evaluation of organic amendments for the management of root-knot nematodes

278 Pages·2017·2.32 MB·English

EVALUATION OF ORGANIC AMENDMENTS FOR THE MANAGEMENT
OF ROOT-KNOT NEMATODES (MELOIDOGYNE SPP.) OF TOMATO
(SOLANUM LYCOPERSICUM L.)

BY
FAYE JERREH MANNEH
NOVEMBER, 2016
EVALUATION OF ORGANIC AMENDMENTS FOR THE MANAGEMENT
OF ROOT-KNOT NEMATODES (MELOIDOGYNE SPP.) OF TOMATO
(SOLANUM LYCOPERSICUM L.)

BY

FAYE JERREH MANNEH
(MPHIL Nematology)

NOVEMBER, 2016
KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY,
KUMASI
SCHOOL OF GRADUATE STUDIES
DEPARTMENT OF CROP AND SOIL SCIENCES

EVALUATION OF ORGANIC AMENDMENTS FOR THE MANAGEMENT
OF ROOT-KNOT NEMATODES (MELOIDOGYNE SPP.) OF TOMATO
(SOLANUM LYCOPERSICUM L.)

BY

FAYE JERREH MANNEH
(MPHIL Nematology)

NOVEMBER, 2016
EVALUATION OF ORGANIC AMENDMENTS FOR THE MANAGEMENT
OF ROOT-KNOT NEMATODES (MELOIDOGYNE SPP.) OF TOMATO
(SOLANUM LYCOPERSICUM L.)

A Thesis submitted to the DEPARTMENT OF CROP AND SOIL SCIENCES.
Kwame Nkrumah University of Science and Technology in partial fulfilment of the
requirements for the Degree of

DOCTOR OF PHILOSOPHY
IN
NEMATOLOGY

BY

FAYE JERREH MANNEH (MPHIL NEMATOLOGY)

NOVEMBER, 2016
DECLARATION
I, hereby, declare that this submission is my own work towards the PhD and that, to
the best of my knowledge, it contains no material previously published by another
person nor material which has been accepted for the award of any other degree of the
University, except where due acknowledgement has been made in the text.

FAYE JERREH MANNEH ……………... ……………..
Student No. 20432611 Signature Date

DR CHARLES KWOSEH ………………. ……………..
(Supervisor) Signature Date

PROF. RICHARD AKROMAH ……………. ……………..
(Supervisor) Signature Date

Certified by:
DR E. A. OSEKRE ……………... ……………..
(Head of Department) Signature Date
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ABSTRACT
Root-knot nematodes are considered as one of the most economically damaging of
plant-parasitic nematodes of tomato. In many cases, crop losses due root-knot
nematodes are reduced by the application of high toxic chemical nematicides. In
Ghana, farmers‟ management practices against root-knot nematodes are in most cases
ineffective. The purposes of the study were to determine farmers‟ perception of root-
knot nematodes, evaluate the efficacy of organic amendments at the laboratory, plant
house and field,assess the effect of fresh sweet orange and cassava combination on
root-knot nematode isolates, confirm the presence of the nematodes (Meloidogyne
spp.) using ITS-PCR molecular analysis,determine the time of application and
appropriate rate for the management of the nematodes on tomato. In determining
farmers‟ perception of root-knot nematodes and their management methods,
multistage sampling technique was used to select the respondents. The survey was
carried out in six district in Ashanti region. Majority (30%) of the tomato farmers
were aware of root-knot nematode problems in their fields. Majority of them
identified root-knot nematodes as the most prevalent pest in tomato. However, none
of them used nematicides in the management of the nematodes. All the farmers used
crop rotation and weeding as a management strategy against the nematodes. For the
evaluation of the efficacy of organic amendments in the management of the
nematodes, experiments were carried out in the laboratory, plant house and research
field of KNUST. In the laboratory, fresh sweet orange peels significantly (P<0.05)
inhibited egg hatching and increased mortality of root-knot nematodes than the rest of
the treatments. Fresh orange peel performed just as well as carbofuran in reducing the
number of root-knot nematodes in the root of tomato and soil, number of eggs, root
galling and reproduction factor of the nematodes in the plant house. In the field, the
application of fresh sweet orange peels followed by cassava peels and the reverse
application of the two materials significantly increased (P<0.05) the fresh shoot
weight, fruit size and yield of tomato than the other treatments. The number of root-
knot nematodes in the roots of tomato and soil, number of eggs and root galling were
significantly reduced (P<0.0) upon the application of carbofuran and its effect was
similar to that of the combined application of fresh sweet orange and cassava peels.
The application of sweet orange peels significantly increased (P<0.05) the number of
colonies of Trichoderma viride than cassava peels, but cassava peels also significantly
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increased (P<0.05) the number of the bacterivorous nematodes,Heterocephalobellus
sp. andEucephalobus sp. and the fungivorous nematodeDitylenchussp.more than
sweet orange peels. Further evaluation of the combined application of fresh sweet
orange and cassava peels on root-knot nematode isolates showed no significant
differences (P>0.05) between the isolates in terms of number of juveniles in the roots
of tomato and soil, number of eggs, root galling and reproduction factor. However,
the pots treated with combined application of fresh sweet orange and cassava peels
significantly increased (P<0.05) the fresh shoot weight and plant height of tomato
more than carbofuran-treated pots. The amplification of the ITS regions of the
nematodes, using multigene loci primers, confirmed the presence of Meloidogyne spp.
The application of fresh sweet orange and cassava peels combination at four weeks
before transplanting significantly reduced (P<0.05) the population of root-knot
nematodes and root galling of tomato than the rest of the treatments. Similarly, the
application of 25g each of both fresh sweet orange and cassava peels per pot
significantly reduced (P<0.05) the nematode population and root galling and inceased
fresh shoot weight and yield of tomato than the other treatments. The study showed
that root-knot nematode infestation was one of the major problems of tomato farmers
and there was no effective management technique. The combined application of fresh
sweet orange and cassava peels at 50g/plant four weeks before transplanting was
found to be the most effective treatment in the management of the nematodes. The
nematode isolates collected from different tomato growings areas were confirmed to
be Meloidogyne spp. Therefore, further evaluation on the efficacy, time and rate of
application of fresh sweet orange and cassava peels combination is recommended.
Also, simple and more reliable method such as isozymes should be used to identify
root-knot nematodes to the species level.

