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UNIVERSITI PUTRA MALAYSIA

BIOLOGY OF COCHLOCHILA BULLITA STAL AS POTENTIAL PEST OF ORTHOSIPHON ARISTATUS (BLUME) MIQ. IN MALAYSIA

TAN LI PENG

FH 2014 2

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BIOLOGY OF Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE), A POTENTIAL PEST OF Orthosiphon aristatus (BLUME) MIQ. (LAMIALES:

LAMIACEAE) IN MALAYSIA

By

TAN LI PENG

Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia,

in Fulfilment to the Requirement for the Degree of Doctor of Philosophy

July 2014

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COPYRIGHT

All material contained within the thesis, including without limitation text, logos, icons, photographs and all other artwork, is copyright material of Universiti Putra Malaysia unless otherwise stated. Use may be made of any material contained within the thesis for non-commercial purposes from the copyright holder. Commercial use of material may only be made with the express, prior, written permission of Universiti Putra Malaysia.

Copyright © Universiti Putra Malaysia

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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment of the requirement for the degree of Doctor of Philosophy

BIOLOGY OF Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE), A POTENTIAL PEST OF Orthosiphon aristatus (BLUME) MIQ. (LAMIALES:

LAMIACEAE) IN MALAYSIA

By

TAN LI PENG

July 2014

Chairman: Prof. Ahmad Said Sajap, PhD

Faculty: Forestry

Cochlochila bullita (Stål) is an importance pest in some Asia countries such as India, Kanpur and Thailand attacking plants form the genus Ocimum, herein its common name, ocimum tingid. Cochlochila bullita is first recorded in Malaysia in the year 2009, attacking one of the important medicinal herbs in this country, the Orthosiphon aristatus (Blume) Miq. Biology of this pest was studied to get a deeper understanding of this bug associated with O. aristatus, which will lead to better integrated management on this pest. Morphology and morphometric of C. bullita were described and measured from 15 samples of all developmental stages of this bug in order to provide the fundamental reference for identification. From here, C. bullita possessed a body length and width ratio for about 1.51 to 1.59 mm and the head capsule width was recommended to distinguish the instars. Life-tables of this pest on O. aristatus and Ocimum basilicum Linnaeus were constructed by analyzing the life-parameters from 10 cohorts on each plant. Results showed that on O. aristatus significantly higher finite rate of increase (λ), 1.07 and intrinsic rate of increase (rm), 0.07 were obtained and thus suggest that C. bullita performed better on O. aristatus. The preferences of C. bullita toward these two plants were also examined, it preferred O. aristatus; with a notable feeding preference and relatively higher oviposition preference. The reason of this phenomenon was presumed related to the trichomes density of the plants; hence a further investigation on the correlation between the trichomes density and the number of eggs deposited was conducted. Strong, positive and significant correlation was found on the O. aristatus. Some behavioural studies were carried out, particularly the mating behaviour and maternal care of this species to comprehend how this bug adapts to produce and ensure the succession of its own kinds. One male was freely mixed around with five females in 7 days. Results showed that the male C. bullita was able to fertilize maximum up to four females within the given periods. In the other hand, females were tested for their maternal care, samples collected from the field were tested by provoking the females

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with a standard protocol; no aggressive approach were shown by the females and hence concluded that this bug possessed only passive guarding behaviour. Injuries caused by this bug were assessed by measuring the injury area, chlorophyll content and faeces production. Results showed a significant higher injury area, faeces production and chlorophyll depletion on O. aristatus. Water content was presumed play a role in the susceptibility of O. aristatus; as O. basilicum having higher water content 91% than O. aristatus with only 88%. After all, O. aristatus is attractive to C. bullita but the tolerance of this plant towards the infestation of the bug is relatively low. Water content and the trichomes density of the plant were believed that contributed to this selectivity and susceptibility. Knowledge of the biology assists in identifying, recognizing and monitoring the insect pest is a important input towards develop a better management strategy for this pest.

