ABSTRACT
Twenty-four Mucuna sloanei accessions were evaluated at Umudike in 2013, while fifteen accessions were evaluated at Amakama and Ishiagu in 2015 with a view to identifying genotypes that are adapted to the environments. The accessions were laid out in a Randomized Complete Block Design (RCBD) with three replications and gross plot size of 16m2. Six selected accessions were used to make crosses which proved unsuccessful because of high rate of flower abortions. Analysis of variance showed that location and accessions effects were significant (P<0.05) for most of the traits. The accessions exhibited significant variability (P<0.01) for most traits evaluated at the localities. There were also non-significant accession x location (GxE) interactions for most traits, depicting stabilities and low environmental effects on the accessions across the locations. Seed yield performances across the two localities revealed highly significant differences (P<0.01), with yield at Amakama (226.3kg/ha) being higher than that of Ishiagu (173.1kg/ha) based on two locations combined data. Heritability and variability studies revealed that the environmental factors exerted greater influence on most of the characters studied in different locations. Those attributes under high genotypic influence had high heritability estimates while those under high environmental influences had lower hertabilities and low genetic advance (GA). Correlation and path-analysis revealed that number of seeds/plant (P<0.001), number of pods/plant at harvest (P<0.001) leaf area/plant (P<0.001) and number of leaves/plant (P<0.001) associated significantly with seed yield and were also the most important direct and indirect contributors to seed yield across the locations, suggesting that these attributes were important indices for seed yield and should be considered in any selection process aimed at improving seed yield in M. sloanei. Analysis of the 21 accessions for nutritional and anti-nutritional compositions revealed that some of the accessions were rich in crude proteins, minerals (Fe and Zn) and vitamins C and other biochemical compounds that are useful to man and his animals if fully exploited. Some of the accessions include, “Obio-Akpor”, Ngwa-North”, “Essien Udim”, Umuahia-South” and “Mbano”. These six selected accessions based on their yield performances and rich nutritional compositions across the two agro-ecological zones are recommended for further evaluations with respect to their stability and adaptation to the zones.
TABLE OF CONTENTS
Title Page
i
Declaration ii
Certification iii
Dedication iv
Acknowledgments v
Table of Contents vii
List of Tables xii
List of Appendices xiv
Abstract xvi
CHAPTER 1: INTRODUCTION 1
CHAPTER 2: LITERATURE REVIEW 7
2.1 Origin and Geographic Distribution 7
2.2 Agro-botanical Features 7
2.3
Ecology and Adaptation 9
2.4 Similarities of Mucuna sloanei with
Other Mucuna Species 10
2.5 Uses and Properties of Mucuna 11
2.6 Cultivation, Agronomic Practices and
Yield of Mucuna 13
2.7 Mucuna as Food and Feed 14
2.7.1 Food 14
2.7.2 Animal
feed 15
2.8 Mucuna
Gene Pools 16
2.9 Mucuna Collections 16
2.10 Evaluation
and Documentation of Mucuna 17
2.11 Earlier Breeding Work on Mucuna 18
2.12 Considerations in Mucuna Breeding 19
2.13 Time Line for Breeding Results in Mucuna 21
2.14
Pest (And Possible Disease) Resistance
in Mucuna 21
2.15 Characterization of Mucuna and other Leguminous Crops Using Morphological
Descriptors –
and Other Methods 23
CHAPTER 3: MATERIALS AND METHODS 26
3.1 Site of the Study 26
3.2 Source of materials Used in the Study 27
3.3 Experiment
1: Assessment of Yield and Associated Traits of 24 Accessions
of Mucuna sloanei 29
3.3.1 Land preparation and design of
experiment 29
3.3.2 Agronomic practices 30
3.3.3 Harvesting 30
3.4 Data
Collection 31
3.5
Experiment 2: Establishment of Crossing Blocks and Hybridization in 2014
and 2015 Cropping
