A COMPARISON OF SUSCEPTIBILITY TO FEEDING BY THE BLACK BEAN

APHID (APHIS FABAE) BETWEEN TWO VARIETIES OF BEANS (PHASEOLUS

VULGARIS) (GLORIA AND NUA 45) COMMONLY GROWN IN ZIMBABWE.

BY
MALVERN TINASHE MUTSIYO (R111557M)

A dissertation submitted in partial fulfilment of the requirements for the Bachelor of

Science Degree in Applied Biosciences and Biotechnology

Department of Applied Biosciences and Biotechnology

Faculty of Science and Technology

November 2017
APPROVAL FORM
This is to certify that the dissertation entitled “A comparison of susceptibility to feeding by

the black bean aphid (Aphis fabae) between two varieties of beans (Phaseolus vulgaris)

(Gloria and Nua 45) commonly grown in Zimbabwe.”, submitted in partial fulfilment of the

requirements for Bachelor of Science Honours Degree in Applied Biosciences and

Biotechnology at Midlands State University, is a record of the original research carried out by

MALVERN T. MUTSIYO R111557M under my supervision and no part of the dissertation

has been submitted for any other degree or diploma.

Name of supervisor……………………………………………………………………

Signature…………………………………………………………………………….

Chairperson signature……………………………………………………………….
ABSTRACT

Gloria and Nua 45 are two of the most commonly grown sugar bean varieties in Zimbabwe.
Beans are a cheap alternative source of protein and have been on high demand due to the
ever-rising meat prices. Gloria and Nua 45 are being affected by the black bean aphid (Aphis
fabae) feeding which is the most problematic insect pest of beans. The aphid feeds on foliage
of plants resulting in stunted growth and reduced yields. A comparison of susceptibility to
feeding by A. fabae was done to determine which one of the two varieties is least affected by
the aphid. Two glasshouses containing both varieties of beans were used and one of the
glasshouses was not inoculated with the aphid and this served as the control. The plant
heights, leaf areas and 100-seed weights of the two varieties were compared after a 90-day
period. The experimental design was Randomised Block Design (RBD) with each block
having three replicates in each glasshouse. It was difficult to create homogeneity due to
confounding factors (temperature and exposure to sunlight) in the glasshouses hence this
choice of experimental design. Data collected on plant height, leaf area and 100-seed weight
were subjected to one-way analysis of variance (ANOVA) for RBD using SPSS version 21.
Dunnett test was done to compare each variety to the control. Multiple comparisons for the
effect of aphid feeding on plant height, leaf area and 100-seed weight was done using Tukey
post hoc tests. Both varieties in the glasshouse that was inoculated with the aphid were
susceptible to feeding. However, the plant heights, leaf areas and 100-seed weights varied
significantly (p<0.05) between the two varieties with the least values of these parameters
being recorded for Gloria. The one-way analysis of variance showed that the plant heights,
leaf areas and 100-seed weights of plants in the control glasshouse did not vary significantly
(p>0.05). Therefore, this study indicated that Nua 45 is the least affected variety between the
two (Gloria and Nua 45). Therefore, this study recommends that in areas infested by the
black bean aphid (Aphis fabae) Nua 45 should be cultivated as it is the more resistant variety.

i
ii
ACKNOWLEDGEMENTS

It is my pleasure to acknowledge the depth l owe to many people who have assisted me in

coming up with this project. I would like to express special gratitude to my academic

supervisor, Mr Bare, for his constant support and mentorship. Mr Bare kindly undertook the

formidable task of reading this whole dissertation and I am grateful to him for his frank and

helpful comments. Special thanks are extended to Mrs R. J. Mbiringa (Associate Research

Scientist: Entomologist) for assisting me in coming up with this project. Her input in terms of

continued assistance throughout my project was greatly appreciated and helpful. Special

thanks are also extended to Dr T. Muteveri for his mentorship on data analysis. I would also

like to thank all research scientists and research associates at DR&SS including Mr. Rwafa,

Mrs. Chihera, Mr. N. Mutema and Dr. G. Chikwenhere for their support and assistance

throughout my project. To all my friends Mr. G.R. Gurupira, Mr. H. T. Takawira, Ms. B.

Chipatiso and other scientists that are not mentioned I owe them my thanks for giving me the

benefit of their expertise and making this project a success.

iii
DEDICATION

This project is dedicated to my late parents. This project is also dedicated to my uncle Dr S.

Kuhudzai and Mrs C. Kuhudzai who have sacrificed everything to give me the most and best

education possible.

iv
CONTENTS

PAGE

LIST OF

FIGURES…………………………………………………………..………….....viii

LIST OF

TABLES……………..………………………………………………………….....ix

LIST OF APPENDICES………..………………………………...........................................x

CHAPTER ONE: INTRODUCTION……………………………………............................1

1.1 Background………………………………………………………………..........................1

1.2 Justification………………………………………………………………..........................3

1.3 Objectives…………………………………………………………………........................4

1.3.1 Main objective……………………………………………………………………….4

1.3.2 Specific objectives……………………………………… ………………………….4

CHAPTER TWO: LITERATURE

REVIEW………………………………………………5

2.1 History of beans (Phaseolus vulgaris)

cultivation………………………………………….5

2.2 Black bean aphid (Aphis fabae)

……………………………………………………………6

2.3 Importance of beans (Phaseolus vulgaris) ………………………………..........................8

v
 2.3.1 Nutritional value…… ………………………………………………………………...8

2.3.2 Medicinal

value………………………………………………………………………..9

2.3.3 Income

generation……………………………………………………………………..9

2.3.4 Improvement of soil

health…………………………………………..........................10

2.4 Current status of sugar bean production in Zimbabwe…………………………………...10

2.5 Factors affecting sugar bean

production…………………………………………………..11

2.5.1 Biotic

factors…………………………………………………………........................11

2.5.2 Abiotic

factors………………………………………………………………………..12

CHAPTER THREE: MATERIALS AND

METHODS…………………………………...13

3.1 Study

site………………………………………………………………………………….13

3.2 Sources of

materials………………………………………………………........................13

3.3 Experimental design……………………………………………………….......................13

vi
3.4 Rearing of

aphids…………………………………………………………........................15

3.5 Preparation of sample

plants……………………………………………………………...16

3.5.1 Pathological seed

analysis…………………………………………………………..16

3.5.2 Soil sampling……………………………………………………………………….16

3.5.3 Watering and fertilisation…………………………………………………………..16

3.5.4 Weed

control………………………………………………………………………..17

3.6 Determination of the effect of the Black bean aphid (Aphis fabae) on the

plant height, leaf area and yield of Gloria and Nua 45………………………........................17

3.6.1 Inoculation of sample plants with the Black bean

aphid (Aphis fabae)

…………………………………………………………………..17

3.6.2 Measuring of plant

height………………………………………………………….17

3.6.3 Determination of leaf

area………………………………………………………….17

3.6.4 Measuring of

yield…………………………………………………........................18

vii
3.7 Data

analysis……………………………………………………………………………...18

CHAPTER FOUR: RESULTS

…………………………………………………………….19

4.1 Effect of A. fabae on the plant heights, leaf areas and 100-seed

weight (yield) of Gloria and Nua

45..…………………………………………………….19

CHAPTER FIVE:

