UNIVERSITI PUTRA MALAYSIA MOUSA KHANI FP 2012 45 ...

UNIVERSITI PUTRA MALAYSIA

MOUSA KHANI

FP 2012 45

INSECTICIDAL EFFECTS OF SELECTED PLANT EXTRACTS ON RICE WEEVIL, Sitophilus oryzae L. AND RICE MOTH, Corcyra cephalonica (St)

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INSECTICIDAL EFFECTS OF SELECTED PLANT EXTRACTS ON RICE

WEEVIL, Sitophilus oryzae L. AND RICE MOTH, Corcyra cephalonica (St)

By

MOUSA KHANI

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

in Fulfilment of the Requirements for the Degree of Doctor of Philosophy

May 2012

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DEDICATION

I dedicate this thesis to my lovely wife Fahimeh, my son Amin and my daughter

Sara for their patience and support during my study in Malaysia

Thank you for your endless support,

Loving you all …

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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment

of the requirements for the degree of Doctor of Philosophy

INSECTICIDAL EFFECTS OF SELECTED PLANT EXTRACTS ON RICE

WEEVIL, Sitophilus oryzae L. AND RICE MOTH, Corcyra cephalonica (St)

By

MOUSA KHANI

may 2012

Chairperson : Professor Rita Muhamad Awang, PhD

Faculty : Agriculture

The present study was conducted with the objectives of extraction and evaluation the

of chemical constituents from black pepper (Piper nigrum L.), peppermint (Mentha

piperita L.), and physic nut (Jatropha curcas L.), and to investigate effects of these

plant extracts on toxicity, mortality, repellency, antifeedant activity, egg hatchability

and adult emergence of the rice weevil and rice moth under laboratory conditions.

Crude extracts were prepared by percolation method and essential oils were prepared

by hydro distillation method. Among the nine extracts screened for toxicity,

petroleum ether and chloroform extract of P. nigrum and petroleum ether extract of

J. curcas showed significant insecticidal potential. The LC50 values of the above

extracts against S. oryzae adults were 1.61, 1.70 and 6.82 µL/mL, and against C.

cephalonica larvae were 12.52, 14.31 and 13.22 µL/mL, respectively. The LC50

values of M. piperita and P. nigrum essential oils against adults of S. oryzae were

85.0 and 288.8 µL/L air, and against C. cephalonica larvae were 343.9 and 530.5

µL/L air, respectively. The GC and GC-MS analysis showed that the major

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components of P. nigrum extracts were piperine (75.5%) and caryophyllene (18.5%).

The major components of J. curcas seed oil were oleic acid (40.7%), linoleic acid

(34.2%) and palmitic acid (18.0%). GC-MS analysis also showed that the major

components of M. piperita essential oils were menthol (47.0%), isomenthone

(19.9%), limonene (7.5%) and cineole (5.4%), while the major components of P.

nigrum essential oils were limonene (33.8%), α-pinene (31.2%) and β-pinene

(23.3%). Mentha piperita and P. nigrum essential oils showed significant

effectiveness against adults of S. oryzae and C. cephalonica larvae compared to the

control. Mentha piperita oil strongly inhibited feeding in S. oryzae compared to P.

nigrum essential oils, but there was an inverse effect on repellency activity. Essential

oils of both plant species showed no repellency activity against 3rd

instar larvae of C.

cephalonica. However, M. piperita essential oils strongly inhibited egg hatchability

in S. oryzae as compared to P. nigrum essential oils. Also M. piperita essential oils

had a stronger inhibitory activity on adult emergence than P. nigrum essential oils in

both insect species.

Nutritional bioassay revealed significant reduction in the relative growth rate (RGR),

relative consumption rate (RCR) and food utilization (ECI) by rice weevil and rice

moth at concentration range between 2 - 10 µL/g of rice kernels with feeding

deterrence indices (FDI) reaching 43.5 and 74.9%, respectively. The plant oils also

showed repellency activity against adults of S. oryzae, while, there was no repellency

activity against 3rd

instar larvae of C. cephalonica. The plant extracts when applied

on rice kernels against adults of S. oryzae at concentrations of 0.5 –2 µL/mL, and C.

cephalonica larvae at 2-10 µL/mL, resulted in significant (P < 0.05) reductions in

egg hatchability. Adult emergence was also drastically reduced by P. nigrum and J.

