Improving Reforestation Success of High-Value and Key Forest Species by Direct Seeding in Southeast Asia and Western Australia
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IMPROVING REFORESTATION SUCCESS OF HIGH-VALUE AND KEY FOREST SPECIES BY DIRECT SEEDING IN SOUTHEAST ASIA AND WESTERN AUSTRALIA By Thea So BSc. (Sylviculture), MSc. (Tropical Forest Resources Management) This thesis is submitted in fulfillment of the requirement for the degree of Doctor of Philosophy School of Biological Sciences and Biotechnology Faculty of Science, Engineering and Sustainability Murdoch University, Perth, Western Australia November 2011 Declaration DECLARATION I declare that all the work described in this thesis is my own account of my research which was undertaken while I was enrolled as a full time research student for the degree of Doctor of Philosophy at Murdoch University, Western Australia, from 2007 to 2010. This work has not previously been submitted for a degree at any tertiary education institution. Thea So November 2011 Murdoch University 2011 ii Acknowledgement ACKNOWLEDGMENTS First of all, I would like to extend my sincere gratitude to my principal supervisor, Professor Bernard Dell, for his tirelessness and inspiration in supervising my study. I appreciate his patience in reading and correcting all my work. My sincere thankfulness to Dr Katinka Ruthrof and Dr Lambert Braü for their guidance, constructive advice and supports in many aspects of this thesis. I would like to thank Mr Chairat Aramsri, Professor Bunvong Thaiutsa and Ms Montathip Sommeechai for their coordination in the establishment of the field trial in Thailand. A/Professor Mike Calver and Professor Ananchai Khuantham are thanked for their guidance on experimental design. Thanks are due to Mr Jason Foster Ralph Sarich for allowing me to use his land for the field trial in Western Australia. Mycorrhizal inocula were provided by Zadco For Quality Gro Pty Ltd and Dr Phakpen Poomipan. The Forest Industry Organization (FIO) of Thailand is thanked for the excellent collaboration in establishment of the trial on its land. Mr Chaiya Junsawang and all FIO staff at the Sakeaw Forest Plantation provided excellent hospitality and support. Dr Jumnian Wongmo is thanked for her assistance in the field trial establishment in Western Australia and procurement of AM inocula in Thailand. Dr Yvette Hill provided instructions on working with N2-fixing bacteria and in setting up glasshouse experiments. Liza Parkinson provided assistance in molecular work. Mr Dy Sophy is thanked for his assistance in monitoring the research trial in Cambodia. Many people provided assistance during my field work in Cambodia, especially Ry Sam El, Suo Hai, So Than, Long Boung, Moy Rotha, Uon Sam Ol, Huot Ainun, Lim Sopheap, Thai Seila Tina, Nuon Pov Ratana, Ken Phan, Lao Sethaphal, Sreang Meng Srun and Preap Sam. I am indebted to colleagues and fellow students at Murdoch University for their support, encouragement and assistance, especially Harry, Tan, Endah and Lily. Special thanks are due to Peter Scott for his support and encouragement. The Australian Leadership Awards (ALA) is gratefully acknowledged for granting a scholarship for this study. The field trials in Cambodia and Thailand were partially financed by Murdoch University and AusAID. A big gratefulness is due to my parents, my wife and son, Veasna, sisters and brother for their encouragement, love and support. Last but not least, I would like to express my sincere thanks to my bosses, H. E. Chheng Kim Sun, and H. E. Chea Sam Ang for their encouragement and support of my study. Murdoch University 2011 iii Abbreviation ABREVIATIONS AM arbuscular mycorrhiza/l ANR assisted natural regeneration ANOVA analysis of variance a.s.l. above sea level BSO breeding seed orchard cm centimetre CITES Convention on International Trade in Endangered Species CRD completely randomized design CSBP Cumming Smith British Petroleum CSO clonal seed orchard CTSP Cambodia Tree Seed Project DBH diameter at breast heigh d.f. degree of freedom DI water distilled deionized water DNA deoxyribonucleic acid DTPA diethylene triamine pentaacetic acid ECM ectomycorrhiza/l EDTA ethylenediaminetetraacetic acid e.g. for example (Latin exemli gratia) EN endangered F (statistic) A value based on a standard statistical test used in ANOVA/MANOVA FA Forestry Administration FAO Food and Agriculture Organization of the United Nations FLD Forest & Landscape Denmark FORRU Forest Restoration Research Unit GoC Government of Cambodia h hour ha hectare i.e. that is (Latin id est) IFSR Independent Forest Sector Review INVAM International Culture Collection of (Vesicular) Arbuscular Mycorrhizal Fungi Murdoch University 2011 iv Abbreviation IUCN International Union for Conservation of Nature L litre MAFF Ministry of Agriculture, Forestry and Fisheries MANOVA multivariate