ASSESSMENT OF DIVERSITY AMONG COWPEA

ACCESSIONS FROM SEMI-ARID AREAS OF KENYA

ROSE KAMBUA MUNYAO

MASTER OF SCIENCE

(Plant Breeding)

JOMO KENYATTA UNIVERSITY

OF

AGRICULTURE AND TECHNOLOGY

2023
 Assessment of Diversity among Cowpea Accessions from Semi-Arid
Areas of Kenya

Rose Kambua Munyao

A Thesis Submitted in Partial Fulfillment of the Requirements for the

Degree of Master of Science in Plant Breeding of the Jomo Kenyatta
University of Agriculture and Technology.

2023
 DECLARATION

This thesis is my original work and has not been presented for a degree in any other
University

Signature ………………………………………. Date…………………………….

Rose Kambua Munyao

This thesis has been submitted for examination with our approval as University
Supervisors

Signature ………………………………………. Date…………………………….

Prof. Edward George Mamati, PhD

JKUAT, Kenya

Signature ………………………………………. Date…………………………….

Prof. Githiri Mwangi, PhD

JKUAT, Kenya

ii
 DEDICATION

I dedicate this work to my late parents Christine Munyao and Charles Mwita, and my
dear husband James Joshua.

iii
 ACKNOWLEDGEMENTS

I wish to convey my sincere gratitude to everyone who contributed to the success in my

studies at Jomo Kenyatta University of Agriculture and Technology. I value every effort
that was made by so many people for me to complete this work.

I thank the Research Production and Extension (RPE) division of Jomo Kenyatta
University of Agriculture and Technology (JKUAT) for the financial support that
enabled me to carry out the research work.

I highly appreciate the National Gene Bank of Kenya for providing cowpea accessions
for the Semi-Arid Areas of Kenya.

I am extremely grateful to my supervisors Prof. Edward Mamati and Prof. Githiri

Mwangi for their invaluable guidance, encouragement and advice throughout my
research work. Thank you for your availability, guidance, patience, guidance during
manuscript preparation and thesis compilation and the mentorship I have received during
my studies.

Special thanks are also extended to Dr. Adelide Mutune and Naomi Nzilani for their
invaluable support and encouragement. I also thank my colleagues Tesfamichael
Semere, John Kariuki and Sylvia Buleti for their assistance. I immensely appreciate the
availability and assistance of Samuel, Brenda, Timothy, Jennifer, Patrick, Mumbi, Rose,
Janet, Joyce, Kimuyu, Wairia and Jemimma during laboratory experiments and data
collection in the field.

I also wish to express my gratitude to Prof. Elijah Ateka for his encouragement and
permitting me to use facilities in the Cassava Diagnostics Laboratory.

My deepest gratitude goes to my husband James Joshua who has supported me

throughout my studies. May the Almighty God bless you abundantly. Special thanks go

iv
to my siblings John, Mary, Verah, Sam and Ken for the undying love and support.
Thanks to all family members who encouraged me continuously throughout my studies.

Above all, I give thanks to the Almighty God for his protection and mercies upon me.

v
 TABLE OF CONTENTS

DECLARATION ............................................................................................................. ii

DEDICATION ................................................................................................................ iii

ACKNOWLEDGEMENTS ........................................................................................... iv

TABLE OF CONTENTS ............................................................................................... vi

LIST OF FIGURES ........................................................................................................ x

LIST OF APPENDICES ............................................................................................... xi

ABSTRACT ................................................................................................................... xii

CHAPTER ONE ............................................................................................................. 1

INTRODUCTION ........................................................................................................... 1

1.1 Background of the study......................................................................................... 1

1.2 Problem Statement ................................................................................................. 3

1.3 Justification ............................................................................................................ 3

1.4 General objective .................................................................................................... 4

1.4.1 Specific objectives ....................................................................................... 4

CHAPTER TWO ............................................................................................................ 5

LITERATURE REVIEW............................................................................................... 5

2.1 Crop botany ............................................................................................................ 5

vi
 2.2 Morphology ............................................................................................................ 6

2.3 Uses of cowpea ....................................................................................................... 7

2.4 Production systems ................................................................................................. 8

2.5 Landraces................................................................................................................ 8

2.6 Released cowpea varieties in Kenya ...................................................................... 9

2.7 Environmental requirements for cultivation of cowpea ....................................... 12

2.8 Cowpea characterization ...................................................................................... 12

CHAPTER THREE ...................................................................................................... 15

MATERIALS AND METHODS ................................................................................. 15

3.1 Experimental site .................................................................................................. 15

3.2 Plant materials ...................................................................................................... 15

3.3 Field Experimental Design ................................................................................... 15

3.4 Data collection ...................................................................................................... 16

3.5 Molecular characterization of cowpea ................................................................. 16

3.6 Data Analysis ....................................................................................................... 20

CHAPTER FOUR ......................................................................................................... 22

RESULTS ...................................................................................................................... 22

4.1 Variation in the Cowpea Accessions for Qualitative Characters ......................... 22

4.2 Variation among accessions for Quantitative Characters..................................... 26

4.3 Molecular characterization based on SSR markers .............................................. 34

vii
CHAPTER FIVE ........................................................................................................... 43

DISCUSSION ................................................................................................................ 43

5.1 Discussion ............................................................................................................ 43

CHAPTER SIX ............................................................................................................. 47

CONCLUSION AND RECOMMENDATIONS ........................................................ 47

6.1 Conclusion ............................................................................................................ 47

6.2 Recommendations ................................................................................................ 47

REFERENCES .............................................................................................................. 48

APPENDICES ............................................................................................................... 57

viii
 LIST OF TABLES

Table 2.1: Improved cowpea varieties in Kenya ............................................................. 10

Table 3.1: SSR Markers used in the study ...................................................................... 19

Table 3.2: PCR amplification conditions used in the study ............................................ 20

Table 4.1: Distribution of accessions among respective categories of the evaluated

morphological characters ............................................................................ 24

Table 4.2: Variation in quantitative morphological traits among cowpea accessions .... 27

Table 4.3: Mean values of 20 highest and 20 lowest accessions based on quantitative
traits ............................................................................................................ 29

Table 4.4: Pearson’s correlation among quantitative traits recorded on cowpea ............ 30

Table 4.5: Eigen vectors and values for five Principal Components .............................. 31

Table 4.6: Distribution of accessions within clusters based on similarity among

quantitative traits......................................................................................... 34

Table 4.7: Amplification status of the SSR markers ....................................................... 37

Table 4.8: Allelic Molecular Variance of the cowpea accessions ................................... 38

Table 4.9: Pairwise population matrix of Nei Genetic distance and Nei Genetic Identity
among cowpea populations ......................................................................... 39

Table 4.10: Estimated genetic diversity of accessions within populations ..................... 40

ix
 LIST OF FIGURES

Figure 4.1: Relationships among cowpea collections from Semi-Arid Areas of Kenya
and commercial lines ..................................................................................... 33

Figure 4.2: Monomorphic band on Agarose gel for primer SSR 6356 ........................... 35

Figure 4.3: Polymorphic bands on Agarose gel for primer SSR 6608 ........................... 35

Figure 4.4: Polymorphic band on Agarose gel for PCR products for SSR 6243 ............ 36

Figure 4.5: Shows the pair wise values means within the populations ........................... 38

Figure 4.6: Variation of band patterns across populations ............................................. 39

Figure 4.7: Clustering of cowpea accessions based on variation in SSR markers.......... 42

x
 LIST OF APPENDICES

Appendix I: Cowpea accessions used in the study and their collection area .................. 57

Appendix II: Data scoring of the traits evaluated during the trial .................................. 63

Appendix III: Ranking based on characteristics associated with performance of cowpea

genotypes .................................................................................................. 65

Appendix IV: Analysis of Molecular Variance .............................................................. 69

Appendix V: Principal coordinates ................................................................................. 70

Appendix VI: Principal component analysis of cowpea genotypes ............................... 71

xi
 ABSTRACT

Cowpea is an important legume crop adapted and widely grown in marginal areas. The
crop is grown mainly from landraces and only a handful of improved varieties have been
developed. Although breeding and identification of superior lines is dependent upon
existence of crop diversity, there is limited information on diversity among the Kenyan
cowpea. The objective of this study was to determine variation among cowpea
accessions from semi-arid areas of Kenya at morphological and molecular levels. One
hundred and ten cowpea accessions obtained mainly from semi arid region of Kenya
were planted in a Randomized Complete Block Design with three replicates.
Quantitative and qualitative morphological data were collected over the growing period
and on harvested seed. For molecular characterization, DNA was extracted from leaves
obtained from two week old seedlings grown in pots. Variation among the genotypes
was determined through amplifying the DNA using twenty pairs of selected SSR
markers. Even distribution of accessions across traits of the characters was recorded for
immature pod color, leaf color, seed shape and testa texture, whereas uneven distribution
was recorded for terminal leaflet shape, raceme position, pod attachment, pod curvature,
mature pod color, flower color and eye color. ANOVA revealed significant differences
(p=0.05) among accessions for number of days to 50% emergence, pod length, number
of pods per plant and number of seeds per pod. The first five principal components
accounted for 19.8, 18, 15.9, 12.4 and 11.22 of the total variation respectively
amounting to 77%. Correlation analysis revealed significant (p=0.05) relationship for
50% emergence to 50% flowering (r= -0.2131), 50 % emergence to number of pods per
plant (r= -0.5258), emergence to terminal leaflet length (r= -0.1881) and emergence to
terminal leaflet width (r= 0.2042); terminal leaflet length to terminal leaflet width (r=
0.5230) and pod length to number of pods per plant (r= 0.5470). Based on
morphological characteristics, the accessions were grouped into two main clusters, with
one cluster having 103 accessions that included all registered varieties while the other
cluster had seven accessions. Molecular characterization of one hundred and ten
accessions (110) was done using eight pairs of the SSR markers that were polymorphic.
Analysis of the molecular variance showed that close to 100% of the variation was
within accessions. Heterozygosity ranged from 0 to 0.5 with a mean value of 0.19. The
molecular data subjected to cluster analysis grouped the accessions into three groups.
Therefore, cowpeas grown in semi rid areas of Kenya are variable and closely related to
the registered cowpea varieties evaluated. The set of accessions could be used for
identification of preferred lines for this region. The morphological data gave significant
variation among the characteristics while molecular characterization showed no
significant variations among the populations.

xii
 CHAPTER ONE

INTRODUCTION

1.1 Background of the study

Cowpea (Vigna unguiculata), is a valuable food legume extensively grown in the tropics
and subtropics of Africa (Lesley, 2005). The area under cowpea production is estimated
to be 14.5 million ha worldwide with an annual production of 6.5 million metric tonnes
(Njonjo, 2018).

In eastern Africa, cowpea is widely grown especially in Tanzania and Kenya. In Kenya,
cowpea is the third most important food legume after common bean and pigeon peas
(Njonjo, 2018). In 2012, there was an estimated area of 214,492 ha under cowpea
production in Kenya, out of which 187,910 ha were from eastern province (Njonjo,
2018, MoALF, 2015). It is mainly grown as an intercrop with maize, sorghum or
cassava (Asiwe, 2009).

Cowpea is cultivated mainly for its green leaves which are used as vegetables, immature
pods used as filler or for mature grains (Bewley et al., 2006). This crop is highly
palatable and nutritious as it contains several minerals including iron, calcium,
phosphorus and zinc, and it is also free from anti-nutritive factors (Njonjo, 2018).

Cowpea is an important food security legume grown in the Arid and Semi Arid Lands
(ASALS) of Kenya especially semiarid lands of eastern Kenya (Gachimbi et al., 2007).
This crop has a deep root system and matures early hence its adaptability to unfavorable
conditions. It is rich in nutrients and hence is good for reducing food and nutrition
insecurity (Joshua et al., 2019). Apart from being very nutritive, cowpea can be a source
of income for household needs as well as enabling farmers to pay for inputs, labor and
maintenance of other later maturing crops, through the sale of its leaves and grains
(Bennett & Jennings, 2013).

Farmers in Kenya experience low cowpea grain yields due to various constraints ranging
from damage by pests and diseases to lack of high yielding varieties (Sariah, 2010).
About 90% of the seed sown in the ASALS are informally produced by Non-
Governmental Organizations (NGOs), farmers, farmer groups or community based
organizations (Muthoni & Nyamongo, 2018). Most farmers often get seeds of variable
quality from their own saved seed, social networks (friends and neighbors) or from local
traders hence recycling seeds.

Improved cowpea varieties are at different levels of adoption among farmers. Certified
seeds are quite expensive and inaccessible compared to local varieties that are sold at a
fraction of the price of the certified seeds (Sperling et al., 2004; Rubyogo et al., 2007;
Njonjo, 2018). The farmers also prefer the local varieties as they are more palatable
compared to the improved varieties. In an effort to improve farmers’ access to improved
seed, governmental organs and NGOs carry out community based seed interventions
such as seed and cultural fairs, community based bulking, seed recovery and bulking
banks (Setimela et al., 2004). The Kenyan government has been distributing large
quantities of seeds to farmers in the Arid and Semi Arid areas since 1992 especially
during emergency situations (Nagarajan, et al., 2008) following drought. Cowpea is
mostly grown by small scale farmers in developing countries and has received low
attention from researchers (Adebowale, 2011; Timko and Singh 2008) hence attracting
less donor support. Information on cowpea use and cultivation is scarce making it
difficult to determine the extent to which farmers use traditional varieties, the adoption
of improved and registered varieties.

Landraces have played a big role in the introduction of improved varieties as they have
rich and complex ancestry with great variations in response to many stresses as well as
2
vast resources for improved varieties (Litchfouse et al., 2011). A large number of
cowpea varieties evolving from local landraces have been commercialized (Pratap &
Kumar, 2011). However, landraces have rarely been used in hybridization to understand
their breeding value in the improvement of grain yield and other traits in cowpea.
Although cowpea is reported to have high protein value, high variability, high
adaptability and drought tolerance capacity (Ashraf et al., 2012), little research has been
done on their landraces hence loss of superior genes that would be useful for genetic
manipulation and improvement of the crop.

1.2 Problem Statement

Cowpea farmers in semi arid parts of Kenya face the challenge of low yields. One of the
reasons for low yields is inadequate access to improved varieties. Although there are
several improved varieties registered in Kenya (KEPHIS), their adoption is not known
(Setimela et al., 2004). Farmers still grow their own seed and other mixed lines whose
overall yields are low (Stoilova & Pereira, 2013). A wide range of accessions have been
collected from marginal areas of the country. The accessions have the potential to be
used as germplasm for breeding, selection and identification of improved and adaptable
accessions for the region. Whereas genetic advance in a crop is dependent on the
existence of heritable variation in the available germplasm, only a few studies have been
conducted to document the extent of this variation and hence this study was conducted.
The objective of the study was to determine variation among cowpea accessions from
semi-arid areas of Kenya using both morphological and molecular markers.

1.3 Justification

Cowpea breeding and identification of superior lines is dependent upon existence of

known crop diversity. However, there is limited information on diversity among the
Kenyan cowpea populations. Development of improved varieties exhibiting early

3
maturity, good grain quality, resistance to diseases and pests can significantly lead to
increased yields (Lesley 2005). Farmers can benefit a lot from improving the local
varieties thus increasing their productivity and reducing poverty. Improvement
programmes introduced can provide information on the genetic diversity within the
accessions.

Genetic characterization is invaluable to gene banks as it makes sampling and use of

available genetic resources easy. In addition, breeders are able to select crops with
superior characteristics from the genetic variations of the various parental lines. Genetic
variation is the raw materials for plant improvement (Doumbia, 2011).

1.4 General objective

To determine variation among cowpea accessions from semi-arid areas of Kenya

1.4.1 Specific objectives

i. To evaluate the diversity among cowpea accessions based on morphological

traits
ii. To determine diversity among cowpea accessions using SSR markers

1.5 Hypotheses

i. There are no morphological differences among cowpea accessions grown in

Kenya
ii. Cowpea accessions grown in Kenya are not genetically different

4
 CHAPTER TWO

LITERATURE REVIEW

2.1 Crop botany

Cowpea (Vigna unguiculata) is a dicotyledonous crop that belongs to the genus Vigna
and family leguminosae. It is a diploid plant with 2n = 22 chromosomes (Timko &
Singh, 2008) and a genome size estimated to cover 620 million base pairs (Timko et al,
2008). The name cowpea was probably coined from the fact that it is a source of hay for
cows in the southern United States of America and other parts of the world. Cowpea is
referred to by different local names around the world. For example, in West Africa, it is
known by the names “niebe”, “wake”, and “ewa” while in Brazil it is “caupi”. Other
names include “southern peas”, “black eyed peas”, “Field Peas”, “pink eyes” and
“crowders” (Timko et al., 2008) in the southern United States. In Kenya, cowpea is
referred to by different local names among the Kenyan communities. For example,
cowpea is known as Kunde (Swahili), Mathoroko (Kikuyu), Nthooko (Kikamba) and
Egesare (Kisii) (Savala et al., 2003).

