Genetic variation of dipterocarps: from molecular phylogenies to the identification of the origin of timber

Reiner Finkeldey, Oliver Gailing, Iskandar Siregar, Ulfah Siregar, Randy Villarin, Cuiping Cao, Nga Phi Nguyen, Hani Nuroniah, Yanti Rachmayanti

Büsgen Institute Forest Genetics and Forest Breeding 1 Structure of presentation

l Background – l Interspecific variation: molecular phylogeny of Dipterocarpaceae l Intraspecific variation of selected spp. l Application: Molecular methods to trace the origin of wood l Conclusions and outlook l Literature

2 Background: Dipterocarpaceae

The pantropical distribution of Dipterocarpaceae

Monotoideae Dipterocarpoideae Pakaraimoideae 3 Genera; 40 13 Genera; 470 Species 1 ; 1 Species 3 Background: Dipterocarpaceae

Dipterocarpoideae in insular Southeast-Asia

Phylogenie der Dipterocarpaceae

Number of endemic species Number of non-endemic species 4 Background: Dipterocarpaceae

Dipterocarpoideae in Southeast-Asia

l Widely distributed – Mainland and insular SE-Asia l Dominating lowland rainforests and monsoon forests in SE-Asia – 50-80% of wood volume – Keystone group l Important timber species – Trade name: Meranti, Balau, Keruing, Kapur l Many endangered taxa Dipterocarp forest on 5 Background: Dipterocarpaceae

Dipterocarp: growth habit

l Medium-sized – emergent – Mainly climax species – Often long, straight boles – Slow to fast growth

Dipterocarpus Shorea selanica (<60 years) 6 condoriensis Background: Dipterocarpaceae

Dipterocarpaceae – Reproductive biology

l Hermaphrodites l Zoogamous (mainly insect- pollinated) – Bees, butterflies – l Self compatible, mixed mating system – t typical 60-90%

7 condoriensis Background: Dipterocarpaceae

Dipterocarp fruits

l Typical 2 (0-5) winged diaspores – Medium-sized to very large l recalcitrant l No efficient means of seed dispersal

8 Background: Dipterocarpaceae

Exploitation of dipterocarps

Local International

9 Hintergrund: Dipterocarpaceae

Use of dipterocarps

By-products Wood

window frames

charcoal

furniture

plywood

10 resin Phylogeny of Dipterocarpaceae

Order: ; Family Dipterocarpaceae

l Blume (1825): related to

l Cronquist (1968) & Takhtajan (1969): Theales

l Dahlgren (1975): Malvales

l Maguire et al. (1977), Dayanandan et al. (1999): Malvales (Sarcolaenaceae)

l Chase et al. (1993); Alverson et al. (1998); Bayer et al., (1999): Malvales (Bombacaceae, Tiliaceae, Sterculiaceae and Malvaceae)

11 Phylogeny of dipterocarps

Malvales; family Dipterocarpaceae

rbcL sequences

12 (nach CHASE et al., 1993) Material

Material for studies on inter- and intraspecific variation

l Over 3000 samples from dipterocarps in SE-Asia – Dried l Silica gel l Long-term storage at -60°C – Extracted DNA – Herbarium material maintained l Mainly with partners l Total of 116 species – All genera of Dipterocarpoideae – From six countries l More than 40 locations l Focus on Indonesia (>2500 samples) – Mainly natural forests (>80%), few plantations, arboreta, botanical gardens

13 Phylogeny of Dipterocarpaceae

Methods: cpDNA polymorphisms (Indrioko et al., 2006)

– PCR-RFLP ofcpDNA regions : trnLF, rbcL, petB, psbA, psaA (Tsumura et al., 1996) – cpSSRs (Microsatellites): ccmp2, ccmp6, ccmp10 (Weising et al., 1999) – Sequences: trnL; trnLF trnL region

