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Many tree planting programmes have long been initiated to increase forest cover to mitigate the effects of global climate change. Successful planting requires careful planning at the project level, including using suitable species with favourable traits. However, there is a paucity of improvement data for tropical tree species. An experimental common garden of Shorea leprosula was established to study traits related to growth performance which are key factors in planting success. Seedlings of S. leprosula were collected from nine geographical forest reserves. To study the effects of genetic variation, seedlings were planted in a common environment following a randomized complete block design. From performance data collected 2017‒2019, one population showed the highest coefficient for relative height growth, significantly higher than most of the other populations. Interestingly, this population from Beserah also exhibited the lowest coefficient for scale insect infestation. This study provides preliminary results on growth performance and susceptibility to scale insect infestation in S. leprosula and the first common garden experiment site conducted on dipterocarp species. It lays a foundation for future genome-wide studies.

Tree diversity in Asia's tropical and subtropical forests is central to nature-based solutions. Species vulnerability to multiple threats, which affect provision of ecosystem services, is poorly understood. We conducted a region-wide, spatially explicit assessment of the vulnerability of 63 socioeconomically important tree species to overexploitation, fire, overgrazing, habitat conversion, and climate change. Trees were selected for assessment from national priority lists, and selections were validated by an expert network representing 20 countries. We used Maxent suitability modeling to predict species distribution ranges, freely accessible spatial data sets to map threat exposures, and functional traits to estimate threat sensitivities. Species-specific vulnerability maps were created as the product of exposure maps and sensitivity estimates. Based on vulnerability to current threats and climate change, we identified priority areas for conservation and restoration. Overall, 74% of the most important areas for conservation of these trees fell outside protected areas, and all species were severely threatened across an average of 47% of their native ranges. The most imminent threats were overexploitation and habitat conversion; populations were severely threatened by these factors in an average of 24% and 16% of their ranges, respectively. Our model predicted limited overall climate change impacts, although some study species were likely to lose over 15% of their habitat by 2050 due to climate change. We pinpointed specific natural areas in Borneo rain forests as hotspots for in situ conservation of forest genetic resources, more than 82% of which fell outside designated protected areas. We also identified degraded areas in Western Ghats, Indochina dry forests, and Sumatran rain forests as hotspots for restoration, where planting or assisted natural regeneration will help conserve these species, and croplands in southern India and Thailand as potentially important agroforestry options. Our results highlight the need for regionally coordinated action for effective conservation and restoration.Especies de Árboles Valoradas y Amenazadas de Asia Tropical y Subtropical Resumen La diversidad de árboles en los bosques tropicales y subtropicales de Asia es un eje central para las soluciones basadas en la naturaleza. La vulnerabilidad de las especies ante las múltiples amenazas, las cuales afectan el suministro de servicios ambientales, es un tema poco comprendido. Realizamos una evaluación regional espacialmente explícita de la vulnerabilidad de 63 especies de árboles de importancia socioeconómica ante la sobreexplotación, incendios, sobrepastoreo, conversión del hábitat y cambio climático. Los árboles se seleccionaron para su evaluación a partir de listas nacionales de prioridades, y las selecciones fueron validadas por una red de expertos de 20 países. Usamos el modelado de idoneidad Maxent para predecir el rango de distribución de las especies, conjuntos de datos espaciales de libre acceso para mapear la exposición a las amenazas y rasgos funcionales para estimar la susceptibilidad a las amenazas. Con base en la vulnerabilidad a las amenazas actuales y al cambio climático, identificamos las áreas prioritarias para su conservación y restauración. En general, el 74% de las áreas más importantes para la conservación de estos árboles quedó fuera de las áreas protegidas y todas las especies estaban seriamente amenazadas en promedio en el 47% de su distribución nativa. Las amenazas más inminentes fueron la sobreexplotación y la conversión del hábitat; las poblaciones estuvieron seriamente amenazadas por estos factores en promedio en el 24% y 16% de su distribución, respectivamente. Nuestro modelo predijo un impacto general limitado del cambio climático, aunque algunas especies estudiadas tuvieron la probabilidad de perder más del 15% de su hábitat para el 2050 debido a este factor. Identificamos áreas naturales específicas en las selvas de Borneo como puntos calientes para la conservación in situ de los recursos genéticos forestales, más del 82% de los cuales estaban fuera de las áreas protegidas designadas. También identificamos áreas degradadas en los Ghats Occidentales, los bosques secos de Indochina y las selvas de Sumatra como puntos calientes para la restauración, en donde la siembra o la regeneración natural asistida ayudarán a conservar estas especies. Además, identificamos campos de cultivo al sur de India y Tailandia como potenciales opciones importantes de agrosilvicultura. Nuestros resultados resaltan la necesidad de acciones regionales coordinadas para la conservación y restauración efectivas.【摘要】保护亚洲热带和亚热带森林的树木多样性是基于自然的解决方案的关注重点。物种面对多种威胁的脆弱性会影响生态系统服务的供给, 但目前人们对此仍知之甚少。我们对63种具有社会经济意义的树种面对过度开发、火灾、过度放牧、栖息地转换和气候变化的脆弱性进行了全区域空间显式评估。我们进行评估的树种来自国家优先保护名录, 树种的选择得到代表 20个国家的专家网络的认可。研究使用Maxent适宜性模型预测了物种分布范围, 并用可获取的空间数据集绘制了威胁暴露情况, 用功能特征估计了威胁敏感性。我们利用暴露地图和敏感性估计结果得到了物种特异的脆弱性地图, 并根据物种对当前威胁和气候变化的脆弱性, 确定了保护和恢复的优先区域。总的来说, 对这些树种的保护最重要的区域有 74%在保护区之外, 所有树种平均在其原生分布区 47%的范围内受到严重威胁。最紧迫的威胁是过度开发和栖息地转换;树木种群平均在其 24%和16%的范围内受到这些因素的严重威胁。我们的模型预测得到的气候变化整体影响有限, 尽管一些物种到2050年可能会因气候变化而失去超过15%的栖息地。我们发现婆罗洲雨林中特定的自然区域应作为森林遗传资源原地保护的热点地区, 该区域82%以上的面积位于已有保护区之外。我们还确定了西高止山、中南半岛旱林和苏门答腊雨林的退化地区为恢复热点地区, 在这些地区种植树木或辅助自然再生将有助于保护树种, 而印度南部和泰国的耕地则是潜在的重要农林地。我们的结果强调了为有效保护和恢复而采取区域性协调行动的必要性。【翻译: 胡怡思;审校: 聂永刚】.