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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Doktor Flasafah

BIOLOGY OF Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE), A POTENTIAL PEST OF Orthosiphon aristatus (BLUME) MIQ. (LAMIALES:

LAMIACEAE) IN MALAYSIA

Oleh

TAN LI PENG

July 2014

Pengerusi: Prof. Ahmad Said Sajap, PhD

Fakulti: Perhutanan

Cochlochila bullita (Stål) merupakan salah satu perosak yang penting di negara-neagara Asia seperti India, Kanpur dan Thailand yang menyerang pokok bawah genus Ocimum, dengan ini, dapat namanya ocimum tingid. Cochlochila bullita juga didapati dan pertama kalinya direkodkan di Malaysia pada tahun 2009. Perosak ini menyerang salah satu pokok herba terkanal di negara ini, iaitu Orthosiphon aristatus (Blume) Miq. Oleh itu, biologi bagi perosak ini telah dikaji untuk mendapat informasi yang lagi mendalam bagi perosak yang menyerang pokok O. aristatus. Morfologi dan morfometrik serangga ini telah digambarkan dan diukur dari 15 sampel serangga bagi setiap instar supaya informasi ini dapat dijadikan satu rujukan untuk pengenalan spesis tersebut. Daripada kajian ini, ratio panjang bahagikan dengan lebar badan serangga ini adalah di antara 1.51 hingga 1.59 mm dan lebar kepala serangga ini adalah parameter yang disyorkan untuk digunakan untuk pengelasan peringakat nympha. Jadual hidup perosak ini juga dikaji dengan menggunakan 10 cohorts atas pokok O. aristatus dan Ocimum basilicum Linneaus. Adalah didapati kadar pertambahan finit harian (λ), 1.07 and kadar pertambahan intrinsik semula jadi (rm), 0.07 bagi C. bullita yang hidup pada O. aristatus adalah lebih tinggi daripada C. bullita yang hidup pada O. basilicum. Dengan ini, serangga ini mempunyai prestasi yang lebih tinggi apabila hidup pada O. aristatus. Selain itu, perosak ini juga didapati lebih cendorong kepada O. aristatus dari segi pemakanan dan pemilihan tempat bertelur. Keadaan ini dijangka adalah disebabkan oleh kepadatan trichomes yang terdapat pada pokok tersebut; oleh itu kajian hubungan antara kepadaptan trichomes dan bilangan telur yang terdapat telahpun dijalankan. Satu hubungan yang rapat, positif dan ketara didapati pada serangga yang bertelur pada the O. aristatus. Tambahan pula, kelakuan perosak ini terutamanya dari segi pengawanan dan penjagaan telur juga dikaji. Keputusan menunjukkan apabila satu jantan dikenalkan dengan lima betina dalam tempoh 7 hari, jantan tersebut dengan maksimum berjaya mengawan dengan 4 betina. Manakala, betina yang dikaji dari

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segi kebolehan menjaga telur tidak mempertahankan telurnya apabila dicabar. Dengan ini disimpulkan betina C. bullita hanya menlindungi telurnya secara pasif. Kecederaan pada pokok yang diakibatkan oleh perosak ini juga dinilai dengan mengukur kawasan kecederaan, pengurangan kandungan klorofil dan penghasilan perkumuhan. Data menunjukkan ketiga-tiga pembolehubah ini adalah lebih tinggi pada O. aristatus. Kandungan air dalam O. aristatus diramal memainkan peranan atas toleransi pokok tersebut terhadap seranggan serangga; ini dapat dijelaskan dengan O. basilicum yang mempunyai kandugan air sebanyak 91% dalamnya dan O. aristatus yang hanya mempunyai 88%. Secara kesimpulan, O. aristatus adalah lebih disukai oleh C. bullita, akan tetapi toleransi pokok ini terhadap serangan C. bullita adalah agak rendah. Akhirnya, kandungan air dan kepadatan trichomes untuk pokok tersebut dijangka merupakan faktor yang menyebabkan kecenderungan C. bullita kepada O. aristatus dan ketahanan pokok ini terhadap serangan C. bullita. Informasi biologi perosak ini penting untuk membina strategi pengurusan perosak yang lebih baik supaya dapat menandangi perosak secara lebih efisien dan selamat.

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ACKNOWLEDGEMENTS

I am very grateful to my supervisors, Prof. Dr. Ahmad Said Sajap and committee members Prof. Dr. Dzolkhifli Omar and Assoc. Prof. Dr. Faizah Abood, for their tireless help, constructive comments, advice and encouragement during this research. This work would not been accomplished without their invaluable input.

I would like to thank Dr. Thomas J. Henry from Agricultural Research Service of USDA in United States for identifying the bugs. Dr. Henry is acknowledged for being the reviewer of my first manuscript and providing important valuable comments.

I would also like thank all my lab mates from the Laboratory of Entomology, Faculty of Forestry and friends for all their support and help. And I wish to express my sincere gratitude to my family members, especially my Mom, Ng Saw Guat; Dad, Tan Kim Wan and last but not least Lee Han Jeen for the encouragement and moral support they provided during the course of my studies.