The exact origin of cowpea is unknown, although, Africa and Asia are discussed as the
domestication areas of this crop (Sariah, 2010). Southern Africa has the highest genetic
diversity of cowpea with the most primitive forms of wild cowpea and it is the most
probable center of cowpea domestication (Acquaah, 2012). The origin and
domestication of cowpea has been determined over time based on morphological and
cytological evidence as well as information on its geographical distribution and cultural
practices (Acquaah, 2012). Cultivated cowpea evolved overtime through domestication
and selection from annual wild cowpea, a process during which, seed dormancy and pod
dehiscence was lost (Anderson & Vicente, 2010).

5
2.2 Morphology

Cowpea accessions have high morphological variation. Cowpea accessions are divided
according to their uses: for grain, forage or dual purpose. Cowpea plant is an
herbaceous, prostate, climbing or sub erect annual plant growing 15-80 cm high
(Omoigui et al., 2018). Leaves are alternating trifoliate with a petiole of 5-25 cm long.
The first pair is simple and opposite. The lateral leaflet is opposite and asymmetrical and
the central one is symmetrical and ovate. Leaves vary in sizes (6-16*4-11cm) and shape
from linear, lanceolate to ovate. The color of the leaves can be pale green to dark green.
The stems are striate, smooth or slightly hairy some with a purple tinge (Oluwakemi et
al., 2021).

The flowers may range from white, yellow, pink, pale blue or purple. The flowers are
arranged in racemose or intermediate inflorescence at the distal ends of 5-60cm long
peduncles. The flowers are in alternate pairs with two flowers per inflorescence. Flowers
are distinct, self pollinating and are produced on short pedicels. Flowers open early in
the morning and close at midday. After the flowers open once, they wilt and collapse
(Doumbia et al., 2013). The resulting fruits which are pods vary in sizes, shapes, colors
and texture. The pods are cylindrical and may be curved or straight growing as long as
22 cm with 8-20 seeds per pod. A mature cowpea seed weight ranges from 8 to 32 mg
(Doumbia et al., 2011). The seeds have different sizes and shapes with the common ones
being kidney shaped, ovoid, crowder, globose or rhomboid. The seed shape correlates
with that of the pod (Oluwakemi et al., 2021). The seed coat can be smooth or wrinkled
with various colors including white, green, brown, black, cream, gray, purple, red,
speckled, blotched, eyed or dotted.

This crop has a determinate or indeterminate growth habit. Most cowpea genotypes have
the indeterminate growth habit. Cowpea has well developed rooting system and thick
stems and branches, some of the qualities which make it adapted to harsh conditions.
6
The early flowering cowpea varieties can mature as early as in 55 days providing the
farmers with the first source of food after the “hunger period” (Hall et al., 2003). On the
other hand, late maturing varieties can take as long as 150 days depending on
photoperiod. Flowers are produced on racemes on 15 to 40 mm peduncles arising from
the leaf axils (Timko et al., 2008). Cowpea plant commonly bears two or three pods per
peduncle and sometimes more than three pods are produced if the conditions are
favorable. Cowpea emergence is epigeal like in common beans where the cotyledons
emerge above the ground during germination. Cowpea is a self pollinating crop but
some out crossing has been recorded of as high as 5% (Timko et al., 2007).

2.3 Uses of cowpea

Cowpea is a very important crop. It is a source of food for human, feed for animals and
also as an income generating commodity for farmers and traders (Singh, 2002). Cowpea
crop is useful at all its stages of growth. This crop has the ability to restore soil fertility
through biological Nitrogen Fixation hence it is very useful in farming systems when
rotated with other crops. The early maturing varieties give current harvest earlier than
other crops and serves to shorten the hunger period as often occurs in before harvesting
the current season’s crop in many farming communities in Africa (Muniu, 2017).

The dry cowpea grains are important for human consumption. The seeds are cooked and
eaten solely or as a side dish mixed with vegetables, spices and oil to make a thick soup
which accompanies the staple foods such as cassava, yam or plantains (Silva et al,
2019). The dry seeds can also be canned for export. In West Africa, the seeds are
decorticated and ground into flour for making cakes. The fresh or dried leaves are also
used as vegetables in many parts of Asia and Africa (Grubben & Denton, 2004). In
addition, fresh peas and green immature pods can also be used as vegetables. Cowpea
leaves are either boiled or fried for eating with porridge (Timko & Singh, 2008). In
addition, the leaves can also be sundried or boiled; sundried for preservation to be used
7
in the dry season. The immature seedless pods can be cooked as vegetables or even
canned for export (Madamba et al., 2006). . The cowpea plant as a whole can be used in
feeding livestock especially in the dry season. As fodder, it can be grazed directly or cut
and mixed with dry cereals for feeding animals (Timko et al., 2007). In United States of
America, cowpea is used as green manure and cover crop (Muniu, 2017). In Nigeria,
some cowpea cultivars are grown for extracting fibre that can be used for making fishing
gear or for paper processing (Zia-ul-haq et al., 2010). Its ability to survive under drought
conditions, mature early and fix nitrogen in the soil makes cowpea crop to grow well in
tropical soils which have low moisture and low soil fertility. Other uses include
medicinal value where leaves and seeds are applied as poultice to treat swellings and
skin infections (Grubben, 2004); leaves are also chewed as a remedy for toothaches.

2.4 Production systems

Dry grain production of cowpea is estimated to be about 6.5 million metric tons annually
under 14.5 million acres worldwide (Njonjo, 2018). However, the amount of leaves and
pods produced is not reflected in any statistical data but it is estimated to be large. Over
70% of worldwide cowpea production takes place in west and central Africa (Timko et
al., 2008). This crop is usually grown as an intercrop with pearl millet, maize, cassava or
sorghum and sometimes as sole crop (Timko & Singh, 2008). Cowpea can fix up to
150kg N/ha under favourable conditions (Woomer et al., 2004; Olal, 2015). Cowpea
productivity in Kenya ranges between 200-500 kg/ha for small scale farmers (Olal,
2015)

2.5 Landraces

Landraces result from a long time of natural and artificial selection by farmers to select
better adapted varieties for the local environments (Casanas et al., 2017). Although
cowpea is reported to have high protein value, high variability, high adaptability and

8
drought tolerant capacity (Ashraf et al., 2012), little research has been done on their
landraces hence loss of superior genes that would be useful in genetic manipulation.

Landraces have played a big role in the introduction of improved varieties as they have
rich and complex ancestry with great variations in response to many stresses as well as
vast resources for improved varieties (Litchfouse et al., 2011). A large number of
cowpea varieties evolved from local landraces have been commercialized (Pratap &
Kumar, 2011). However, landraces have rarely been used in hybridization to understand
their breeding value in the improvement of grain yield and other traits in cowpea.

2.6 Released cowpea varieties in Kenya

The list of released cowpea varieties in Kenya is shown on Table 1. These varieties are
grown either for their leaves, seeds or both (dual purpose).

9
Table 2.1: Improved cowpea varieties in Kenya

Variety Year Owner(s) Maturity Target areas of Grain yield Special characteristics
released (days) production (Masl) (t/ha)
1. KVU HB 48 E 10 1987 KARI 85-95 0-1200 1.2-1.4 Tolerant to viral diseases; grown for
vegetable use
2. KVU 27-1 1989 KARI 70-90 600-1200 1.5-1.8 Dual purpose; dark red seeds;

3. ICV 11 1992 ICIPE 75 1-1500 2.2 Pest tolerant;

4. MTW 610 1998 IITA 60 1-1500 2.5 Large seeds

5. MTW 63 1998 IITA 60 1-1500 2.5 Pest tolerant;

6. Kunde 1 ND Western 75-90 Below 2000 1.2-2.5 Dual purpose;

Seed Co.
7. Machakos 66 (M66) 1998 KARI 85-95 1200-1500 1.5-1.8 Dual purpose; creamy brown seeds and deep
green midribs; tolerant to cowpea yellow
mosaic virus and scab
8. Katumani 80 (K80) 2000 KARI 75-85 0-1500 1.8-2.0 Dual purpose; creamy brown seeds; resistant
to aphids
9. KVU-419 (Kunde 419) 2000 KARI 65-72 0-1200 1.2-1.5 Drought tolerant, extra early maturity

10. KCP 022 2000 KARI 60-75 0-1200 1.2-2.5 Super early maturity drought tolerant

11. Kunde Mboga 2014 Simlaw 120-140 Low and mild altitude Seed yield Vegetable use.
seeds Co. 1.6-2.2
Drought resistant.
12. Simlaw Kunde 2014 Simlaw 75-90 Low and mild altitude 1.8-2.6 Large seeds
seeds Co.
Drought tolerant

High yielding
 Variety Year Owner(s) Maturity Target areas of Grain yield Special characteristics
released (days) production (Masl) (t/ha)
13. 1002/1005/3 (Kunde 2017 KALRO 70-80 Low-high altitudes(5- 1.5-2.13 Large seeds
Faulu) 2000 msl) coast,
eastern central and Early maturity
western
Alectra tolerant

Dual purpose
14. 1005/1002/1 (Kunde 2017 KALRO 70-80 Low-high altitudes 5- 1.5-2.0 Early maturity
Tamu) 2000 msl
Alectra vogelii tolerant

Dual purpose
15. 1005/1003/3 (KAT 2017 KALRO 80-90 Low-high altitudes 1.4-2.0 Alectra vogelii tolerant
Kunde) ranging from 5-2000
msl Dual purpose
16. 1005/1002/1/1/1 (Kunde 2017 KALRO 80-90 (Coastal eastern and 1.4-1.9 Large seeds
Soko) western)
Alectra vogelii tolerant

Dual purpose
17. 1005/1004/3 (Kunde 2019 KALRO 80-90 Altitudes 600-1500 msl 1.5-2 Drought tolerant
Tumaini)
AEZ: LM 4-5, LM 3-4 Tolerant to Alectra vogelii parasitic weed

Dual purpose

White grain with brown eyes

National Crop Variety List (KEPHIS-2019)

11
2.7 Environmental requirements for cultivation of cowpea

Cowpea thrives well under a wide range of environmental conditions. In Kenya, cowpea
does well in arid and semi arid areas but is also recommended for medium and higher
altitudes between 1200-1500 metres above sea level (Karanja, 2016). This crop can
survive high temperatures and drought conditions but is easily affected by frost. It
requires a temperature range of 15ºC-30ºC (Karanja, 2016) but does best at 36.1ºC
(Muniu, 2017). Cowpea germinates rapidly under warm temperatures while the cold
temperatures slow germination. Cowpea can grow well in well drained soils including
sandy, clay or loamy soils but does not tolerate waterlogging (Karanja, 2016). The soils
should have a PH range of 5.5-6.5 (Nkouannessi 2005; Olal, 2015).

2.8 Cowpea characterization

In many areas, cowpea yields are low because the environments where they are
produced have various abiotic and biotic stresses (Makari, 2022). The yields may also
vary due to differences in the growth and development of each plant. Knowledge of the
extent, distribution and nature of the variation would help in the development of cowpea
genotypes with high yield potential and improved adaptation to environmental stresses
(Sheidu, 2023)

In the past, genetic diversity in plants was evaluated by studying the differences between
quantitative characters and qualitative traits (Kameswara, 2004). It has been important in
classifying cultivars and in the study of taxonomic status (Doumbia, 2011).

Morphological characterization is still the first step in the studies of genetic relationships
in many breeding programmes. However, evaluation of genetic relationships among
germplasm is lengthy and expensive (Doumbia, 2011). Morphological characters are

12
believed to be controlled by complex genes that are subject to environmental
modification and interactions including epistatic interactions (Doumbia 2011).

Most of the best cultivated and breeding materials have limited number of observable
morphological markers; most of which have deleterious effects on agronomic
performance (Doumbia, 2011). Therefore, morphological characterization cannot
adequately describe cultivars without many replications over a long time (Malek et al,
2014). Comparisons can only be made for morphological characteristics taken from the
same location at the same time.

Determination of genetic diversity in cowpea genotypes is very important in the

development of superior cultivars. Traditionally, the estimation of genetic diversity
made use of morphological markers. However, the limited number of morphological
markers, their poorly known genetic control and environmental influence on phenotypic
expression at different stages of growth has limited their reliability over time (Sariah,
2010). Development of molecular markers such as restriction fragment length
polymorphism (RFLP) (Lambrides et al., 2000), random amplified polymorphic DNAs
(RAPDs) (Betal et al., 2004), amplified fragment length polymorphism (AFLP) (Zong et
al, 2003) and microsatellites (Li et al., 2001; Wang et al 2004) have greatly improved
the analysis of plant genomes and the genetic structure and variations among cowpea
accessions (Sariah, 2010). In the past RAPDs have been used to study genetic diversity
between cowpea cultivars (Ba et al., 2004). Studies using SSR markers to characterize
diversity among cowpea accessions collected from different agroecological zones have
been carried out in Kenya, (Kuruma, et al, 2008, Wamalwa et al., 2016). Studies show
that simple sequence repeats (SSR) markers which are single locus with multiple alleles
are better than other markers and are effective in differentiating genotypes (Doumbia,
2011). They are highly polymorphic, codominant and easily reproducible (Asare et al.,
2010; Mafakheri et al., 2017). SSRs have been widely used in genotyping, seed purity
13
checks and protection of varieties (Asare et al., 2010). They have also been used for
pedigree analysis and genetic mapping of simple and quantitative characteristics (Asare
et al., 2010).

14
 CHAPTER THREE

MATERIALS AND METHODS

3.1 Experimental site

The experiments were conducted in the laboratory and experimental farm of Jomo
Kenyatta University of Agriculture and Technology in Juja. Juja is located in central
Kenya at 1 º 11′ 0′′ south, 37 º 7' 0" East. This area has semi-arid conditions under AEZ
IV (Jaetzold and Schmidt 1983) with two distinct rain seasons. The long rains fall from
March to May while the short rains fall between October and December; with an
average annual rainfall of 989 mm. The daily temperature ranges from 10-30ºC
depending on the season. The area has rich black cotton soils.

3.2 Plant materials

One hundred and ten accessions, (Appendix I) were used for this study. Collections from
farmers in semi arid region of Kenya comprising of 82 accessions were procured from
the National Gene Bank of Kenya; Muguga Kenya. These comprised of accessions from
Machakos (74), Makueni (3) and Kitui (5). Twenty-three landraces from Machakos (14)
and Baringo (9) were collected directly from farmers and five commercial lines (K80,
M66, Kenkunde, KAR 1 and KVU-27-1) were obtained from registered seed companies.

3.3 Field Experimental Design

The trial was laid down in a Randomized Complete Block Design (RCBD) with three
replications. Blocking of the replicates was based on the gradient of the field. Each
replicate measured 12m by 24m comprising of 110 plots. Cowpea seed was sown in
three lines per plot at inter- and intra-row spacing of 0.6m and 0.3m respectively. Two

15
cowpea seeds were planted per hole and later thinned to a single plant per hole after two
weeks from date of germination. Irrigation was done immediately after planting and
whenever it became necessary. Weeding was done manually three weeks from date of
planting followed by rouging of weeds whenever they emerged.

3.4 Data collection

Qualitative and quantitative agronomic data was collected and recorded from five plants
in the middle row of each plot as described by IBPGR (1983). Data collected at
emergence was the number of days it took for 50% of the plants to emerge. Data for
vegetative stage of development recorded at six weeks; growth habit, growth pattern,
twining tendency, pigmentation, terminal leaflet shape, leaf color, terminal leaflet
length, terminal leaflet width and number of main branches. At flowering stage of
development, data recorded included raceme position, pod attachment, immature pod
pigmentation, pod curvature, flower color, pod length, number of seeds per pod and pods
per plant. At harvesting, data was obtained for mature pod color, seed shape, texture of
testa, seed color, eye color and 100 seed weight.

3.5 Molecular characterization of cowpea

This experiment was carried out at the Cassava Diagnostics Laboratory in Jomo
Kenyatta University of Agriculture and Technology in Juja Kenya.

