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Conservation Genetics and Ecology of Two Rare Grevillea Species

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Conservation Genetics and Ecology of Two Rare Grevillea Species University of Wollongong Research Online University of Wollongong Thesis Collection 1954-2016 University of Wollongong Thesis Collections 2004 Conservation genetics and ecology of two rare grevillea species Tanya M. Llorens University of Wollongong Follow this and additional works at: https://ro.uow.edu.au/theses University of Wollongong Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: This work is copyright. Apart from any use permitted under the Copyright Act 1968, no part of this work may be reproduced by any process, nor may any other exclusive right be exercised, without the permission of the author. Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong. Recommended Citation Llorens, Tanya M, Conservation genetics and ecology of two rare grevillea species, PhD thesis, Department of Biological Sciences, University of Wollongong, 2004. http://ro.uow.edu.au/theses/374 Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: [email protected] NOTE This online version of the thesis may have different page formatting and pagination from the paper copy held in the University of Wollongong Library. UNIVERSITY OF WOLLONGONG COPYRIGHT WARNING You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Conservation genetics and ecology of two rare Grevillea species A thesis submitted in fulfilment of the requirements for the award of the degree DOCTOR OF PHILOSOPHY from the UNIVERSITY OF WOLLONGONG by Tanya M. Llorens B. Sc. (Hons) DEPARTMENT OF BIOLOGICAL SCIENCES 2004 Certification I, Tanya M. Llorens, declare that this thesis, submitted in fulfilment of the requirements for the award of Doctor of Philosophy, in the Department of Biological Sciences, University of Wollongong, is wholly my own work unless otherwise referenced or acknowledged. The document has not been submitted for qualifications at any other academic institution. Tanya Llorens 10 September 2004 Table of Contents List of Tables…………………………………………………………………………… vi List of Figures………………………………………………………………………….. viii Abbreviations and conventions………………………………………………………… xi Abstract………………………………………………………………………………… xii Acknowledgements…………………………………………………………………….. xv Chapter 1 General Introduction…..……………………………………………………. 1 1.1 The impact of habitat fragmentation on natural populations..…………………………….. 1 1.1.1 Ecosystem fragmentation………….…………………………...………………….… 1 1.1.2 Viability of fragmented populations – mechanisms for declines…….…………...…. 1 1.1.3 Clarification of some important processes……………………...…….…………...…. 4 1.1.4 Genetics or demography / environment?……………………….…….…………...…. 5 1.1.5 Empirical evidence for fragmentation effects on viability……...…….…………...…. 7 1.2 Fragmentation of terrestrial plants…………………………………...…………………….. 9 1.3 Habitat losses and the decline of Australian plants……………………………………...…10 1.4 This study……………………………………………………………………………….… 11 1.5 Grevillea in Australia…………………………...……………………………………...… 12 1.6 Grevillea caleyi and G. longifolia……………....……………………………………...… 14 1.6.1 Biology and ecology……...…….…………....……....……....……....……..…….… 14 1.6.2 Species distributions, fragmentation and current threats.......……....……....………. 17 1.7 Thesis aims and outline.………………………...……………………………………...… 21 1.7.1 Predictions……………………………………………..........……....……....…….… 22 1.7.2 Questions……………………………………………..........……......……....……..... 24 1.7.3 Genetic markers...……………………………………..........……....……....……..... 24 1.7.4 Thesis outline...………………………………………..........……....……....…….... 25 Chapter 2 Genetic subdivision among recently fragmented populations of Grevillea caleyi…………………………………...………………………….. 27 2.1 Introduction…………………………………...……………………………………...….. 27 2.2 Methods…………………………………...……………………………………...……… 29 2.2.1 Biology…………………………………...……………………………………...… 29 2.2.2 Population sampling…………………...………………………………………...… 30 2.2.3 DNA extraction………...………………...……………………………………...… 31 2.2.4 AFLP procedure…………………...