Finally, I would like to thank God for giving me the strength and guidance to complete my studies on time.

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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfillment of the requirement for the Degree of Doctor of Philosophy. The members of the Supervisory Committee were as follows:

Ahmad Said Sajap, PhD Professor Faculty of Forestry Universiti Putra Malaysia (Chairman) Dzolkhifli Omar, PhD Professor Faculty of Agriculture Universiti Putra Malaysia (Member) Faizah Abood, PhD Associate Professor Faculty of Forestry Universiti Putra Malaysia (Member)

________________________________ BUJANG KIM HUAT, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia

Date:

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Declaration by graduate student I hereby confirm that: • this thesis is my original work; • quotations, illustrations and citations have been duly referenced; • this thesis has not been submitted previously or concurrently for any other degree

at any other institutions; • intellectual property from the thesis and copyright of thesis are fully-owned by

Universiti Putra Malaysia, as according to the Universiti Putra Malaysia (Research) Rules 2012;

• written permission must be obtained from supervisor and the office of Deputy Vice-Chancellor (Research and Innovation) before thesis is published (in the form of written, printed or in electronic form) including books, journals, modules, proceedings, popular writings, seminar papers, manuscripts, posters, reports, lecture notes, learning modules or any other materials as stated in the Universiti Putra Malaysia (Research) Rules 2012;

• there is no plagiarism or data falsification/fabrication in the thesis, and scholarly integrity is upheld as according to the Universiti Putra Malaysia (Graduate Studies) Rules 2003 (Revision 2012-2013) and the Universiti Putra Malaysia (Research) Rules 2012. The thesis has undergone plagiarism detection software.

Signature: _______________________ Date: ______________________ Name and Matric No.: _________________________________________

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Declaration by Members of Supervisory Committee This is to confirm that: • the research conducted and the writing of this thesis was under our supervision; • supervision responsibilities as stated in the Universiti Putra Malaysia (Graduate

Studies) Rules 2003 (Revision 2012-2013) are adhered to. Signature: __________________ Signature: _________________ Name of Name of Chairman of Member of Supervisory Supervisory Committee: Ahmad Said Sajap, PhD Committee: Dzolkhifli Omar, PhD Signature: __________________ Name of Chairman of Supervisory Committee: Faizah Abood, PhD

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TABLE OF CONTENTS Page ABSTRACT ii ABSTRAK iv ACKNOWLEDGEMENTS vi APPROVAL vii DECLARATION ix LIST OF TABLES xiv LIST OF FIGURES xvi LIST OF ABBREVATIONS xxiv CHAPTER 1 INTRODUCTION 1

2 LITERATURE REVIEW 4 2.1 Cat’s Whiskers Plant, Orthosiphon aristatus 4

(Blume) Miq. 2.1.1 Importance of Orthosiphon aristatus 5

2.1.2 Insect Pest Associated with 6 Orthosiphon aristatus

2.2 Basil plant, Ocimum basilicum Linneaus 6 2.2.1 Importance of Ocimum basilicum 7 2.2.2 Insect Pest Associated with 8 Ocimum basilicum 2.3 Hemiptera, Suborder Heteroptera 8 2.4 Tingidae 9 2.5 Cochlochila bullita (Stål) 10 (Heteroptera: Tingidae) 2.5.1 Morphology 10

2.5.2 Adaptation 11 2.5.3 Distribution, Host Plants and 11

Pest Status 2.5.4 Life-table 11

2.5.5 Feeding and Oviposition Preferences 12 2.5.6 Injury Causes by Lace Bugs 13 3 GENERAL MATERIALS AND METHODS 15 3.1 Host Plants Cultivation 15

3.2. Maintenance of Insect Colony 16

4 MORPHOLOGICAL RE-DESCRIPTION 17 OF Cochlochila bullita (STÅL) (HETEROPTERA: TINGIDAE), A POTENTIAL PEST OF Orthosiphon aristatus (BLUME) MIQ. (LAMIALES: LAMIACEAE) IN MALAYSIA

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4.1 Introduction 17 4.2 Materials and Methods 17 4.2.1 Morphology and Morphometric 17 Study