One hundred and ten accessions; (Appendix 1) were planted in pots filled with sand in
the green house. The leaf samples of all the accessions were obtained from each
genotype two weeks after planting. The leaf samples (0.4g) were freeze dried and then
stored at -70o C until used for DNA extraction.

16
About 0.1g of young leaf tissue of each accession was taken and DNA extracted
according to the CTAB DNA extraction protocol (Doyle & Doyle, 1990). Briefly, each
0.1g of leaf sample was ground in a mortar with a pestle with the aid of acid washed
sand to break the cell walls and membranes. 1500µl CTAB buffer was added and the
slurry was transferred to a 2,000µl Eppendorf tube. The solution was mixed thoroughly
to suspend the cellular material and incubated for 30 minutes at 65ºC in a water bath.
The slurry was then centrifuged at 13,000 rpm for ten minutes. 750µl of the supernatant
was transferred to a fresh Eppendorf tube and mixed with an equal amount of
chloroform: Isoamyl alcohol; 24:1 v:v and the solution vortexed for three minutes. The
solution was span in a centrifuge at 13,000 rpm for 10 minutes. The aqueous layer was
transferred (400µl) into a fresh tube Eppendorf tube using a pipette. An equal volume of
ice cold isopropanol was added and the tubes incubated in a freezer for 10 minutes at -20
ºC and then centrifuged at 13,000 rpm for ten minutes. The supernatant was decanted
carefully leaving DNA pellet at the bottom of the tube. The pelleted DNA was washed
with 500µl 70% ethanol and centrifuged at 13,000rpm for 5minutes. DNA was dried at
room temperature for 20 minutes or until the ethanol evaporated and then dissolved in
50 µL of deionized water. The DNA samples were then stored at -20°C until used.

The quality of the extracted DNA samples was tested on 1% Agarose. 1g of agarose was
weighed and mixed with 100 mL Tris/Borate/EDTA (TBE) buffer. This solution was
poured in a flask and heated in a microwave for three minutes or until the agarose was
fully dissolved. The solution was allowed to cool and 2µl of Ethidium Bromide added.
About 5µl of the extracted DNA sample were mixed with 2µl dye loaded on to the gel
and run for 30 minutes.

Molecular characterization of one hundred and ten accessions (110) was done using 20
primer pairs of Simple Sequence Repeat (SSR) markers to analyze the variations and
relatedness among the genotypes. Initially, 20 pairs of SSR markers (Table 1) were
17
screened for polymorphisms using mixed DNA samples. These primers were similarly
used by Li et al. (2001), Asare et al. (2010) and Doumbia et al. (2013). PCR
amplification was carried out in 0.2-mL PCR tube with final volume of 12.5µL,
comprising of a master mix containing 2.5µ 10 ×PCR buffer, 0.25µM of each primer,
0.5 U Taq DNA polymerase. For each of these 2 µL sample DNA template DNA was
added. The tubes were placed in a Gene-Amp PCR system 2720 (Applied Bio systems,
USA) with an initial DNA denaturation at 94 °C for 5 minutes followed by 35 cycles of
30 seconds at 94 °C, 30 seconds at 45 °C to 65 °C for annealing temperature depending
on the primer pair, 1 minute at 72 °C and a final incubation at 72 °C for 10 minutes
(Table 4). The PCR products were then analyzed on 2% agarose gels using 0.5 ×TBE
buffer stained with 2mg/mL ethidium bromide to establish polymorphism. The gel
associated with each marker was photographed under a UV trans illuminator. The
amplified bands were scored for each accession as present (1) or absent (0).

18
Table 3.1: SSR Markers used in the study

Name Primer sequence No bases Source

SSR-6265F 5’-CAG AAG CGG TGA AAA TTG AAC -3’ 21
Doumbia/Asare
SSR-6265R 5’- GCA TGT TGC GAC AAT GG-3’ 17
SSR-6258F 5’- GGT TTC CTA GTT GGG AAG GAA-3’ 21
Doumbia/Asare
SSR-6258R 5’-ATT ATG CCA TGG AGG GTT CA-3’ 20
SSR-6243F 5’-GTA GGG AGT TGG CCA CGA TA-3’ 20
Doumbia/Asare
SSR-6243R 5’-CAA CCG ATG AAA AAG TGG ACA-3’ 21
SSR-6218F 5’-GTG GAA GGA ATG GGT CCA G-3’ 19
Doumbia/Asare
SSR-6218R 5’-AGG AAA TTT GCA TTC CCT TGT-3’ 21
SSR-6217F 5’-GGG AGT GCT CCG GAA AGT-3’ 18
Doumbia/Asare
SSR-6217R 5’-TTC CCT ATG AAC TGG GAG ATC-3’ 21
SSR-6353F 5’-TCA TGG GTT AAA TTT GCT TCA A-3’ 22
Doumbia/Asare
SSR-6353R 5’-AAA CCA TGT GGT TGT TGC AC-3’ 20
SSR-6352F 5’-GTT GTG AGC TTC CCC AGA TG-3’ 20
Doumbia/Asare
SSR-6352R 5’-ATT TTT GAA CCC ACC ACC AG-3’ 20
SSR-6336F 5’-TGA AAA CAA CGA TAT GCA GAA-3’ 21
Doumbia/Asare
SSR-6336R 5’-TCA GTC TTA GAA TTG AGT TTT C-3’ 22
SSR-6323F 5’-CAA AGG GTC ATC AGG ATT GG-3’ 20
Doumbia/Asare
SSR-6323R 5’-TTT AAG CAG CCA AGC AGT TGT-3’ 21
SSR-6451F 5’-AAA GAG ATA CAC ATG CCT AAC-3’ 21
Doumbia/Asare
SSR-6451R 5’-GAC CAA CAG CGA CTT TGA GC-3’ 20
SSR-6277F 5’-CAC CCC CGT ACA CAC ACA-3’ 18
Doumbia/Asare
SSR-6277R 5’-CAC TTA AAT TTC CAC CAG GCA T-3’ 22
SSR-6436F 5’-CAG AAT CCT TGT GAA CCT G-3’ 21
Doumbia/Asare
SSR-6436R 5’-TTT CGC AAT ATG CCC TTT TC-3’ 21
SSR-6375F 5’-GCT CGG ATA TGG TCC TGA AA-3’ 20
Doumbia/Asare
SSR-6375R 5’-TCA GTG TCA GCA CCA TCC C-3’ 19
SSR-6371F 5’-TGC TCA TCG TGC TTT GTC TT-3’ 21
Doumbia/Asare
SSR-6371R 5’-CAC TTC AGA CTT AGA GCG AAG-3’ 21
SSR-6370F 5’-CAA CTT CAC AGC CCT CAA-3’ 18
Doumbia/Asare
SSR-6370R 5’-TTG AAG GTA TGG CCT TTT GTT T-3’ 22
SSR-6356F 5’-TGC AAT ATG GAC CAG AAG AAA-3’ 21
Doumbia/Asare
SSR-6356R 5’-ATG CCC CAA CAA CAA CAT TT-3’ 20
SSR-6613F 5’-CTA TTG GAA TCT TGC CGT TG-3’ 20
Doumbia/Asare
SSR-6613R 5’-CTT TAC CTT TAT GCA AAC CAA T-3’ 22
SSR-6608F 5’-CTA AAT TAT AAT ATT CGT CGG T-3’ 21
Doumbia/Asare
SSR-6608R 5’-GGT TAA GGA AAA GAG GGT AGG-3’ 21
SSR-6603F 5’-GAG AAC TTC ACG CAC AAT AG-3’ 20
Doumbia/Asare
SSR-6603R 5’-CGC GGT AGC ATG ATT GAA TTT-3’ 21
SSR-6587F 5’-GAT ATA GAA TAG CAT ATT TAA C-3’ 22
Doumbia/Asare
SSR-6587R 5’-GTT GAA AGT TTG ATA GTA AAG-3’ 21

Asare et al (2010) and Doumbia, (2011)

19
Table 3.2: PCR amplification conditions used in the study

Stages Temperature Time (s) Number of cycles

Initial Denaturation 94 300 1
Denaturation 94 30
Annealing 55 45 35
Extension 72 60
Final Extension 72 600 1
End 4 ∞

3.6 Data Analysis

The field data was recorded in data sheets and thereafter entered, organized and
managed in an excel sheet. These data were analyzed using GENSTAT program version
14. Qualitative data was used to assess the distribution of accessions in different traits of
the respective characteristics expressed as a percentage of the total number of
accessions; number of accessions possessing a certain attribute of character divided by
the total number of accessions multiplied by 100. Quantitative data was subjected to
ANOVA to determine the variation in the respective traits. The contribution of the
respective characters to the variation of the different traits was assessed using Principal
Component Analysis. The accessions were classified into groups using cluster analysis
while correlation among traits was determined using Pearson correlation analysis.
Unweighted ranking of the characters positively associated with yield and productivity
was done by ranking five traits; pod length, number of pods per plant, number of seeds
per plant and number of branches per plant, individually and summing up the ranks for
each accession. The sum of ranks was used to estimate the accessions potential
providing ranking of greatest performance and least performance.

20
For the molecular data, the bands that were not polymorphic with at least one of the
samples were not scored. The polymorphic bands were scored as present or absent (1/0).
This was then used as raw data to generate a matrix which was subjected to Principal
Component Analysis and analysis of molecular variance (AMOVA).

21
 CHAPTER FOUR

RESULTS

4.1 Variation in the Cowpea Accessions for Qualitative Characters

Variation among the accessions based on each character (Appendix II) was evaluated by
determining the distribution of these accessions in the respective traits. The accessions
could be evenly distributed across the traits for a character or skewed in favor of one of
the traits within a character. The distribution of accessions among the traits of 15
qualitative morphological characters is presented in Table 4.1. Distribution of accessions
among the traits were evenly distributed for immature pod color, leaf color, seed shape
and twining tendency and skewed for terminal leaflet shape, raceme position, pod
attachment, pod curvature, mature pod color, flower color, testa texture and eye color.

The accessions could be placed into three groups based on growth habit: acute erect,
semi erect and intermediate. Over half of the accessions (73%) were semi erect, acute
erect (23%) and 5% intermediate. Two classes of growth pattern were observed;
determinate (27%) and indeterminate (73%). It was also recorded that the accessions had
three distinct groups based on the raceme position; above canopy (24%), upper canopy
(56%) and throughout the plant (20%). Based on pod color, both mature and immature
pods showed a wide range of variation. There were accessions that had no pigmentation
(green) on immature pods to those with uniform pigmentation; 31% of the accessions
did not have pigmented pods, 34% of the lines had pigmented valves and green sutures,
splashes of pigment were observed in 28% of the accessions and 6% of the lines had
pigmented tips. Only 1% of the accessions had uniformly pigmented immature pods.
The mature dried pods also showed uneven distribution of accessions; straw (65%), dark
brown (34%), dark purple (1%). The accessions also showed uneven distribution for

22
flower color and terminal leaflet shape. The observed flower colors included; white
(1%), white-purple (4%), purple (95%) and terminal leaflet shape: Globose (99%) and
hastate (1%). 88% of the lines under study produced slightly curved pods while the
remaining 12% had straight pods.

Twining tendency of the accessions showed even distribution with 6% of the accessions
with no twining, 39%, 42% and 13% of the accessions were observed to show slight,
intermediate and pronounced twining respectively.

23
Table 4.1: Distribution of accessions among respective categories of the
evaluated morphological characters

Crop Characteristic Distribution of the accessions in respective traits (%)

1. Growth Habit Acute erect 22.7
Semi erect 72.7
Intermediate 4.5
2. Growth pattern Determinate 27
Indeterminate 73
3. Twinning tendency None 6
slight 39
Intermediate 42
Pronounced 13
4. Pigmentation None 43.6
very slight 36.3
Moderate 4.5
Intermediate 10.9
Extensive 4.5
5. Terminal leaflet shape Globose 99
Hastate 1
6. Raceme position Above canopy 24
upper canopy 56
Throughout 20
7. Pod attachment Pedant 81
30-90º 16
Erect 3
8. Immature pod color None 31
pigmented valves green sutures 34
splashes of pigment 28
uniformly pigmented 1
pigmented tip 6
9. Pod curvature Straight 12
Slightly curved 88
10. Mature pod color Straw 64.5
Dark brown 33.6
Black/dark purple 1.8
11. Flower color White 1
white purple 4
purple 95
12. Leaf color Pale green 1
intermediate green 54.5
Dark green 44.5
13. Seed shape Kidney 2
ovoid 47
Globose 1
Rhomboid 50
14. Eye color Absent 81
Brown splashes 2
Tan brown 3
Blue to black 4
speckled 10
15. Testa texture Smooth 89
Smooth to rough 7
Rough to wrinkled 4
16. Seed coat colour White 4
Cream 13
Brown 23
Red 44
Purple 0
Black 10
Other 6

24
Eighty accessions had semi erect growth habit, 25 had acute erect growth habit and 5
accessions portrayed intermediate type of growth habit. Eighty accessions had
indeterminate growth pattern while the remaining 30 accessions were determinate.
Fourty three genotypes had slight twining tendency; 46 accessions showed
intermediate twining tendency while 14 accessions had pronounced twining
tendency. GBK 003650, GBK 003651, GBK 003674, GBK 003713, GBK 003726,
GBK 003780 and GBK 003796 did not show any twining tendency.

GBK 003713, GBK 003816, KOL 6, KOL 8 and MBL were pigmented moderately
at the base and tips of petioles; MAR. 5, Kenkunde and GBK 034722 showed
extensive pigmentation; GBK 003657, GBK 003658, GBK 003663, GBK 003670 B,
GBK 003687 B, GBK 003696, GBK 003699, GBK 003701, GBK 003709, KIP 1,
KIP 2 and LAM 4 had intermediate pigmentation. 40 accessions had very slight
pigmentation while the remaining 48 accessions showed no pigmentation. All
accesssions had globose shaped terminal leaflets except GBK 003804 that had
hastate shaped terminal leaflets. Sixty two accessions had raceme on the upper
canopy. Twenty six had their racemes mostly above the canopy while 22 genotypes
had raceme spread throughout the canopy. Eighty nine genotypes had their pod
attachment to penducle to be pendant. Eighteen accessions had their pods to peduncle
attachment between 30 and 90 down from erect. GBK 003685 and KOL 2 had erect
pod to peduncle attachment. Pigmented valves and green sutures were found on
immature pods of 37 accessions. Thirty one accessions had splashes of pigment on
their pods. Seven accessions showed pigmented tip on their immature pods. No
pigmentation was found on 34 accessions and only GBK 003705 from Machakos
showed uniformly pigmented immature pods. Ninety seven accessions had slightly
curved pods while thirteen accessions had straight pods. Three pod colours were
obtained at maturity. Seventy one varieties were straw (65%) coloured pods, 37
accessions dark brown podcolor at maturity and 2 accesions showed black to dark
purple color. Three flower colours were observed in this study. A hundred and five
accessions showed purple coloured flowers. Only GBK 003650 produced white
flowers while GBK 003675 A, GBK 003727, GBK 003916 and GBK 046540 had
purple-white flowers. Fourty nine cowpea accessions had dark green leaf color, 60

25
had intermediate green leaves and only GBK 003985 had pale green leaves. Fifty
five genotypes had rhomboid shaped seeds. Only GBK 003651 had globose shaped
seeds. Two accessions had kidney shaped seeds and they were GBK 003717A and
GBK 046540. The seeds of 52 accessions had ovoid shape. Eighty nine accesions
had no (0) eye colour; Eleven accessions had speckled (7) eye colour; four
accessions had brown splashes; three had tan brown eye colour. The testa texture of
98 accessions were smooth while 8 were smooth to rough. Four genotypes had rough
to wrinkled testa texture.

4.2 Variation among accessions for Quantitative Characters

The means and ranges among accessions for respective quantitative characteristics
are presented in Table 4.2. The accessions were significantly different for number of
days to 50% emergence, pod length, number of pods per plant and number of seeds
per pod. There were no significant differences among the accessions for one hundred
seed weight, number of branches per plant, number of days to 50% flowering,
terminal leaflet length and terminal leaflet width (Table 4.2).

From the day of planting, the accessions germinated between 4 to 8 days with an
average of 6.06 days. Seven accessions germinated after 4 days, 45 accessions
emerged after 5 days, 35 genotypes after 6 days and the remaining accessions
emerged after 8 days.

The accessions attained 50% flowering between 65 and 75 days after emergence with
a mean of 70 days. KOL 5 and M66 were the earliest genotypes to flower at 65 days.
On the other hand, KOL 2, GBK 046540, GBK 003717A, GBK 003676, GBK
003675 and GBK 003652 were late flowering genotypes flowered at 75 days.

