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A Protocol for Floral Odour Analyses of Schismatoglottis Calyptrata (Araceae: Schismatoglottideae)

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A Protocol for Floral Odour Analyses of Schismatoglottis Calyptrata (Araceae: Schismatoglottideae) Faculty of Resource Science and Technology A PROTOCOL FOR FLORAL ODOUR ANALYSES OF SCHISMATOGLOTTIS CALYPTRATA (ARACEAE: SCHISMATOGLOTTIDEAE) NUR ADILAH BINTI MUHAMMAD ARIF (24521) Bachelor of Science with Honours (Plant Resources Science and Management) 2012 A Protocol for Floral Odour Analyses of Schismatoglottis calyptrata (Araceae: Schismatoglottideae) Nur Adilah Binti Muhammad Arif (24521) This Report is submitted in partial fulfillment of the requirements of the Degree of Bachelor of Science with Honours (Plant Resource Science and Management) Plant Resource Science and Management Programme Department of Plant Science and Environmental Ecology Faculty of Resource Science and Technology Universiti Malaysia Sarawak 2012 APPROVAL SHEET Name of Candidate : Nur Adilah Binti Muhammad Arif Title of Dissertation : A Protocol for Floral Odour Analyses of Schismatoglottis calyptrata (Araceae: Schismatoglottideae) ………………………………………….. (Dr. Wong Sin Yeng) Supervisor …………………………………….. (Dr. Siti Rubiah Binti Zainudin) Coordinator Plant Science and Management Program Department of Plant Science and Environmental Ecology Faculty Resource Science and Technology Universiti Malaysia Sarawak i DECLARATION I hereby declare that the report is based on my original work except for quotation and citations which have been duly acknowledgment. I also declare that it has not been previously or concurrently submitted for any degrees at UNIMAS or any other institutions of higher learning. …………………………… (Nur Adilah Binti Muhammad Arif) Plant Resource Science and Management Program Department of Plant Science and Environmental Ecology Faculty of Resource Science and Technology Universiti Malaysia Sarawak. ii ACKNOWLEDGEMENTS In the name of Allah the Almighty, with grateful pleasure and profound sense of gratitude, I, Nur Adilah Binti Muhammad Arif would like to express my most cordial and humble thanks to my respected supervisor Dr. Wong Sin Yeng for her valuable guidance, keen interest, inspiration, encouragement, time spent and moral support throughout completion of my study. Special appreciation and gratitude is also addressed to MSc candidate, Mr Hoe Yin Chen for his useful information, guidance, materials provided and valuable suggestion in this study. My sincere thank is also extended to my mentor, Prof. Dr. Cheksum @ Supiah Binti Tawan for her continuous moral support and kindness. Special thanks to the lab assistants for their kind co-operations. Lastly, I wish to express my sincere appreciation especially to Nurul Fatini Shahirah Suharizan, Zulaikha Zainal, Nurul Atiqah Abdul Rahman, Khairul Afiq Mohamed Noordin and Muhammad Firdaus Latip who gave a lot of support and motivation. Also not forgotten, to all my colleagues, who had assisted me in conducting this study either directly or indirectly. From the deepest depth of my heart, I express my thanks to my beloved mother and father, and also the rest of my family members who kept supporting. iii TABLE OF CONTENTS Page Approval Sheet i Declaration ii Acknowledgements iii Table of Contents iv List of Tables and Figures x Abstract 1 1.0 Introduction 1.1 Problem Statements 5 1.2 Objectives 5 2.0 Literature Review 2.1 Bornean Araceae 6 2.2 Tribe Schismatoglottideae 9 2.3 Schismatoglottis 12 2.4 Schismatoglottis calyptrata complex 15 2.5 Floral Odours and Pollination Biology 19 3.0 Materials and Methods 3.1 Sampling Location 21 3.2 Floral Odour Trapping Method 22 3.3 Sampling of Floral Odours 24 3.4 Gas Chromatography 25 3.5 Mass Spectrometry Analysis 30 iv 4.0 Results and Discussions 4.1 Compounds 32 4.2 Chromatograms 34 4.3 Discussions 44 5.0 Conclusions and Recommendations 54 References 56 v A Protocol for Floral Odour Analyses of Schismatoglottis calyptrata (Araceae: Schismatoglottideae) Nur Adilah Muhammad Arif Plant Resource Science and Management Programme Department of Plant Science and Environmental Ecology Faculty of Resource Science and Technology Universiti Malaysia Sarawak ABSTRACT The Schismatoglottis calyptrata complex comprises about 30 species of herbs originating from the Asian humid tropics belonging to the plant family Araceae. Inflorescences of Schismatoglottis flower by a series of functional ‘phases’, beginning with female flower activity (the ‘pistillate phase’) and ending with pollen release (‘staminate phase’), with these phases separated by a transitional period. The production of primary floral attractants was in the form of powerful esteric-like odours. Several species of the Calyptrata complex frequently occur together in a habitat, with the different species often flowering at the same time yet nowhere are there recorded interspecific