4.2.2 Feeding and Ovipositing Patterns 19 4.2.3 Statistical Analysis 19

4.3 Results 19 4.3.1 Morphologies and Morphometrics 19

4.3.2 Feeding and Ovipositing Patterns 28 4.4 Discussion 31

4.5 Conclusion 32

5 LIFE-TABLE OF Cochlochila bullita (STÅL) 34 (HEMIPTERA: TINGIDAE) ON Orthosiphon aristatus (BLUME) MIQ. AND Ocimum basilicum LINNAEUS 5.1 Introduction 34 5.2 Materials and Methods 34 5.2.1 Maintenance of Test Colony 35 5.2.2 Preparation of Test Host Cutting 36 5.2.3 Experimental Design 36 5.2.4 Development and Survivorship 36 5.2.5 Fecundity and Adult Longevity 38 5.2.6 Life Table Analysis 39 5.3 Results 40 5.3.1 Development and Survivorship 40 5.3.2 Fecundity and Adult Longevity 45 5.3.3 Life Table Analysis 48 5.4 Discussion 49 5.5 Conclusion 51 6 MATING BEHAVIOUR AND OVIPOSITIONAL 52 PATTERN OF LACE BUG, Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE) ON ITS HOST PLANTS, Orthosiphon aristatus (BLUME) MIQ. AND Ocimum basilicum LINNAEUS 6.1 Introduction 52 6.2 Materials and Methods 54 6.2.1 Mating Behaviour 54 6.2.2 Mating Success 54 6.2.3 Oviposition Behaviour 55 6.2.4 Oviposition Preference 56 6.2.5 Oviposition-site Preference 56 6.2.6 Trichomes Density 57 6.2.7 Oviposited Leaf Cross-section 58

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6.2.8 Maternal Care 58 6.2.9 Statistical Analysis 59 6.3 Results 59 6.3.1 Mating Behaviour 59 6.3.2 Mating Success 64 6.3.3 Oviposition Behaviour 66 6.3.4 Oviposition Preference 67 6.3.5 Oviposition-site Preference 67 6.3.6 Trichomes Density 68 6.3.7 Oviposited Leaf Cross-section 75 6.3.8 Maternal Care 78 6.4 Discussion 80 6.5 Conclusion 86 7 FEEDING PREFERENCE AND INJURY 88 ASSESSMENTS OF LACE BUGS, Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE) ON Orthosiphon aristatus (BLUME) MIQ. AND Ocimum basilicum LINNAEUS 7.1 Introduction 88 7.2 Materials and Methods 90 7.2.1 Preparation of Host Plants 90 7.2.2 Injured area and Excrement 91 Production 7.2.3 Chlorophyll Content Measurement 93 7.2.4 Leaf Water Content 93 7.2.5 Histological Examination on Feeding 95 Injury and Ovipositional Injury 7.2.6 Feeding Preference 95 7.2.7 Statistical Analysis 96 7.3 Results 96 7.3.1 Injured Area and Excrement 96 Production 7.3.2 Chlorophyll Content Measurement 103 7.3.3 Leaf Water Content 104 7.3.4 Histological Examination on Feeding 105 Injury and Ovipositional Injury 7.3.5 Feeding Preference 108 7.4 Discussion 109 7.5 Conclusion 113 8 CONCLUSIONS AND RECOMMENDATIONS 115 8.1 Conclusions 115 8.2 Recommendations 117 REFERENCES/BIBLIOGRAPHY 118

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BIODATA OF STUDENT 136 LIST OF PUBLICATIONS 137

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LIST OF TABLES

Table Page 4.1 Morphological characteristics of 20 C. bullita (n = 15) (mean ± SE, mm). 4.2 Morphometric characteristics of 26 C. bullita (n = 15) (mean ± SE, mm). 4.3 Linear regression of the morphometric 26 relationship in C. bullita nymphs. 4.4 Length to width ratio of C. bullita 28 (n = 15) (mean ± standard error, mm). 4.5 Site of egg oviposition of C. bullita at 31 O. aristatus. 5.1 Development times of C. bullita immature 43 O. basilicum leaves stages fed either on O. aristatus or O. basilicum leaves under laboratory conditions. 5.2 Mortality rate of C. bullita immature 43 stages fed either on O. aristatus or O. basilicum leaves under laboratory conditions. 5.3a Pooled life table of C. bullita on 44 O. aristatus. 5.3b Pooled life table of C. bullita on 44 O. basilicum. 5.4 Adult longevity of C. bullita fed on 48 O. aristatus or O. basilicum.

