.88 166.2 Chaemacrista rotundifolia C031 1.96 127.6 Cynometra megalophylla H043 1.99 75.8 Cynometra megalophylla C032 1.93 91.5 Danielii oliveri C033 2.00 35.9 Danielii oliveri C034 2.01 116.1 Danielii oliveri C089 1.91 143.7 Danielii oliveri C090 1.99 231.4 Delonix regia C037 2.00 320.3 Delonix regia C038 1.99 211.1 Detarium macrocarpum C035 1.86 337 Detarium macrocarpum C036 1.89 166.3 Detarium macrocarpum C088 1.91 191.2 Dialium guineensis C039 1.97 89.1 Isoberlina doka C040 1.95 164.2 Isoberlina doka C041 2.00 458.4 Isoberlina doka C091 1.91 1252.7 Isoberlina doka C092 1.92 622.1 Isoberlina tomentosa C042 1.95 344.7 Isoberlina tomentosa C093 1.88 451 Isoberlina tomentosa C094 1.97 63.1 Piliostigma reticulata C043 2.00 149.1 Piliostigma reticulata C044 1.97 103.6

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Piliostigma reticulata C045 1.93 151.6 Piliostigma reticulata C046 1.95 130.6 Piliostigma reticulata C095 1.82 79.3 Piliostigma reticulata C096 1.88 68.8 Piliostigma thonningii C047 1.92 113.1 Piliostigma thonningii C048 1.82 76.6 Piliostigma thonningii C049 1.98 142.5 Piliostigma thonningii C050 1.86 138.9 Cynometra megalophylla C097 1.94 1037.8 Cynometra megalophylla C098 1.95 956.1 Senna alata C051 1.92 671.1 Senna alata C052 2.00 635.7 Senna hirsuta C053 2.01 352.8 Senna hirsuta C054 1.91 318.7 Senna obtusifolia C063 1.94 499.1 Senna obtusifolia C064 1.88 501.6 Senna obtusifolia C065 1.82 679.3 Senna obtusifolia C066 1.82 1509.6 Senna obtusifolia H082 1.97 186.7 Senna obtusifolia H083 1.94 494.9 Senna obtusifolia H084 1.95 228.5 Senna obtusifolia H085 1.91 266.8 Senna occidentalis C055 1.79 555.8 Senna occidentalis C056 1.85 678.2 Senna occidentalis C105 1,94 757 Senna occidentalis C106 1.86 452 Senna occidentalis H076 1.95 459.8 Senna occidentalis H077 1.96 132.3 Senna occidentalis H079 1.79 335.1 Senna occidentalis H080 1.96 176.6 Senna occidentalis H081 1.97 234.6 Senna occidentalis H078 1.88 183 Senna rotundifolia C107 1.83 435.7 Senna rotundifolia C108 1.99 415.9 Senna siamae C067 1.87 243.9 Senna siamae C068 1.99 555.0 Senna siamae C101 1.96 528.6 Senna siamae C102 1.86 296.1 Senna sieberiana C060 1.88 241.4 Senna sieberiana C061 1.98 448.2 Senna sieberiana C062 1.84 280.6 Senna sieberiana C099 1.95 1328.9 Senna sieberiana C100 1.98 500.8 Senna singuena C057 1.85 253.6 Senna singuena C058 1.81 197.7 Senna singuena C059 1.82 183.4