hybrids. It thus seems very probable that aspects of the species’ pollination maintain distinct lineages of the various species, even in situations of multi-species flowering. A protocol for floral odour analyses of S. calyptrata was established in this study. The main compound of the floral scent trapped was examined by using gas-chromatography mass-spectrometry (GCMS) techniques with the aim of identifying the compounds emitted. Four compounds were identified and characterised, representing 73.74-93.16% of the total GC area. The major volatile representing about 50% was the Methyl 3-methyl-3-butenoate. This compound was found in higher concentrations from female anthesis of S. Calyptrata (F3). Generally, a significant difference in the composition of volatile compounds between the compounds from the same antheses and from different geographic regions was recorded. The results suggest that climatic factors, latitude and longitude affect the formation of volatiles. Key terms: Araceae – Schismatoglotideae – Schismatoglottis calyptrata –floral odour – GCMS ABSTRAK Kompleks Schismatoglottis calyptrata terdiri daripada kira-kira 30 spesies herba yang berasal dari kawasan lembap tropika Asia, keluarga tumbuhan Araceae. Pendebungaan bunga Schismatoglottis oleh satu siri 'fasa' berfungsi, bermula dengan aktiviti bunga betina ('fasa pistilat') dan berakhir dengan pembebasan debunga ('fasa staminat'). Fasa-fasa ini dipisahkan oleh tempoh peralihan. Pengeluaran penarik utama bunga ini dalam bentuk bau esterik yang kuat. Beberapa spesies daripada kompleks Calyptrata sering berlaku bersama-sama di dalam habitat, dengan spesies berbeza yang sering berbunga pada masa yang sama tetapi mana yang ada direkodkan sebagai kacukan antaraspesies. Berkemungkinan besar bahawa aspek pendebungaan spesis mengekalkan keturunan yang berbeza daripada pelbagai spesies, walaupun dalam keadaan berbunga multi-spesies. Satu protokol untuk analisis bau bunga S. calyptrata telah diwujudkan dalam kajian ini. Sebatian utama aroma bunga yang terperangkap telah diperiksa dengan menggunakan teknik kromatografi gas-spektrometri jisim (GCMS) bertujuan untuk mengenal pasti sebatian dikeluarkan. Empat sebatian telah dikenal pasti dan ciri-ciri, mewakili 73.74-93.16% daripada jumlah kawasan GC. Sebatian ruapan utama yang mewakili kira-kira 50% adalah Methyl 3-metil-3-butenoate. Sebatian ini didapati dalam kepekatan yang lebih tinggi dari bunga putik betina S. Calyptrata (F3). Secara umumnya, perbezaan yang signifikan dalam komposisi sebatian meruap antara sebatian dari putik-putik bunga yang sama dan dari kawasan geografi yang berbeza telah direkodkan. Hasil kajian mencadangkan bahawa faktor cuaca, latitud dan longitud memberi kesan kepada pembentukan meruap. Key terms: Araceae – Schismatoglotideae – Schismatoglottis calyptrata –floral odour – GCMS 1 1.0 INTRODUCTION The Araceae is a family of herbaceous monocots. The family members are highly diverse in life form from submerged to free-floating aquatics. The leaves can vary considerably from species to species. Species in Araceae are often rhizomatous or tuberous and are often found to contain calcium oxalate crystals or raphides. Inflorescences of some plants in this family are thermogenic (heat-producing) as the temperature of the inflorescences can reach up to 45 degrees Celsius even when the surrounding air temperature is much lower (Gibernau, et al. 2005). Boyce et al. (2010) reported that the total aroid flora for Borneo quite likely exceeds 1,000 species. In short, Borneo is an aroid habitat of global significance; and arguably one of the richest and most diverse on the planet. Barabe & Lacroix (2001) explained the lack of floral developmental studies in the Araceae family results in part from the difficulty in obtaining enough material to adequately document the early stages of development. An alpha-taxonomic revision of the Malesian species, one of the results of which was the recognition of six informal species grouping based accordingly on short architechture and the senescence mechanics of the upper spathe (Hay & Yuzammi, 2000). Tribe Schismatoglottideae is a predominantly Indomalayan taxon comprising 13 genera of terrestrial, lithophytic or rheophytic herbs with a primary distribution centered on Borneo but with the total distribution ranging to South West Myanmar, South West China, and Vanuatu (Hay & Yuzammi, 2000). Schismatoglottis Zoll. & Moritzi is the largest genus in Schismatoglottideae with over 200 species. Spadix structure of Schismatoglottis is complex and it is divided into a lower female flower zone and a middle male flower zone with interstice 2 (sterile female and male flowers) and often with a sterile appendix. Schismatoglottis inflorescences are generally rather small and inconspicuous, characterized by a complex spadix with up to three different kinds of staminode present, the fertile elements segregated
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