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5.5 Reproductive parameters of C. bullita 48 fed on O. aristatus or O. basilicum. 5.6 Life table parameters of C. bullita 49 fed on O. aristatus or O. basilicum. 6.1a Activities displayed by the male on 60 different host plants during day times. 6.1b Activities displayed by the female on 60 different host plants during day times. 6.2 Mating duration (mean ± SE) of C. bullita 60 on different host plants. 6.3a – c Mean number of females mated and eggs 65 fertilized by single male fed on different host plants. 6.4 Lodging Preference of C. bullita in 67 choice tests: the number of adults and percent preference (mean ± SE) on selected plant for everyday. 6.5 Oviposition preferences of C. bullita 67 in choice tests: responses in egg numbers and percent preference (mean ± SE) within 21 days of oviposition. 6.6 Oviposition preferences of C. bullita 68 in no-choice tests: the mean number of eggs and percent preference (mean ± SE) in selected plant’s sites of O. aristatus and O. basilicum leaf. 6.7 Trichomes density (mean ± SE) 69 associated with different plant’s sites on O. aristatus and O. basilicum.

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6.8 Linear regression of the trichomes 75 density in relation with eggs deposition preference. 6.9 Mean number of maternal care 80 responses of C. bullita on different. host plants. 7.1 Percentage of leaf injury and number 97 of faeces spots (mean ± SE) produced by C. bullita. 7.2 Linear regression of injury area and 99 faeces production. 7.3 Chlorophyll content (mean ± SE) of 103 control and infested O. aristatus and O. basilicum after 3 days of test had been started. 7.4 Percentage of leaf water content 105 (mean ± SE) of O. aristatus and O. basilicum. 7.5a Linear regression of the water content in 105 relation with injury area and faeces production on O. aristatus. 7.5b Linear regression of the water content in 105 relation with injury area and faeces production on O. basilicum. 7.6 Stage-specific Feeding preferences of 109 C.bullita nymphs in choice tests: the average number of nymphs (mean ± SE) on selected plant for everyday. 7.7 Sides-specific Feeding preferences of 109 C. bullita nymphs in choice tests: the average number of nymphs (mean ± SE) on selected plant for everyday.

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LIST OF FIGURES Figure Page 2.1 Orthosiphon aristatus plants in the nursery. 5 2.2 Ocimum basilicum plants in the nursery. 7 3.1a – b O. aristatus plant (a) and O. basilicum 15 plant (b) planted in polystyrene bag. 3.2 Wooden cage used to mass rear C. bullita. 16 4.1 Five characters measured in C. bullita. 18 4.2 Eggs of C. bullita. 21 4.3 Five nymphal stages of C. bullita. 22 4.4 First instar of C. bullita. 22 4.5 Second instar of C. bullita. 23 4.6 Third instar of C. bullita. 23 4.7 Fourth instar of C. bullita. 24 4.8 Fifth instar of C. bullita. 24 4.9 C. bullita adult female ventral (a) 25 and dorsal views (b). 4.10 C. bullita adult male ventral (a) and 25 dorsal views (b).

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4.11a – d Growth patterns of four biometric 27 of C. bullita in relation to the development stage. 4.12a – d Feeding and damage of C. bullita. 28 4.13 The eggs were inserted into the stems in 29 clusters. 4.14 Eggs inserted singly into the leaf 30 veins and margin. 4.15 Eggs inserted singly into the leaf 30 margin. 5.1 Acrylic cages used to rear C. bullita 35 on O. aristatus (a) and O. basilicum (b). 5.2 O. aristatus and O. basilicum 36 cuttings rooted inside a small plastic cups. 5.3 A plastic container contained 3 pots 37 of plants for ovipositing. 5.4 A plastic container used to observe 38 the development and survivorship of nymphs. 5.5a Survival rates (lx) of C. bullita on 41 O. aristatus in 3 different cohorts. 5.5b Survival rates (lx) of C. bullita on 42 O. basilicum in 3 different cohorts. 5.6a Survivorship curve (lx) of C. bullita 45 on O. aristatus.

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5.6b Survivorship curve (lx) of C. bullita 45 on O. basilicum. 5.7a Daily age-specific survival (lX) and 47 fecundity (mX) of female C. bullita fed on O. aristatus. 5.7b Daily age-specific survival (lX) and 47 fecundity (mX) of female C. bullita fed on O. basilicum. 6.1 Plastic container with a small pot of 55 O. aristatus cutting. 6.2 Arenas used for choice test in both 56 host and feeding preferences tests. 6.3 Arenas used for no-choice test in 57 eggs distribution study. 6.4 Leaf parts where trichomes densities 57 were measured. 6.5 Male C. bullita riding on top of the 61 female, grasp the margin of the female’s wings with its mid- and hind-legs and pronotum with its front legs. 6.6 Male (Left) in positioning to mate 61 with the female (Right) by placing the female in between its wings and body. 6.7 Male (Right) trying to disengage 62 himself from the female by walking to the opposite direction from the female (Left).