DISCUSSION…………………………………………………………..24

5.1 Effect of Aphis fabae feeding on the plant heights of Gloria and Nua

45………………..24

5.2 Effect of Aphis fabae feeding on the leaf areas of Gloria and Nua 45…………………...24

5.3 Effect of Aphis fabae feeding on the 100-seed weight of Gloria

and Nua

45….………………………………………………………………….......................25

5.4 Conclusion………………………………………………………………………………..26

5.5 Recommendations…………………………………………………………......................26

6.0 References ..……………………………………………………………………………...27

7.0 Appendices……………………………………………………………………………….35

viii
LIST OF FIGURES PAGE

1.1: Arrangement of blocks in Glasshouse 1 where the black bean

aphid (Aphis fabae) was inoculated……………………………………………….. 15

1.2: Arrangement of blocks in Glasshouse 2 which served as the control....…………... 16

2: Effect of Aphis fabae feeding on Gloria and Nua 45 compared to the control……..23

ix
LIST OF TABLES

PAGE

1: Effect of A. fabae on the mean plant heights, leaf areas and 100-seed weight (yield) of

Gloria and Nua

45…………………………………………………………………….21

2: Mean plant heights, leaf areas and 100-seed weights of the control plants (where the

aphid was not inoculated)…………………..………………………………………..22

x
LIST OF APPENDICES

PAGE

1: Pathological seed analysis using Blotter test……..……..………………………….34

2: Multiple comparisons for effect of A. fabae on the plant heights, leaf areas and 100-

seed weights of the inoculated plants and the control plant..………………………35

3: Means plot of the plant heights of the inoculated plants and the control

plants………………………..………………………………………………………37

4: Means plot of the leaf areas of the inoculated plants and the control plants.

…….……………………………………………………………………….……….38

5: Means plot of the 100-seed weights of the inoculated plants and the control plants.

…………….…….………………………………………………………………….39

xi
xii
 CHAPTER 1: INTRODUCTION

1.1 Background

The common sugar beans (Phaseolus vulgaris) are of a very important economic status

(FAO, 2006). Dry beans were grown on 27.7 million ha in 148 countries in 2004 and total

production was 18.7 million metric tons (MT) with a market value of more US$10.7 billion

(FAO, 2004). Sugar bean originated in Central South America and is one of the most

important grain legume for human direct consumption in the world (Jones and Corlett, 1992).

Developing countries produce 86 % of the worldwide production of beans. The per capita

consumption is the highest in East Africa in countries such as Burundi, Rwanda and Uganda

(Chester, 1969). However, the leading bean producer and consumer is Latin America, where

beans are a traditional, significant food, especially in Brazil, Mexico, the Andean Zone,

Central America, and the Caribbean (Schwartz, Brick, Nuland and Franc, 1996).

Beans are nutritionally rich because they are a good source of protein, folic acid, dietary fibre

and complex carbohydrates. Sugar beans are also known to be one of the best non-meat

sources of iron, providing 23% to 30% of daily recommended levels from a single serving

(Pachico,1993). Beans are mostly consumed especially in developing countries because they

are a cheap alternative source of protein and they are a means of keeping malnutrition at bay.

Qualitatively, bean seed proteins provide essential amino acids such as lysine, but are poor in

methionine, thus complementing cereals in this respect (Bressani, 1983; Gepts and

Bliss,1985). Beans are also the third most important major source of calories (Pachico,1993).

In addition, sugar beans play an important role as a source of minerals, especially iron and

zinc (Chester, 1969), for which it also complements cereals. Genetic variation for seed

content of these minerals has been demonstrated (Lynch, Lauchli and Epstein, 1991).

1
Constraints to common bean production are poor production practices, pests and diseases,

low soil fertility, adverse weather conditions, weed infestations and plant populations

(Wortman and Allen, 1994; Gridley and Danial, 1995). According to the Ministry of

Agriculture, Mechanization and Irrigation, the Department of Research and Specialist

Services Gloria and Nua 45 are the two varieties of sugar beans that are commonly grown in

Zimbabwe and the most problematic pest of these two varieties is the black bean aphid (Aphis

fabae). Wilkinson and Douglas (2003) state that Aphis fabae causes economic damage

mainly due to direct feeding. Aphids tap into the phloem, and to a lesser extent the xylem,

with their stylets and ingest large quantities of soluble nutrients.

Beans suffer damage to flowers and pods which may fail to develop properly. Early-sown

crops may avoid significant damage if they have already flowered before the number of

aphids builds up in the spring (Berim,2009). For the aphids to obtain enough protein they

need to suck large volumes of sap. They secret excess sugary fluid, honeydew, which adheres

to plants where it promotes growth of sooty moulds. These reduce the surface area of the

plant available for photosynthesis causing stunted growth, reduced yields and may reduce the

value of the crop (Banks and Macaulay,1967). These aphids are also vectors of about thirty

plant viruses, mostly of the non-persistent variety (Blane et al.,1990). The aphids may not be

the original source of infection but are instrumental in spreading the virus through the crop

(Rothamsted Research, 2003). Various chemical treatments are available to kill the aphids

and organic growers can use a solution of soft soap (Godfrey and Trumble, 2009).

While substantial bean improvement work has been done by breeding improved cultivars,

response to soil fertility and disease control (Makini and Danial, 1995; Tyagi et al., 1996;

Maiuki, 1998) the susceptibility to feeding by the black bean aphid between Gloria and Nua

45 varieties has not been given due attention. No study has been carried out to determine

which one of the two varieties of beans is least affected by A. fabae. Any advances in

2
scientific research that benefit bean yields, particularly in developing countries, help to feed

the hungry and give hope for the future. Identifying and minimizing yield limiting factors is

an ongoing concern for many bean improvement programs. In addition, given the prevalence

of beans in these diets, modifying the nutrient content in general to make it a more balanced

and nutritious food source is also receiving emphasis. Hence this study is going to bridge that

gap.

1.2 Justification

There has been an increased demand for beans due to the economic hardships that we are

currently going through. The ever- rising prices of meat, which is also a source of protein has

led to reliance on beans for subsistence (Katungi et al., 2017). Institutions such as

universities, boarding schools, prisons and hospitals rely on beans for their diets. Beans is a

cheap alternative source of protein and a crop of economic importance. Other than proteins

beans also contain minerals and fibre. Beans contain iron, about 95mg per kg and zinc, about

38 mg per kg (Norman, Pearson and Searle, 1995). This makes beans a key player in the

reduction of anaemia in pregnant women and also helpful in individuals living with

HIV/AIDS (Kaplan, 2008).

Knowing which variety between Gloria and Nua 45 is affected less by A. fabae will go a

long way in helping farmers make a better decision when buying seed. Farmers will invest

less in pesticide procurement, some of which are harmful to the environment. For instance,

over 98% of sprayed pesticides reach a destination other than their target species, because

they are sprayed or spread across entire agricultural fields. The knowledge from this study

will also help in boosting the farmers‟ yields. Higher yields will help cater for the unfulfilled

demand of the legume crop. The United Nations declared 2016 The International Year of

Pulses to celebrate the growing importance of beans.

3
Gloria and Nua 45 are affected by the black bean aphid and this has been a challenge to

farmers. It is unknown which variety between the two is least affected. The aphid populations

cause a significant loss in the yield by decreasing flower and pod production. Yield loss in

beans is as a result of fewer pods per plant and fewer and smaller seeds per pod. Seed

viability and value is also reduced due to aphid feeding (Cammell and Way, 1983).