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TABLE OF CONTENTS
DECLARATION..........................................................................................................ii
ABSTRACT .................................................................................................................iii
TABLE OF CONTENTS ............................................................................................v
LIST OF TABLES .....................................................................................................xv
LIST OF PLATES .....................................................................................................xv
LIST OF FIGURES ..................................................................................................xix
ABBREVIATIONS ....................................................................................................xx
ACKNOWLEDGEMENTS .....................................................................................xxi
DEDICATION..........................................................................................................xxii
CHAPTER ONE ..........................................................................................................1
1.0 GENERAL INTRODUCTION .............................................................................1
CHAPTER TWO .........................................................................................................4
2.0 LITERATURE REVIEW .....................................................................................4
2.1 Economic Importance of Tomato ........................................................................ 4
2.2 Constraints in Tomato Production ....................................................................... 4
2.3 Economic Importance of Root-knot Nematodes.................................................. 5
2.4 Means of Survival of Root-knot Nematodes (Meloidogyne spp.) ....................... 6
2.5 Reproduction and Development of Root-knot Nematodes .................................. 7
2.6 Damage Symptoms of Root-knot Nematode Species in Tomato Plant ............... 8
2.7 Control Methods of Root-knot Nematodes .......................................................... 9
2.7.1 Use of Synthetic Nematicides in Controlling Root-knot Nematodes ........... 9
2.7.2 Use of Contact Nematicides (Non-volatile)................................................ 10
2.7.3 Use of Carbofuran in the Management of Nematodes................................ 11
2.7.4 Use of Fumigants (Volatile Nematicides)................................................... 11
2.7.5 Soil Solarization in Management of Root-knot Nematodes ....................... 12
2.7.6 Use of Crop Rotation in the Management of Root-knot Nematodes .......... 13
2.7.7 Biological Control of Root-knot Nematodes .............................................. 14
2.7.7.1 Potential of T. viride in the Management of Root-knot Nematodes ..... 16
2.7.8 Use of Organic Soil Amendments .............................................................. 17
2.7.9 Potential of Cassava Peels in Root-knot Nematode Management ........... 18
2.7.9.1 Cyanide Content in Cassava Peel ......................................................... 19
2.7.9.2 Mode of Action of Cyanogens on Nematodes ...................................... 19
2.7.10 Potential of Sweet Orange Peels in Root-knot Nematode Management20
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2.7.10.1 Mode of Action of Lemonene on Nematodes ..................................... 21
2.7.11 Effect of Rice Husk on Nematodes........................................................ 21
2.7.11.1 Mode of Action of Rice Husk on Nematodes ..................................... 22
2.7.12 Effect of Poultry Manure in the Management of Root-knot Nematodes
............................................................................................................ 22
2.7.12.1 Effect of Soil Nutrients and pH on Root-knot Nematodes ................. 23
2.8 Identification of Root-knot Nematodes ............................................................. 24
2.8.1 Morphological Identification ...................................................................... 25
2.8.2 Biochemical Method ................................................................................... 26
2.8.2.1 Identification of Root-knot Nematodes using Isozyme ........................ 26
2.8.3 Molecular Identification.............................................................................. 27
CHAPTER THREE ...................................................................................................28
3.0 PRODUCTION PRACTICES AND PEST AND DISEASE PROBLEMS OF
TOMATOES IN ASHANTI REGION OF GHANA ..............................................28
3.1 Introduction ........................................................................................................ 28
3.3 Methodology ...................................................................................................... 29