14 PCR RFLPs cpSSRs Sequences Outgroup: Outgroup: Monotes Upuna borneensis lanceolatum Monotes kerstingii bantamensis Upuna borneensis Vatica granulata costata 99 99 Anisoptera reticulata 68 Vatica bantamensis 95 Vatica venulosa Vatica bella Vatica granulata Anisoptera marginata 99 Vatica pauciflora 99 Anisoptera reticulata 83 Vatica rassak 100 94 Dipterocarpus grandiflorus Vatica venulosa Dipterocarpus oblongifolius Dipterocarpus grandiflorus 100 Dipterocarpus oblongifolius Dipterocarpus rigidus Dipterocarpus retusus 71 100 Dipterocarpus rigidus aromatica Dipterocarpus tempehes 99 bancana Dryobalanops lanceolata 99 Hopea bancana Hopea celebica

80 92 Hopea odorata Hopea dryobalanoides 85 Hopea sangal <50 Hopea nigra Hopea dryobalanoides <50 <50 Hopea griffithii Hopea nigra Hopea mengarawan <50 Hopea griffithii 67 Parashorea lucida Hopea mengarawan

100 66 globosa Parashorea globosa 94 Shorea blumutensis Shorea blumutensis Shorea montigena Shorea guiso Shorea scaberrima Shorea montigena Shorea seminis Shorea scaberrima Shorea acuminata Shorea seminis Shorea andulensis Shorea acuminata Shorea javanica Shorea andulensis

58 Shorea johorensis Shorea javanica Shorea leprosula 67 Shorea johorensis Shorea macroptera Shorea leprosula Shorea macroptera Shorea mecistopteryx 98 95 69 Shorea mecistopteryx Shorea ovalis 69 63 Shorea ovalis Shorea palembanica 63 Shorea palembanica Shorea parvifolia Shorea parvifolia Shorea platyclados Shorea platyclados Shorea xanthophylla 67 <50 Shorea xanthophylla Shorea balangeran Shorea balangeran Shorea selanica 50 Shorea selanica 59 Shorea splendida Shorea splendida Shorea macrophylla Shorea macrophylla 58 Shorea pinanga 55 Shorea pinanga 50 Shorea stenoptera <50 Shorea stenoptera Shorea acuminatissima Shorea acuminatissima

Shorea dasyphylla Shorea dasyphylla Shorea faguetiana Shorea faguetiana 77 82 Shorea multiflora Shorea multiflora 15 Shorea fallax Shorea fallax Shorea materialis Shorea materialis 98 98 Shorea virescens Shorea virescens 60 61 Phylogeny of Dipterocarpaceae

Diagnostic characters of endemic species

Monotes kerstingii Cotylelobium lanceolatum Upuna borneensis Anisoptera costata Upuna borneensis Anisoptera marginata 68 99 Anisoptera reticulata Vatica bantamensis Vatica bella 99 Vatica granulata Vatica pauciflora 83 94 Vatica rassak Vatica venulosa Dipterocarpus grandiflorus Dipterocarpus oblongifolius 71 Dipterocarpus retusus 100 Dipterocarpus rigidus Dipterocarpus tempehes Dryobalanops aromatica 99 Dryobalanops lanceolata Hopea bancana Hopea celebica Hopea odorata 85 Hopea sangal <50 Hopea dryobalanoides Hopea nigra <50 Hopea griffithii 66 Hopea mengarawan Anisoptera reticulata 100 Parashorea lucida Parashorea globosa Shorea blumutensis Shorea guiso Shorea montigena Shorea scaberrima Shorea seminis Shorea acuminata Shorea andulensis Shorea javanica 67 Shorea johorensis Shorea leprosula Shorea macroptera 95 69 Shorea mecistopteryx 63 Shorea ovalis Shorea palembanica Shorea parvifolia Shorea platyclados <50 Shorea xanthophylla Shorea balangeran 59 Shorea selanica Shorea splendida Shorea macrophylla 55 Shorea pinanga <50 Shorea stenoptera Shorea acuminatissima Shorea dasyphylla 77 Shorea faguetiana Shorea multiflora 16 Shorea fallax Shorea fallax Shorea materialis 98 60 Shorea virescens Phylogeny of Dipterocarpaceae