Terminal leaflet length for the accessions ranged between 3.4cm and 5.8 cm with an
average of 4.39 cm. GBK 003663 and K80 had the shortest and longest terminal
leaflets respectively.

26
The terminal leaflet width ranged from 1.8 to .1 cm with an average of 2.8cm. GBK
003701 and GBK 003674 had the shortest and longest width across the leaf of the
accessions.

The average number of branches counted per plant was 4.38 with a range of 3.4 to
5.2. Most accessions had 4 branches per plant.

The number of pods per plant was observed to range between 6.67 and 30 pods per
plant with an average of 21.95 pods per plant. GBK 003698 had the highest number
of pods per plant while GBK 003713 had least number of pods per plant.

The length of pods of the accessions was observed to range from 9.01 cm to 13.96
cm with a mean length of 11.56 cm. GBK 003650, GBK 003651, GBK 003660,
GBK 003663, GBK 003675 A, GBK 003682, GBK 003685, GBK 003689, GBK
003693, GBK 003694, GBK 003697 and GBK 003701 were some of the accessions
that gave long pods on average.

The number of seeds per pod averaged at 8.36 with a range of 5.07 to 11.07 cm.
GBK 003682, GBK 003693, GBK 003694, GBK 003697, GBK 003701, GBK
003876, GBK 027089, K80, M66, and MAR.2 are some of the accessions that were
observed to give high number of seeds per pod.

Table 4.2: Variation in quantitative morphological traits among cowpea

accessions

Trait P Mean Range SD CV (%)

Emergence -50% (days) ‹0.001 6.06±1.24 4.3-8.3 0.966 20.4
Flowering- 50% (days) 0.28 70.87±3.27 65-75.7 1.952 4.6
Terminal leaflet length (cm) 0.487 4.39±0.83 3.4-5.8 0.486 19
Terminal leaflet width(cm) 0.42 2.81±0.72 1.8-4.11 0.428 25.6
No. of branches per plant 0.155 4.38±0.69 3.4-5.2 0.42 15.7
No. of pods per plant ‹0.001 21.95±5.31 6.67-30 4.54 24.2
Pod length(cm) 0.006 11.56±1.51 9.01-13.96 1.051 13.1
No. of seeds per pod 0.005 8.36±1.92 5.07-11.07 1.34 22.9
100Seed Weight (g) 0.398 8.8±2.05 5.9-11.4 1.21 23.2

27
The average 100 seed weight was 8.8 g with a range between 5.9g and 11.4g. Some
of the accessions with large seeds included GBK 003654, GBK 003666, GBK 3676
B, GBK 003687, GBK 003701 and GBK 003723.

Ranking based on pod length, number of pods per plant, number of seeds per plant
and number of branches per plant, was used to identify accessions with high and low
potential yields. The accessions that showed high potential (top 20) and least
potential (bottom 20) in productivity based on overall unweighted ranking among
characters positively associated with productivity are presented in Table 4.3. These
results indicate that the top 5% of the lines that showed high productivity potential
are GBK 003662, GBK 003663, GBK 003676, GBK 003723, GBK 003650 and
GBK 003642 which superseded the registered varieties. K80 was ranked number 20
whereas Kenkunde and M66 were ranked 91 and 104 respectively. The overall
ranking for all accessions is presented in Appendix 2.

28
Table 4.3: Mean values of 20 highest and 20 lowest accessions based on
quantitative traits

ACC PL SPP 100SW PPP BP Ranks

High Rank
GBK 003662 11.8 10.9 8.7 26.3 4.7 1
GBK 003663 12.6 10.2 6.7 27.0 4.1 2
GBK 003676 10.5 8.1 8.7 26.7 4.3 3
GBK 003723 11.9 8.6 11.2 23.0 4.2 4
GBK 003650 12.9 9.6 9.5 26.7 5.1 5
GBK 003642 9.9 6.2 9.3 10.7 4.3 6
GBK 003669 11.4 10.8 9.8 22.0 4.7 7
GBK 003668 D 13.0 9.5 8.3 25.3 4.3 8
GBK 003780 12.9 9.1 7.1 19.7 5.1 9
GBK 003709 10.3 7.8 6.2 7.0 4.9 10
GBK 003685 12.0 8.5 8.3 26.0 3.9 11
KAB 1 11.9 9.3 10.8 22.3 4.8 12
GBK 003985 12.6 9.5 10.2 23.0 5.2 13
GBK 003796 11.5 6.3 9.1 23.3 4.1 14
GBK 003645 11.5 6.5 8.9 27.0 4.1 15
GBK 003676 11.7 9.6 9.3 20.7 4.2 16
GBK 003676 B 11.0 7.0 11.4 20.7 4.4 17
GBK 003687 11.6 6.3 6.9 25.0 4.0 18
GBK 003654 9.5 5.1 10.0 19.7 4.8 19
K80 13.0 10.9 8.4 28.3 4.5 20
Low Rank
Kenkunde 12.7 8.2 8.9 21.3 4.1 91
GBK 003698 12.3 9.3 9.2 30.0 4.5 92
GBK 003694 12.5 10.3 8.5 21.3 3.9 93
GBK 003711 11.0 9.2 9.7 22.7 3.8 94
KOL 8 12.0 8.0 8.0 19.0 4.3 95
KOL 2 11.3 8.6 8.3 18.3 4.5 96
KAT 1 11.9 9.4 8.1 21.7 4.2 97
MBL 10.6 7.8 11.4 18.7 3.9 98
MAC 1 12.3 7.1 9.4 24.3 4.3 99
GBK 003682 13.2 10.2 10.0 25.3 4.5 100
GBK 003687 B 10.1 8.5 10.0 24.7 4.7 101
MAR.5 12.6 6.7 10.1 10 4.9 102
MAR.3 11.2 7.0 7.8 7.0 4.6 103
M66 13.1 10.4 9.7 19.7 4.1 104
MAC 3 11.9 8.5 9.6 18.0 4.3 105
K0L9 C 10.1 7.5 11.4 25.3 4.7 106
GBK 005173 B 10.4 7.7 6.9 26.7 4.5 107
KIP 2 11.3 6.9 9.3 24.7 4.3 108
GBK 027089 11.2 10.9 7.8 19.3 5.1 109
GBK 034722 11.1 7.8 8.7 23.0 4.9 110

29
Ranking of accessions based un-weighted indices of five traits; pod length, number of pods per plant,
number of seeds per plant and number of branches per plant

ACC=accession number PL=Pod length SPP=Number of seeds per pod 100SW=a hundred
seed weight PPP= number of pods per plant BP=Number of main branches per plant

Pair wise correlation values among the quantitative traits are presented in Table 4.4.
Significant positive correlation among the traits was found for number of days to
50% emergence and terminal leaflet length (r= 0.188), number of days to 50%
emergence and terminal leaflet width (r=0.204), terminal leaflet length and terminal
leaflet width(r=0.523), and pod length and number of seeds per pod (r=0.547).
Significant negative correlation was recorded for number of days to 50% emergence
and number of days to 50% flowering (r=-0.21) and number of days to 50%
emergence and number of pods per plant were (r=-0.53).

Table 4.4: Pearson’s correlation among quantitative traits recorded on cowpea

Character E (50%) BP TLL TLW F PPP PL SPP 100SW

(50%)
E (50%) 1
BP -0.0273 1
TLL 0.1881* -0.0242 1
TLW 0.2042* 0.0880 0.5230** 1
F (50%) -0.2131* 0.0510 -0.0787 0.0890 1
PPP - -0.1200 -0.0611 0.0742 0.1376 1
0.5258**
PL 0.0510 -0.1324 -0.0221 -0.0328 -0.1147 0.0986 1
SPP 0.0749 0.0789 -0.0091 0.0560 -0.0295 0.1742 0.5470** 1
100SW 0.0085 0.0392 0.0424 0.0140 0.0782 0.1053 0.0411 0.0429 1

30
The quantitative characters evaluated were reduced to five major principal components
that accounted for 19.8, 18.0, 15.91, 12.4 and 11.2 respectively accounting for 77% of
the total variation (Table 4.5). PC1 was attributed mainly to number of days to 50%,
emergence, number of pods per plant, terminal leaflet length and terminal leaflet width.
PC 2 was attributed to pod length and seeds per pod. PC 3 was associated with number
of days to 50% emergence, number of days to 50% flowering, number of pods per plant,
terminal leaflet length and terminal leaflet width (Table 4.5).

Table 4.5: Eigen vectors and values for five Principal Components

PC 1 PC 2 PC 3 PC 4 PC 5
Eigen value 1.782 1.620 1.431 1.116 1.010
% variance 19.80 18.00 15.91 12.40 11.22
Cumulative Variance 19.80 37.80 53.71 66.11 77.33
Vector Loadings
% 100Seed Weight 0.05686 0.05984 0.01451 0.54445 0.73494
No. of Branches/Plant -0.07734 -0.10131 0.07847 -0.65861 0.60621
Emergence (50%) -0.57700 0.09694 -0.29519 0.00341 0.02544
Flowering (50%) 0.21671 -0.14965 0.38164 -0.37066 -0.12195
Pod Length 0.07599 0.65918 -0.17454 -0.08388 -0.16279
No. of Pods/Plant 0.46836 0.21126 0.44474 0.20598 0.00878
No. of Seeds/Pod 0.06598 0.66252 -0.03755 -0.24503 0.16710
Terminal Leaflet Length -0.46927 0.11945 0.44362 0.15119 -0.13919
Terminal Leaflet Width -0.40149 0.14837 0.57826 0.00644 0.05451

The quantitative data for nine characters evaluated were subjected to multivariate cluster
analysis generating a dendrogram classifying the accessions in groups based on
similarity. Figure 1 shows the relationship among the 110 accessions that were evaluated

31
based on quantitative characters. The accessions are initially divided into two major
clusters (1 and 2), 61% at similarity level. Cluster 1 is subdivided into two sub-clusters
A and B. Sub-cluster A had five accessions while sub-cluster B had two accessions.
Cluster 1 is comprised of seven accessions (Appendix 1, Table 4.6) obtained from
Machakos except one, KIP2, from Baringo. Cluster 2 had 82 accessions obtained from
Machakos.

Cluster 2 also had two sub-clusters C and D which are divided further into four groups;
I, II, III and IV respectively. Sub-cluster C group I had 84 accessions while sub cluster C
group II had 3 accessions. Sub-cluster D group III and sub-cluster D group IV each had
8 accessions. The commercial varieties were observed to have been grouped in one
cluster; Cluster 2 sub-cluster C group I. Collections from the National Gene Bank of
Kenya procured from Machakos were found in all clusters with majority in Cluster 2,
sub-cluster C; I (Figure 1)

Collections obtained directly from farmers in Kola in Machakos were also observed to
be evenly distributed across all clusters with Cluster 1 A having one accession, cluster C
I has four accessions, cluster D III has one accession and cluster D IV having two
accessions. Accessions from Makueni were distributed into two groups; cluster 2 C I had
two accessions and cluster 2 D IV had one accession. Accessions from Kitui were
grouped into one cluster; Cluster 2 C I except for one accession that was grouped in
Cluster 2 D IV (Figure 4.1). Accessions that were sourced from the central Rift Valley;
Baringo county were spread out throughout the clusters; cluster 1 A (1), cluster 2 C I (6)
and 2 C II (2).

32
 A
1
B

I
1

II

III
D
IV

Figure 4.1: Relationships among cowpea collections from Semi-Arid Areas of

Kenya and commercial lines

33
Table 4.6: Distribution of accessions within clusters based on similarity among
quantitative traits

Cluster Similarity No. of Name of accessions

of accessions
coefficient
Cluster 1 0.61
A 0.64 5 KOL 5, GBK 003645, GBK 003694, KIP 2, GBK 003714
B 0.64 2 GBK 003651, GBK 003687
Cluster 2 0.61
CI 0.64 84 GBK 003688, GBK 003780, GBK 027089, GBK 003699,
GBK 003816, KOL 6, GBK 003709, KOL 2, KAR 1, GBK
003654, GBK 003670, GBK 003695, GBK 003687, GBK
003718, GBK 003670, GBK 003675, GBK 036582, GBK
003680, GBK 003685, GBK 026941 B, GBK 026958, GBK
003642, GBK 034722, GBK 003720, GBK 003705, GBK
003820, GBK 003727, GBK 003796, GBK 003669,
LAM4,KVU-27-1, KAT 1, GBK 003676, GBK 003707,
GBK 003700, K80, GBK 003674, GBK 003724, GBK
026941, GBK 003717, KOL 2, GBK 003916, GBK
026958,KOL 8, GBK 003689, MAC 2, MAR.3, GBK
003650, GBK 003682, GBK 003701, GBK 003985, GBK
003696, GBK 003723, KAB 1, KAT 3, MAR.2, GBK
003660, GBK 003693, GBK 003666, GBK 005173, GBK
027036, MAC 3, GBK 003662, GBK 003698, GBK 003663,
GBK 003668, GBK 003685, GBK 027079, GBK 003697,
GBK 003876, GBK 003701, KAB 3, GBK 003706, GBK
003717, MAC 1, GBK 003711,GBK 00394, GBK 003711,
GBK 003713, GBK 003804, GBK 003726, KENKUNDE,
M66, MAR.5
C II 0.70 3 KIP 1, MBL,GBK 003642
DIII 0.67 8 GBK 003657, GBK 003676, GBK 005173, GBK 003690,
GBK 034722, KOL 1, GBK 003816, GBK 003814
DIV 0.71 8 GBK 003642, GBK 046540, GBK 003658, GBK 003888,
KOL 9B, GBK 003667, KOL 9, GBK 003676

4.3 Molecular characterization based on SSR markers

Twenty primer pairs (forward and reverse) were used in genetic characterization of 110
cowpea accessions from semi arid region of Kenya. Seven of the SSR primer pairs did
not amplify any fragment and five more generated monomorphic allelic amplifications
across all the tested accessions; all these were excluded in the analysis as they did not

34
show any variations between the genotypes. Figure 4.2 shows the monomorphic bands
while Figure 4.3 and Figure 4.4 show polymorphic bands.

M 1 2 3 4 5 6 7 8 9 10 11
12

Figure 4.2: Monomorphic band on Agarose gel for primer SSR 6356

M 1 2 3 4 5 6 7 8 9 10 11
12

Figure 4.3: Polymorphic bands on Agarose gel for primer SSR 6608

35
 M 1 2 3 4 5 6 7 8 9 10 11
12

Figure 4.4: Polymorphic band on Agarose gel for PCR products for SSR 6243

Key:
[Link] 003642 [Link] 003652
[Link] 003642 A [Link] 003654
[Link] 003645 [Link] 003657
[Link] 003650 [Link] 003658
[Link] 003651 [Link] 003660
[Link] 003642 [Link] 003662

36
Out of the twenty primers screened, seven pairs of primers were monomorphic, five did
not amplify and eight were polymorphic as shown on Table 4.7.