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6.8 Female lifted-up its wing when a 63 male approached. 6.9 Two male (Upper and Bottom) 64 competing to mate with the female (Middle) by placing the female in between its wings and body. 6.10 Female C. bullita drawing out it 66 ovipositor in order to probe into the plant stem and lay its egg. 6.11 Pubescences or trichomes the entire 69 young leaves of the O. aristatus (a) and O. basilicum (b). 6.12a Trichomes on the midrib of O. aristatus 70 (adaxial side). 6.12b Trichomes on the midrib of O. basilicum 70 (adaxial side). 6.13a Trichomes on the midrib of O. aristatus 71 (abaxial side). 6.13b Trichomes on the midrib O. basilicum 71 (abaxial side). 6.14a Trichomes on the vein of O. aristatus 72 (adaxial side). 6.14b Trichomes on the vein of O. basilicum 72 (adaxial side). 6.15a Trichomes on the vein of O. aristatus 73 (abaxial side). 6.15b Trichomes on the vein of O. basilicum 73 (abaxial side).

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6.16a Trichomes on the margin of O. aristatus. 74 6.16b Trichomes on the margin of O. basilicum. 74 6.17 Cross section of eggs inserted 45° into 76 the midrib of a leaf. 6.18 Closed-up of an egg inserted 76 horizontally into the leaf margin. 6.19 Closed-up of an egg inserted 77 horizontally into the leaf margin of O. aristatus. 6.20 Longitudinal section of a cluster of eggs 77 inserted perpendicularly into a fresh plant stem. 6.21 Longitudinal section of a cluster of eggs 78 inserted perpendicularly into a plant stem. 6.22 A female found covering the egg 79 mass laid on O. basilicum. 6.23 A female found staying close to the 79 egg mass laid on O. aristatus, as the egg mass was over pack with no trichomes around the eggs. 7.1 Rooted plant cuttings transplanted 90 into a small cup of soil. 7.2 Test subjects that were caged in a 91 cylinder cage. 7.3 LI-3100 Leaf Area Meter 92 (Lincoln NE, USA).

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7.4 Leaf injured area measurement by 92 using grid graph. 7.5 Minolta Chlorophyll Meter 94 SPAD-502. 7.6 Six SPAD reading along the halfway 94 between leaf margin and leaf midrib. 7.7 A control plate (Left) and test subject 95 hosted four C. bullita adults (Right). 7.8 Arenas used for choice test in feeding 96 preferences. 7.9 Pattern of feeding injury on 98 O. aristatus from Day 1 – 4 (Left – Right) with increasing of white patches. 7.10 Pattern of feeding injury on 98 O. basilicum from Day 1 – 4 (Left – Right) with increasing of white patches. 7.11 Faeces produced by C. bullita that 99 fed on O. aristatus (Left) and O. basilicum (Right). 7.12 The appearance of O. aristatus (a) 100 and O. basilicum (b) on Day 1 or before infestation, turgid and healthy with fresh green leaves. 7.13 The appearance of O. aristatus (a) 100 and O. basilicum (b) on Day 2 or 24 hr after infestation.

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7.14 O. aristatus (a) with one leaf 101 dropped and O. basilicum with one leaf became less turgid (b) on Day 3 or 48 hr after infestation. 7.15 O. aristatus wilted (a) and 101 O. basilicum still intact (b) on Day 4 or 72 hr after infestation. 7.16 Physical appearance of infested 102 O. aristatus leaf on day 2, 3 and 4 (left to right). 7.17 Physical appearance of infested 102 O. basilicum leaf on day 2, 3 and 4 (left to right). 7.18 Cumulative chlorophyll changes in 104 control plants. 7.19 Cumulative chlorophyll changes in 104 infested plants. 7.20 O. aristatus leaf blade cross section, 106 healthy leaf (Upper) and injured leaf (Lower). 7.21 O. basilicum leaf blade cross section, 106 with a brown spot of injury at midrib. 7.22 Horizontal cross section of a healthy 107 plant stem (a) and a stem with a cluster of eggs inserted into (b). 7.23 Ovipositional wound damage due to 107 egg insertion. 7.24 Feeding Preference of C. bullita nymphs: 108 average number of total 4 stages of nymphs (mean ± SE) on both tested plants.