Phylogeny of tribe Dipterocarpae

l Based on sequence variation of two cpDNA regions – trnL Intron – trnL-F intergenic spacer

17 (Nga, 2009) Phylogeny of Dipterocarpaceae

Molecular phylogeny based on AFLPs

Dipterocarpus retusus Dipterocarpus oblongifolius Dipterocarpus rigidus Dipterocarpus tempehes Dipterocarpus glandiflorus 1 Dipterocarpus glandiflorus 2 Anisoptera reticulata Anisoptera marginata Anisoptera costata 1 Anisoptera costata 2 Cotylelobium lanceolatum Upuna borneensis Vatica bantamensis 1 Vatica bantamensis 2 Vatica venulosa Vatica bella Vatica granulata Vatica rassak Vatica pauciflora 1 Vatica pauciflora 2 Hopea celebica Hopea bancana 1 Hopea bancana 2 Hopea sangal 1 The phylogeny based on morphology Hopea sangal 2 Hopea griffithii Hopea nigra Hopea dryobalanoides 1 and cpDNA variation is also reflected at Hopea dryobalanoides 2 Hopea mengarawan 1 Hopea mengarawan 2 Parashorea lucida AFLPs, indicating genome-wide Parashorea globosa Hopea odorata 1 Hopea odorata 2 Shorea seminis differentiation pattern congruent with Shorea macroptera Shorea montigena Shorea andulensis Shorea balangeran variation at cpDNA Shorea selanica 1 Shorea selanica 2 Shorea xanthophylla 1 Shorea xanthophylla 2 Shorea materialis Shorea macrophylla 1 Shorea macrophylla 2 Shorea splendida 1 Shorea splendida 2 Shorea mecistopteryx 1 Shorea mecistopteryx 2 Shorea mecistopteryx 3 Shorea scaberrima Shorea palembanica 1 Shorea palembanica 2 Shorea leprosula 1 Shorea leprosula 2 Shorea parvifolia 1 Shorea parvifolia 2 Shorea guiso 1 Shorea guiso 2 Shorea platyclados 1 Shorea platyclados 2 Shorea ovalis 1 Shorea ovalis 2 Shorea faguetiana Shorea acuminatissima Shorea javanica Shorea johorensis 1 Shorea johorensis 2 Shorea blumutensis 1 Shorea blumutensis 2 Shorea acuminata 3 Shorea acuminata 1 Shorea acuminata 2 Dryobalanops aromatica Dryobalanops lanceolata 1 Dryobalanops lanceolata 2 Shorea virescens 1 Shorea virescens 2 18 1 (from CAO et al., 2006a) Intraspecific diversity in Indonesian Shorea spp.

Sampling locations

Indonesia

Sumatra SLB (7, 8, 9, 10, 11) Borneo

NS (1, 2, 3, 4, 5, 6)

Java

19 Diversity within Shorea species

Genetic diversity within populations of nine Shorea species in Indonesia at AFLP loci

Pop. ID Distribution Sample size Polymorphic loci PPL na ne He I 1. Spar_NS common 26 38 44.71% 1.447 1.176 0.110 0.174 2. Sacu_NS common 32 42 49.41% 1.494 1.159 0.100 0.162 3. Sdas_NS scattered 20 47 55.29% 1.553 1.273 0.164 0.251 4. Sblu_NS rare 21 53 62.35% 1.624 1.266 0.165 0.257 5. Slep_NS common 16 36 42.35% 1.424 1.224 0.134 0.204 6. Smac_NS common 26 45 52.94% 1.529 1.259 0.155 0.238

Mean 24 44 51.18% 1.512 1.226 0.138 0.214 7. Spar_SLB common 31 44 51.76% 1.518 1.199 0.122 0.193 8. Slep_SLB common 26 39 45.88% 1.459 1.192 0.115 0.178 9. Spal_SLB common 25 52 61.18% 1.612 1.245 0.149 0.232 10. Splat_SLB common 27 56 65.88% 1.659 1.235 0.144 0.230 11. Sjoh_SLB common 24 47 55.29% 1.553 1.183 0.115 0.186

Mean 27 48 56.00% 1.560 1.211 0.129 0.204 Notes: Information of species distribution on islands was obtained from Newman et al. (1996a; 1996b).