Table 4.7: Amplification status of the SSR markers

Primer Name Primer sequence Status

Set
1 SSR-6265F 5’-CAG AAG CGG TGA AAA TTG AAC -3’ Polymorphic
SSR-6265R 5’- GCA TGT TGC GAC AAT GG-3’
2 SSR-6258F 5’- GGT TTC CTA GTT GGG AAG GAA-3’ Did not amplify
SSR-6258R 5’-ATT ATG CCA TGG AGG GTT CA-3’
3 SSR-6243F 5’-GTA GGG AGT TGG CCA CGA TA-3’ Polymorphic
SSR-6243R 5’-CAA CCG ATG AAA AAG TGG ACA-3’
4 SSR-6218F 5’-GTG GAA GGA ATG GGT CCA G-3’ Monomorphic
SSR-6218R 5’-AGG AAA TTT GCA TTC CCT TGT-3’
5 SSR-6217F 5’-GGG AGT GCT CCG GAA AGT-3’ Polymorphic
SSR-6217R 5’-TTC CCT ATG AAC TGG GAG ATC-3’
6 SSR-6353F 5’-TCA TGG GTT AAA TTT GCT TCA A-3’ Monomorphic
SSR-6353R 5’-AAA CCA TGT GGT TGT TGC AC-3’
7 SSR-6352F 5’-GTT GTG AGC TTC CCC AGA TG-3’ Polymorphic
SSR-6352R 5’-ATT TTT GAA CCC ACC ACC AG-3’
8 SSR-6336F 5’-TGA AAA CAA CGA TAT GCA GAA-3’ Monomorphic
SSR-6336R 5’-TCA GTC TTA GAA TTG AGT TTT C-3’
9 SSR-6323F 5’-CAA AGG GTC ATC AGG ATT GG-3’ Monomorphic
SSR-6323R 5’-TTT AAG CAG CCA AGC AGT TGT-3’
10 SSR-6451F 5’-AAA GAG ATA CAC ATG CCT AAC-3’ Polymorphic
SSR-6451R 5’-GAC CAA CAG CGA CTT TGA GC-3’
11 SSR-6277F 5’-CAC CCC CGT ACA CAC ACA-3’ Did not amplify
SSR-6277R 5’-CAC TTA AAT TTC CAC CAG GCA T-3’
12 SSR-6436F 5’-CAG AAT CCT TGT GAA CCT G-3’ Polymorphic
SSR-6436R 5’-TTT CGC AAT ATG CCC TTT TC-3’
13 SSR-6375F 5’-GCT CGG ATA TGG TCC TGA AA-3’ Polymorphic
SSR-6375R 5’-TCA GTG TCA GCA CCA TCC C-3’
14 SSR-6371F 5’-TGC TCA TCG TGC TTT GTC TT-3’ Did not amplify
SSR-6371R 5’-CAC TTC AGA CTT AGA GCG AAG-3’
15 SSR-6370F 5’-CAA CTT CAC AGC CCT CAA-3’ Did not amplify
SSR-6370R 5’-TTG AAG GTA TGG CCT TTT GTT T-3’
16 SSR-6356F 5’-TGC AAT ATG GAC CAG AAG AAA-3’ Monomorphic
SSR-6356R 5’-ATG CCC CAA CAA CAA CAT TT-3’
17 SSR-6613F 5’-CTA TTG GAA TCT TGC CGT TG-3’ Monomorphic
SSR-6613R 5’-CTT TAC CTT TAT GCA AAC CAA T-3’
18 SSR-6608F 5’-CTA AAT TAT AAT ATT CGT CGG T-3’ Polymorphic
SSR-6608R 5’-GGT TAA GGA AAA GAG GGT AGG-3’
19 SSR-6603F 5’-GAG AAC TTC ACG CAC AAT AG-3’ Monomorphic
SSR-6603R 5’-CGC GGT AGC ATG ATT GAA TTT-3’
20 SSR-6581F 5’-GAT ATA GAA TAG CAT ATT TAA C-3’ Did not amplify
SSR-6581R 5’-GTT GAA AGT TTG ATA GTA AAG-3’

37
Figure 4.5: Shows the pair wise values means within the populations

There were five populations based on the sources of accessions. Evaluation was carried
out to determine variation among these different classes of the materials. Analysis of
molecular variance (AMOVA) was done to determine the total genetic variation among
and within the populations as shown in Table 4.8 and Appendix 7. It was established that
there is no variation among the populations. All the variation (100%) was accounted for
within the population (among the genotypes). The genetic distance among the accessions
ranged from 0.006 to 0.105

Table 4.8: Allelic Molecular Variance of the cowpea accessions

Source of variation Df SS MS Est. Var. Genetic variation (%)

Among Pops 4 6.079 1.520 0.005 0%
Within Pops 102 150.483 1.475 1.475 100%
Total 106 156.561 1.480 100%

38
Figure 4.6: Variation of band patterns across populations

Table 4.9: Pairwise population matrix of Nei Genetic distance and Nei Genetic
Identity among cowpea populations

Makueni Machakos Kitui Commercial Baringo

Makueni - 0.955 0.918 0.901 0.931
Machakos 0.079 - 0.991 0.927 0.994
Kitui 0.130 0.023 - 0.963 1.003
Commercial 0.147 0.087 0.061 - 0.964
Baringo 0.111 0.015 0.018 0.054 -

39
The genetic variation observed among the different populations was low with the least
distance genetic distance of 0.015 observed between Baringo and Machakos while the
greatest genetic distance of 0.147 observed between Commercial seeds and Makueni.
The lowest genetic distance among semi arid regions was observed between Kitui and
Machakos with genetic distance of 0.023 while the highest genetic distance was
observed between Kitui and Makueni with genetic distance of 0.130 (Table 4.9).

An average of 1.86 alleles was produced by Machakos population compared to 0.79

generated by Makueni population. uHe values are higher in all regions compared to he
He values. Machakos population recorded the highest He (0.246) and uHe (0.247) values
respectively while Commercial seeds recorded the lowest He (0.15) and uHe (0.167)
respectively.

Table 4.10: Estimated genetic diversity of accessions within populations

Population N Na Ne I He uHe P%
Makueni 2.000 0.793 1.268 0.229 0.157 0.209 37.93
Machakos 86.000 1.862 1.387 0.384 0.246 0.247 93.1
Kitui 5.000 1.103 1.316 0.278 0.185 0.206 51.72
Commercial 5.000 1.207 1.227 0.237 0.150 0.167 51.72
Baringo 9.000 1.517 1.315 0.323 0.205 0.217 72.41
Total 107
Mean 0.189 0.209 61.38
0.015 0.017
N= Number of individuals

Na = No. of Different
Alleles
Ne = No. of Effective Alleles = 1 / (p^2 + q^2)
I = Shannon's Information Index = -1* (p * Ln (p) + q *
Ln(q))
He = Expected Heterozygosity = 2 * p * q
uHe = Unbiased Expected Heterozygosity = (2N / (2N-1))
* He
Where for Diploid Binary data and assuming Hardy-Weinberg Equilibrium, q = (1 - Band Freq.)^0.5 and p
= 1 - q.

P%= percentage of polymorphic loci

40
The molecular data generated from the informative SSR markers clustered the 107
accessions into three main clusters; I, II and III (Fig 6). Cluster I had the highest number
of accessions (54), Cluster II had 44 accessions while cluster II had the least number of
accessions (9). It was observed that the accessions did not cluster based on their place of
origin rather they were distributed across the clusters.

41
Figure 4.7: Clustering of cowpea accessions based on variation in SSR markers

42
 CHAPTER FIVE

DISCUSSION

5.1 Discussion

Genetic diversity is an important aspect in plant breeding programmes. It informs on

whether progress can be made in selection for desirable attributes from a population. It
also provides an indication of richness and distribution of the alleles available in the
population. The accessions in this study comprised of landraces originally collected
from the semi arid region of the country and conserved in the National Gene Bank of
Kenya, collections from farmers’ fields in semi arid region and commercial lines listed
by the Registrar of plant varieties (KEPHIS). Skewed or uneven distribution of
accessions among the qualitative characters was recorded for terminal leaflet shape,
raceme position, pod attachment, pod curvature, mature pod colour, flower colour and
eye colour. The low occurrence of accessions in some categories of the respective traits
of the characters indicated that some of the alleles were rare in the population or were
recessive. Even distribution of accessions in traits was observed for immature pod
colour, leaf colour, seed shape and testa texture. Such even distribution of the accessions
among the various categories of traits could be attributed to non-selective pressure
among such characters.

In this study, 95% of the accessions produced purple flowers which corroborated with
Doumbia et al. (2013), who similarly established that most accessions produced purple
flowers, followed by white flowers and the least were white-purple flowered accessions.
In a similar study, Cobbinah et al. (2011) also found that majority of accessions gave
purple flowers. Unlike Cobbinah et al. (2011) and Doumbia et al., (2013), this study
showed that the accessions with white flowers were less than the white-purple flowered
accessions. This could be attributed to ecological and climatic conditions or the farmers’
preferences that may not favor accessions with white flowers. In another study, Gibbon
and Pain (1985) showed that there were additional flower colours of cowpea such as
pale-blue, yellow and pink though they were not observed in this study (Doumbia et al,
2014). Sangwan and Lodhi (1998) indicated that purple flower colour is dominant over
white which has a monogenic recessive nature of inheritance.

Five classes of immature pod pigmentation (IBPGR, 1983) were found in this study.
This compared favorably with Nkouannessi (2005) who reported four classes and
Doumbia (2011) who had six groups of immature pod pigments.

Raceme position plays a very important role in the harvesting of mature pods. When
racemes are on the same level as the canopy or within the canopy, harvesting becomes
difficult as most pods are hidden. Pandey and Ngarm (1985) and Bennett-Lartey and
Ofori (1999) indicated that accessions which bear racemes above the canopy are easier
and cheaper to harvest compared to racemes borne throughout the canopy. Above
canopy raceme accessions encourage use of mechanical harvesting while those borne
within or throughout the canopy require uprooting of the whole plant (Cobbinah et al.,
2011). Cobbinah et al. (2011), observed that 59.7% of the accessions had above canopy
racemes, 29.8% had the same level as the plant racemes and 10.4% had racemes within
the canopy. In contrast, this study showed that 56% of the accessions produced upper
canopy racemes, 24% of the accessions had a raceme position above canopy and 20%
had raceme throughout the canopy. Grain cowpea farmers in the marginal areas prefer
cowpea which has raceme position above the canopy.

Nkoiannessi (2005), showed that seed testa texture ranged from rough to wrinkled. On
the other hand, Adebowale et al., (2011) reported accessions with smooth to rough seed
texture. This study showed that 89% of the accessions had smooth textured seeds, 7%
smooth to rough textured seeds and 4% rough to wrinkled textured seeds. Smooth seed
coat texture is preferred in Eastern Africa, unlike in West Africa where preference for

44
rough seed coat allows for easy removal of the seed coat that is important for indigenous
food preparations (Singh & Ishiyaku, 2000).

Commercial lines; K80, Kenkunde, M66, Kvu-27-1 and KAR 1 were classified together
in group two by cluster analysis. This could be attributed to breeding and selection of
these varieties for the region where they have been supplied as government interventions
following drought as a mitigation measure (Recha et al., 2012). Accessions obtained
from farmers in Kola of Machakos County were evenly distributed in all clusters of the
accessions. The even distribution of accessions across groups was also observed for
materials procured from farmers in Baringo and those from Machakos counties. The
even distribution is an indication of sharing and exchange of seed among the farmers.
Therefore, there is no clear pattern in distribution of the accessions associated with areas
of origin. The farmers also seem to have similar preferences in the attributes of the
cowpea.

Based on variation in SSR markers, the results indicated a low level of genetic diversity
among the cowpea genotypes as noted from principal coordinates analysis which did not
cluster the accessions in any specific groupings. Asare et al. (2010), used 25 pairs of
SSR markers. Of these, 20 pairs of SSR markers gave reproducible polymorphism.
These primer combinations gave a total of 74 alleles at 20 loci with 3.8 alleles per locus
on average. According to Ogunkanmi et al. (2014), 12 SSR markers generated 37 alleles
with the number of alleles per locus ranging from 2 to 5 and an average of 2.92 alleles
per locus. Ali et al. (2015) studied the genetic diversity and phylogenetic relationships
among 252 cowpea accessions collected in six geographical regions of Sudan using 18
pairs of SSR markers. Sixteen primers gave reproducible polymorphism and a total of
129 alleles were detected with an average of 8.1 alleles per locus. In this study, a total of
eight markers generated a total of 29 alleles with the number of alleles per locus ranging
from 2 to 5 and an average of 3.63 alleles per locus.

45
Asare et al. (2010) reported low genetic variability among Ghanaian cultivated
genotypes. Doumbia (2011), reported low level of similarity between and within the
accessions. Kuruma et al. (2008) reported similar results as the current study; low level
of genetic diversity among cowpea genotypes. The high similarity among the accessions
indicates high levels of geneflow among the populations (Doumbia, 2011). Ali et al,
(2015) found out that there was low genetic diversity among Sudanese cowpea
population with more variations within individuals.

46
 CHAPTER SIX

CONCLUSION AND RECOMMENDATIONS

6.1 Conclusion

The accessions used in the trial were variable in the characters that they were evaluated
in. Accessions similarly were evenly distributed across the traits of the various
characteristics; an indication of a wide range of alleles in the accessions used in the
study. The study showed that there was a high level of variation among the cowpea
accessions with respect to qualitative and quantitative traits. In terms of a performance
index derived from yield associated attributes; GBK 003662, GBK 003663, GBK
003676, GBK 003723, GBK 003650 and GBK 003642 were among the top 5%
accessions in the trial. The study showed that there was a high level of similarity among
the cowpea accessions with respect to SSR markers. The SSR markers used in this study
showed no variation among the cowpea accessions.

6.2 Recommendations

All the variations were found within the individual accessions. Accessions GBK 003662,
GBK 003663, GBK 003676, GBK 003723, GBK 003650 and GBK 003642 were among
the top 5% high yielding accessions in the trial. These superior accessions could be
adopted widely for cultivation by farmers. Future studies should explore using more
SSR primers in order to detect the variation among accessions in this species. Moreover,
a larger sample of cowpea accessions should be included in studies.

47
 REFERENCES

Acquaah, G. (Ed) (2012). Principles of Plant Genetics and Breeding. MA: Wiley-
Blackwell Publishing.

Adebowale, B.D., Adeigbe, O.O. & Aremu C.O. (2011): Genetic distance and diversity
among some cowpea (Vigna unguiculata L. Walp) genotypes: International
Journal of Research in Plant Science. 1(2), 9-14

Ali, Z.B., Yao, K.N., Odeny, D.A, Kyalo, M., Skilton, R. & Eltahir I.M. (2015).
Assessing the genetic diversity of cowpea (Vigna unguiculata (L.) Walp.)
accessions from Sudan using simple sequence repeat (SSR) markers. African
Journal of Plant Sciences. 9(7) 293-304.

Andersson, M.S. & Vicente, M.C. (2010). Gene flow between crops and their wild
relatives. Evolutionary Applications. 3(4), 402-403

Aremu, M. O., Ogunlade, I., & Olonisakin, A. (2007). Fatty acid and amino acid
composition of protein concentrate from cashew nut (Anarcadium
occidentale) grown in Nasarawa State, Nigeria, Pakistan Journal of
Nutrition 6(5), 419–423.

Asare A.T, Gowda B.S, Galyuon I.K.A, Aboagye L.M, Takrama J.F and Timko M.P.
(2010) Assessment of the genetic diversity in cowpea (Vigna unguiculata
(L)Walp) germplasm from Ghana using simple sequence repeat markers.
Plant Genetic Resources -C 8(2), 142-150

Ashraf, M.Y., Mahmood, K., Ashraf M., Akhter, J. & Hussain, F. (2012). Optimal
Supply of micronutrients improves drought tolerance in legumes. In: Crop
production for agricultural improvement1, (eds.) Ashraf, M., Ozturk, M,

48
 Ahmad, M. S. A. DOI 10.1007/978-94-007-4116-4-25, New York: Springer.

Asiwe, J.A.N. (2009). Insect mediated outcrossing and gene flow in cowpea Vigna
unguiculata (L.) Walp: implication for seed production and provision of
containment structures for genetically transformed cowpea. Africa Journal of
Biotechnology, 8(2), 226-230.

Ba, F.S., Pasquet, R.E. & Gepts, P. (2004). Genetic diversity in cowpea [Vigna
unguiculata (L.) Walp.] as revealed by RAPD markers. Genetic Resources
and Crop Evolution. 51, 539-550.

Bennet-Lartey, S.O. & Ofori, K. (1999). Variability studies in some qualitative

characters of cowpea (Vigna unguiculata (L) walp) accessions from four
cowpea growing regions in Ghana. Ghana journal of Agricultural Science,
32, 3-9.

Bennett, D.J. & Jennings, R.C. (Eds) (2013). Successful Agricultural Innovation in
Emerging Economies: New Genetic Technologies or Global Food
Production. New York: Cambridge University Press.

Betal, S., Chowndry, P.R., Kundu, S. & Raychaunduri, S.S. (2004). Estimation of
genetic variability of vigna radiate cultivars by RAPD analysis. Biological
Planarum 48, 205-209.

Bewley, J.D., Black, M. & Halmer, P. (2006). The Encyclopedia of Seeds: Science,
Technology and Uses. Wallingford, UK: CABI Publishing Series

Casanas, F., Simo J., Casals J. & Prohens J. (2017). Towards an evolved concept of
landrace. Frontiers in Plant Science, 8, 145.

49
Cobbinah, F. A., Addo-Quaye A.A. & Asante, I. K. (2011). Characterization, evaluation
and selection of cowpea (Vigna unguiculata (L.) Walp ) Accessions with
desirable traits from eight regions of Ghana. ARPN Journal of Agricultural
and biological science, 6(7), 21–32.

Doumbia, I. Z., Akromah, R. & Asibuo, J.Y. (2014). Assessment of cowpea germplasm
from Ghana and Mali using simple sequence repeats (SSR) markers.
International Journal of Agriculture and Forestry, 4(2), 118-123.