Seasons 34
3.5.1 Crossing
blocks and hybridization in 2014 34
3.5.2 Crossing
blocks and hybridization in 2015 35
3.6 Experiment
3: Analysis of Nutrient and Anti-Nutrient Contents of the
Mucuna sloanei
Accessions 37
3.6.1 Preparation
of seed materials 37
3.6.2 Biochemical
analysis 37
3.7 Data Analysis 58
CHAPTER 4: RESULTS
4.1 Soil
Characteristics of the Experimental Sites 60
4.2
Weather Data of the Experimental Sites 62
4.3
Crosses 64
4.4 Vegetative
Attributes of Mucuna sloanei
Accessions 68
4.4.1 Days
to 50% Emergence 68
4.4.2
Leaf Area per Plant 68
4.5 Reproductive
Attributes of Mucuna sloanei Accessions 72
4.5.1
Number of days to first flowering 72
4.5.2
Number of days to first pod appearance 72
4.5.3
Percentage flower abortion 76
4.5.4
Number of flowers per inflorescence 76
4.5.5
Number of days to maturity 76
4.5.6
Days to 50% flowering 77
4.6 Yield
and associated traits of Mucuna sloanei
at the three locations 79
4.6.1
Seed yield per hectare 78
4.6.2
100- seed weight (seed size) 78
4.6.3
Pod weight at harvest 82
4.6.4
Individual seed weight ( Individual
seed size) 82
4.6.5
Number of pods per plant 83
4.6.6
Number of seeds per plant 83
4.6.7 Pod length at harvest 84
4.6.8 Pod filling period 84
4.6.9
Seed weight per pod at harvest 84
4.7 Estimates
of Genetic Parameters in Mucuna sloanei Accessions 85
4.8 Inter-Relationship
between Seed Yield and Associated Traits in Mucuna
sloanei Accessions
at the Three Locations 89
4.8.1 Correlation studies 89
4.8.2 Path co-efficient analysis in Mucuna sloanei under Umudike condition 95
4.8.2.1 Number of pods per plant and seed yield 95
4.8.2.2 Pod weight and seed yield 95
4.8.2.3 Pod length and seed yield 99
4.8.2.4 Number of seeds per/plant and
seed yield 99
4.8.2.5 Seed weight per pod and seed
yield 99
4.8.2.6 100-seed weight (seed size) and seed yield 100
4.8.2.7 Number of leaves per plant
and seed yield 100
4.8.2.8 Leaf area per plant and seed
yield 100
4.9 Effect
of Location and Accession on Seed Yield and Associated Traits of
Mucuna Sloanei 106
4.10 Nutritional
and Anti-Nutritional Compositions of Mucuna
sloanei Accessions 108
4.10.1 Proximate analysis of the
accessions of M. sloanei 108
4.10.2 Mineral profile analysis of
the accessions of M sloanei 111
4.10.3 Vitamins content of different accessions of M. sloanei 114
4.10.4 Anti-nutritional contents of the accessions of Mucuna sloanei 117
CHAPTER
5: DISCUSSION, CONCLUSION AND RECOMMENDATIONS
5.1 Discussion 120
5.2 Conclusion 137
5.3 Recommendations 138
References 139
Appendices 153
LIST OF TABLES
|
3.1
|
Areas of collection and
some of the seed characteristics of the accessions used.
|
28
|
|
3.2
|
Morphological
characteristics of the six selected accessions used for hybridization
|
36
|
|
|
|
|
|
4.1
|
Selected properties of the surface soil of the
experimental sites at three locations during the period of study
|
61
|
|
4.2
|
Comparison of the Agro-meteorological data of the
experimental sites at three locations (Umudike, Amakama and Ishiagu)
|
63
|
|
4.3
|
Crossing Matrix of six selected parential genotypes
used for hybridization
|
65
|
|
4.4
|
Number of crosses made at Umudike in 2014, number of
successful crosses and percentage of successful crosses
|
66
|
|
4.5
|
Number of crosses made at Amakama and Ishiagu in
2015, number of successful crosses and percentage of successful crosses
|
67
|
|
4.6
|
Number of days to 50% emergence of Mucuna sloanei accessions at different
locations
|
69
|
|
4.7
|
Leaf area/plant of Mucuna sloanei accessions at different locations
|
70
|
|
4.8
|
Number of leaves/plant of Mucuna sloanei accessions at different locations
|
71
|
|
4.9
|
Reproductive
attributes of Mucuna sloanei
accessions grown at Umudike
|
73
|
|
4.10
|
Reproductive attributes of Mucuna
sloanei accessions grown at Amakama
|
74
|
|
4.11
|
Reproductive
attributes of Mucuna sloanei
accessions grown at Ishiagu
|
75
|
|
4.12
|