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LIST OF ABBREVATIONS

USDA United States Department of Agriculture IPM Integrated Pest Management GRIN Germplasm Resources Information Network

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CHAPTER ONE

INTRODUCTION

Orthosiphon aristatus (Blume) Miq. is a well known medicinal plant from the family Lamiaceae. It can be found throughout Southeast Asia and also tropical Australia with several common names. The most widely used common name, cat’s whiskers plant, is derived from the appearance of its flower with two-lipped and protruding stamens. Orthosiphon aristatus is a fast growing perennial herbaceous plant, 1 – 2 m tall, with multiple branches stem and oppositely growing ovate leaves (Dzulkarnain et al. 1999). Apart from its medicinal uses, the plant is planted ornamental as it has attractive solid-white or lavender flowers.

As an important plant with medicinal properties, the plant has been widely used among many communities. It has been trusted for many centuries for treating ailments of the kidney, kidney stone, urinary tract infection, liver and bladder problems, diabetes and many other ailments (Indubala and Ng 2000). Its effectiveness for treating bladder and kidney stone had given it a Chinese name “hua shi cao”, which means stone dissolving plant. In Malaysia, it is a traditional herb often consumed as herbal tea. In a recent study, the extract form this plant has high anti-oxidant content and can be used to prevent and control the growth of cancer cells (Chin et al. 2009).

There are currently many products of O. aristatus marketed as herbal tea, supplement capsule and even soap. In Okinawa, Japan this plant is even systematically cultivated to produce and consume as a healthy tea (Awale et al. 2002). Presently, there is a shortage in this herbal resource as the demand of Malaysian consumers exceeds the supply of the product (Farhana et al. 2007). Although no mass planting of this plant is practiced in Malaysia, the potential of this plant to be widely produced is a certainty. Therefore, proper cultivation and management of this plant is required, especially when dealing with pests and diseases.

Since its establishment as an important crop, the herb has been notably free from serious insect problems until recently. In August 2009, the herb, grown in a backyard garden at Subang Jaya, Selangor, Malaysia, was heavily attacked by an unknown lace bug. A research station in Kelantan also reported that there is a similar insect pest infestation that kills the plant. Several photographs of the lace bug were sent to Dr. Thomas J. Henry, a research entomologist from United States Department of Agriculture (USDA), Systematic Entomology Laboratory, Washington, DC and Dr. Masaki Tomokuni, a researcher at the National Science Museum, Tokyo. Both scientists identified the lace bug, Cochlochila bullita (Stål), commonly known as the ocimum tingid.

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The economic importance of the lace bug has been increasing since the last four decades (Neal and Schaefer 2000). The bugs once believed as a monophagous pest (Drake and Ruhoff 1965; Livingstone 1977; Cobben 1978) were found to attack other plants and indicating that many species may be oligophagous (Tomokuni 1983; Wheeler 1989; Qi, et al. 1991). With a relatively specific host range, lace bug, Teleonemia scrupulosa Stål was used as biological control agents for controlling weeds, such as Lantana camara (Linnaeus) (Harley and Kassulke 1971).

As a pest, C. bullita had caused outbreak in India in 1950 and 1983 respectively (Mohanasundram and Rao 1973; Palaniswami and Pillai 1983). To date no study has been conducted elsewhere on account of the quality and efficacy of any herbal functional product or medicine that is highly dependent on the source - the plant material, proper managements on this pest become a necessity. Thus a biological study was needed to clearly define the morphology, growth and development, damage caused, and behaviour of this pest in order to manage the pest.

The morphology of the insect pest needs to be studied thoroughly with pictures provided for correct identification. External appearance of the insect has to be described qualitatively (shape or colour) and quantitatively (measurement). Apart from morphological studies, the life cycle and growth pattern of an insect on a particularly host, represented by its life table, need to be investigated. By constructing a life table of the pest, a comprehensive description of the insect growth and survival could be gathered. This is useful information for estimating the insect population.

In order to accomplish an effective pest management, knowledge on the behaviour of the insect is very pertinent. Among the behaviours that allow an insect successfully thrive as a pest are reproductive behaviour and host preferences. These behaviours are a collective response to maximise the fitness of one species and an ultimate success throughout the evolution. This is vital information required for developing a management strategy to minimize the number of this pest under threshold level and prevent it from outbreak.

Feeding and oviposition behaviours have been shown to cause injury on the host plants. These injuries are reflected on the damages inflicted by the insect on the plants. Many methods have been used to appraise the level of damages of a particular insect on a crop. The result of the assessment provides fundamental information for developing the decision making guidelines such as the Economic Injury Level (EIL) and Aesthetic Injury Level (AIL) (Klingeman et al. 2001).