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curcas extracts when applied to rice kernels. The plant extracts strongly suppressed

adult emergence of both insects at equivalent dose levels.

The results revealed significant insecticidal effects of the selected plant extracts.

There is therefore immense potential for development of novel insecticides from

these plant species.

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Abstrak tesis yang dikemukakan lepada Senat Universiti Putra Malaysia sebagai

memenuhi keperluan untuk Ijazah Doktor Falsafah

SIFAT INSEKTISIDAL EKSTRAK TANAMAN TERPILIH TERHADAP

KUMBANG BERAS, Sitophilus oryzae L. DAN KUPU KUPU BERAS, Corcyra

cephalonica (St)

Oleh

MOUSA KHANI

mei 2012

Pengerusi : Profesor Rita Muhamad Awang, PhD

Fakulti : Pertanian

Oleh sebab itu, kajian ini dijalankan dengan objektif untuk mengekstrak dan menilai

konstituen kimia lada hitam (Piper nigrum L.), pepermin (Mentha piperita L.), dan

kacang fizik (Jatropha curcas L.), dan mengkaji kesan ekstrak tumbuhan ini ke atas

ketoksikan, mortaliti, repelensi, aktiviti antimakanan, kebolehtetasan telur dan

kemunculan beras kumbang dewasa dan beras rama-rama dewasa di bawah keadaan

makmal.

Ekstrak mentah disediakan dengan menggunakan kaedah perkolasi dan minyak

esential disediakan dengan menggunakan kaedah distilasi hidro. Antara sembilan

ekstrak yang diskrin untuk ketoksikan, eter petroleum dan ekstrak kolorofom P.

nigrum dan ekstrak eter petroleum J. curcas menunjukkan potensi insektisidal yang

signifikan. Nilai LC50 bagi ekstrak tersebut terhadap S. oryzae dewasa ialah 1.61,

1.70, dan 6.82 µL/mL, dan terhadap larva C. cephalonica ialah masing-masing

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12.52, 14.31 dan 13.22 µL/mL. Nilai LC50 M. piperita dan minyak esential P. nigrum

terhadap S. oryzae dewasa ialah 85.0 dan 288.8 µL/L air dan terhadap larva C.

cephalonica ialah masing-masing 343.9 dan 530.5 µL/L air. Analisis GC dan GC-

MS menunjukkan bahawa komponen utama ekstrak P. nigrum ialah piperin (75.5%)

dan kariofilena (18.5%). Komponen utama minyak biji J. curcas ialah asid oleik

(40.7%), asid linoleik (34.2%) dan asid palmitik (18.0%). Analisis GC-MS juga

menunjukkan bahawa komponen utama minyak esential M. piperita ialah mentol

(47.0%), isomenton (19.9%), limonena (7.5%) dan sineol (5.4%), manakala

komponen utama minyak esential P. nigrum ialah limonena (33.8%), α-pinena

(31.2%), dan β-pinena (23.3%). Minyak esential Mentha piperita dan P. nigrum

menunjukkan keberkesanan yang signifikan terhadap S. oryzae dewasa dan larva C.

cephalonica berbanding kawalan. Minyak M. piperita menghalang kuat suapan S.

oryzae berbanding minyak esential P. nigrum, tetapi terdapat kesan yang sebaliknya

ke atas aktiviti repelensi. Minyak esential bagi ketiga-tiga spesis menunjukkan

ketiadaan aktiviti repelensi terhadap larva C. cephalonica instar ketiga. W alau

bagaimanapun, minyak esential M. piperita menghalang kuat kebolehtetasan telur S.

oryzae berbanding minyak esential P. nigrum. Tambahan pula, minyak esential M.

piperita mempunyai aktiviti penghalang yang amat kuat terhadap kemunculan

serangga dewasa berbanding minyak esential P. nigrum bagi kedua-dua spesis

serangga.