Farmers are investing too much capital in buying pesticides some of which are harmful to the

environment. The pesticides DDT, methyl parathion and especially pentachlorophenol have

been shown to interfere with legume-rhizobium chemical signalling. Reduction of this

symbiotic chemical signalling results in reduced nitrogen fixation and thus reduced crop

yields. This study helped to determine which one of the two varieties (Gloria and Nua 45) is

less affected by the black bean aphid (Aphis fabae) hence reducing the environmental

footprint of pesticides.

1.3 Objectives

1.3.1 Main objective:

• to compare the effect of aphid feeding between Gloria and Nua 45.

1.3.2 Specific objectives:

• to compare the effect of aphid feeding on the plant height of Gloria and Nua 45

over a 90-day period,

• to compare the effect of aphid feeding on the leaf area of Gloria and Nua 45 ,over a

90-day period, and

• to compare the effect of aphid feeding on the yield of Gloria and Nua 45 over a 90-

day period.

4
 CHAPTER TWO: LITERATURE REVIEW

2.1 History of beans (Phaseolus vulgaris) cultivation

Beans have been known to be cultivated since long ago (Kaplan, 2008). Seeds that had the

size of a small fingernail were initially gathered in their wild state in Afghanistan and the

Himalayan foothills (Kaplan, 2008). Cubero (1974) postulated a Near Eastern centre of

origin, with four radii to Europe along the North African coast to Spain, along the Nile to

Ethiopia, and from Mesopotamia to India (Hawtin and Hebblethpiait, 1983). Secondary

centres of diversity are postulated in Afghanistan and Ethiopia.

Chester (1969) reported that beans were grown in Thailand as early as the seventh

millennium (B.C). It is believed that the dead deposited them in ancient Egypt. Large seeded

broad beans then appeared in the Aegean and transalpine Europe during the second

millennium (B.C) (Daniel and Maria, 2000). However, Hajjar and Hodgkin (2007) reported

the origin to be Central Asia. The Chinese used them for food almost 5,000 years ago, and

they were cultivated by the Egyptians 3,000 years ago, by the Hebrews in biblical times, and

a little later by the Greeks and Romans (Mihailovic et al., 2005; Singh and Bhatt, 2012).

Probably, the Europeans introduced it as a garden crop into India during the Sultanic period

(1206–1555), during which its cultivation has been mentioned (Naqvi, 1984; Akbar, 2000).

The oldest domesticated beans were found in Guitarrero cave, which is an archaeological site

in Peru in the Americas. These beans dated to around the second millennium (B.C) (Chazan,

2000). Even back then beans were an important source of protein as they still are this modern

age. The United States alone is responsible for the cultivation of more than 4,000 cultivars of

beans. However, cultivars of beans that are commonly eaten fresh were first discovered in

what is believed to be the Bahamas by a European by the name of Christopher Columbus

5
during his exploration. Some tribes that dwelled on the East Coast of what is now known as

the United States would grow maize, beans and squash intermingled them together in a

system that had originated from Mexico (Everett, 2009).

2.2 Black bean aphid (Aphis fabae)

The black bean aphid, A. fabae, is widespread in temperate regions, where it is an important

economic pest of sugar beans (Phaseolus vulgaris). Since the 1960s, A. fabae has become an

important pest of Phaseolus vulgaris in East Africa (Eastop, 1977). The aphid causes great

economic damage through direct feeding and is also responsible for indirect damage where it

serves as a vector of plant viruses. It is of major economic importance because it causes

direct feeding damage on beans, and feeding and virus transmission on Beta vulgaris.

However, A. fabae is of relatively minor importance as a virus vector on other crops.

Dense aggregations of aphids on actively growing plants results in severe yield losses.

Aphids tap their stylets deep into the phloem, and to a lesser extent the xylem, and ingest

large quantities of soluble nutrients (Wilkinson and Douglas, 2003). Winged aphids may

ingest more from the xylem to counter dehydration during flight (Powell and Hardie, 2002).

Their saliva contains chemicals that may also disrupt plant physiology. Large infestations of

aphids stunt plant growth, reduce seed formation, and may eventually cause premature plant

death.

A. fabae has been known to cause serious losses in beans for a very long time (Curtis, 1960;

Davidson, 1921; Cammell, 1981). Similar studies have shown that the aphids prefer the

growing parts on Vicia faba (field bean, broad bean, faba bean). If young plants are severely

attacked by dense infestations they may become stunted and may die early. Since these

aphids suck on the sap which contains the plant nutrients, infestation slows the rate of stem

elongation, leaf growth and root system development. An A. fabae nymph can ingest 3.5-4.5

6
mg of sap during its 7-day development period, while each adult consumes about 30 mg

during its lifetime (Banks and Macaulay, 1964).

Aphid aggregations become less pronounced as the plant grows and the colonies become

distributed around the plant. Populations on older plants, before or during flowering, can still

cause significant crop losses, however, by decreasing flower and pod production. Yield loss

in beans is primarily due to fewer pods per plant, and fewer and smaller seeds per pod.

Damage also reduces seed viability and food value (Cammell and Way, 1983).

A 3-year study that was conducted in the former East Germany in the late 1970s estimated

the yield losses caused by A. fabae in pot and field experiments. Yields were calculated in

terms of the weight of 1000 seeds. The yields of uninfested beans were higher than that of the

infested beans. Uninfested beans were around 615 g, while the lowest yields in plots heavily

attacked by A. fabae at the time of flowering were around 380 g. It was observed that the

levels of injuriousness caused by the A. fabae are dependent on the time and intensity of

colonization. If plants are infested before flower buds form usually the plants totally perish

before pods form. When aphid attack coincided with the onset of flowering, yields declined

by 60-65% in pot experiments, compared with uninfested controls, while yields declined by

around 50% in plot experiments. Later attacks, at the time of pod setting, caused yields to

decline by 6% at most in equivalent experiments (Hinz and Daebeler, 1981).

In similar studies that were carried out in Britain, the beans that were sown in March were

attractive to primary colonists, while crops sown in late April were susceptible to secondary

colonists migrating from other bean crops. However, late autumn-sown or 'winter' beans were

rarely damaged by A. fabae as their critical growth period occurs at a time when aphids are

not present in the crop (Cammell and Way, 1983).

7
In experiments carried out in France, A. fabae and the pea aphid (Acyrthosiphon pisum)

reduced yields (quantified as weight of 1000 seeds and number of seeds per pod) of V. faba

by up to 30%, dependent on the physical condition of the plants (Bouchery, 1977). Higher

yield losses due to A. fabae on V. faba were reported in a field study in Iraq, using artificial

infestations. Pod number, pod weight and dry weight of seeds were negatively correlated with

aphid numbers, the reductions in these parameters reached 70, 76 and 64%, respectively

(Mohammad and Abdulla, 1988).

In a field study conducted in South America the length of time that A. fabae is present on V.

faba has been correlated with yield loss. Early infestations (before the flowering period)

resulted in considerable damage, dependent on the population levels attained in the aphid

colonies. Early stage high infestations decreased yields (total weight of seeds) by up to 42%

compared with controls. Infestation at the beginning of the flowering period has little effect

on flower formation and development, although high populations at this stage may have an

adverse effect on pod formation. Any infestations later than this stage had no effect on yield

(Salazar, 1984).