Doumbia, I. Z., Akromah, R., & Asibuo, J.Y. (2013). Comparative study of cowpea
germplasms diversity from Ghana and Mali using morphological
characteristics. Journal of Plant Breeding and Genetics, 01(03), 139–147.

Doumbia, I.Z. (2011). Comparative Study of Cowpea Germplasm from Ghana and Mali
using Morphological and Molecular markers. Unpublished MSc Thesis,
Kumasi: Kwame Nkrumah University of Science and Technology.

Doyle, J.J. & Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus 12, 13-
15

Gachimbi, L.N., Kamoni, S.N., Macharia, P.N. & Gicheru, P.T. (2007). Using farmer
field school approaches to overcome land degradation in agro-pastoral areas
project: Land use practices in Mbeere District: Biophysical and economic
challenges, copping strategies and opportunities: A baseline survey report.
Retrievedfrom [Link]

Gómez, C. (2004). Cowpea: Post-harvest Operations. In: Mejia (Ed.), Post-Harvest

Compedium, AGST: FAO.

Grubben, G.J.H & Denton, O.A. (2004). Plant Resources of Tropical Africa 2.

50
 Vegetables., Wagenigen: PROTA Foundation.

Hall, A.E., Cisse, N., Thiaw, S., Elawad, H.O.A., Ehlers, J.D., Ismail, A., Fery, R., … &
McWatters, K.H. (2003), “Development of cowpea cultivars and germplasm
by the bean/cowpea CRSP”, Field Crops Research, 82(2-3), 103-134,

IBPGR. (1983). Descriptors for Cowpea. Zhurnal Eksperimental’noi I Teoreticheskoi

Fiziki, 1–30.

Jaetzold, R., & Schmidt, H. (1983). Farm Management Handbook of Kenya. Ministry of
Agriculture, Kenya, in Cooperation with the German Agricultural Team
(GAT), German Agency for Technical Cooperation (GTZ) 2, 245-285

Joshua O.O., Abong G., Okoth M. & Mwang’ombe A.W. (2019). A review of the
contribution of cowpea leaves to food and nutrition security in East Africa.
Food Science and Nutrition; 8(1), 36-47.

Kameswara, R.N. (2004). Biotechnology for Plant Resources Conservation and Use.
Principles of Seed Handling in Genebanks Training Course, Kampala,
Uganda. African Journal of Biotechnology 3 (2), 136-145

Karanja, D. (2016). Pulses crops grown in Ethiopia, Kenya and United Republic of
Tanzania for local and Exports Market. Tanzania: International Trade
Centre, Eastern Africa Grain Council.

Kenya Plant Health Inspectorate Service (KEPHIS) (2019). National Crop Variety List-
Kenya. Retrieved from [Link]/images/uploads/[Link].
KEPHIS.

Kuruma, R.W., Kiplagat, O., Ateka, E. & Owuoche G. (2008). Genetic diversity of

51
 Kenyan cowpea accessions based on morphological and microsatellite
markers. East African Agricultural and Forestry Journal, 76, 3-4.

Lambrides, C.J., Lawn, R.J., Godwin, I.D., Manners, J. & Imrie, B.C (2000). Two
genetic linkage maps of mungbean using RFLP and RAPD markers.
Australian Journal of Agricultural Resources, 51, 415-425.

Lesley D.W. (2005). Characterization and Evaluation of Cowpea (Vigna unguiculata

[L.]Walp) Germplasm. Unpublished MSc Thesis. Dharwaad: University of
Agricultural Sciences.

Li, CD, Fatokun, CA, Ubi, B, Singh, BB, & Scoles, GJ (2001) Determining genetic
similarities and relationships among cowpea breeding lines and cultivars by
microsatellite markers. Crop Science Journal, 41, 189-197.

Madamba, R., Grubben, G.J.H., Asante, I.K. & Akromah, R. (2006). Vigna unguiculata
(L) Walp. In: Brink, M. and Belay, G.I. (eds). PROTA (Plant Resources of
Tropical Africa/ Resources vegetales de I’Afrique tropicale) Wagenigen
PROTA.

Mafakheri, K., Mohammed, R.B. & Ali, R.A. (2017). Assessment of genetic diversity in
Cowpea (vigna unguiculata) germplasm using morphological and molecular
characterization. Congent Food and Agriculture.
3.10.1080/23311932.2017.1327092

Makari, M.C. (2022). Growth and Yield Responses of Beans, Cowpea and Bambara nuts
to Phosphorus Fertilization in Kakamega County. Unpublished MSc. thesis,
Nairobi: Kenyatta University.

Malek, M.A., Rafii, M., Afroz, M., Nath, U. & Mondal, M. (2014). Morphological

52
 characterization and assessment of genetic variability, character association
and divergence in soybean mutants. The Scientific World Journal. 2014.
968796. 10. 1155/2014/968796.

Ministry of Agriculture, Livestock and Fisheries (2015). Economic Review for

Agriculture. Nairobi: Ministry of Agriculture, Livestock and Fisheries.

Muniu, F.K. (2017). Characterization and Evaluation of Local Cowpea Accessions and
Their Response to Organic and Inorganic Nitrogen Fertilizers in Coastal
Kenya, Unpublished MSc. Thesis, Nairobi: University of Nairobi.

Muthoni J. & Nyamongo, D. O. (2008). 'Traditional food crops and their role in food and
nutritional security in Kenya, Journal of Agricultural & Food Information,
11(1), 36-50.

Muthoni, J. and Nyamongo D.O (2008). Seed systems in Kenya and their relationship to
on farm conservation of food crops. Journal of New Seeds, 9(4) 330-342

Nagarajan, L., Audi, P., Jones, R. & Smale, M. (2008). Seed Provision and Dryland
Crops in the Semi-arid Areas of Eastern Kenya, IFRI Discussion Paper No.
78

Njonjo, M.W (2018). Quality of Cowpea Seed used by Farmers in Makueni and Taita
Taveta Counties and its Effect on Crop Performance. Unpublished MSc
Thesis. Nairobi: University of Nairobi.

Nkouannessi, M. (2005). The Genetic, Morphological and Physiological Evaluation of

African Cowpea Genotypes. MSc. thesis, Bloemfontein: University of the
Free State.

53
Ogunkanmi, L.A., Ogundipe, O.T. & Fatokun, C.A. (2014). Molecular characterization
of cultivated cowpea (Vigna unguiculata [Link]) using simple sequence
repeat markers. African Journal of Biotechnology. 13(34), 3464-3472.

Olal, D.A (2015). Determining Quantity of Cowpea (Vigna unguiculata) Leaf Yield
under Different Manure Application Regimes and Cropping Systems in
Western Kenya. Unpublished MSc. Thesis, Eldoret: University of Eldoret.

Oluwakemi, M.O., Olatunji, F. and owoade, F.M. (2021). Response of cowpea varieties
to sources and rates of Phosphorus in Ogbomoso. Global Scientific Journals.
Vol. 9(10), 2091-2143

Omoigui, L.O., Kamara, A.Y., Batieno, J., Iorlamen, T., Kouyate, Z., Yirzagla, J.,
Diallo, S. & Garba, U. (2018). Guide to Cowpea Production in West Africa,
Ibadan: IITA.

Pratap, A. & Kumar, J. (Eds) (2011). Biology and Breeding of Food legumes.
Wallingford: CABI.

Raikhel, N.V. (2001). One Year later. The state of the journal Plant Physiology, 126 (1)
3-4.

Recha, J., Kinyangi, J. & Omondi, H. (2012). Climate Related Risk and opportunities for
Agricultural adaptation and mitigation in semi arid eastern Kenya. Climate
Change Agriculture and Food Security. CGIAR. Retrieved from
[Link]
org/sites/default/files/assets/doc/climate_related_risk_and_opportunities.pdf.

Rubyogo, J.C., Sperling, L. & Assefa, T. (2007). A new approach for facilitating access
to bean seed. LEISA Magazine, 23(2), 27-29.

54
Sariah, J. E. (2010). Enhancing Cowpea (Vigna unguiculata L.) Production Through
Insect Pest Resistant Lines in East Africa. Unpublished PhD Thesis,
Denmark: University of Copenhagen.

Savala C.E.N., Omare M.N. & Woomer P.L. (Eds). (2003). Organic resource
management in Kenya: perspectives and guidelines. Forum for organic
resource management and agricultural technologies, Nairobi, Kenya. 184.

Setimela P.S, Monyo E. & Banziger M. (Eds). (2004). Successful Community-Based

Seed Production Strategies. Mexico, D.F.: CIMMYT.

Sheidu, A. & Igyuve, T.M. (2023). Correlatin analysis on yield components of cowpea
genotypes (Vigna unguiculata L. Walp). EAS Journal of Anaestheiology and
critical care. 5(1), 7-10.

Singh, B.B & Ishiyaku, M. F. (2000). Genetics of rough seed coat texture in cowpea.
Journal of Heredity, 91, 170-174.

Singh, B.B. (2002). Recent genetic studies in cowpea. In challenges and opportunities
for enhancing sustainable cowpea production. IITA Annual Report.
Retrieved from [Link]

Sperling, L., Remington, T., Haugen, J.M. & Nagoda, S. (eds). (2004). Addressing seed
security in disaster response: Linking relief with development, overview.
International Centre for Tropical Agriculture.

Stoilova, T., & Pereira, G. (2013). Assessment of the genetic diversity in a germplasm
collection of cowpea (Vigna unguiculata (L.) Walp.) using morphological
traits. African Journal of Agricultural Research, 8(2), 208–215.

55
Timko, M.P & Singh, B.B. (2008). Cowpea, a Multifunctional Legume, in: Moore, P.H.,
Ming R. (Eds), Genomics of Tropical Crop Plants., New York: Springer

Timko, M.P., Elhers, J.D., & Roberts, P.A., (2007). Cowpea. In: Kole, C. (eds) Pulses,
Sugar and Tuber crops. Genome mapping and molecular breeding in plants.
Vol.3., Berlin, Heidelberg: Springer.

Wamalwa, N.E, John, M. & Clabe, W (2016). Genetic diversity of cowpea (Vigna
Unguiculata (L.) Walp.) accessions in Kenya Genebank Based on Simple
Sequence Repeat Markers. International Journal of Genomics, 2016, 2016,
8956412.

Wang, X.W, Kaga, A., Tomooka, N. & Vaughan, D.A (2004). The development of SSR
markers by a new method in plants and their application to gene flow studies
in azuki bean (Vigna angularis (wild.) ohwi & ohashi). Theoretical Applied
Genetics 109, 352-360.

Woomer, P.L., Langat, M., & Tungani, J.O., (2004). Innovative maize–legume
intercropping results in above- and below-ground competitive advantages for
understory Legumes. West African Journal of Applied Ecology. 6, 85–94.

Zia-ul-haq, M., Ahmad, S., Chiavro, E.,Mehjabeen & Ahmed, S. (2010). Studies of oil
from cowpea (Vigna unguiculata (L) walp) cultivars commonly grown in
Pakistan. Pakistan Journal of Botany, 42(2), 1333-1341

Zong, W.X., Li, C., Hatzivassiliou, G., Lindsten, T., Yu, Q.C., Yuan, J., & Thompson,
C.B. (2003). Bax and Bak can localize to the endoplasmic reticulum to
initiate apoptosis. Journal of Cell Biology, 162, 59–69.

56
 APPENDICES

Appendix I: Cowpea accessions used in the study and their collection area

Source
Accession Area/locality Latitude Longitude
County
GBK-003642 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003642 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003645 Machakos KDFS 0º 35´S 37º 15´E
GBK-003650 Machakos KDFS 0º 35´S 37º 15´E
GBK-003651 Machakos KDFS 0º 35´S 37º 15´E
GBK-003652 Machakos KDFS 0º 35´S 37º 15´E
GBK-003654 Machakos KDFS 0º 35´S 37º 15´E
GBK-003657 Machakos KDFS 0º 35´S 37º 15´E
GBK-003658 Machakos KDFS 0º 35´S 37º 15´E
GBK-003660 Machakos KDFS 0º 35´S 37º 15´E
GBK-003662 Machakos KDFS 0º 35´S 37º 15´E
GBK-003663 Machakos KDFS 0º 35´S 37º 15´E
GBK-003666 Machakos KDFS 0º 35´S 37º 15´E
GBK-003667 Machakos KDFS 0º 35´S 37º 15´E
GBK-003668 D Machakos KDFS 0º 35´S 37º 15´E
GBK-003669 Machakos KDFS 0º 35´S 37º 15´E
GBK-003670 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003670 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003674 Machakos KDFS 0º 35´S 37º 15´E
GBK-003675 Machakos KDFS 0º 35´S 37º 15´E
GBK-003676 A Machakos KDFS 0º 35´S 37º 15´E

57
 Source
Accession Area/locality Latitude Longitude
County
GBK-003676 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003680 Machakos KDFS 0º 35´S 37º 15´E
GBK-003682 Machakos KDFS 0º 35´S 37º 15´E
GBK-003685 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003685 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003687 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003687 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003688 Machakos KDFS 0º 35´S 37º 15´E
GBK-003689 Machakos KDFS 0º 35´S 37º 15´E
GBK-003690 Machakos KDFS 0º 35´S 37º 15´E
GBK-003693 Machakos KDFS 0º 35´S 37º 15´E
GBK-003694 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003694 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003695 Machakos KDFS 0º 35´S 37º 15´E
GBK-003696 Machakos KDFS 0º 35´S 37º 15´E
GBK-003697 Machakos KDFS 0º 35´S 37º 15´E
GBK-003698 Machakos KDFS 0º 35´S 37º 15´E
GBK-003699 Machakos KDFS 0º 35´S 37º 15´E
GBK-003700 Machakos KDFS 0º 35´S 37º 15´E
GBK-003701 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003701 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003705 Machakos KDFS 0º 35´S 37º 15´E
GBK-003706 Machakos KDFS 0º 35´S 37º 15´E
GBK-003707 Machakos KDFS 0º 35´S 37º 15´E

58
 Source
Accession Area/locality Latitude Longitude
County
GBK-003709 Machakos KDFS 0º 35´S 37º 15´E
GBK-003711 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003711 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003713 Machakos KDFS 0º 35´S 37º 15´E
GBK-003714 Machakos KDFS 0º 35´S 37º 15´E
GBK-003717 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003717 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003718 Machakos KDFS 0º 35´S 37º 15´E
GBK-003720 Machakos KDFS 0º 35´S 37º 15´E
GBK-003723 Machakos KDFS 0º 35´S 37º 15´E
GBK-003724 Machakos KDFS 0º 35´S 37º 15´E
GBK-003726 Machakos KDFS 0º 35´S 37º 15´E
GBK-003727 Machakos KDFS 0º 35´S 37º 15´E
GBK-003780 Machakos KDFS 0º 35´S 37º 15´E
GBK-003796 Machakos KDFS 0º 35´S 37º 15´E
GBK-003804 Machakos KDFS 0º 35´S 37º 15´E
GBK-003814 Machakos KDFS 0º 35´S 37º 15´E
GBK-003816 A Machakos KDFS 0º 35´S 37º 15´E
GBK-003816 B Machakos KDFS 0º 35´S 37º 15´E
GBK-003820 Machakos KDFS 0º 35´S 37º 15´E
GBK-003876 Machakos KDFS 0º 35´S 37º 15´E
GBK-003888 Machakos KDFS 0º 35´S 37º 15´E
GBK-003916 Machakos KDFS 0º 35´S 37º 15´E
GBK-003985 Machakos KDFS 0º 35´S 37º 15´E

59
 Source
Accession Area/locality Latitude Longitude
County
GBK-005173 A Machakos KDFS 0º 35´S 37º 15´E
GBK-005173 B Machakos KDFS 0º 35´S 37º 15´E
Ndalani Loc. 1º 45’8’S 37º 19’5´E
(Ndalani
GBK-027036 Machakos Maktano)
Ndalani Loc. 2º 37’25´S 37º 15´E
(Ndalani
GBK-027079 Machakos Maktano)
Mukuyuni, 2º 23’38´S 37º 15´E
GBK-027089 Machakos Kibwezi
Kanyangi Loc. 1º 36’12´S 37º 54’35´E
GBK-026941 A Kitui Wangata
Kanyangi Loc. 1º 36’12´S 37º 54’35´E
GBK-026941 B Kitui Wangata
GBK-026958 A Kitui Mutha Loc. 1º 48’30´S 38º 25’0´E
GBK-026958 B Kitui Mutha Loc. 1º 48’30´S 38º 25’0´E
GBK-046540 Kitui Maliku – Kitui 1º 34.986´S 37º 54.946´E
Sakai; 13km 1º 39´S 37º 35´E
Kilata Tawa
GBK-034722 A Makueni Road
Sakai; 13km 1º 39´S 37º 35´E
Kilata Tawa
GBK-034722 B Makueni Road
Sakai; 13km 1º 39´S 37º 35´E
GBK-036582 Makueni Kilata Tawa