Mean seed yield and associated traits of Mucuna sloanei accessions at Umudike
|
79
|
|
4.13
|
Mean seed yield and associated traits of Mucuna sloanei accessions at Amakama
|
80
|
|
4.14
|
Mean seed yield and associated traits of Mucuna sloanei accessions at Ishiagu
|
81
|
|
4.15
|
Genetic Parameters of Mucuna sloanei at Umudike, Amakama and Ishiagu
|
87
|
|
4.16
|
Correlation
co-efficients (r) between seed yield of Mucuna
sloanei and other traits at Umudike
|
90
|
|
4.17
|
Correlation co-efficients (r) between seed yield of Mucuna sloanei and other traits at
Amakama
|
92
|
|
4.18
|
Correlation co-efficients (r) between seed yield of Mucuna sloanei and other traits at
Ishiagu
|
94
|
|
4.19
|
Path analysis showing direct and indirect influences
of 10 yield traits on seed yield per plant of Mucuna sloanei accessions at Umudike (2013)
|
96
|
|
4.20
|
Path Analysis showing direct and indirect influences
of 11 physiological traits on seed yield per plant of M. sloanei accessions in Umudike (2013)
|
102
|
|
4.21
|
Main effects of location and
accessions on yield and yield components of fifteen (15) accessions of M. sloanei grown in Ishiagu and
Amakama in 2015
|
107
|
|
4.22
|
Proximate analysis of different accessions of M. sloanei
|
109
|
|
4.23
|
Result of mineral profile analysis for different
accessions of M. sloanei
|
112
|
|
4.24
|
Vitamins
content analysis for different accessions of Mucuna. sloanei
|
115
|
|
|
Anti-nutrient components analysis for different
accessions of Mucuna. sloanei
|
118
|
APPENDICES
1. Appendix
1: Co-efficient of determination of seed yield (Y) by 10 attributes and the
residual of M. sloanei accessions
evaluated in Umudike (2013). 153
2. Appendix
2: Summary output for path analysis at Umudike in 2013
(for 10 yreproductive traits) 154
3.
Appendix 3: Direct and indirect
effects of yield components on seed yield of Mucuna sloanei at Umudike in 2013 155
4.
Appendix 4: % Direct and indirect contribution of yield
components to seed
yield of Mucuna slonaei at Umudike in 2013 156
5.
Appendix 5: Correlation
coefficients (γ) between M. sloanei
10 reproductive
traits
and seed yield/plant at Umudike in 2013 157
6. Appendix 6: Path diagram showing direct
and indirect influences of 10 yield
traits
in seed/plant of Mucuna sloanei
accessions in Umudike (2013) 158
7. Appendix
7: Coefficient of determination of seed yield (Y) of M. sloanei
accessions by 11 physiological attributes and the
residual in Umudike (2013) 159
8.
Appendix 8: Summary output for
path analysis at Umudike in 2013 (for 11
physiological traits) 160
9.
Appendix 9: Direct and
Indirect effect of 11 vegetative
attributes to seed yield of Mucuna
sloanei at Umudike in 2013 161
10. Appendix 10: Percentage direct and
indirect contribution of 11 vegetative attributes
to seed yield of Mucuna
sloanei at Umudike in 2013 162
11. Appendix 11: Correlation
coefficients (γ) between M. sloanei
11 vegetative
traits and seed yield/plant at Umudike in 2013 163
12. Appendix
12: Path diagram showing direct and indirect influences of 11
physiological attributes on seed yield of Mucuna sloanei accessions in
Umudike 2013 164
13.
Appendix 13: ANOVA Table of Some
Attributes at Umudike 2013 165
14.
Appendix 14: ANOVA Table of Some
Attributes at Amakama 2015 167
15.
Appendix 15: ANOVA Table of Some
Attributes at Ishiagu 2015 169
CHAPTER 1
INTRODUCTION
Mucuna
(Family Fabaceae) is the third
largest genus among the flowering plants. It consists of approximately 650
genera and 20,000 species (Doyle, 1994), and is the second most important
source for human and animal nutrition (Vietmeyer, 1986).
Mucuna
sloanei is synonymous with Mucuna urens auct.
(L) medik (Jansen, 2005). Its common names in English language are; Horse
–Eye bean, Deer-Eye bean, Ox-Eye bean and True sea bean. In Igbo dialects, it
is known as Ukpo, Oruru, Ukpotoro, Ibaa, Ukweregbe, Okobo and in Cross River
and Akwa Ibom States, it is known as Ibaba and Anyen Enang.