analysis of variance min minute mL millilitre mm millimetre n.a. not available NFP National Forest Programme NGO non-governmental organization nM nanomole nom. ined unpublished name (Latin nomen ineditum) OM organic matter p probability PCR polymerase chain reaction RCBD randomized complete block design rpm revolution per minute rRNA ribosomal ribonucleic acid s second S.E. standard error Sig. significance TAE Tris, acetic acid and EDTA TY-medium tryptone peptone yeast extract-medium UNEP United Nations Environment Programme UTM Universal Transverse Mercator V volt VU vulnerable v:v volume:volume WA Western Australia WCMC World Conservation Monitoring Centre w:v weight:volume w:w weight:weight µL microlitre μm micrometre Murdoch University 2011 v Definition of terms DEFINITION OF TERMS Buffer (solution) An aqueous solution containing a weak acid in its conjugate base (Chapter 6). Effective Significant improvement in establishment, survival and growth (height, diameter or dry weight), brought about by a treatment (as compared to the control) inoculated to seeds or seedlings under field or glasshouse conditions. Taq DNA A thermostable emzyme wich replicate DNA at 74°C. The ability of polymerase the enzyme to survive multiple rounds of temperature cycling make it extremely useful in PCR and cycle sequencing (Fisher Biotech Australia, www.fisherbiotech.com). Survival The state of seedlings that have survived after trial establishment (seedling) until the time of monitoring. Treatment A single or a set of materials that were applied or inoculated to seeds or seedlings under field or glasshouse conditions in an expectation that they promote establishment, survival and growth of the seedlings. In the experimental Chapters 3 and 5-7, the term treatment is frequently abbreviated by the letter T followed by a number (e.g., T1, T2, …,T8). Note: The species specific epithet is used when a species is cited the first time. However, for those species used in experiments, the full names are given in each Chapter the first time they are cited. Murdoch University 2011 vi Abstract ABSTRACT Worldwide, natural forests have been decreasing in area at an alarming rate. In Cambodia, the annual deforestation rate was 127 000 ha year-1 from 2005 to 2010 and this seriously threatens biodiversity and the livelihoods of rural communities. Therefore, there is an urgent need to reforest or establish forest plantations to meet two main objectives: economic development and biodiversity conservation. This thesis concerns the promotion of early survival and growth of planted or direct-seeded seedlings to overcome the harsh conditions of reforestation sites in tropical and mediterranean-type ecosystems, with special attention given to threatened high-value timber species of Southeast Asia. Worldwide, there is an increasing effort to reforest degraded forests and old agricultural lands. However, reforestation of degraded lands is often difficult and is usually expensive; therefore, direct seeding is an alternative to conventional tree planting. Some of the major constraints facing reforestation efforts were reviewed, and opportunities that could be useful for promotion of early establishment and growth of seedlings were explored. These were then used to define reforestation field studies in Australia, Thailand and Cambodia. In a harsh mediterranean-type ecosystem in Western Australia, effects of microorganisms (mycorrhizal fungi and N2-fixing bacteria) and planting material (seed and seedling) on survival and growth of two key post-disturbance colonizing species, Eucalyptus gomphocephala and Acacia saligna, were investigated. For E. gomphocephala, survival at 13 months was higher for out-planted seedlings (81%) than from direct seeding (7.5%). Inoculation with ectomycorrhizal fungal spores was not beneficial. For A. saligna, survival at 13 months was also higher for seedlings (84%) than for seeding (42.5%). Nitrogen-fixing bacteria from crushed root nodules of A. saligna did not promote survival or growth of the species nor did a mixed commercial mycorrhizal inoculum. In Southeast Asia, the leguminaceous rosewoods, Afzelia xylocarpa and Dalbergia cochinchinensis, are threatened throughout their range by habitat loss and over exploitation for their extremely highly-prized timber. The species have been promoted for reforestation in Cambodia for economic development and genetic conservation. The Murdoch University 2011 vii Abstract current conservation status of A. xylocarpa and D. cochinchinensis in Cambodia was examined, and information on silviculture, trade and current conservation measures applied in that country was drawn together. Some important steps in the development of domestication strategies, including testing and improving silvicultural practices and increasing the supply of genetically superior seeds from seed production