2.3 Importance of beans (Phaseolus vulgaris)

2.3.1 Nutritional value

Sugar beans are characterized as a near perfect food by nutritionists because they contain a

high protein content (20%-24%), complex carbohydrates and other much needed dietary

necessities (Bliss,1980; Lincoln,1987). Sugar beans also provide the recommended daily

levels of certain trace minerals that include iron (25%-30%), magnesium and copper (25%)

and potassium and zinc (15%) (Peters,1993). Dry beans (22% protein content) have a wide

range of dishes which include simply boiling them in water and even more sophisticated

8
preparations which provide us with products such as salads, soups, baked beans, pastes, chips

and creams (Hosfield, 2000).

Dry leaves, threshed pods, and stalks are fed to animals and used as fuel for cooking,

especially in Africa and Asia. In Peru and Bolivia, where high altitudes prolong cooking

times and fuel costs, the ancient tradition of toasting grains comparable to corn and peanuts

may be the reason why popping or "toasted" beans have been developed. They are cooked

similarly to popcorn. Dry beans are mostly eaten whole in cooked recipes. Some

manufactured products use bean flour. Roasted beans can be pin-milled to produce whole

flour or cracked by corrugated rollers for easy removal of hulls by air aspiration. Hulls may

be ground as high fibre (40 percentage) flour to desired particle size (Sperling,1996).

2.3.2 Medicinal value

Beans have demonstrated impressive health benefits that include lowering cholesterol levels,

improving diabetics' blood glucose, reducing risk of many cancers, lowering blood pressure,

regulating functions of the colon, preventing and curing constipation, preventing piles and

other bowel problems.

A lesser-known benefit of beans, though, is their high levels of isoflavones, compounds that

are similar in structure to estrogen produced by the body (which is why they are also called

phytoestrogens). These isoflavones may ease the symptoms of menopause, prevent some

form of cancer, reduce the risk of heart disease and improve bone and prostate health, among

other benefits.

2.3.3 Income generation

Despite the production being low the price of beans has been trending upwards on the local

market. Current production levels in Zimbabwe are relatively low compared to other

countries like Malawi and Tanzania. The margins of growing sugar beans are larger than that

9
of growing any other food crop for example maize (World Vision, 2003). The key to getting

profitable with bean farming is choosing a variety that does not have high input costs such as

costly seed and is not highly affected by pests such as the black bean aphid (Aphis fabae).

Farmers would prefer to invest less capital in pesticides procurement some of which are also

harmful to the environment.

2.3.4 Improvement of soil health

Beans are nitrogen fixing and can improve the fertility of soil. Good soils help to grow

healthier, more resilient crops and improve crop yields. Beans use less water to grow, they

are water-efficient.

2.4 Current status of sugar bean production in Zimbabwe

Currently in Zimbabwe the national yield is still significantly low such that the domestic

market is failing to meet local demands. These local demands are attributed to the high

consumption of beans in prisons, hospitals, boarding schools, universities and restaurants.

The Agricultural Development and Market Corporation of Malawi sells about 100 MT to 150

MT of their beans to their domestic market and 80-85% of their produce is exported to

Zimbabwe and the rest to South Africa (World Vision,2003).

However, it is not known which variety is least susceptible to aphid feeding between the two

sugar bean varieties (Gloria and Nua 45). This study will go a long way on educating the

farmers about the variety that is least affected by the black bean aphid (Aphis fabae). This

will help farmers have better yields especially in areas where there is a high occurrence of the

aphid.This study will also help farmers to invest less in pesticide procurement some of which

are harmful to our environment.

10
2.5 Factors affecting sugar bean production

2.5.1 Biotic factors

Pests

Besides the black bean aphid which was used in this study a number of other pests attack

beans. These include the bean stem maggot, bean stem maggot, bean bug, common cutworm,

bean flower thrips, bean weevil and the bean gail weevil.

Diseases

A wide range of diseases are known to attack beans and these include Fusarium species, Halo

blight, Powdery mildew, Common blight, Bean Common Mosaic Virus(BCMV), Angular

leaf spot, Bacterial brown spot, Pythium species and Ascochyta. All the listed diseases cause

significant damage in beans which affects the crop and may cause reduction in yields

(Norman et al,1995).

Weeds

Weeds reduce crop yields. Every living plant requires a certain amount of space for the

circulation of air, moisture and sunlight. When plants are crowded and this much-needed

space is occupied by weeds then they cannot grow and produce well. The reproductive

behaviour and the growth rate of the plant is affected and returns from the plant will be

correspondingly less (Vital, 2001).

11
2.5.2 Abiotic factors

Use of uncertified seed

The production of beans is inhibited by the use of uncertified and retained seed. Some

farmers tend to use seed of the beans they harvested the past season and stored under

traditional conditions where they can easily get infested by post-harvest bean weevils

resulting in poor emergence (Dube and Nyoka,1993).

12
 3.0 MATERIALS AND METHODS

3.1 Study site

This study was carried out at the Department of Research and Specialist Services in Harare

which is under the Ministry of Agriculture, Irrigation and Mechanization. The experiments

were carried out in the glasshouses, Entomology and Pathology laboratories.

3.2 Sources of materials

The Gloria and Nua 45 bean seeds used in this study were provided by the Plant Protection

department after having been checked for their viability and after being confirmed to be

disease-free in the Pathology laboratory. The seeds were planted into their respective pots

containing soil which had been collected from the DRSS fields. The aphids (black bean

aphid) used in this research were reared on a potted bean plant after being collected from

infested fields in Zaka. To avoid injuring the aphids, an instrument called the aphid gun was

used. Two glasshouses were used to carry out the experiment. The first glasshouse contained

plants exposed to the treatment (presence of the aphid). The second glasshouse was the

control glasshouse and this had plants growing under normal conditions free from the

influence of the treatment.

3.3 Experimental design

Randomized block design (RBD) was used during this experiment. The blocks were

randomly assigned to block out the effect of temperature and exposure to sunlight (Figure 1.1

and 1.2). These two were found to be confounding factors in the glasshouses and it was

13
difficult to create homogeneity among our experimental units hence this choice of

experimental design. The first block contained 10 plants of Gloria and the second block

contained 10 plants of Nua 45. Each of the two blocks had three replicates thus giving a total

of four blocks containing each bean variety. Each glasshouse thus had a total of 8 randomly

assigned blocks. The other glasshouse acted as the control and Aphis fabae was not

inoculated in this one.

Nua 4 Nua 2

Nua 3 Glo 1

Glo 4 Glo 3

Nua 1 Glo 2

Figure 1.1: Arrangement of blocks in Glasshouse 1 where the black bean aphid (Aphis fabae)

was inoculated.

14
 Nua 3 Nua 4

Glo 2 Glo 1

Nua 1 Glo 3

Nua 2 Glo 4

Figure 1.2: Arrangement of blocks in Glasshouse 2 which served as the control.

Key

Glo- Gloria Nua- Nua 45

3.4 Rearing of aphids

A single mother of apterous adult aphid collected from infested fields in Zaka was used to

initiate black bean aphid culture on the bean plant. Prior to the experiment, the black bean

aphid culture had already been running for 4 weeks. New bean seedlings were provided

continuously to replace old ones for the maintenance and continuous growth of the black

bean aphid cultures.