Even though, ocimum tingid, C. bullita has the potential to rise as a serious pest of O. aristatus. Heretofore, there were no studies on the life table parameters, development, fecundity and behaviours for this pest associated with O. aristatus. Thus this study

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was aimed of gathering information on the full life table parameters and behaviours of C. bullita on O. aristatus with O.basilicum as a compare model. This is to provide a better understanding of the biology of the pest for the purpose of pest management. Therefore, the objectives of this study were to analyse the biology of C. bullita, in order to establish continuous rearing for biological control purposes.

The specific objectives of this thesis were:

i. to describe the external morphology of C. bullita.

ii. to establish the life table of C. bullita on O. aristatus and O.basilicum.

iii. to examine the mating behaviour and maternal care of C. bullita.

iv. to determine the host preference of C. bullita on O. aristatus and O. basilicum in response to the plants’ trichomes.

v. to assess the injury pattern of C. bullita on O. aristatus and O. basilicum.

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BIODATA OF STUDENT

Tan Li Peng was born in Kuala Lumpur, the capital of Malaysia on 20th July 1986. She received her primary education at Sekolah Rendah Jenis Kebangsaan (C) Chung Kwok, Kuala Lumpur. She completed her secondary education in 2003 at Sekolah Menengah Kebangsaan (P) Titiwangsa followed by her Sijil Tinggi Pelajaran Malaysia (STPM) in 2005 at the same school. In 2006, she pursued a Bachelor’s degree at Universiti Putra Malaysia and completed her three years programme with Bachelor Science of Forestry, minor in Urban Forest Management. She then furthers her study at the same university as a full time Doctor of Philosophy (PhD) student in Faculty of Forestry in 2009. In 2011, she has successfully passed her comprehensive examination as a PhD student. Her researches are mainly in the areas of Entomology and Biological control by using Entomopathogenic Fungi.

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LIST OF PUBLICATIONS Sajap, A.S. and T.L. Peng. 2010. The lace bug, Cochlochila bullita (Stål) (Heteroptera: Tingidae), a potential pest of Orthosiphon stamineus Bentham (Lamiales: Lamiaceae) in Malaysia. Insecta Mundi, pp. 654.

Peng, T.L., A.S. Sajap and L.H. Jeen. 2012. First report and morphological re-description of Cochlochila bullita (Stål) (Heteroptera: Tingidae), a potential pest of Orthosiphon stamineus Bentham (Lamiales: Lamiaceae) in Malaysia. Paper presented at International Symposium on Insects, Dec 3–5, 2012. Mines Wellness Hotel, Kuala Lumpur, Malaysia, p. 27. Lee, S.H., P.S. H’ng, T.L. Peng and W.C. Lum. 2013. Response of Coptotermes curvignathus (Isoptera: Rhinotermitidae) to formaldehyde catcher-treated particleboard. Pakistan Journal of Biological Sciences. 16 (21): 1415 – 1418. Peng, T.L., A.S. Sajap L.H. Jeen, S.H. Lee and W.C. Lum. 2013. Morphological re-description of Cochlochila bullita (Stål) (Heteroptera: Tingidae), a potential pest of Orthosiphon aristatus Blume Miq. (Lamiales: Lamiaceae) in Malaysia. Pakistan Journal of Biological Sciences. 16 (23): 1786 – 1790. Peng, T.L., A.S. Sajap L.H. Jeen and K.H. Yen. 2013. Evaluation of Metarhizium anisopliae Metschnikoff and Isaria fumosorosea Wize in Controlling Cochlochila bullita Stål. Paper presented at 2nd Global Conference on Entomology, Nov 8–12, 2013. Kuching, Malaysia, p. 14. Sajap, A.S., T.L. Peng, J.J. Jessica and A.S. Syazwan. 2014. Efficacy of Formulated Metarhizium anisopliae on Selected Termites Species. Proceeding of 8th International Conference on Plant Protection in Tropics, Apr 8-10, 2014. Kuala Lumpur, Malaysia, p. 182.

BIOLOGY OF Cochlochila bullita (STÅL) (HEMIPTERA: TINGIDAE), APOTENTIAL PEST OF Orthosiphon aristatus (BLUME) MIQ. (LAMIALES:LAMIACEAE) IN MALAYSIAAbstractTABLE OF CONTENTSCHAPTER 1REFERENCES