Bioassai pemakanan menunjukkan penurunan yang bererti terhadap kadar

pertumbuhan relatif (RGR), kadar konsumsi relatif (RCR) dan utilisasi makanan

(ECI) oleh beras kumbang dan beras rama-rama pada julat konsentrasi antara 2-10

µL/g kernel padi dengan indeks deteren suapan (FDI), masing-masing mencapai 43.5

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dan 74.9%. Minyak tumbuhan juga menunjukkan aktiviti repelensi terhadap S.

oryzae dewasa, manakala, tidak terdapat aktiviti repelensi terhadap larva C

cephalonica instar ketiga. Ekstrak tumbuhan apabila diaplikasikan ke atas kernel

padi terhadap S. oryzae dewasa pada konsentrasi 0.5-2 µL/mL, dan larva C.

cephalonica pada 2-10 µL/mL, menghasilkan penurunan yang signifikan (P<0.05)

pada kebolehtetasan telur.Kemunculan serangga dewasa juga menurun secara drastik

oleh ekstrak P. nigrum dan J. curcas apabila diaplikasikan pada kernel beras. Ekstrak

tumbuhan menghambat kuat kemunculan kedua-dua serangga dewasa pada paras dos

yang setara.

Hasil dapatan kajian menjelaskan sifat insektisidal yang signifikan pada ekstrak

tumbuhan yang terpilih. Oleh sebab itu, terdapat potensi yang besar untuk

pertumbuhan racun serangga yang menggalakkan dari spesis tumbuhan tersebut.

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ACKNOWLEDGEMENTS

I would like to express my heartfelt gratitude and appreciation to my dear supervisor,

Prof. Dr. Rita Muhamad Awang for her encouragement, available support, patience,

invaluable advice, understanding that made this study possible and intellectual

guidance throughout the studies. I am also greatly indebted to my supervisory

committee members, Prof. Dr. Dzolkhifli Omar and Prof. Dr. Mawardi Rahmani, for

their enlightening comments, advice and help throughout my study and

encouragement in the completion of this thesis.

My gratitude is also due to the authorities of Iranian Institute of Medicinal Plants and

Academic Centre for Education, Culture and Research (ACECR) in Iran for the

official permission to pursue a Ph.D. study at Universiti Putra Malaysia (UPM),

Malaysia.

I would like to thank all the staff from Department of Plant Protection for their

cooperation in the preparation of test equipments.

I would like to express my gratitude to my dear friend Mohammad Ebrahim

Farashiani for him comments and helps.

Finally, I wish to express my deepest appreciation to my wife Fahimeh for her

patience, hardness, understanding support and inspiration given to me during the

period of my study in Malaysia and my loving children Amin and Sara for patience

and hardships.

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I certify that a Thesis Examination Committee has met on 4 May 2012 to conduct the

final examination of MOUSA KHANI on him thesis entitled “Insecticidal effects of

selected plant extracts on rice weevil, Sitophilus oryzae L. and rice moth, Corcyra

cephalonica (St)” in accordance with Universities and University Colleges Act 1971

and the Constitution of the Universiti Putra Malaysia [P.U.(A) 106] 15 March 1998.

The committee recommends that the student be awarded the degree of Doctor of

Philosophy.

Members of the Thesis Examination Committee were as follows:

Kamaruzaman Sijam, PhD

Associate Professor

Department of Plant Protection

Faculty of Agriculture

Universiti Putra Malaysia

(Chairman)

Nur Azura binti Adam, PhD

Senior Lecturer

Department of Plant Protection



(Internal Examiner)

Faizah bt Abood, PhD

Associate Professor

Department of Forest Management

Faculty of Forestry


(Internal Examiner)

Ghulam Hussain Abro, PhD

Y. Bhg. Professor

Department of Entomology

Faculty of Crop Protection

Sindh Agriculture University

Tandojam Pakistan

(External Examiner)

SEOW HENG FONG, PhD

Professor and Deputy Dean

School of Graduate Studies


Date: 12 July 2012

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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been

accepted as fulfilment of the requirement for the degree of Doctor of Philosophy.