PPL, percentage of phenotypically polymorphic loci; na, observed number of alleles per locus; ne, effective number of alleles per locus; ’ He, Nei s (1973) gene diversity; I, Shannon's information index [Lewontin (1972)]. 20 (from Cao et al., 2009) Intraspecific variation

Intraspecific diversity – common, widespread species

Shorea leprosula and S. parvifolia

BB IB

SmB BaB TJS KB TB BkB

TS PS SB MtB WkB NS AS

HJ, DJ

CJ 21 Intraspecific variation

Variation within species

Species N PPL He Ι Ht Hs Gst (%) S. 117 75 0,1973 0,3062 0,1973 0,1362 0,3121 parvifolia

S. 152 94,64 0,2404 0,3788 0,2404 0,1814 0,2355 leprosula

N: sample size PPL. Percentage of polymorphic loci

He: Nei's (1973) gene diversity I: Shannon's Information index

Ht: Total variation HS : Variation within populations GST: Differentiation among populations 22 (Cao et al, 2006b) Intraspecific variation

Genetic differentiation among populations

G st for AFLP markes in Shorea leprosula

0,8

0,6

0,4

0,2

0 1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55

G st for AFLP markers in Shorea parvifolia

1 0,8 0,6 0,4 0,2 0 23 1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 Intraspecific variation

Conversion of AFLP bands to SCAR markers 10 20 30 40 50 60 70 80 90 100 ....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....| SUMATRA 427 GATACAGGGCAGCAATCACATGGATAGACTCAGCAGCTAATGGATAATTTTAGGCTTAAAGGGGGCTATCACATACCAAAAGTTCAGCAAGTAAGGCCGG BORNEO 421 GATACAGGGCAGCAATCACATGGATAGACTCAGCAGCTAATGGATAATTTTAGGCTTAAAGGGGGCTATCACATACCAAAAGTTCAGCAAGTAAGGCCGG BORNEO 418 GATACAGGGCAGCAATCACATGGATAGACTCAGCAGCTAATGGATAATTTTAGGCTTAAAGGGGGCTATCACATACCAAAAGTTCAGCAAGTAAGGCCGG BORNEO 409 GATACAGGGCAGCAATCACATGGATAGACTCAGCAGCTAATGGATAATTTTAGGCTTAAAGGGGGCTATCACATACCAAAAGTTCAGCAAGTAAGGCCGG

110 120 130 140 150 160 170 180 190 200 ....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....| SUMATRA 427 AGCTGGCAGGTTTGGGGGAGAGCCGGCGGGAAGTGCTGGGAAATTTAGGGGAATCTACCGAAAATTGGGWGGAACCAAGAGAAATTAGAGTAAACCCAAG BORNEO 421 AGCTGGCAGGTTTGGGGGAGAGCCGGCGGGAAGTGCTGGGAAATTTAGGGGAATCTACCGAAAATTGGGAGGAACCAAGAGAAATTAGAGTAAACCCAAG BORNEO 418 AGCTGGCAGGTTTGGGGGAGAGCCGGCGGGAAGTGCTGGGAAATTTAGGGGAATCTACCGAAAATTGGGAGGAACCAAGAGAAATTAGAGTAAACCCAAG BORNEO 409 AGCTGGCAGGTTTGGGGGAGAGCCGGCGGGAAGTGCTGGGAAATTTAGGGGAATCTACCGAAAATTGGGAGGAACCAAGAGA------AAACCCAAG