15
3.5 Preparation of sample plants

This study was carried out in two separate glasshouses and each glasshouse had a total of 80

plants, that is, 40 for each variety under study. The seeds were planted in pots that were filled

with fine loam soil at a density of three seeds per pot and were placed 2.5 to 5.0 cm below the

soil surface. Seeds were planted in plastic pots (20cm diameter) and seven days after

emergence, seedlings were thinned to a single plant for each pot to avoid plants competing

for nutrients.

3.5.1 Pathological seed analysis

The seeds were tested to see if they were free from disease and healthy using the blotter test

method (Zad,1987). Prior to the blotter test, seeds were sterilised in 1% sodium hypochlorite

for 5 minutes to free or reduce superficial microorganisms (Sharma et al., 1997). The seeds

were then placed on layers of water soaked paper in petri dishes and incubated for 7 days at

20℃.

3.5.2 Soil sampling

Soil used in this study was collected from the DRSS fields using a spade at a depth of 0-

50cm. The soil was air dried under shade and ground with a wooden roller and then passed

through a 2-mm sieve. The pH of the soil used in this study was tested using a pH test kit.

The soil in the pots had a depth of 90 cm. All the soil used in this research was made as free

as possible of clumps of earth or sod by manually destroying lumps of soil using hands.

3.5.3 Watering and fertilisation

16
The plants were drenched with water every two days to ensure that they received adequate

water supply for photosynthesis and to maintain soil moisture. Shortly after emergence the

plants were fertilised with a 5:10:10 blend of synthetic fertilizer with four granules per plant.

Fertiliser was applied to all the plants in both glasshouses including the one which served as

the control.

3.5.4 Weed control

Seeds were planted in soils that were already free from weeds as beans are slow growing

plants and do not easily overshadow weeds. Weeds that emerged during the experiment were

removed manually by pulling them out using hands.

3.6 Determination of the effect of the Black bean aphid (Aphis fabae) on the plant

height, leaf area and yield of Gloria and Nua 45.

3.6.1 Inoculation of sample plants with the Black bean aphid (Aphis fabae)

Since young plants are very susceptible to aphid feeding, the Aphis fabae was inoculated 14

days after plant emergence. Ten 3 to 5 days old nymphs (apterous) were introduced on each

plant and covered with clear PVC tubes (90 cm high by 30 cm in diameter) and ventilated

using muslin cloth. Aphids were only inoculated in the first glasshouse whilst no aphids were

inoculated at all in the other glasshouse which served as the control. To avoid injuring the

aphids an aphid gun was used to transfer the aphids from the rearing unit onto the sample

plants.

3.6.2 Measuring of plant height

At day 90 (maturity), 6 randomly selected plants from each block had their height measured

in cm. Sampled plants were measured from the ground level to the tip of the plant using a

ruler and a tape measure.

17
3.6.3 Determination of leaf area

The areas of 5 randomly selected leaves on each of the 6 randomly sampled plants in each

block were assessed at maturity at day 90 in the glasshouse. Leaf area measurements were

performed early in the morning, when there was more diffuse and less direct solar radiation.

The leaf areas were measured using leaf area metre (L.I 3100, Li-Cor.inc, Lincoln, NE). Dead

and senescent leaves were disregarded in determinations of leaf area.

3.6.4 Measuring of yield

At maturity, the 100-seed weight was recorded for each of the 6 randomly selected plant from

each block.

3.7 Data analysis

Analysis of variance was conducted using one-way ANOVA for RBD in SPSS. There was

one predictor variable (factor) that is, bean variety with two factor levels (Gloria and Nua 45)

and three response variables that is leaf area, plant height and yield (100-seed weight). A one

-way ANOVA was conducted to compare the susceptibility to feeding by the black bean

aphid (Aphis fabae) between two varieties of beans commonly grown in Zimbabwe, that is,

Gloria and Nua 45. Dunnett test was used to compare the susceptibility to feeding by the

black bean aphid (Aphis fabae) between two varieties of beans (Gloria and Nua 45) to the

control.

18
 CHAPTER FOUR: RESULTS

4.1 Effect of A. fabae on the plant heights, leaf areas and 100-seed weight (yield) of

Gloria and Nua 45 plants

Aphis fabae feeding had an effect on the plant heights, leaf areas and 100-seed weight of

Gloria and Nua 45 (p<0.05). Generally, Gloria was more susceptible to feeding by the black

bean aphid (Aphis fabae) more than Nua 45 (Table 1) (p<0.05). The sample plants in the

glasshouse where the aphid was inoculated had reduced plant heights, leaf areas and 100-seed

weight (yield) compared to control plants where the aphids were not inoculated (p<0.05).

Table 1 shows the means for the plant height, leaf area and 100-seed weight of the 6 plants

that were randomly selected from each block at maturity.

19
Table 1: Effect of A. fabae on the mean plant heights, leaf areas and 100-seed weight (yield)

of Gloria and Nua 45.

The mean plant height ranged from 26.27 cm to 45.22 cm for both varieties and the mean leaf

areas ranged from 29.80 cm2 to 48.94 cm2. The 100-seed weights ranged from 34.83g to

46.23g (Table 1).

20
Table 2: Mean plant heights, leaf areas and 100-seed weights of the control plants

(where the aphid was not inoculated)

The mean plant heights in the control glasshouse ranged from 61.24 cm to 64.24cm for both

varieties and the mean leaf area ranged from 54.97cm2 to 59.01cm2. The mean 100-seed

weight ranged from 58.44g to 62.68g. The mean plant height, leaf area and 100-seed weight

21
were significantly different (p<0.05) from those of plants in the glasshouse where the aphid

was inoculated.

Figure 2: Effect of Aphis fabae feeding on Gloria and Nua 45 compared to the control

22
Figure 2 is showing the mean plant height, leaf area and the 100-seed weights of the sample

plants that were in the glasshouse where the Aphis fabae was inoculated compared to those of

the plants that were in the control glasshouse where the aphid was not inoculated.

The results in Table 1 and Figure 2 show that the least mean plant height (26.27cm) in the

inoculated glasshouse was recorded for Gloria. The least mean leaf area (35.16cm2) was

recorded for Gloria and the least mean 100-seed weight (34.83g) was also recorded for

Gloria.

The results in Table 2 and Figure 2 show that the mean plant height ranged from 61.24 cm to

64.24 cm for both varieties. The mean leaf area ranged from 55.81 cm2 to 59.01 cm2 and the

mean 100-seed weight for both varieties ranged from 58.44g to 63.37g.

Tukey post hoc test revealed that there was a significant difference in the plant heights, leaf

areas and 100-seed weights between the Gloria and Nua 45 plants that were in the glasshouse

which was inoculated with the aphid (ANOVA: p<0.05), confirming differences shown in

Fig 2.

Dunnett post hoc pairwise comparisons showed no significant differences between the plant

heights, leaf areas and 100-seed weight of the Gloria control plants and Nua 45 control plants

(ANOVA: p>0.05). However, all the sample plants in the inoculated glasshouse were

significantly different (ANOVA: p<0.05) from the control.