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Dialium guineense C103 1.83 269.5 Dialium guineense C104 1.75 13.8 Tamarindus indica C069 2.01 293.5 Tamarindus indica C070 2.00 196.6 Tamarindus indica C071 1.94 296.8 Tamarindus indica C072 1..84 295.2 Abrus precatorious H007 2.00 105.9 Abrus precatorius P001 2.01 328.6 Abrus precatorius P002 1.98 247 Abrus precatorius P103 1.93 269.8 Abrus precatorius P104 2.00 206.9 Adenidolichos paniculatum P105 1.94 1020.5 Adenidolichos paniculatum P106 1.86 178.6 Aeschyonomene abyssinica H014 1.96 251.8 Aeschyonomene indica H013 1.99 469 Aeschyonomene uniflora H015 2.00 626.8 Eriosema psoraloides P003 2.01 98.4 Eriosema psoraloides P004 1.98 145 Alysicarpus glumaceus P005 2.00 168.7 Alysicarpus glumaceus P006 1.86 207.1 Alysicarpus glumaceus P107 1.82 126 Alysicarpus glumaceus P108 1.86 303.9 Alysicarpus glumaceus H025 1.86 100.4 Alysicarpus glumaceus H026 1.89 61.8 Alysicarpus glumaceus H027 1.86 111.2 Alysicarpus ovalifolius H028 1.99 124.7 Alysicarpus ovalifolius H029 1.85 43.7 Alysicarpus ovalifolius H030 1.91 105.5 Alysicarpus ovolifolius P009 1.96 198.9 Alysicarpus ovolifolius P010 2.00 231.8 Alysicarpus rugosus P109 2.01 235.5 Alysicarpus rugosus P110 2.00 203.6 Alysicarpus rugosus H035 1.97 57.4 Alysicarpus rugosus H036 1.93 139 Alysicarpus rugosus H037 1.87 87.3 Alysicarpus rugosus H038 1.97 124.5 Alysicarpus rugosus H039 2.00 147.2 Alysicarpus rugosus H040 1.80 208.9 Alysicarpus rugosus H041 1.95 110.7 Alysicarpus vaginalis P007 1.94 207.5 Alysicarpus vaginalis P008 1.84 268.6 Alysicarpus vaginalis H031 1.83 58.8 Alysicarpus vaginalis H032 1.93 55.2 Alysicarpus vaginalis H033 1.93 25.5 Alysicarpus vaginalis H034 2.00 90.8 Alysicarpus vaginalis H052 1.93 186.8

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Arachis hypogea P011 1.89 135.2 Arachis hypogea P012 1.91 375.9 Arachis hypogea P111 1.94 210.2 Arachis hypogea P112 1.81 240.8 Cajanus cajan P013 1.80 110 Cajanus cajan P014 1.90 77.2 Cajanus cajan P015 2.00 242.9 Cajanus cajan P115 1.80 917.2 Cajanus cajan P116 1.90 1129.1 Calopogonum mucunoides P016 2.00 386.3 Calopogonum mucunoides P017 2.01 282.9 Calopogonum mucunoides P113 1.90 262.7 Calopogonum mucunoides P114 1.96 206.3 Crotalaria arenaria P018 1.95 158.3 Crotalaria arenaria P019 1.97 189.4 Crotalaria comosa P020 2.00 139.8 Crotalaria comosa P021 1.91 376.2 Crotalaria falcata P029 2.00 278.4 Crotalaria falcata P030 1.95 182.7 Crotalaria falcata P123 1.93 58.1 Crotalaria falcata P124 1.91 106.1 Crotalaria falcata P125 1.86 720.9 Crotalaria falcata P126 1.96 315.8 Crotalaria glucoides P027 1.87 665.6 Crotalaria glucoides P028 1.92 580.9 Crotalaria hyssopifolia P119 1.98 39.9 Crotalaria hyssopifolia P120 1.81 102.8 Crotalaria lachnosema P022 2.00 137.8 Crotalaria lachnosema P023 1.98 169 Crotalaria lachnosema P024 2.01 262.4 Crotalaria lachnosema P025 1.95 233.9 Crotalaria lachnosema P026 1.83 233.2 Crotalaria macrocalyx P117 1.91 347.8 Crotalaria macrocalyx P118 2.00 543.9 Crotalaria mucronata P036 1.99 532.5 Crotalaria mucronata P037 1.81 298.4 Crotalaria mucronata P038 2.00 427.9 Crotalaria naragutensis P033 1.88 83.3 Crotalaria naragutensis P034 2.01 486.6 Crotalaria naragutensis P035 2.01 352.6 Crotalaria retusa P039 1.91 525 Crotalaria retusa P040 1.94 350.6 Crotalaria retusa P041 1.98 309.1 Crotalaria retusa P042 1.97 295.7 Crotalaria retusa P121 1.93 27.1 Crotalaria retusa P122 1.86 29.1