The members of the Supervisory Committee were as follows:

Rita Muhamad Awang, PhD

Professor



(Chairperson)

Dzolkhifli Omar, PhD

Professor



(Member)

Mawardi Rahmani, PhD

Professor

Faculty of Science


(Member)

Shamsali Rezazadeh, PhD

Iranian Institute of Medicinal Plants

Karaj, Iran

(Member)

BUJANG BIN KIM HUAT, PhD

Professor and Dean

School of Graduate Studies


Date:

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DECLARATION

I declare that the thesis is my original work except for quotations and citations which

have been duly acknowledged. I also declare that it has not been previously, and is

not concurrently, submitted for any other degree at Universiti Putra Malaysia or any

other institutions.

MOUSA KHANI

Date: 4 May 2012

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TABLE OF CONTENTS

Title Page

ABSTRACT iii

ABSTRAK vi

ACKNOWLEDGEMENT ix

APPROVAL x

DECLARATION xii

LIST OF TABLES xvi

LIST OF PLATES xviii

LIST OF FIGURES xx

LIST OF APPENDICES xxi

LIST OF ABBREVIATIONS xxiii

CHAPTER

1 INTRODUCTION 1

2 LITERATURE REVIEW 5

2.1 Rice weevil, Sitophilus oryzae L. 5

2.1.1 Biology and life history 5

2.1.2 Distribution and habits 6

2.2 Rice moth, Corcyra cephalonica St. 7

2.2.1 Biology and life history 7

2.2.2 Distribution and habits 10

2.3 Control methods 10

2.3.1 Physical methods 11

2.3.2 Chemical methods 12

2.4 Botanical insecticide 13

2.4.1 Piper nigrum L. (Piperaceae) 17

2.4.2 Mentha piperita L. (Lamiaceae) 23

2.4.3 Jatropha curcas L. (Euphorbiaceae) 28

2.4.4 Effect of plant extracts on the rice weevil, Sitophilus

oryzae and rice moth, Corcyra cephalonica 33

3 EXTRACTION AND ANALYSIS OF CHEMICAL

CONSTITUENTS IN Piper nigrum, Mentha piperita AND

Jatropha curcas 39

3.1 Introduction 39

3.2 Materials and Methods 40

3.2.1 Preparation of the plant materials 40

3.2.2 Preparation of plant extracts by percolation method 42

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3.2.3 Extraction of essential oils by hydrodistillation method

44

3.2.4 Identification of chemical components 46

3.3 Results and Discussion 47

3.3.1 GC analysis 47

3.3.2 GC-MS analysis 51

3.4 Conclusion 62

4 TOXICITY OF PLANT EXTRACTS AND ESSENTIAL OILS

FROM BLACK PEPPER, PEPPERMINT AND JATROPHA

AGAINST RICE WEEVIL AND RICE MOTH 63

4.1 Introduction 63


4.2.1 Insects 64

4.2.2 Preparation of plant materials, solvent extracts and

essential oils 65

4.2.3 Toxicity bioassay 66

4.2.4 Data analysis 68


4.4 Conclusion 78

5 EFFECT OF BLACK PEPPER AND JATROPHA EXTRACTS

AND BLACK PEPPER AND PEPPERMINT ESSENTIAL OILS

ON SOME BIOLOGICAL RESPONSES OF RICE WEEVIL

AND RICE MOTH 79

5.1 Introduction 79


5.2.1 Insects 80

5.2.2 Botanical preparations (extracts and essential oils) 80

5.2.3 Evaluation of antifeedant activity of plant extracts and

essential oils in S. oryzae adults and C. cephalonica

larvae 81

5.2.3.1 Extracts 81

5.2.3.2 Essential oils 81

5.2.4 Evaluation of repellency activity 85

5.2.5 Effect of plant extracts and essential oils on egg

hatchability 87

5.2.6 Effect of plant extracts and essential oils on adult

emergence 88

5.2.7 Data analysis 89


5.3.1 Evaluation of antifeedant activity of plant extracts and

essential oils against S. oryzae adults and C.

cephalonica larvae 90

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5.3.2 Evaluation of repellency activity 99

5.3.3 Effect of plant extracts and essential oils on egg

hatchability 104

5.3.4 Effect of plant extracts and essential oils on adult

emergence 109

5.4 Conclusion 115

6 GENERAL CONCLUSION AND RECOMMENDATION 118

REFERENCES 124

APPENDICES 149

BIODATA OF STUDENT 167

LIST OF PUBLICATIONS 168

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