210 220 230 240 250 260 270 280 290 300 ....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....| SUMATRA 427 CTAATTCAAGGAAGAACAAAGGAATTAGTGGCTGTTTCTTACCCAAAAATTTGAAGAATTCACACCCACACAATGAAGAGCAGTRGATAGAAATAGAAGT BORNEO 421 CTAATTCAAGGAAGAACAAAGGAATTAGTGGCTGTTTCTTACCCAAAAATTTGAAGAATTCACACCCACACAATGAAGAGCAGTAGATAGAA------GT BORNEO 418 CTAATTCAAGGAAGAACAAAGGAATTAGTGGCTGTTTCTTACCCAAAAAT---AAGAATTCACACCCACACAATGAAGAGCAGTAGATAGAA------GT BORNEO 409 CTAATTCAAGGAAGAACAAAGGAATTAGTGGCTGTTTCTTACCCAAAAAT---AAGAATTCACACCCACACAATGAAGAGCAGTAGATAGAA------GT EcoRI

310 320 330 340 350 360 370 380 390 400 ....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....|....| SUMATRA 427 AACAGAGCAGAGCAGATCCTGAATTTCTCTGCAGGGGGGAGGTTTCAAGGCTCCAAATCTTGTTCTTGAACAAGAAAAGAAAGGAAAAATCAAGCTCTCT BORNEO 421 AACAGAGCAGAGCAGATCCTGAATTTCTCTGCAGGGGGGAGGTTTCAAGGCTCCAAATCTTGTTCTTGAACAAGAAAAGAAAGGAAAAATCAAGCTCTCT BORNEO 418 AACAGAGCAGAGCAGATCCTGAATTTCTCTGCAGGGGGGAGGTTTCAAGGCTCCAAATCTTGTTCTTGAACAAGAAAAGAAAGGAAAAATCAAGCTCTCT BORNEO 409 AACAGAGCAGAGCAGATCCTGAATTTCTCTGCAGGGGGGAGGTTTCAAGGCTCCAAATCTTGTTCTTGAACAAGAAAAGAAAGGAAAAATCAAGCTCTCT

410 420 430 ....|....|....|....|....|....|. SUMATRA 427 TACCGTTTCTTGAGGGTGGGTGCTGTGTCGA BORNEO 421 TACCGTTTCTTAAGGGTGGGTGCTGTGTCGA BORNEO 418 TACCGTTTCTTAAGGGTGGGTGCTGTGTCGA BORNEO24 409 TACCGTTTCTTAAGGGTGGGTGCTGTGTCGA (Nuroniah et al in prep.) MseI Intraspecific variation

Shorea leprosula: Allele frequency at a co-dominant SCAR marker

1 2 BB 3

1 1

2 2 3 IB 3

1 2 3 BORNEO

1 2 3 TjS KB SmB BkB

1 1 2 2 1 TS 3 3 2 3 4 5 6 7

SB 1 TB 2 3 PS 1 1 2 2 3 3 4 5 NS AS MtB WkB

1

427 418 1 2 2 421/ 421 3 3 427 4 418/ 5 418/ 6 427 421 7 409/ 409/ 8 JAVA 421 418 25 (Nuroniah et al., 2010; Finkeldey et al., 2010) Application: Timber identification

Genetic fingerprints to trace the origin of tropical timber

l Wood from tropical forests is (and will continue to be) traded on international markets – origin l illegal logging l sustainably managed forests – certified forests l How to trace the origin of wood and wood products? – Confirm or reject the origin from particular enterprises, regions, countries,... Only the best is good enough! 26 Application: Timber identification

The chain of custody

Transport Processing

Harvest

End user Further Export & Import 27 processing Application: Timber identification

Material for DNA extraction from wood: Dipterocarps

l N = 382 – 181 from countries of origin l Indonesia l l Philippines l – 151 processed wood l Germany l Indonesia

28 Application: Timber identification

DNA Extraction method

l Based on DNeasy mini kit (Qiagen) with modifications – 50-100 mg of wood shavings 27-a. first eluat 27-a. second eluat – Addition of PVP l RACHMAYANTI et al. (2006, 2009)