60
 Source
Accession Area/locality Latitude Longitude
County
Road
K80 Commercial Commercial
KENKUNDE Commercial Commercial
KVU-27-1 Commercial Commercial
M66 Commercial Commercial
KAR 1 Commercial KALRO
KAT 1 Machakos Katumani
KAT 3 Machakos Katumani
KOL 1 Machakos Kola
KOL 2 A Machakos Kola
KOL 2 B Machakos Kola
KOL 5 Machakos Kola
KOL 6 Machakos Kola
KOL 8 Machakos Kola
KOL 9 A Machakos Kola
KOL 9 B Machakos Kola
KOL 9 C Machakos Kola
MAC 1 Machakos Machakos
MAC 2 Machakos Machakos
MAC 3 Machakos Machakos
MAR.2 Baringo Margat
MAR.3 Baringo Margat
MAR.5 Baringo Margat
MBL Baringo Mbili mbili
LAM 4 Baringo Lambwe

61
 Source
Accession Area/locality Latitude Longitude
County
KIP 1 Baringo Kipsarum
KIP 2 Baringo Kipsarum
KAB 1 Baringo Kabartonjo
KAB 3 Baringo Kabartonjo

GBK=Gene Bank of Kenya; KDFS=Katumani Dryland Farming Station; A, B,

D=Selections within the accession

62
Appendix II: Data scoring of the traits evaluated during the trial

TRAITS Acronym SCORING

Growth habit GH 1. Acute erect 2. Erect 3. Semi erect 4. Intermediate 5.
Semi prostate 6. Prostate 7. Climbing
Growth pattern GP 1. Determinate 2. Indeterminate
Twinning TT 0 None 3 Slight 5 Intermediate 7 Pronounced
tendency
Pigmentation P 0 None 1 Very slight 3 Moderate at the base and tips
of petioles 5 Intermediate 7 Extensive 9 Solid
Terminal leaflet TLS 1 Globose 2 Sub-globose 3 Sub-hastate 4 Hastate
shape
Raceme position RP 1 Mostly above canopy 2 In upper canopy 3
Throughout canopy
Pod attachment PA 3 Pedant 5 30-90 down from erect 7 Erect
Immature pod 0 None 1 Pigmented tip 2 Pigmented sutures 3
color Pigmented valves, green sutures 4 Splashes of pigment
5 Uniformly pigmented 6 Other
Pod curvature PC 0 Straight 3 Slightly curved 5 Curved 7 Coiled
Mature pod color MPC 1 Pale tan/ straw 2 Dark tan 3 Dark brown 4 Black or
dark purple 5 Other
Flower color FC 1 Purple 2 white-purple 3 White
Leaf color LC 3 Pale green 5 Intermediate green 7 Dark green
Seed shape SS 1 Kidney 2 Ovoid 3 Crowder 4 Globose 5Rhomboid
Eye color EC 0 Absent 1 Brown splash/gray 2 Tan brown 3 Red 4
Green 5 Blue to black 6 Blue to black spots/mottle 7
Speckled 8 Mottled 9 Mottled and speckled 10 Other
Seed coat color SCC 1 White 2 Cream 3 Brown 4 Red 5 Purple 6 Black 99

63
TRAITS Acronym SCORING
Other
Testa texture Tt 1 Smooth 3 Smooth to rough 5 Rough 7 Rough to
wrinkled 9 Wrinkled
Days to 50% E (50%) From planting to the time when 50% of the seeds have
emergence germinated
Days to 50% F (50%) From planting date to when 50% of the plants have
flowering produced flowers
Terminal leaflet TLL Mean length of 10 terminal leaflets from 10 randomly
length selected plants
Terminal leaflet TLW Mean width of 10 terminal leaflets measured on the
width broadest part of 10 randomly selected plants
Pod length PL Mean length of 10 mature pods from 10 randomly
selected plants
Seeds per pod SPP Mean number of the randomly selected pods
100Seed Weight 100SW Mean weight of 100 seeds with moisture content of
12%
Number of pods PPP Mean number of mature pods from 10 randomly
per plant selected plants
Number of main BP At 8 weeks after planting. Mean of 10 randomly
branches per plant selected plants

Adapted from IBPGRI (1983)

64
Appendix III: Ranking based on characteristics associated with performance of
cowpea genotypes

ACC PL SPP 100SW PPP BP Selection Index Ranks

GBK 003662 98 110 106 101 71 486 1
GBK 003663 103 91 77 98 103.5 472.5 2
GBK 003676 108 99 90 88.5 62.5 448 3
GBK 003723 89 88 100 55 109.5 441.5 4
GBK 003650 104 106.5 41 107 71 429.5 5
GBK 003642 79 82.5 70 110 71 412.5 6
GBK 003669 110 106.5 64 95.5 28 404 7
GBK 003668 D 65 108 52 95.5 80.5 401 8
GBK 003780 70 81 103.5 51.5 90 396 9
GBK 003709 54.5 86.5 101.5 66.5 71 380 10
GBK 003685 93.5 96 75 83.5 28 376 11
KAB 1 97 97.5 53.5 24.5 103.5 376 12
GBK 003985 87 46.5 101.5 40.5 96 371.5 13
GBK 003796 48 105 88.5 49 80.5 371 14
GBK 003645 90.5 85 80.5 108 5 369 15
GBK 003676 105 89 35 88.5 45 362.5 16
GBK 003676 B 71 104 31 88.5 62.5 357 17
GBK 003687 51 54 105 83.5 62.5 356 18
GBK 003654 50 72 66 103.5 62.5 354 19
K80 106 102.5 83 28 33.5 353 20
GBK 003696 46 71 68 75 85.5 345.5 21
GBK 003718 75 102.5 85 60 19 341.5 22
GBK 003720 68 67 107 55 38.5 335.5 23
GBK 003675 A 66.5 75.5 78.5 91 19 330.5 24
GBK 003660 88 100 6 101 33.5 328.5 25
GBK 003690 A 13 59 92 79 85.5 328.5 26
KOL 6 90.5 51 83 12.5 90 327 27
GBK 026958 A 92 91 55.5 35.5 52 326 28
GBK 027036 83.5 74 29.5 35.5 103.5 326 29
GBK 003680 15 31 110 88.5 80.5 325 30
KOL 2 72 82.5 92 15 62.5 324 31
GBK 046540 102 75.5 10 28 107.5 323 32
GBK 003685 76 54 63 83.5 45 321.5 33

65
ACC PL SPP 100SW PPP BP Selection Index Ranks
GBK 003705 109 97.5 18.5 66.5 28 319.5 34
GBK 003670 B 59 64.5 98.5 93 3 318 35
GBK 003689 82 78.5 69 79 8.5 317 36
GBK 003674 5 68 47 93 103.5 316.5 37
GBK 003814 49 94 53.5 49 71 316.5 38
MAC 2 85 93 25 6 103.5 312.5 39
GBK 003642 A 41 10.5 60.5 109 85.5 306.5 40
GBK 003697 81 26.5 76 75 45 303.5 41
KIP 1 40 52 103.5 18 90 303.5 42
GBK 003694 B 47 73 15 75 90 300 43
GBK 026941B 62 91 72 35.5 38.5 299 44
GBK 003701 56 28 72 66.5 76 298.5 45
KAR 1 37 109 21.5 24.5 103.5 295.5 46
GBK 003820 101 70 17 44.5 62.5 295 47
MAR.2 86 13.5 94 4 96 293.5 48
GBK 003816 83.5 101 45 44.5 19 293 49
GBK 003652 64 95 12 105.5 13.5 290 50
GBK 003701 1 22 98.5 66.5 100 288 51
GBK 003707 77 44 95.5 66.5 1 284 52
GBK 003916 10.5 54 83 40.5 96 284 53
GBK 003888 93.5 50 62 44.5 33.5 283.5 54
KOL 5 69 61 80.5 15 52 277.5 55
GBK 003670 73 57 35 93 19 277 56
GBK 003688 45 32 20 83.5 96 276.5 57
GBK 003876 95 64.5 26 44.5 45 275 58
KVU-27-1 96 42.5 48 10 76 272.5 59
GBK 003724 33 37 50.5 55 96 271.5 60
GBK 034722 60.5 64.5 5 30.5 109.5 270 61
GBK 003816 66.5 84 28 47 38.5 264 62
GBK 003699 28 61 21.5 71 80.5 262 63
GBK 003693 43.5 19 74 79 45 260.5 64
GBK 003666 19.5 46.5 50.5 98 45 259.5 65
GBK 003706 60.5 40 46 66.5 45 258 66
GBK 003726 31 78.5 86 53 8.5 257 67
GBK 003700 25 46.5 60.5 71 52 255 68
GBK 003658 52 9 59 101 33.5 254.5 69
GBK 026958 32 21 108.5 35.5 56 253 70
66
ACC PL SPP 100SW PPP BP Selection Index Ranks
GBK 003717 A 38.5 78.5 41 60 33.5 251.5 71
GBK 003657 14 10.5 41 103.5 80.5 249.5 72
LAM 4 107 46.5 55.5 9 28 246 73
GBK 003717 B 63 29 78.5 60 13.5 244 74
GBK 003651 58 26.5 38.5 105.5 13.5 242 75
KOL 9 B 99 69 43 11 19 241 76
GBK 003714 34 78.5 4 60 56 232.5 77
GBK 003667 17 34.5 8 98 71 228.5 78
GBK 003804 18 34.5 29.5 49 96 227 79
GBK 005173 80 49 16 40.5 38.5 224 80
GBK 003695 12 17 95.5 75 23.5 223 81
KOL 1 42 64.5 35 18 62.5 222 82
GBK 003711 2 15 88.5 60 52 217.5 83
GBK 036582 6 1 92 28 90 217 84
GBK 003713 54.5 6 67 60 28 215.5 85
GBK 003727 9 7 35 51.5 107.5 210 86
KAT 3 74 41 27 22 45 209 87
GBK 026941 10.5 2 97 35.5 62.5 207.5 88
KAB 3 26 61 7 24.5 85.5 204 89
GBK 027079 78 17 65 35.5 6 201.5 90
KENKUNDE 22 36 108.5 20.5 13.5 200.5 91
GBK 003698 19.5 24.5 14 71 71 200 92
GBK 003694 100 12 3 75 2 192 93
GBK 003711 24 24.5 57.5 60 23.5 189.5 94
KOL 8 29 86.5 49 12.5 8.5 185.5 95
KOL 2 23 33 32 15 80.5 183.5 96
KAT 1 43.5 39 13 24.5 62.5 182.5 97
MBL 53 57 57.5 1 13.5 182 98
MAC 1 30 57 23 7 62.5 179.5 99
GBK 003682 57 5 9 83.5 23.5 178 100
GBK 003687 B 21 17 44 83.5 8.5 174 101
MAR.5 36 23 24 2.5 76 161.5 102
MAR.3 16 38 2 2.5 96 154.5 103
M66 3 42.5 87 8 4 144.5 104
MAC 3 8 4 72 5 52 141 105
KOL 9 C 4 30 38.5 20.5 45 138 106
GBK 005173 B 7 20 35 40.5 33.5 136 107
67
ACC PL SPP 100SW PPP BP Selection Index Ranks
KIP 2 38.5 13.5 1 18 56 127 108
GBK 027089 27 3 18.5 32 23.5 104 109
GBK 034722 35 8 11 30.5 13.5 98 110

68
Appendix IV: Analysis of Molecular Variance

69
Appendix V: Principal coordinates

70
 Appendix VI: Principal component analysis of cowpea genotypes

Axis No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
EigenValue 5.61 3.95 3.88 3.42 3.13 2.71 2.64 2.24 2.16 2.07 1.87 1.68 1.51 1.35 1.19 1.01
GBK 034722 0.12 0.44 -0.16 -0.01 0.03 -0.07 0.15 -0.17 -0.07 -0.03 0.02 -0.01 -0.11 0.05 0.03 0.02
GBK 036582 0.17 -0.11 0.29 0.00 -0.30 0.15 -0.16 0.02 0.30 -0.27 0.05 -0.07 -0.02 -0.08 0.10 -0.02
GBK 003642 A -0.25 -0.20 -0.29 0.00 0.02 -0.05 -0.17 0.07 -0.01 -0.13 -0.07 -0.15 -0.26 0.04 -0.09 0.14
GBK 003642 B 0.08 0.19 -0.10 -0.35 -0.32 -0.27 -0.20 0.14 0.08 -0.10 -0.02 0.14 0.06 -0.06 -0.13 0.11
GBK 003645 0.24 -0.02 0.07 0.19 -0.45 0.02 -0.20 -0.13 -0.17 0.16 0.02 0.25 -0.05 0.02 -0.09 -0.06
GBK 003650 0.19 -0.17 0.35 0.05 -0.29 -0.03 -0.19 0.00 0.33 0.07 0.16 0.00 -0.10 0.21 0.14 0.02
GBK 003651 0.32 -0.12 -0.09 0.12 0.11 -0.15 -0.09 0.02 -0.09 -0.20 0.07 0.08 0.10 0.03 -0.01 0.05
GBK 003652 0.17 -0.11 -0.07 0.13 0.01 -0.15 -0.15 0.22 0.14 -0.28 0.14 0.00 0.08 0.14 0.12 -0.06
GBK 003654 0.01 -0.15 -0.26 -0.31 -0.19 0.03 0.06 0.18 -0.09 0.05 0.13 0.00 0.18 0.03 0.04 -0.05
GBK 003657 -0.38 0.06 -0.26 0.31 -0.18 -0.18 -0.08 -0.29 0.06 0.12 0.10 -0.05 0.06 0.01 0.04 -0.05
GBK 003658 0.24 0.05 -0.09 -0.01 0.35 0.15 -0.02 -0.03 0.25 -0.04 0.05 0.19 0.07 -0.03 -0.20 -0.10
GBK 003660 0.04 0.08 0.17 -0.11 0.18 0.25 -0.04 0.33 -0.13 -0.09 0.06 -0.01 0.04 -0.10 0.12 -0.07
GBK 003662 0.24 0.09 -0.17 0.26 -0.18 0.08 0.03 0.00 0.19 -0.30 -0.12 -0.07 0.08 0.08 -0.10 0.07
GBK 003663 0.10 0.26 -0.02 -0.16 0.14 0.17 0.21 0.12 -0.12 0.21 -0.06 -0.15 -0.05 0.00 -0.06 -0.05
GBK 003666 0.24 0.15 0.07 0.15 -0.29 0.15 0.13 0.14 -0.15 -0.04 -0.19 0.26 -0.09 -0.14 -0.14 -0.02
GBK 003667 0.27 -0.23 -0.10 0.01 -0.08 0.03 -0.15 -0.13 -0.12 0.00 -0.11 -0.17 0.04 -0.01 0.12 0.01
GBK 003668 D -0.14 -0.16 0.18 0.34 0.16 0.20 0.07 0.04 0.14 -0.10 -0.08 -0.10 0.19 0.14 -0.08 0.13
GBK 003669 0.23 0.14 -0.07 0.05 0.18 -0.02 0.14 -0.19 0.13 -0.26 0.08 0.07 -0.05 0.08 0.05 0.17
GBK 003670 A 0.06 0.01 -0.06 0.17 0.00 0.31 -0.15 -0.18 -0.09 0.03 -0.08 -0.16 -0.04 -0.20 0.03 -0.04
Axis No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
GBK 003670B -0.17 -0.08 -0.04 0.09 0.04 0.15 -0.18 -0.40 0.04 0.04 -0.21 0.11 0.06 -0.06 0.19 0.14
GBK 003674 -0.30 0.20 0.40 0.04 -0.19 -0.05 0.10 0.18 0.23 -0.20 -0.13 -0.01 -0.05 -0.18 -0.14 -0.20
GBK 003675 A -0.43 0.05 -0.14 0.19 -0.02 0.17 0.10 0.22 0.06 -0.09 -0.08 -0.22 -0.18 -0.05 0.00 -0.04
GBK 003676 A -0.03 0.01 0.12 0.12 -0.01 0.10 -0.19 0.23 -0.13 0.41 -0.15 0.04 0.12 -0.10 -0.12 0.12
GBK 003676 B 0.20 0.22 -0.06 -0.01 -0.11 0.03 -0.26 0.25 0.01 0.29 0.19 0.18 -0.07 0.16 0.03 0.08
GBK 003676 B -0.19 -0.15 0.02 0.19 -0.01 -0.08 0.04 0.08 0.00 0.16 -0.21 -0.17 0.18 0.22 -0.11 0.05
GBK 003680 0.21 -0.12 -0.17 0.21 -0.06 -0.12 0.15 0.22 0.04 -0.07 -0.21 0.00 0.23 0.08 0.04 -0.18
GBK 003682 -0.05 -0.02 -0.13 -0.43 -0.11 -0.09 -0.21 -0.07 -0.03 -0.01 -0.01 -0.17 0.08 0.02 0.14 -0.02
GBK 003685 A 0.08 -0.10 -0.09 -0.11 -0.07 0.23 0.36 -0.15 -0.03 0.02 -0.02 -0.26 -0.09 0.09 -0.01 0.09
GBK 003685 B 0.33 -0.14 0.33 -0.05 -0.15 0.19 0.22 -0.07 0.15 0.01 -0.02 0.05 -0.09 -0.13 0.04 0.10
GBK 003687 A -0.25 0.20 0.19 0.01 -0.29 -0.16 -0.30 0.01 -0.05 0.09 -0.10 0.02 -0.17 0.02 -0.09 0.18
GBK 003687 B -0.31 0.12 -0.06 0.15 0.07 -0.21 -0.20 -0.19 -0.19 0.16 -0.15 0.01 -0.08 0.00 0.02 0.06
GBK 003688 -0.16 0.24 0.31 -0.12 0.21 -0.04 0.12 -0.01 -0.21 0.08 0.07 -0.02 0.22 -0.08 -0.02 0.08
GBK 003689 -0.18 -0.06 -0.32 0.08 -0.21 0.20 -0.01 -0.07 0.24 -0.23 0.14 -0.20 0.04 0.07 -0.10 0.00
GBK 003690 -0.04 -0.21 -0.13 0.08 0.26 -0.24 0.31 0.04 0.08 0.10 0.14 0.14 -0.13 -0.11 -0.12 0.12
GBK 003692 0.05 0.02 -0.35 0.05 0.20 0.16 -0.16 -0.01 0.11 -0.02 -0.13 0.20 0.10 -0.16 -0.09 0.07
GBK 003693 0.32 0.08 0.01 0.30 0.15 -0.23 0.05 0.08 0.11 0.04 0.25 -0.07 -0.07 -0.02 0.05 -0.09
GBK 003694 A -0.15 -0.39 0.29 -0.01 -0.14 0.00 0.03 -0.12 -0.10 0.16 0.00 0.09 0.06 0.05 -0.28 -0.03
GBK 003694 B -0.35 -0.20 -0.03 0.16 -0.03 -0.25 0.13 -0.20 0.14 0.07 0.07 0.15 0.26 0.01 0.11 -0.04
GBK 003695 0.31 -0.12 -0.29 -0.09 -0.06 0.03 -0.17 -0.05 -0.04 0.07 0.04 0.02 0.23 -0.05 0.03 -0.03
GBK 003696 -0.02 0.07 0.22 0.32 0.24 0.21 -0.22 -0.10 -0.14 -0.02 0.02 0.01 0.02 0.00 -0.08 0.07
GBK 003697 -0.18 0.15 0.10 0.01 -0.08 0.20 0.05 0.18 -0.30 -0.01 -0.02 -0.07 -0.02 0.18 0.07 0.05

