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Lydia Rubiang-Y Alambing Doctor of Philosophy

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Lydia Rubiang-Y Alambing Doctor of Philosophy Aibika (a green leafy vegetable in PNG): Biodiversity and its effect on micronutrient composition Lydia Rubiang-Y alambing A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Food Science and Technology School of Chemical Engineering Faculty of Engineering March 2014 PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thesis/Dissertation Sheet Surname or Family name: Rubiang-Yalambing First name: Lydia Other name/s: Abbreviation for degree as given in the University calendar: PhD School: Chemical Sciences and Engineering Faculty: Engineering Title: Aibika (a green leafy vegetable in PNG): Biodiversity and effect on micronutrient composition Abstract 350 words maximum: (PLEASE TYPE) Over twenty different varieties of aibika or Abe/moschus manihot (L.) which is a commonly consumed green leafy vegetable in Papua New Guinea were studied with two main objectives. Firstly to determine the extent of genetic diversity between the accessions currently held at the National Agricultural Research Institute (NARI) in PNG which would aid effective management of the aibika germplasm and secondly to analyse micronutrients (total folate and minerals) in all accessions and identify any relationships between the nutrient contents and genotypes. Total folate contents ranged from 34 - 132 f..Lg/1 00 g on a fresh weight basis over two years indicating a wide range and a significant difference (p<0.05) between the two years. The mineral contents (mg/lOOg fresh weight) were in the following ranges for the 3 year period; iron, 0.8 -8.7; zinc, 0.32- 2.31; calcium, 197- 635; potassium, 265- 630; sodium, 1.0- 41; magnesium, 79-264; manganese, 0.42 - 2.09 and copper, 0.13 - 1.7. A significant (p<0.05) variation in the mineral contents was observed reflecting on variations in growing conditions between the collections. There was no significant effect of genotype on micronutrient content in all accessions. UPGMA analysis was performed to determine the clusters of accessions of aibika according to concentrations of all minerals and folate in all 3 years. Generally, the clusters of accessions were different in each of the 3 years. Environmental and other factors seem to have a greater impact on the micronutrient data compared to the genotype. The 23 accessions studied were grouped into five main groups according to the techniques of random amplification of polymorphic DNA (RAPD) and directed amplification of minisatellite region DNA (DAMD). Sequencing results from two chloroplast DNA regions studied, the psbM-trnD and the trnL-trnF intergenic spacer regions did not show any variation in the accessions. The nuclear encoded ITS region was also studied which showed that sequences from all accessions were identical with the exception of LAL Am 22 1 where the sequence data from the reverse primer suggested that there were 11 differences from the rest of the accessions. Data from this sequence has been submitted to the Genbank (accession number KC48173). Declaration relating to disposition of project thesis/dissertation I hereby grant to the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or in part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all property rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstracts International (this is applicable to doctoral lhe•oso"~). ~ .. .J75./.9..3.i_~QJ~ ··· ........~~ ·2:~ · ········· ·· ·· ·· Witness ;fate The University recognises that there may be exceptional circumstances requiring restrictions on copying or conditions on use. Requests for restriction for a period of up to 2 years must be made in writing. Requests for a longer period of restriction may be considered in exceptional circumstances and require the approval of the Dean of Graduate Research. FOR OFFICE USE ONLY Date of completion of requirements for Award: THIS SHEET IS TO BE GLUED TO THE INSIDE FRONT COVER OF THE THESIS ORIGINALITY STATEMENT 'I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.' Signed ....~~~ ·· · ····· · · ·· · ·········· Date ........\ .t .l.. .9. ..:?. /... ~ . o. .t .tT-················· COPYRIGHT STATEMENT 'I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). I have either used no substantial portions of copyright material in my thesis or I have obtained permission to use copyright material; where permission has not been granted I have applied/will apply for a partial restriction of the digital copy of my thesis or dissertation.' Signed .. j$l.'\- bL~ .. ......................... ........... Date ..... \~ . /. ..C? . .3 . ..f . .~ .. I.T-··· ·· ·· ··· ·· ········· ····· ··· ·· · AUTHENTICITY STATEMENT 'I certify that the Library deposit digital copy is a direct equivalent of the final officially approved version of my thesis. No emendation of content has occurred and if there are any minor variations in formatting, they are the result of the conversion to digital format.' Signed ..... ~."~· "'%·· ············· ·· ··· ·········· ··· ··· Date ...... ( . ~ . t.0 .3. ...J .. ;)o.l ~ ... ···· ···· ·· ··· ······· ···· ··· ACKNOWLEDGEMENTS I wish to thank the following individuals and organisations, without whose support I would never have done or completed this PhD study: Assoc. Professor Jayashree Arcot, my supervisor – I am grateful and fortunate to have a supervisor like you. You have assisted me in so many ways not just academically. Thank you for your patience, understanding and for the exposure you have given me in the area of food composition. The Australian Government (Australian Development Scholarship) – for the full scholarship to study in Australia The Papua New Guinea University of Technology, Lae, PNG – for allowing me to take up the study scholarship. The PNG National Agricultural Research Institute (NARI), for the MOA to use the aibika germplasm at Laloki. Rosa Kambuou, Janet Paofa and their staff at NARI Laloki, for the assistance with sample collection, sample exportation and information when needed. The assistance of other staff at Bubia is also acknowledged. Assoc. Professor Paul Holford, Hawkesbury Campus, University of Western Sydney, Richmond. I would never have done the DNA work without the assistance of someone so helpful, and understanding. I started the DNA work with no background knowledge on molecular biology and have learnt a lot in this study. Thank you. Professor Heather Greenfield, my co-supervisor, for her professional advice and guidance. Mr. Camillo Taraborrelli, for his assistance in the importation and quarantine procedures for the aibika samples, and the technical help in the running of my experiments. Karrie, Nisha, Veronica and Yang, thank you for the friendship and assistance during my study. My husband and son for their support, understanding and sacrifices made so I could complete my studies. i ABSTRACT Aibika or Abelmoschus manihot as it is scientifically known is the most popular and commonly-consumed indigenous green leafy vegetable in Papua New Guinea (PNG). Many different varieties of aibika can be found in the country and the PNG National Agricultural Research Institute (NARI) is overseeing the collection and maintenance of these varieties in a field gene bank at Laloki, PNG. At this stage the accessions have only been morphologically characterised and there is no data on genetic differences between the accessions. Over twenty different varieties of aibika were imported into Sydney over a three year period. The two (2) main objectives for this study were: Firstly to determine the extent of genetic diversity between the accessions with the aim of assisting PNGNARI in identifying core genotypes in the current collection which would aid its effective management. Second aim was to analyse micronutrients (total folate and minerals) in all accessions and identify any relationships between the nutrient contents and genotypes. It is known that micronutrients do vary within varieties of same species so that the identification of a nutrient rich variety or
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