29 Results: Extraction

Success of DNA extraction and amplification

l Main impact – Species – Age and processing of wood – Inhibitory substances – Location on stem disk (outer – inner) – Size and genomic origin of PCR fragment

30 Results: Extraction

Impact of fragment size and position on disk

PCR success

1

0.9

0.8 a m i 0.7 0.6

0.5

0.4 first eluat of a. 0.3 second eluat of a. 0.2 first eluat of m. 0.1 second eluat of m. first eluat of i. 0 second eluat of i. Ccmp2 trnL trnLF 0.15 0.6 (150 bp) (0.6 kbp) (1.1 kbp) 1 fragment length (kb)

31 (Rachmayanti et al., unpublished) Results: Extraction

Inhibitory tests: PVP and position on disk

High-quality DNA (from leaves) and diluted or undiluted wood extracts as PCR template

Wood extract dilution (x10 time) inhibitory first eluat rate 1

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

2. 1. 3. 0.1

0 DNA extraction : 1. without PVP addition 0 1 2 4 8 16 32 wood extract ( x10 times dilution) 2. with 2.6% PVP inner heartwood (i) rings transition wood (m) outer sapwood (a) 3. with 5.0% PVP

32 (Rachmayanti et al., unpublished) Results: Extraction

Summary: Success of extraction and amplification

PCR analysis of dipterocarp wood DNA

short fragment (ccmp2, 150 bp; PS,100-200 bp; G3,119-133 bp) middle length fragment (trnL, ca. 600 bp) long fragment (trnLF, ca. 1.1 kb) 1

0.9

0.8

0.7

0.6

0.5

0.4 Amplification success rate success Amplification 0.3

0.2

0.1

0 Vietnam Indonesia Thailand Philippine Enterprises Tectona Eusideroxylon Prunus StrombosiaTriplochiton Populus sp. Costa Rica Taxus Salix spp. Pinus Picea abies (40) (38) (53) (50) (151) grandis (12) zwageri (2) arborea (1)ceylanica (1)scle rox ylon (25) wood (3) baccata (2) (1) sylve stris (12) L (8) (7)

Dipterocarp wood Non-dipterocarp tropic wood non-tropic wood

33 (Rachmayanti et al., unpublished) Conclusion and outlook

Conclusions and outlook I

l Molecular genetic tools are in principle suitable to infer the 27-a. first eluat 27-a. second origin of tropical wood eluat – DNA extraction from wood is feasible l but: limitations (age, inhibitors, processing,...)

Monotes kerstingii trnL region Cotylelobium lanceolatum Upuna borneensis – Anisoptera costata Development of informative Anisoptera marginata 68 99 Anisoptera reticulata Vatica bantamensis Vatica bella 99 Vatica granulata markers is demanding Vatica pauciflora 83 94 Vatica rassak Vatica venulosa Dipterocarpus grandiflorus Dipterocarpus oblongifolius 71 Dipterocarpus retusus l 100 Problems are taxa specific Dipterocarpus rigidus Dipterocarpus tempehes Dryobalanops aromatica 99 Dryobalanops lanceolata Hopea bancana Hopea celebica l Different marker types often Hopea odorata 85 Hopea sangal <50 Hopea dryobalanoides Hopea nigra <50 Hopea griffithii necessary Hopea mengarawan 66 100 Parashorea lucida Parashorea globosa Shorea blumutensis Shorea guiso Shorea montigena – Development of sufficient Shorea scaberrima Shorea seminis Shorea acuminata Shorea andulensis Shorea javanica 67 Shorea johorensis information (data bases) is Shorea leprosula Shorea macroptera 95 69 Shorea mecistopteryx 63 Shorea ovalis Shorea palembanica costly, time consuming Shorea parvifolia Shorea platyclados <50 Shorea xanthophylla Shorea balangeran 59 Shorea selanica Shorea splendida Shorea macrophylla 55 Shorea pinanga <50 Shorea stenoptera Shorea acuminatissima Shorea dasyphylla 77 Shorea faguetiana Shorea multiflora Shorea fallax Shorea materialis 98 34 60 Shorea virescens Conclusion and outlook