23
 CHAPTER FIVE: DISCUSSION

5.1 Effect of Aphis fabae feeding on the plant heights of Gloria and Nua 45

All the plants of the two varieties used in this study were susceptible to feeding by the black

bean aphid (Aphis fabae). The plant heights of Gloria and Nua 45 were reduced but up to

varying degrees. This was because the aphids feed by sucking plant juices and remove sap

which creates a lack of vigour in the plant and reduces plant height. Gloria was the most

susceptible because it had the most reduced plant heights. This is because the aphid preferred

feeding on the Gloria plants more than the Nua 45 plants. This could have been due to

different plant quality between the two varieties. Aphids tend to change feeding locations

more often when feeding on poor food sources compared with higher quality food sources

(Dixon, 1998). Previous work has shown that on preferred host plants, aphids have short

walking times and prolonged probing periods, in this case this could have been the Gloria

plants whereas the opposite behaviour is observed on non-preferred host plants, for instance,

Nua 45 plants in this study (Traicevski and Ward, 2002).

5.2 Effect of Aphis fabae feeding on the leaf areas of Gloria and Nua 45

Gloria also had the most reduced leaf areas compared to Nua 45 but both varieties were

susceptible to aphid feeding and had their leaf areas reduced. However, Nua 45 seemed more

resistant to aphid infestation better than Gloria. Reduction in leaf area was because of the

toxins found in the saliva that is injected into plants by the black bean aphid, which causes

leaves to curl and reduce their surface area. Aphids produce two different types of saliva

(Miles, 1999). The first type is dense and proteinaceous, and, jellifying around the stylets

(stylet sheaths), it constitutes an intercellular path towards the phloem for the piercing stylets,

24
isolating plant tissues from the mouthparts, thus preventing plant reaction at the site of

feeding (Felton and Eichenseer, 1999). When the stylets have reached the phloem flow,

aphids start to produce the second type of saliva, referred to as „watery‟, that is injected

directly into the vascular system of the plant (Douglas, 2006).

The aphids also secrete excess sugary fluid called honeydew which adheres to plants where it

promotes growth of sooty moulds. This also reduces the surface area of the plant available for

photosynthesis and may reduce the value of the crop. This is the reason why the plants of

both varieties in the control had much increased leaf areas.

5.3 Effect of Aphis fabae feeding on the 100-seed weight of Gloria and Nua 45

Gloria had much reduced 100-seed weights than Nua 45 and this is because Nua 45 had a

better compensatory effect where it managed to compensate for the loss that was being

caused by aphid feeding. The aphids were more localised in Nua 45 plants than in Gloria and

this allowed compensation to occur. Aphid feeding causes damage to flowers and pods which

may not develop properly. Flower and pod formation may abort due to the action of the toxic

saliva injected by the aphid to improve the flow of sap. Present findings were close to those

of studies carried out in France, where A. fabae reduced yields (quantified as weight of 1000

seeds and number of seeds per pod) of V. faba (another variety of Phaseolus vulgaris) by up

to 30% (Bouchery, 1977). Higher yield losses due to A. fabae on V. faba were also reported

in a field study in Iraq, using artificial infestations. Pod number, pod weight and dry weight

of seeds (yield) were negatively correlated with aphid numbers, the reductions in these

parameters reached 70, 76 and 64%, respectively (Mohammad and Abdulla, 1988).

A similar study by Salazar (1984) showed that the length of time that A. fabae is present on

V. faba has been correlated with yield loss in a field study in South America. Early

25
infestations (before the flowering period) resulted in considerable damage, dependent on the

population levels attained in the aphid colonies. High infestations at this early stage decreased

yields (total weight of seeds) by up to 42% compared with controls. Infestation at the

beginning of the flowering period has little effect on flower formation and development,

although high populations at this stage may have an adverse effect on pod formation. Any

infestations later than this stage had no effect on yield.

5.4 Conclusion

Gloria was more susceptible to aphid feeding, with Nua 45 showing to be more resistant over

a 90-day period. Trends from the results revealed that Gloria plants that were in the

glasshouse inoculated with A.fabae had the most reduced plant heights, leaf areas and 100-

seed weights. However, both varieties were susceptible to feeding by A.fabae although in

varying degrees because the control plants where the aphis was not inoculated had much

increased plant heights, leaf areas and 100-seed weights. Hence between the two varieties

that were under study, Nua 45 is the least affected by the A.fabae.

5.5 Recommendations

This study recommends that in areas where there is occurrence of A.fabae farmers cultivate

the Nua 45 variety as it is more resistant to the aphid feeding. Moreover, farmers will have

higher yields and invest less in pesticides procurement some of which are harmful to the

environment and may cause pest outbreaks. Damage and economic loss caused by A. fabae

can also be avoided through the use of agricultural practices such as intercropping the bean

varieties with maize.

26
REFERENCES

Akbar, H. G (2000). Seed availability, an ignored factor in crop varietal adoption studies: a

case study of beans in Tanzania. Journal of Sustainable Agriculture. 21:5-20.

Banks, C. J. and Macaulay, E. D. M. (1964). The feeding, growth and reproduction of Aphis

fabae Scop. on Vicia faba under experimental conditions. Annals of Applied Biology, 53:229-

242.

Banks, C. J. and Macaulay, E. D. M. (1967). “Effects of Aphis fabae and of its attendant ants

and insect predators on yields of field beans (Vicia faba).” Annals of Applied Biology. 60 (3):

445-453.

Berim, M. N. (2009). “Aphis fabae Scopoli – Black bean aphid. Interactive Agricultural

Ecological Atlas of Russia and Neighbouring Countries. AgroAtlas.

Bhatt, M. and Chanda, S. V. (2003). Prediction of leaf area in Phaseolus vulgaris by non-

destructive method. Bulgarian Journal of Plant Physiology,29(2):96-100.

Blane, W. M., Simmonds, M. S. J., Ley, S. V., Anderson, J. C and Toogood, P. L. (1990).

Antifeedant effects of Azaradirachtin and structurally related compounds on Lepidoptera

larvae. Entomological Experiments et Applicata. 55:149-160.

Bliss, M.S. (1980) Effects of insect-vector preference for healthy or infected plants on

pathogen spread: insights from a model. Journal of Economic Entomology. 101:1–8.

Bouchery, H. S. (1977). "Origin of the Common Bean, Phaseolus vulgaris". Economic

Botany. New York: New York Botanical Garden Press. 23 (1): 55–69.

27
Bressani, J. S. (1983). "Host alternation in Aphis fabae Scop. I. feeding preferences and

fecundity in relation to the age and kind of leaves". Annals of Applied Biology. 38 (1): 25–64.

Cammell, M. E. (1981). The black bean aphid, Aphis fabae. Biologist, 28 (5):247-258.

Cammell M. E. and Way M. J. (1983). Aphid pests. In: Hebblethwaite PD, ed. The Faba

Bean. Cambridge, UK: Cambridge University Press, pp. 315-346.

Chazan, M. (2000). World Prehistory and Archaeology: Pathways through Time. Pearson

Education, Inc.

Chester, F. G. (1969). Hohabinhian: a pebble tool complex with early plant association in

South Asia.

Cubero, J. I. (1974). On the evolution of Vicia faba. Theor. Appl. Genet. 45:47-51

Curtis, J. (1960). Farm insects: being the natural history and economy of the insects injurious

to the field crops of Great Britain and Ireland, and also those which infest barns and granaries

with suggestions for their destruction. Glasgow, UK: Blackie.

Daniel, Z. and Maria, H. (2000). Domestication of plants in the old world.

Daughtry, C. (1990). Direct measurements of canopy measurements of canopy structure.