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Crotalaria senegalensis P031 1.98 209.3 Crotalaria senegalensis P032 1.87 163.9 Cynometra megalophylla P127 2.00 94.7 Cynometra megalophylla P128 2.01 64.3 Dalbergia sisso P043 1.82 188 Dalbergia sisso P044 1.87 230.6 Dalbergia sisso P045 1.73 227.1 Dalbergia sisso P129 1.89 490.2 Dalbergia sisso P130 1.85 427.2 Desmodium gangeticum P046 1.81 577.9 Desmodium gangeticum P047 1.98 272.6 Desmodium gangeticum P048 1.86 722.1 Desmodium gangeticum P133 1.88 543.7 Desmodium gangeticum P134 2.00 455.3 Desmodium scorpiurus P049 2.01 449.9 Desmodium scorpiurus P050 2.00 430.3 Desmodium scorpiurus P051 2.01 412.2 Desmodium tortuosum P052 2.01 647.9 Desmodium tortuosum P053 2.00 462.7 Desmodium tortuosum P054 1.83 794.4 Desmodium tortuosum P131 1.84 318.4 Desmodium tortuosum P132 1.92 359.4 Desmodium velutinum P055 1.98 276.6 Desmodium velutinum P056 2.01 277.8 Desmodium velutinum P057 1.83 709.9 Desmodium velutinum P058 2.00 186.6 Erythrina senegalensis P059 1.82 193 Erythrina senegalensis P060 1.93 433.8 Erythrina senegalensis P061 1.82 353.3 Erythrina senegalensis P135 1.98 294.2 Erythrina sigmoidea P136 2.00 384.7 Erythrina sigmoidea P062 1.82 224.5 Erythrina sigmoidea P063 1.91 242.5 Gliciridia sepium P067 1.95 584.1 Gliciridia sepium P068 2.01 492.3 Glycine max P064 1.79 745.1 Glycine max P065 2.00 559.1 Glycine max P066 2.00 531.2 Glycine max P137 1.94 611.8 Glycine max P138 1.91 1178.1 Indigofera arrecta P072 1.77 735 Indigofera arrecta P073 1.80 722.4 Indigofera arrecta P074 1.77 855.2 Indigofera nummulariifolia P145 1.84 96 Indigofera nummulariifolia P146 2.00 207.7 Indigofera nummulariifolia H053 1.95 533.1

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Indigofera leprieurii H054 1.98 442.4 Indigofera conferta P141 2.01 319.9 Indigofera conferta P142 2.01 210.2 Indigofera dendroides H057 2.00 423.6 Indigofera leprieurii P069 2.01 295.6 Indigofera hirsute P070 1.92 325.6 Indigofera hirsute H055 1.79 146.3 Indigofera macrocalyx H058 1.95 144.4 Indigofera nigritana P071 1.92 343.8 Indigofera paniculata H056 1.97 90.6 Indigofera spicata P143 1.91 325.7 Indigofera spicata P144 1.90 425.2 Indigofera suffruiticosa P139 1.93 668.4 Indigofera suffruiticosa P140 2.00 348.8 Lonchocarpus cyanescens P147 1.81 192.2 Lonchocarpus cyanescens P148 1.91 292.1 Lonchocarpus sericeus P075 1.99 302.5 Lonchocarpus sericeus P076 1.82 361.5 Mucuna pruriens P077 1.95 148.1 Mucuna pruriens P078 2.00 551.1 Mucuna pruriens P079 1.97 225.3 Mucuna pruriens P149 2.00 315.7 Mucuna pruriens P150 2.01 335.6 Pericopsis laxiflora P153 2.00 311.4 Pericopsis laxiflora P154 2.00 190.8 Phaseolus vulgaris P083 1.83 90.8 Phaseolus vulgaris P084 1.88 126.6 Phaseolus vulgaris P151 1.97 491.6 Phaseolus vulgaris P152 1.93 552.3 Pterocarpus erinaceus P080 1.96 544.8 Pterocarpus erinaceus P081 1.97 249 Pterocarpus erinaceus P082 1.83 864.3 Sesbania dalzielii P155 1.98 737.3 Sesbania dalzielii P156 1.82 547.1 Sesbania bispinosa P085 1.91 1579.7 Sesbania bispinosa P086 1.94 1530.8 Stylosanthes eracta P159 1.91 638.1 Stylosanthes eracta P160 1.84 346.1 Stylosanthes eracta H020 1.75 290.3 Swartzia madagascriensis P087 1.97 288.1 Swartzia madagascriensis P088 1.81 281 Swartzia madagascriensis P157 1.81 570.6 Swartzia madagascriensis P158 1.96 297.7 Tephrosia bracteolata P091 1.93 431.9 Tephrosia bracteolata P092 2.00 428.5 Tephrosia bracteolata P163 2.00 699.6