Conclusions and outlook II

l The dipterocarps are a suitable model group to further develop and to apply molecular methods to infer the origin of wood – Ecologically relevant keystone species – Conservation need () – Important timber species (national, international) – Some knowledge on molecular genetic variation

35 See also: http://www.uni-goettingen.de/en/82738.html http://aunp.forst.uni-goettingen.de/outputs/teaching_material.shtml

Selected relevant literature from the group

Tsumura, Y., Kado, T., Yoshida, K., Abe, H., Ohtani, M., Taguchi, M., Fukue, Y., Tani, N., Ueno, S., Yoshimura, K., Kamiya, K., Harada, K., Takeuchi, Y., Diway, B., Finkeldey, R., Na’iem, M., Indrioko, S., Ng, K.K.S., Muhammad, N., Lee, S.L. (2011). Molecular database for classifying Shorea species (Dipterocarpaceae) and techniques for checking the legitimacy of timber and wood products. Journal of Plant Research 124: 35-48 Nuroniah, H.S., Gailing, O, Finkeldey, R. (2010). Development of SCAR markers for species identification in the genus Shorea (Dipterocarpaceae). Silvae Genetica 59: 249-256 Finkeldey, R., Leinemann, L., Gailing, O. (2010). Molecular genetic tools to infer the origin of forest and wood. Applied Microbiology and Biotechnology 85: 1251-1258 Open Access review: http://www.springerlink.com/content/u65730127511k770/ Cao, C.P., Gailing, O., Siregar, I.Z., Siregar, U., Finkeldey, R. (2009) Genetic variation in nine Shorea species revealed by AFLPs. Tree Genetics and Genomes 5: 407-420 Rachmayanti, Y., Leinemann, L., Gailing, O., Finkeldey, R. (2009) DNA from processed and unprocessed wood: factors influencing the isolation success. Forensic Science International, Genetics 3: 185-192 Cao, C.P., Finkeldey, R., Siregar, I.Z., Siregar U.J., Gailing, O. (2006). Genetic diversity within and among populations of Shorea leprosula Miq. and Shorea parvifolia Dyer (Dipterocarpaceae) in Indonesia detected by AFLPs. Tree Genetics and Genomes 2: 225-239 Cao, C.P., Gailing, O., Siregar, I., Indrioko, S., Finkeldey, R. (2006). Genetic variation at AFLPs for the Dipterocarpaceae and its relation to molecular phylogenies and taxonomic subdivisions. Journal of Plant Research 119: 553-558 Indrioko, S., Gailing, O., Finkeldey, R. (2006). Molecular phylogeny of Dipterocarpaceae in Indonesia based on chloroplast DNA. Plant Systematics and Evolution 261: 99-115 Rachmayanti, Y., Leinemann, L., Gailing, O., Finkeldey R. (2006). Extraction, 36 amplification and characterization of wood DNA from Dipterocarpaceae. Plant Molecular Biology Reporter 24: 45-55 Acknowledgements

l The main sponsors

l The main partners – Dr. Paciencia Milan, Dr. Tony Quimio, Leyte State University, Philippines – Prof. Le-Cong Kiet, Vietnam Natl. University – Dr, Suchitra Changtragoon, RFD Thailand – Dr. Nyan Htun, UoF, Burma

Thank You for Your attention!

37 Results: Marker development

Intraspecific Variation at cpDNA (cpSSRs and PCR-RFLPs)

Haplotype distribution in Shorea parvifolia 38 (Indrioko, 2005) Results: Marker development

Poor geographic structure at cpDNA markers – Why?

Sea level in SE-Asia approx. 14580 years ago

39 (SATHIAMURTY and VORIS, 2006)