72
Axis No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
GBK 003698 0.15 -0.01 0.20 0.24 0.13 -0.06 -0.23 0.00 -0.27 0.02 0.07 -0.03 0.03 0.10 0.05 0.00
GBK 003699 -0.30 0.05 0.12 0.08 0.22 0.10 -0.01 0.11 -0.02 -0.16 -0.09 0.13 -0.15 -0.24 0.25 0.02
GBK 003700 0.25 -0.13 -0.20 0.10 -0.02 -0.06 0.11 0.16 -0.16 -0.03 -0.14 0.07 0.14 0.03 0.05 0.09
GBK 003701 A -0.29 0.05 0.25 -0.24 0.08 -0.06 0.05 0.22 0.32 -0.08 -0.01 -0.10 0.14 -0.20 -0.10 0.12
GBK 003701 B 0.21 -0.27 0.04 -0.11 0.28 0.02 -0.12 -0.13 0.19 0.23 0.16 -0.10 -0.02 -0.16 -0.06 0.04
GBK 003705 0.29 0.00 -0.05 0.11 0.15 0.06 -0.02 -0.04 -0.04 -0.30 0.05 0.10 0.12 0.00 -0.02 0.07
GBK 003706 -0.17 -0.05 0.14 0.02 0.17 -0.24 -0.16 -0.22 -0.07 -0.03 -0.05 -0.02 0.10 0.14 -0.10 0.15
GBK 003707 0.21 -0.02 0.34 -0.22 -0.14 0.27 -0.21 -0.04 0.25 0.07 0.18 0.15 -0.04 0.09 0.03 0.11
GBK 003709 -0.34 0.02 -0.15 0.14 -0.23 0.33 0.12 0.01 -0.14 0.02 0.22 -0.01 0.03 0.07 -0.06 0.08
GBK 003711 -0.15 -0.02 -0.16 -0.39 0.05 -0.21 -0.20 0.03 -0.06 -0.10 -0.25 0.11 -0.13 0.08 0.00 -0.25
GBK 003711 0.10 0.31 0.05 0.08 -0.33 -0.33 0.28 0.10 0.14 0.16 -0.08 0.01 -0.07 -0.02 0.00 -0.01
GBK 003714 -0.04 -0.34 -0.12 0.11 0.13 0.00 0.01 0.07 -0.09 0.04 -0.43 0.16 -0.13 -0.06 -0.08 -0.17
GBK 003717 -0.08 -0.37 -0.14 0.20 -0.03 0.02 0.02 0.00 0.14 0.14 -0.23 -0.11 0.06 -0.14 0.17 -0.08
GBK 003718 0.17 -0.09 0.07 -0.25 0.24 -0.11 0.17 0.13 -0.03 0.14 0.04 -0.10 0.03 0.09 -0.09 -0.01
GBK 003720 0.25 -0.14 0.12 0.11 0.01 -0.03 -0.16 -0.10 -0.21 -0.06 0.09 -0.11 0.01 -0.18 0.07 -0.32
GBK 003723 0.00 0.03 0.21 0.18 -0.12 -0.04 0.28 0.04 0.09 -0.22 -0.03 0.15 0.31 0.21 -0.22 -0.18
GBK 003724 -0.47 0.17 0.04 0.10 0.09 0.00 0.09 0.03 -0.09 -0.09 0.13 0.31 0.00 -0.03 0.10 0.05
GBK 003726 -0.24 -0.40 0.13 0.09 0.16 0.03 -0.13 0.15 -0.07 0.00 -0.10 0.29 -0.13 0.14 0.05 0.01
GBK 003727 0.05 0.27 0.14 0.07 -0.32 -0.33 -0.11 -0.01 0.03 0.00 -0.21 -0.21 -0.06 -0.08 -0.04 -0.02
GBK 003780 -0.01 -0.02 -0.18 -0.27 0.14 0.00 -0.17 0.13 0.13 0.15 -0.01 -0.14 -0.16 0.14 -0.13 0.05
GBK 003796 -0.10 0.22 0.35 0.12 0.11 0.07 0.11 0.12 0.12 0.00 0.04 0.00 -0.03 0.21 0.35 -0.04
GBK 003804 -0.09 0.34 0.22 -0.12 0.19 -0.02 -0.14 -0.15 -0.18 -0.01 0.06 -0.02 0.18 0.05 -0.04 -0.01

73
Axis No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
GBK 003814 -0.38 0.21 -0.38 0.04 -0.02 -0.20 -0.18 0.13 0.18 -0.01 0.07 -0.06 -0.04 -0.07 -0.02 -0.08
GBK 003816 A 0.05 0.43 -0.14 -0.18 0.08 0.01 0.29 0.02 -0.06 0.15 -0.01 0.05 0.06 -0.03 0.02 -0.02
GBK 003816 B 0.22 -0.28 0.10 -0.17 0.09 -0.14 0.09 -0.25 -0.05 0.05 0.17 -0.13 -0.21 0.07 -0.10 -0.09
GBK 003820 0.00 -0.33 0.21 -0.22 -0.18 -0.10 0.09 0.14 -0.07 -0.05 0.20 -0.13 -0.03 -0.14 0.02 0.12
GBK 003876 -0.19 -0.40 -0.20 0.11 0.04 -0.12 0.25 -0.09 -0.01 -0.03 0.23 0.02 0.01 -0.24 -0.14 0.11
GBK 003888 0.26 0.06 -0.09 -0.11 0.15 0.19 -0.17 0.03 -0.04 -0.06 -0.20 -0.09 0.13 -0.01 -0.03 0.06
GBK 003916 0.18 0.06 -0.06 0.37 0.18 -0.12 -0.04 0.05 0.20 0.08 0.16 -0.02 -0.08 0.00 -0.03 -0.13
GBK 003985 -0.23 -0.31 -0.43 -0.03 -0.17 0.02 -0.10 -0.11 0.00 -0.11 0.09 0.08 -0.24 0.04 -0.13 -0.05
GBK 005173 0.00 -0.12 0.07 -0.17 0.27 0.26 -0.28 0.10 0.09 -0.18 -0.05 0.09 -0.06 0.01 -0.15 -0.07
GBK 005173 A -0.03 -0.19 -0.02 -0.23 -0.15 -0.12 0.16 -0.19 -0.22 0.03 -0.23 -0.08 0.02 0.04 0.07 -0.08
GBK 005173 B 0.00 0.18 -0.16 -0.40 -0.27 0.06 -0.11 -0.07 0.09 -0.11 0.09 0.07 0.14 -0.09 0.03 0.08
GBK 027036 -0.13 -0.31 0.18 -0.33 -0.05 0.06 -0.05 0.07 -0.17 0.09 0.14 0.06 0.11 0.22 -0.02 -0.14
GBK 027079 0.16 0.14 -0.10 -0.05 0.11 -0.19 -0.12 0.09 -0.21 -0.02 0.25 -0.01 0.13 -0.10 -0.19 -0.15
GBK 027089 -0.39 0.00 0.15 -0.29 0.29 0.08 0.07 -0.03 0.14 0.10 -0.05 0.30 -0.13 0.18 -0.01 0.00
KAT 1 0.13 0.09 -0.14 0.12 0.15 -0.25 0.08 -0.04 -0.10 0.03 0.24 0.09 -0.23 0.00 0.00 -0.10
KAT 3 0.01 0.33 0.09 0.03 -0.45 0.00 -0.03 -0.31 -0.10 0.02 0.05 0.14 0.08 -0.06 -0.04 0.09
KOL 1 -0.25 0.27 0.09 -0.21 0.22 0.06 -0.20 0.02 0.00 -0.13 0.08 -0.07 -0.03 -0.13 -0.05 -0.04
KOL 5 -0.43 0.24 -0.14 -0.04 -0.06 0.13 0.13 -0.03 0.26 0.30 0.14 0.15 0.06 0.03 0.20 -0.13
KOL 6 0.34 0.27 -0.17 0.18 0.07 0.14 0.06 -0.02 -0.06 0.00 0.03 0.03 -0.13 0.03 0.05 0.00
KOL 8 -0.39 -0.03 -0.01 0.24 -0.12 0.03 -0.02 0.25 -0.05 0.10 0.00 -0.26 -0.03 0.22 -0.06 0.02
KOL 9 B -0.11 -0.09 0.13 -0.23 0.05 -0.23 0.08 0.13 -0.13 -0.21 -0.03 -0.07 0.01 0.07 0.14 0.07
MAC 1 -0.32 -0.13 -0.34 -0.28 -0.08 -0.08 0.02 0.03 0.07 0.10 0.00 0.16 0.10 -0.15 0.12 0.00

74
Axis No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
MAC 2 -0.07 -0.14 0.11 -0.08 0.12 -0.30 0.24 -0.02 -0.14 -0.16 -0.01 -0.20 0.04 -0.05 -0.05 0.21
MAC 3 -0.30 -0.07 0.23 0.36 0.22 -0.25 0.00 0.03 0.04 0.16 -0.07 0.04 0.07 -0.15 0.18 0.02
MAC 3 -0.10 0.03 0.00 -0.11 -0.18 0.26 0.12 0.08 -0.24 -0.05 0.04 -0.20 -0.06 -0.05 0.15 -0.08
GBK 026941 A 0.25 0.22 -0.17 0.21 0.01 0.14 -0.05 0.01 -0.18 -0.13 -0.03 0.02 0.02 -0.06 0.02 0.00
GBK 026942 B 0.34 -0.15 -0.24 -0.14 -0.01 0.08 0.16 0.04 0.04 0.28 0.05 0.06 0.14 0.01 0.13 -0.02
GBK 026958 A 0.12 -0.12 -0.07 -0.14 0.14 0.37 0.11 -0.06 0.15 0.26 -0.13 -0.06 0.06 -0.10 0.02 0.03
GBK 026958 B 0.14 -0.14 -0.20 0.01 -0.08 -0.01 0.04 0.31 -0.11 -0.11 -0.06 0.09 0.05 0.09 0.14 0.09
GBK 046540 -0.29 0.14 0.23 0.17 0.02 0.03 -0.09 -0.35 0.06 0.00 0.09 -0.17 0.15 0.00 0.04 -0.11
K80 0.07 -0.06 0.28 -0.26 -0.06 0.01 0.37 -0.31 0.05 -0.11 -0.14 0.10 -0.12 0.04 -0.04 -0.24
KAR 1 0.37 -0.25 0.06 -0.02 0.11 -0.02 -0.03 -0.19 0.03 0.05 0.11 -0.11 -0.09 0.05 0.16 -0.02
KENKUNDE 0.31 0.19 0.03 0.06 -0.02 -0.06 -0.18 0.18 0.16 0.30 -0.07 -0.19 -0.07 -0.02 0.00 -0.09
KVU-27-1 0.09 0.21 -0.15 -0.12 0.03 -0.05 0.39 -0.08 0.00 -0.04 -0.11 0.06 0.03 0.11 0.05 0.13
M66 0.28 0.14 0.09 -0.10 0.01 0.04 0.03 -0.23 0.08 -0.10 -0.35 0.09 -0.24 0.12 0.02 0.04
KAB 1 -0.06 0.25 -0.09 -0.15 0.20 -0.01 0.00 -0.14 0.35 0.10 -0.20 -0.21 0.05 0.07 -0.09 0.00
KAB 3 0.00 0.33 0.13 -0.19 0.21 -0.04 -0.08 -0.10 -0.14 -0.11 0.02 -0.11 0.09 -0.12 -0.04 -0.07
KIP 1 0.21 0.32 -0.19 0.07 0.03 0.05 0.14 -0.07 -0.11 0.00 0.09 -0.05 -0.14 0.08 -0.08 -0.02
KIP2 0.28 -0.31 0.29 -0.02 -0.26 -0.04 0.02 0.03 -0.03 -0.02 -0.04 0.02 0.06 -0.23 0.01 0.05
LAM 4 -0.35 -0.03 -0.32 0.10 -0.16 0.26 0.17 0.04 -0.07 -0.07 0.16 0.01 -0.14 -0.17 0.02 -0.01
MAR.2 0.32 -0.02 0.37 0.11 -0.03 -0.14 0.16 0.19 -0.02 0.16 0.04 0.00 -0.24 -0.21 -0.04 0.12
MAR.3 0.40 -0.01 -0.25 0.08 0.06 -0.09 0.00 0.00 0.01 0.03 -0.03 0.04 -0.07 0.15 0.04 0.09
MAR.5 -0.14 -0.07 0.00 -0.18 0.11 -0.31 -0.14 -0.02 -0.06 -0.26 0.00 0.01 -0.14 0.03 0.18 0.04
MBL -0.40 -0.15 0.19 0.05 -0.06 0.34 0.06 -0.06 -0.18 -0.04 0.20 -0.03 -0.13 0.10 -0.13 -0.08

75
Publications

1. Munyao R. K, Mamati E. G, Githiri S.M, Ateka E.M 2019. Genotypic Diversity among Cowpea Genotypes from the
Lower Eastern Region of Kenya. International Journal of Agronomy and Agriculturaral Research14(5), 9-19.

76