3.4 Data Collection and Analysis............................................................................. 30
3.5 Results ................................................................................................................ 31
3.5.1 Production practices .................................................................................... 31
3.5.2 Pests and Diseases Associated with Tomato Production in the Ashanti
Region ........................................................................................................ 33
3.6 Management Practices Carried out by Respondents.......................................... 34
3.6.1 Insecticides and Fungicides Used in the Management of Pests and Diseases
on Tomato by the Farmers in the Surveyed Areas..................................... 35
Anchor Allied Factory Ltd. .................................................................................. 35
3.6.2 Crop Rotation Systems Practiced by Respondents in the Surveyed Areas. 36
3.6.3 Effectiveness of management practices of tomato farmers in the surveyed
areas ........................................................................................................... 37
3.7 Discussion .......................................................................................................... 38
3.8 Conclusions and Recommendations .................................................................. 40
CHAPTER FOUR......................................................................................................42
4.0 EFFICACY OF ORANGE AND CASSAVA PEELS IN THE
MANAGEMENT OF ROOT-KNOT NEMATODES IN VITRO AND IN VIVO42
4.1 Introduction ........................................................................................................ 42
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4.2 Experiment 1: Evaluation of aqueous extracts of sweet orange and cassava peel
for their nematicidal potentials on root-knot nematodes in vitro ...................... 43
4.3 Materials and Methods....................................................................................... 44
4.3.1 Location of the Experiment......................................................................... 44
4.3.2 Experimental Design................................................................................... 44
4.3.3 Sources of the Organic Amendment Materials ........................................... 45
4.3.4 Analysis of Nutrient Content of the Organic Amendments ........................ 45
4.3.4.1 Determination of pH of the Organic Amendments ............................... 45
4.3.4.2 Preparation and Dry Ash Digestion of Organic Amendments for
Elemental Analysis ............................................................................. 46
4.3.4.3 Determination of Phosphorus (P).......................................................... 46
4.3.4.4 Determination of Potassium (K) and Sodium (Na)............................... 47
4.3.4.5 Determination of calcium (Ca) and magnesium (Mg) .......................... 48
4.3.4.6 Determination of Calcium..................................................................... 49
4.3.4.7 Determination of Magnesium ............................................................... 49
4.3.5 Determination of Hydrogen Cyanide Content in Fresh and Dry Cassava
Peels.................................................................................................... 50
4.3.6 Preparation of the Aqueous Extracts of the Organic Amendments ............ 51
4.3.7 Sterilization of Soil for Culturing of Root-knot Nematodes....................... 52
4.3.8 Culturing and Extraction of Root-knot Nematode Juveniles and Eggs ...... 52
4.3.9 Assessment of Root-knot Nematode Juvenile Mortality ............................ 53
4.3.10 Assessment of Root-knot Nematode Egg Hatch Inhibition in vitro ......... 53
4.3.11 Data Collection ......................................................................................... 54
4.3.12 Data analysis ............................................................................................. 54
4.4 Results ................................................................................................................ 54
4.4.1 Root-knot Nematode Eggs Hatched at 3, 6 and 9 Days after Inoculation in
Different Aqueous Extracts in vitro ........................................................... 54
4.4.2 Mortality of Root-knot Nematode Juveniles in the different Aqueous
Extracts in vitro .......................................................................................... 57
4.5 Experiment 2: Evaluation of Aqueous Extracts of fSweet Orange and Cassava
Peel or Their Nematicidal Potentials on Root-Knot Nematodes of Tomato in
the Plant House (In vivo)................................................................................... 58
4.5.1 Materials and Methods................................................................................ 58
4.5.1.1 Condition in the Plant House ................................................................ 58
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