universiti putra malaysia mousa khani fp 2012 45
TRANSCRIPT
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
Faculty of Agriculture
Universiti Putra Malaysia
(Internal Examiner)
Faizah bt Abood, PhD
Associate Professor
Department of Forest Management
Faculty of Forestry
Universiti Putra Malaysia
(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
Universiti Putra Malaysia
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
Faculty of Agriculture
Universiti Putra Malaysia
(Chairperson)
Dzolkhifli Omar, PhD
Professor
Faculty of Agriculture
Universiti Putra Malaysia
(Member)
Mawardi Rahmani, PhD
Professor
Faculty of Science
Universiti Putra Malaysia
(Member)
Shamsali Rezazadeh, PhD
Iranian Institute of Medicinal Plants
Karaj, Iran
(Member)
BUJANG BIN KIM HUAT, PhD
Professor and Dean
School of Graduate Studies
Universiti Putra Malaysia
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 Materials and Methods 64
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
4.3 Results and Discussion 69
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 Materials and Methods 80
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 Results and Discussion 90
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