Remote Sens.Rev, 5:45-60.

Davidson, J. (1921). Biological studies of Aphis rumicis Linn. Bulletin of Entomological

Research, 12:81-89.

Dixon, A.F.G. (1987). The way of life of aphids: host specificity, speciation and distribution.

In: Minks A. K, Harrewijn, P. editors. Aphids. New York: Elsevier. p. 197-207.

Douglas, M. (2006). Collins Field Guide to the Insects of Britain and Northern Europe.

Harper Collins.

28
Dube, R. and Nyoka, B. I. (1993). Past, present and future status of cowpea research in

Zimbabwe. In: Trends in cowpea research in Zimbabwe.

Eastop, V.F. (1977). Aphis fabae. In: Kranz J, Schmutterer H, Koch W, eds. Diseases, Pests

and Weeds of Tropical Crops. Berlin & Hamburg, Germany: Verlag Paul Parey, pp. 326-327.

Everett, H. (2009) Bickley Collection, Archives Center, National Museum of American

History, Smithsonian Institution.

FAO (2004). Databases, http://www.fao.org/waicent/Agricul.htm (accessed 04/09/17)

FAO (2006). Food and agriculture organization of the United Nations - statistical database.

http://www.fao.org/faostat. (accessed 05/08/17).

Felton, J. S. and Eichenseer, C. O. (1999). "Host alternation in Aphis fabae Scop. I. feeding

preferences and fecundity in relation to the age and kind of leaves". Annals of Applied

Biology. 38 (1): 25–64.

Gentry, H. S. (1969). "Origin of the Common Bean, Phaseolus vulgaris". Economic Botany.

New York: New York Botanical Garden Press. 23 (1): 55–69.

Gepts, P. and Bliss, D. (1985). Phaseolin as an evolutionary marker. Genetic resources of

genes for seed proteins, isozymes, and morpho- logical traits in common bean (Phaseolus

vulgaris). pp 215-241.

Godfrey, L. D. and Trumble, J. T. (2009) “UC IPM Pest management guidelines: Celery”.

UC IPM.

Gridley, S. and Daniel, G. (1983). The epidemiology of some aphid-borne viruses in

Australia. Plant Virus Epidemiology (ed. by RT Plumb & JM Thresh). Blackwell, Oxford,

UK.

29
Hajjar, R. and Hodgkin, T. (2007). The use of wild relatives in crop improvement: a survey

of developments over the last 20 years. Euphytica 156:1-13.

Hardwick, R. C. (1988). Review of recent research on navy beans (Phaseolus vulgaris) in the

United Kingdom.

Harper, J. L. (1977). Population Biology of Plants, Academic Press, Oxford, UK.

Hawtin, G. C. and Hebblethpiait, P. D. (1983). Background and history of faba bean

production. In: The Faba Bean (Vicia faba L.) (Hebblethwaite, P.D., ed.). Buttenvorths,

London, U.K, pp 3-22.

Hinz, B. and Daebeler, F. (1981). Harmful effects of the black bean aphid (Aphis fabae

Scop.) on field beans. Nachrichtenblatt fur den Pflanzenschutz in der DDR, 35(9):175-178.

Hosfield, G. L., Vebersax, M. A. and Occena, L. G. (2000). Technological and genetic

improvements in dry bean quality and utilization. In: Singh Bean Research. Production and

Utilization. Proc Idaho Bean Workshops. University of Idaho, Moscow, pp 135-152.

Hurej, M. and Werf, W van der.(1993). The influence of black bean aphid, Aphis fabae Scop.,

and its honeydew on leaf growth and dry matter production of sugar beet. Annals of Applied

Biology, 122(2):201-214

Johnes, H. G. and Corlett, J. E. (1992). Current topics in drought physiology. Journal of

Agricultural Science, 119: 291-296.

Kaplan, L. (2008). Legumes in the history of human nutrition. The world of soy.

Katungi, E., Sperling, L., Karanja, D., Farrow, A. and Beebe S.( 2011). Relative importance

of common bean attributes and variety demand in the drought areas of Kenya. Journal of

Development and Agricultural Economics. 3(8):411-422.

30
Limonard, T. (1966). A modified blotter test for seed health. Government seed testing station,

Wageningen.

Lincoln, C.P. (1987). Vegetable characteristics, production and marketing. Printed in the

U.S.A.

Lynch, J., Lauchli, A. and Epstein, E. (1991). Vegetative growth of the common bean in

response to phosphorus nutrition. Crop Science, 3:380-387.

Makini, F. W. and Danial, D. L. (1995). Bean production in Kenya with emphasis on

diseases, Proceedings of a regional workshop for Eastern, Central and Southern Africa held at

Njoro, Kenya. Wageningen Agricultural University: Wageningen, 104-109.

Marshall, J. K. (1968). “Methods of leaf area measurement of large and small leaf samples.”

Photosynthetica, 2:41-47.

Mathur, S. B. and Jorgensen, J. (1998). Different types of damages in seeds caused by seed

borne fungi. Proceedings of CTA seminar, Copenhagen, Denmark.

Mauki, S. J. N (1988). The evaluation of pesticides and control of bean flower thrips in the

common bean (Phaseolus vulgaris) at Thika, Kenya: ACTA Horticulture, 218.

Mihailovic, H., Powell, G. and Tosh, C.R (2005) Host plant selection by aphids: behavioral,

evolutionary, and applied perspectives. Annual Review of Entomology. 51:309–330.

Miles, F. H. (1999). Food Composition and Nutrition Tables. Medpharm Scientific

Publishers Stuttgart.

Mohammad, M. A. and Abdulla, S. A. (1988). Study on the effect of the black bean aphid,

Aphis fabae Scop. (Homoptera, Aphididae) on the green and dry product yield of broad beans

in the Mosul region. Mesopotamia Journal of Agriculture, 20(2):293-300.

31
Naqvi, H. K. (1984). Cultivation under the Sultans of Delhi c. 1206–1555. Indian J. History.

Sci. 19:329–340.

Nelson, B. D. (1999). Fusarium blight or wilt, root rot and pod and collar rot. In:

Compendium of soybean diseases (3rd ed.

). American Phytopathological Society, St. Paul,

MN.

Norman, M. J. T., Pearson, C. J. and Searle, P. G. E. (1995). Tropical food crops in their

environment (2nd edition). University of Cambridge, New York, U.S.A.

Nyakwende, E., Paull, C. J. and Atherton, J. G. (1997). Non-destructive determination of leaf

area in tomato plants using image processing. J.Hort.Sci.72: 225-262.

Pachico, D. (1993). The demand for bean technology. Trends in CIAT Commodities.

Working document 128:60-74.

Peksen, E. (2007). Non-destructive leaf area estimation model for faba bean (Vicia faba L.).

Sci.Hort.11(3): 322-328.

Peters, A. (1993). Michigan Dry Bean Digest, China. 17 (4): 18-20.

Powell, G. and Hardie, J. (2002). Xylem ingestion by winged aphids. Entomologia

Experimentalis et Applicata.104(1):103-108; 22 ref.

Razia, A. (2000). Nuskha Dar Fanni-Falahat: The Art of Agriculture, Persion manuscript

compiled in the 17th century by the Mughal Prince Dara Shikoh, Agri-History pp. 98.

Rothamstead Research (2003). Breeding Better Beans. Rothamstead Agricultural Research

magazine.