In
terms of origin and geographical distribution, M. sloanei is very wide
spread in Africa, starting from Sierra Leone to Democratic Republic of Congo in
the West and to Angola in the South. It is also wide spread in the Caribbean
region, tropical America and Islands of the Pacific Ocean (Jansen, 2005). It is
described as a self pollinated species (Duke, 1981).
Mucuna
sloanei is a relatively under-utilized legume,
used as a soup thickner among the Igbos, Efik,and Kalabari of South Eastern
Nigeria in the West African sub region (Ene-Obong and Carnovale, 1992; Versteeg
et al., 1998). It is cracked by hitting it with hard object before
boiling. A black dye is obtained from all parts of M. sloanei, which is
used in Nigeria to dye fibre and leather black.
Cooked
young fruits are eaten as vegetable. Oil extracted from the seed can be used in
the preparation of resin, paint, polish, wood vanish, skin cream and liquid
soap (Jansen, 2005). The Edo people in Nigeria use the leaf sap to stop
diarrhoea. In tropical America, the seed is used as a diuretic (drugs that
cause increased passing of urine), and in Gabon and tropical America, a seed
decoction (preparation of teas from the seed) is used as a soothing medicine to
relief the discomfort of haemorrhoids. The seeds are also used for decoration
and in games (Jansen, 2005).
Mucuna
sloanei has been used in ethno-medicinal
preparations in some parts of West Africa. They have been applied as anti-helminthic
(drugs that expel parasitic worms from the system) (Farida and Van der Macsen,
1996), as expectorant for cough and asthma (Prakash and Misra, 1987). The pod
hairs have been used as anti snake agents (Houghton and Skari, 1994) and as
aphrodisiac (Siddhuraji et al., 1996). Extracts from the seeds are used
as uterine stimulants (Lorrenthi et al., 1998). All parts of the plant possess valuable medicinal
properties (Caius, 1989; Pandey, 1999), and there is heavy demand for Mucuna in India drug market. After the
discovery that Mucuna seeds contain
L-Dopa, an anti parkinson’s disease drug, its demand in International market
has increased many folds (Faroogi et al.,1999)
and the demand has motivated Indian farmers to start commercial cultivation. It
has been reported that the dried seed of M. sloanei contains 3% of the
amino acid, L-Dopa, which stimulates the formation of the neurotransmitter
dopamine in the brain (Jansen, 2005).
Mucuna sloanei is
found in wet localities, in swamp forests, at boarders of rivers and lakes, in
Savannah wood land and secondary vegetation. In Nigeria, cultivation of M. sloanei is done using tall poles and
stakes just like climbing types of common bean. It is usually cultivated around
homes and gardens mostly for immediate culinary or other subsistence purposes.
Consumers of improperly processed Mucuna seeds are known to suffer from
gastro-intestinal disturbances notably, nausea, vomiting and anorexia (loss of
appetite for food), as well as more serious effects such as paranoid delusions,
hallucinations,delirium and unmasking of dementia(Caius,1989). Mucuna’s high content of L-Dopa is seen as the greatest impediment to its
increased utilization as food and feed (Carsky et al., 1998; Flores et al., 2002).
Several
studies have focused on L_dopa quantification in plant parts of various Mucuna species and in their proximate
analysis. Bell and Jansen (1971), in surveying six accessions found a range of
L-dopa from 5.9 – 9.0% in the seed, while Daxenblichler et al., (1971, 1972) in the same survey observed variability
between 3.1 and 6.7%. The ranges of 2.2 to 7.2% were found in a survey of 36
accessions (Lorrenthi et al., 1998)
and from 1.9 to 7.6% in a survey of 38 accessions (ST-Laurent et al., 2002).
Mucuna sloanei
is a source of dietary protein as it is a legume. It is however, sliding into
an endangered species list because its utility and cultivation have been
neglected over the years, especially in the Southeastern part of Nigeria where
it is used as a soup condiment. In many instances, accessions are described
only in terms of where they were grown (e.g. Mucuna sp. Var. Ghana) or by the many popular names under which
they came to be known in various places such as Mucuna conchinchinensis in South East Africa or M. deeringian in Florida. It is
difficult to confirm that the name given to a species is a representative of
its genotype. Extensive exchange of seeds over the years probably led to some
species being given different names according to the locality where they are
grown. On the other hand, it is also highly plausible that species given the
same name in two or more areas might in fact be different original stock or
germplasm. This lack of information on the taxanomy of Mucuna has impeded the effective utilization of its genetic
resources. At the same time, the wide geographical and climatic distribution of
the crop is likely to result into a tremendous genetic diversity which needs to
be estimated before any cultivar development (Capo-Chichi et al., 2003).