……………………………………...……….. 32 2.2.5 Data analysis…………………...……………………………………...…………… 32 i 2.3 Results…………………...……………………………………...………………………... 34 2.3.1 AFLP markers…………………...……………………………………...…………. 34 2.3.2 Population differentiation…………………...……………………………………... 35 2.3.3 Effects of population size and isolation…………………...………………………. 37 2.4 Discussion…………………...………………………………………………………....… 38 2.4.1 Genetic structure within G. caleyi…………………...…………………………….. 38 2.4.2 Within-population genetic diversity…………………...…………………………... 40 Chapter 3 Genetic diversity and structure across different spatial scales, and inferred past mating system, in Grevillea caleyi and G. longifolia...... 42 3.1 Introduction…………………...……………………………………...…………………... 42 3.2 Methods…………………...……………………………………...……………………… 44 3.2.1 Population sampling…………………...……………………………………...…… 44 3.2.2 Microsatellite primers – transfer from G. macleayana and G. iaspicula………….. 48 3.2.3 Microsatellite procedure…………………...…………………………………….… 50 3.2.4 Testing of microsatellite marker structure, independence and power…...………… 52 3.2.4.1 Microsatellite allele structure…………………...………………………… 52 3.2.4.2 Independence of microsatellite loci…………………...………………….. 52 3.2.4.3 Power of the microsatellite markers…………………...…..…………...… 53 3.2.5 Data analysis…………………...……………………………………...…………... 53 3.2.5.1 Genetic diversity within populations…...……...……...……...……...……. 53 3.2.5.2 Population differentiation...……...……...……...……...………………….. 53 3.2.5.3 Fine-scale diversity and structure within G. caleyi populations.…...……... 54 3.2.5.4 Indirect estimates of the mating system for G. caleyi and G. longifolia….. 54 3.3 Results...……...……...……...……...……..…...……...……...……...………………….... 55 3.3.1 Basis of the microsatellite variation...……...……...……...……...………………... 55 3.3.2 Genetic diversity within populations...……...……...……...……...……………….. 55 3.3.3 Population differentiation...……...……...……...……...…………………………... 64 3.3.4 Fine-scale diversity and structure within G. caleyi populations...……...……...….. 68 3.3.5 Indirect estimates of the mating system for G. caleyi and G. longifolia…………... 70 3.4 Discussion……...……...……...……...…………………………………………………... 71 3.4.1 Microsatellite diversity in G. caleyi and G. longifolia……...……...……...…….… 71 3.4.2 Genetic effects of population size and isolation……...……...……...…….……….. 72 3.4.3 Genetic structure among populations……...……...……...……...………………… 74 3.4.4 Fine-scale genetic structure in G. caleyi……...……...……...……...……………... 77 3.4.5 Mating system estimates……...……...……...……...……………………………... 79 ii Chapter 4 Mating system and pollinators of Grevillea caleyi …………………... 82 4.1 Introduction…...……...……………………………...……………………..…………….. 82 4.2 Methods…...……...……………………………...………………………………………. 84 4.2.1 Flowering characteristics of Grevillea caleyi…...……...………………..………… 84 4.2.2 Mating system - Autogamy…...……...…………………………….……………… 85 4.2.3 Mating System - Outcrossing rates…...……...…………………………….……… 85 4.2.3.1 Collection and genotyping of samples………………………….………… 85 4.2.3.2 Analysis…………….………………….………………….…………….… 86 4.2.4 Pollinators………….………………….…………….…………………………….. 88 4.2.4.1 Bird observations………….………………….…………….…………….. 88 4.2.4.2 Honeybee observations………….………………….…………….………. 89 4.2.4.3 Characteristics of sites and observation plots………….…………………. 90 4.2.4.4 Statistical tests………….………………….…………….……………….. 91 4.3 Results………….………………….…………….……………………………………….. 92 4.3.1 Mating system………….………………….…………….………………………… 92 4.3.1.1 Autogamy………….………………….…………….……………………. 92 4.3.1.2 Outcrossing rates………….………………….…………….…………….. 92 4.3.2 Pollinators………….………………….…………….…………………………….. 95 4.3.2.1 Characteristics of sites and observation plots..….………………………... 95 4.3.2.2 Flower visitors………….………………….…………….……………….. 96 4.3.2.3 Foraging behaviour………….………………….……………………….…. 98 4.3.2.4 Temporal foraging patterns………….………………….…………….…… 100 4.3.2.5 Visitation rate……….………………….……………………….………... 102 4.3.2.6 Bird species……….………………….……………………….………….. 103 4.3.2.7 Pollinator movements……….………………….………………………... 104 4.4 Discussion……….………………….……………………….………………………….
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