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Tephrosia bracteolata P164 2.01 726.9 Tephrosia linearis P089 2.00 454.2 Tephrosia linearis P090 1.82 568.1 Tephrosia linearis P167 1.90 424.5 Tephrosia linearis P168 1.99 537.9 Tephrosia pedicellata P161 1.94 330.7 Tephrosia pedicellata P162 1.97 378.3 Tephrosia platycarpa P169 1.94 344.9 Tephrosia platycarpa P170 1.81 419 Tephrosia elegans P165 2.00 572.5 Tephrosia elegans P166 2.00 598.6 Vigna ambacensis P093 2.00 400.9 Vigna ambacensis P094 1.85 227.2 Vigna gracilis P095 1.98 204.1 Vigna gracilis P096 1.95 154.1 Vigna gracilis H021 2.00 216.9 Vigna racemosa P097 1.94 232.6 Vigna racemosa P098 1.86 215.4 Vigna racemosa P099 1.90 136.9 Vigna racemosa P100 1.88 255.6 Vigna reticulata P171 2.00 295.4 Vigna reticulata P172 1.96 305.2 Vigna subretrena P173 1.85 519.7 Vigna subretrena P174 1.89 482.9 Vigna unguiculata P101 1.99 363.8 Vigna unguiculata P102 1.87 381.9 Afzelia bipidensis H001 1.96 302.2 Vigna reticulata H002 2.01 266.8 Milletia bateri H003 1.93 617.5 Milletia bateri H004 1.93 178.3 Ostryoderris stuhlmannii H005 1.89 265.1 Ostryoderris stuhlmannii H006 1.81 734.6 Abrus precatorious H007 2.00 105.9 Alysicarpus ovalifolius H008 2.00 349.4 Afzelia bella H009 1.88 284 Afzelia pachyloba H010 1.96 323.5 Afzelia pachyloba H011 1.92 526.7 Afzelia africana H012 1.86 99 Aeschyonomene indica H013 1.99 469 Aeschyonomene indica H014 1.96 251.8 Aeschyonomene lateritia H015 2.00 626.8 Aeschyonomene lateritia H016 2.01 250.5 Acacia dudgeonii H017 2.00 131.3 Acacia dudgeonii H018 1.96 383 Acacia senegal H019 1.97 196.5 Stylosanthes eracta H020 1.95 290.3