Sarutayophat
et al. (2007) characterized 13 cowpea
accessions based on growth habit, days to 50% flowering, pod colour, pod
length, number of pods per plant, seed yield per plant. Stoilova and Pereira
(2013) has used 24 different morphological descriptors in order to identify
accessions with specific behavior that could be exploited by plant breeders and
they found that descriptors like pod length, number of seeds per pod, seed
thickness and 100-seed weight were the most suitable traits and they concluded
that these characters can be used in characterization. Increasing major
components of grain yield such as pods/plant, pod length, seeds/pod and seed
size will allow improving M. sloanei
yield potentials. Understanding the genetic variability and genetic
inter-relationship present among germplasm collections is valuable to avoid
redundancy and allow plant breeders to select potential parents with desirable
traits for cultivar development (Chaudhary and Singh. 1982; Yoshida, 2004).
Various
methods are available to measure genetic variation among crop genetic
resources. These methods rely on the availability of data based on pedigree,
plant morphology, agronomic performance and molecular analysis (Mohammadi and
Prasanna, 2003). Field phenotyping is a common method to determine genetic
variation between and within genotypes (Yoshida, 2004). Phenotypic markers
reflect crop ideotypes, which are relatively cheap and easy to use, depending
on prior knowledge of such traits and their expressions (Elameen et al., 2011). Phenotypic traits are however
highly affected by environmental factors (Elameen et al., 2011). When using phenotypic traits to characterize germplasm, it was recommended
to test a set of genotypes across seasons and locations with sufficient
replications for a meaningful comparison and selection (Jacoby et al., 2003).
Combined
use of morphological, biochemical and molecular (DNA) markers has been proposed
for genetic diversity studies (Elameen et
al., 2011). Plant morphological characters have been recognized as the
universally undisputed descriptors for protection and varietal characterization
of crop varieties. Use of morphological descriptors in sequential fashion is
useful and convenient to discriminate the different varieties (Joshi et al, 2011). Characterization can be done by using morphological
characteristics or molecular markers or both. Morphological descriptors have
traditional significance and one can immediately be accessible on the spot
without the need of equipment. Although,
it has its limitations like environmental influence and time consuming but this
has been universally adopted as classical taxonomic approach.
The
study of morphological variability is the classical way of assessing genetic
diversity. For many species, especially minor crops, it is still the only
approach. There has been a great deal of confusion regarding the status of Mucuna types within the genus. Some
authors have classified them as separate species within the genus, while others
have considered them as sub-species or varieties of the species Mucuna pruriens. A major finding of the
work of Capo-chichi is that the evaluated accessions (many of which are
commonly utilized) can all be considered as mere varieties of Mucuna pruriens( Capo-Chichi, 2002).
Apart
from the work done by Capo-Chichi (2001) on taxonomy and genetic improvement of
Mucuna, particularly in the areas of;
i.
Improved understanding of
the variability in nutritional characters among different accessions of Mucuna
pruriens;
ii.
Genetic diversity among Mucuna accessions; and
iii.
Assessment of the role of
genotype and environment on the production of L-Dopa in Mucuna; no breeding work has been done on the characterization and
genetic improvement of Mucuna sloanei in the South-East agro-ecology of
Nigeria till date. Since M. sloanei and other Mucuna species represent a potential food and feed source for human
and animal nutrition, which could relieve critical food shortages as well as
have medicinal properties if given adequate promotion and research attention,
breeding efforts should be initiated for improved nutritional quality,
productivity and acceptability.
The
objectives of this study therefore, are as follows:
(i) To
determine the yield and associated traits in Mucuna sloanei.
(ii) ascertain the genetic variability among
the accessions.
(iii) determine inter-relationships between
yield and associated traits.
(iv) investigate the hybridization potentials
of the selected accessions.
(v) carry out meiotic analysis of the selected Mucuna sloanei
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