32
Salazar, V. J. (1984). Efecto de diferentes epocas de infestacion por Aphis fabae Scop. en el

rendimiento de Vicia faba L. (The effect of different periods of infestation by Aphis fabae

Scop. on the yield of Vicia faba L.). Agronomia Tropical, 34 (4-6):21-33.

Schmutz, J., McClean, P. E., Mamidi, S., Wu, G. A., Cannon, S. B., Grimwood, J., Jenkins,

J., Shu, S., Song, Q. and Chavarro, C. (2014). A reference genome for common bean and

genome-wide analysis of dual domestications. Nature Genetics 46 (7):707-713.

Schwartz, H. F., Brick, M. A., Nuland, D. S. and Franc, G. D. (1996). Dry bean production

and pest management. Regional bulletin 562, pp 106. Universities of Colorado, Nebraska,

U.S.A.

Sharma, J. K., Ali, M. I. M. and Sutherland, J. R. (1997). Seed pathology of tropical

hardwoods. In: Prochazkova, Z(ed) Proc.ISTA Tree seed pathology meeting, Opocno, Czech

Republic, pp. 74-81.

Singh, S. P. (2002). Broadening the genetic base of common bean cultivars. Crop science:

41, 1659-1675.

Sperling, L., Scheidegger, U. and Buruchara, R. (1996). Designing seed systems with small

farmers: principles derived from bean research in the Great Lakes region of Africa.ODI

Network, pp 14.

Traicevski, V. and Ward, S.A (2002) Probing behaviour of Aphis craccivora Koch on host

plants of different nutritional quality. Ecological Entomology27:213–219.

Tyagi, O. A., Pokhariyal, G. P. and Buruchena, R. A. (1996). Character association for yield

and its components in field beans (Phaseolus vulgaris). Discovery and Innovations, 8: 47-51.

33
Tylwoska, K. (1997). Seed health testing in Eastern and Central European countries- the

present and prospects. In: Hutchins, J. D. and Reeves, J. C.(eds). Seed health testing, CAB

International, Wallingford, UK, pp 21-26.

Vital, S. (2001). The trends that are shaping our future. World Watch Institute, Washington

D.C.

Wood, A. J. and Roper, J. (2000). A simple and non-destructive technique for measuring

plant growth and development. American Biology Teacher.

Wortman, C. S and Allen, D. J. (1994). African bean production: their definitions,

characteristics and constraints. Network on bean research in Africa.

Wilkinson T. L. and Douglas A. E. (2003). Phloem amino acids and the host plant range of

the polyphagous aphid, Aphis fabae. Entomologia Experimentalis et Applicata, 106(2):103-

113.

World vision (2003). Malawi communications department “Chingale ADP Update,” World

Vision, Blantyre, Malawi. Wortmann, C. S. and Allen, D. J. (1994). Africa bean production

environments: their definition, characteristics and constraints. Network on bean research in

Africa.

Zad ,S.J. (1987). Soybean seed-borne diseases. Med. Fac. Landbouww. Rijksuniv. Gent 52

(3a): 825-829.

34
APPENDICES

Appendix 1: Pathological seed analysis using Blotter test

1. Sterilise seeds in 1% sodium hypochlorite for 5 minutes.

2. Soak blotter paper in distilled water then place it in a petri dish in the laminar flow.

3. Place 5 seeds on the soaked blotter paper and close the petri dish.

4. Incubate for 7 days at 20℃.

35
Appendix 2: Multiple comparisons for effect of A.fabae on the plant heights, leaf areas and

100-seed weights of the inoculated plants and the control plants.

Multiple Comparisons

Dependent Variable (I) (J) Mean Std. Sig. 95%

beanvariety beanvariety Difference (I- Error Confidence
J) Interval

Lower
Bound
*
Nua -15.93417 .74438 .000 -17.8819
*
Gloria gloriacontrol -34.35958 .74438 .000 -36.3073
*
nuacontrol -34.94208 .74438 .000 -36.8898
*
Gloria 15.93417 .74438 .000 13.9864
*
Nua gloriacontrol -18.42542 .74438 .000 -20.3732
*
nuacontrol -19.00792 .74438 .000 -20.9557
Tukey HSD *
Gloria 34.35958 .74438 .000 32.4118
*
plantheight gloriacontrol Nua 18.42542 .74438 .000 16.4777

nuacontrol -.58250 .74438 .862 -2.5302

*
Gloria 34.94208 .74438 .000 32.9943
*
nuacontrol Nua 19.00792 .74438 .000 17.0602

gloriacontrol .58250 .74438 .862 -1.3652

*
Gloria nuacontrol -34.94208 .74438 .000
Dunnett t *
b
Nua nuacontrol -19.00792 .74438 .000
(<control)
gloriacontrol nuacontrol -.58250 .74438 .416
*
leafarea Tukey HSD Gloria Nua -9.43333 1.01048 .000 -12.0774

36
 *
gloriacontrol -19.86583 1.01048 .000 -22.5099
*
nuacontrol -21.46292 1.01048 .000 -24.1070
*
Gloria 9.43333 1.01048 .000 6.7893
*
Nua gloriacontrol -10.43250 1.01048 .000 -13.0765
*
nuacontrol -12.02958 1.01048 .000 -14.6736
*
Gloria 19.86583 1.01048 .000 17.2218
*
gloriacontrol Nua 10.43250 1.01048 .000 7.7885
nuacontrol -1.59708 1.01048 .395 -4.2411
*
Gloria 21.46292 1.01048 .000 18.8189
*
nuacontrol Nua 12.02958 1.01048 .000 9.3855
gloriacontrol 1.59708 1.01048 .395 -1.0470
*
gloria nuacontrol -21.46292 1.01048 .000
Dunnett t *
b
nua nuacontrol -12.02958 1.01048 .000
(<control)
gloriacontrol nuacontrol -1.59708 1.01048 .136
*
Nua -8.90250 .88224 .000 -11.2110
*
gloria gloriacontrol -25.05917 .88224 .000 -27.3677
*
nuacontrol -25.64333 .88224 .000 -27.9518
*
Gloria 8.90250 .88224 .000 6.5940
*
nua gloriacontrol -16.15667 .88224 .000 -18.4652
*
nuacontrol -16.74083 .88224 .000 -19.0493
Tukey HSD *
Gloria 25.05917 .88224 .000 22.7507
Seedweig *
gloriacontrol Nua 16.15667 .88224 .000 13.8482
ht
nuacontrol -.58417 .88224 .911 -2.8927
*
Gloria 25.64333 .88224 .000 23.3348
*
nuacontrol Nua 16.74083 .88224 .000 14.4323

gloriacontrol .58417 .88224 .911 -1.7243

*
gloria nuacontrol -25.64333 .88224 .000
Dunnett t *
b
nua nuacontrol -16.74083 .88224 .000
(<control)
gloriacontrol nuacontrol -.58417 .88224 .469

*. The mean difference is significant at the 0.05 level.

b. Dunnett t-tests treat one group as a control, and compare all other groups against it.

37
Appendix 3: Means plot of the mean plant heights of the inoculated plants and the control

plants.

38
Appendix 4: Means plot of the mean leaf areas of the inoculated plants and the control

plants.

39
Appendix 5: Means plot of the mean 100-seed weights of the inoculated plants and the

control plants.

40