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Vigna gracilis H021 2.00 216.9 Acacia karoo H022 1.96 171.7 Acacia karoo H023 1.90 143.7 Caesalpinia pulcherrima H024 2.00 246.8 Zornia latifolia H025 2.01 100.4 Zornia latifolia H026 2.00 61.8 Zornia latifolia H027 1.86 111.2 Alysicarpus ovalifolius H028 2.00 124.7 Alysicarpus ovalifolius H029 2.00 43.7 Alysicarpus ovalifolius H030 1.81 105.5 Alysicarpus vaginalis H031 1.83 58.8 Alysicarpus vaginalis H032 1.93 55.2 Alysicarpus vaginalis H033 1.83 25.5 Alysicarpus vaginalis H034 1.85 90.8 Alysicarpus rugosus H035 1.97 57.4 Alysicarpus rugosus H036 1.93 139 Alysicarpus rugosus H037 1.87 87.3 Alysicarpus rugosus H038 1.87 124.5 Alysicarpus rugosus H039 1.84 147.2 Alysicarpus rugosus H040 1.88 208.9 Alysicarpus rugosus H041 1.95 110.7 Chamaecrista mimosoides H042 1.85 135.7 Chamaecrista mimosoides H043 1.99 75.8 Chamaecrista mimosoides H044 1.89 15.6 Chamaecrista mimosoides H045 2.01 16 Chamaecrista mimosoides H046 1.93 80.1 Chamaecrista mimosoides H047 1.97 72.2 Cassia occidentalis H048 1.92 200.1 Cassia occidentalis H049 1.85 171.3 Cassia occidentalis H050 1.88 113.5 Cassia occidentalis H051 1.84 44.1 Alysicarpus vaginalis H052 1.79 186.8 Indigofera pulchra H053 1.95 533.1 Indigofera pulchra H054 1.98 442.4 Indigofera hirsuta H055 1.79 146.3 Indigofera paniculata H056 1.97 90.6 Indigofera dendroides H057 2.00 423.6 Indigofera macrocalyx H058 1.95 144.4 Cassia tora H059 1.96 399 Cassia tora H060 1.96 142.5 Cassia mimosoides H061 1.94 44 Cassia mimosoides H062 2.01 44.6 Cassia sieberiana (seed) H063 1.90 313.8 Cassia singeana H064 2.00 405.5 Cassia italica H065 1.83 131.2 Cassia italica H066 2.01 232.6

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Cassia italica H067 1.84 200.8 Cassia italica H068 2.00 670.6 Cassia obtusifolia H069 1.99 304.9 Cassia obtusifolia H070 1.81 128.6 Cassia obtusifolia H071 1.91 403.7 Cassia obtusifolia H072 1.95 220.3 Cassia obtusifolia H073 2.01 150.4 Cassia obtusifolia H074 1.73 277.1 Cassia obtusifolia H075 1.84 240 Senna occidentalis H076 1.95 459.8 Senna occidentalis H077 1.96 132.3 Senna occidentalis H079 1.79 335.1 Senna occidentalis H080 1.96 176.6 Senna occidentalis H081 2.00 234.6 Senna obtusifolia H082 1.97 186.7 Dolichos stenophyllus H083 1.99 494.9 Dolichos stenophyllus H084 1.95 228.5 Dolichos stenophyllus H085 1.91 266.8 Acacia nilotica H086 2.01 164.2 Acacia nilotica H087 1.95 341.2 Acacia senegal H088 1.93 329.4 Acacia ataxacantha H089 2.01 179.5 Acacia senegal H090 1.97 160.4 Albizia lebbeck H091 1.88 10.1 Albizia lebbeck H092 1.81 6.9 Albizia lebbeck H093 1.78 19.9 Dichrostachys cinerea H094 1.77 306.7 Dichrostachys cinerea H095 1.95 311.7 Albizia lebbeck H096 2.01 271.5 Entada abyssinica H097 2.01 261.3 Entada abyssinica H098 1.86 356.1 Leucaena leucocephala H099 1.84 552.5 Leucaena leucocephala H100 1.94 467 Leucaena leucocephala H101 1.91 191.7 Leucaena leucocephala H102 1.99 675.3 Leucaena leucocephala H103 1.99 349.7 Mimosa pudica H104 2.01 380.8 Mimosa pudica H105 2.00 139.5 Mimosa pudicasd H106 1.87 61.8 Mimosa pudica H107 2.00 168 Mimosa pigra H108 1.88 319.5 Mimosa pigra H109 2.01 204.6 Mimosa pigra H110 1.87 157.6 Mimosa pigra H111 1.84 207.9 Neptunia oleraceae H112 1.96 66.8 Pithecellobium dulce H113 1.96 239.4

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