Worldwide, many mangrove species are experiencing significant population declines, including Rhizophora apiculata, which is one of the most widespread and economically important species in tropical Asia. In Malaysia, there has been an alarming decline in R. apiculata populations driven primarily by anthropogenic activities. However, the lack of genetic and demographic information on this species has hampered local efforts to conserve it. To address these gaps, we generated novel genetic information for R. apiculata, based on 1,120 samples collected from 39 natural populations in Peninsular Malaysia. We investigated its genetic diversity and genetic structure with 19 transcriptome and three nuclear microsatellite markers. Our analyses revealed a low genetic diversity (mean He: 0.352) with significant genetic differentiation (FST: 0.315) among populations of R. apiculata. Approximately two-third of the populations showed significant excess of homozygotes, indicating persistent inbreeding which might be due to the decrease in population size or fragmentation. From the cluster analyses, the populations investigated were divided into two distinct clusters, comprising the west and east coasts of Peninsular Malaysia. The western cluster was further divided into two sub-clusters with one of the sub-clusters showing strong admixture pattern that harbours high levels of genetic diversity, thus deserving high priority for conservation.

Southeast Asian rainforests at upper hill elevations are increasingly vulnerable to degradation because most lowland forest areas have been converted to different land uses. As such, understanding the genetics of upper hill species is becoming more crucial for their future management and conservation. Shorea platyclados is an important, widespread upper hill dipterocarp in Malaysia. To elucidate the genetic structure of S. platyclados and ultimately provide guidelines for a conservation strategy for this species, we carried out a comprehensive study of the genetic diversity and demographic history of S. platyclados. Twenty-seven populations of S. platyclados across its range in Malaysia were genotyped at 15 polymorphic microsatellite loci and sequenced at seven noncoding chloroplast DNA (cpDNA) regions. A total of 303 alleles were derived from the microsatellite loci, and 29 haplotypes were identified based on 2892 bp of concatenated cpDNA sequences. The populations showed moderately high genetic diversity (mean HE = 0.680 for microsatellite gene diversity and HT = 0.650 for total haplotype diversity) and low genetic differentiation (FST = 0.060). Bayesian clustering divided the studied populations into two groups corresponding to western and eastern Malaysia. Bottleneck analysis did not detect any recent bottleneck events. Extended Bayesian skyline analyses showed a model of constant size for the past population history of this species. Based on our findings, priority areas for in situ and ex situ conservation and a minimum population size are recommended for the sustainable utilization of S. platyclados.

Ensuring the authenticity of raw materials used in herbal manufacturing is a key step prior to material processing. As species authentication is fundamental in the confirmation of herbal product quality, DNA barcoding techniques represent an efficient method for detecting plant-based adulterants in traded herbal products. Through this study, we established a DNA barcoding authentication system for 112 common herbal plant species in the tropics, which can be used for species identification and authentication. The DNA barcode reference database for the authentication system was generated using rbcL for primary differentiation, and trnH-psbA for secondary differentiation. The performance of the barcodes in resolving species was evaluated using similarity BLAST, phylogenetic tree reconstruction and by estimating the barcoding gap. In this study, the multigene tiered approach for DNA barcoding is proven robust with high species-level resolution (96.4%). Upon completion of the DNA barcoding authentication system, 30 herbal products from the local market were tested for their authenticity using this approach. Recovery of DNA barcodes from the herbal products was 73.4%, of which 56.7% of the products tested were authentic, whereas 10% of the herbal products were substituted with other plant taxa and 6.7% were contaminated. To this end, authentication of herbal products is challenging, but with the establishment of a new DNA barcoding authentication system for common herbal plants in the tropics, the existing quality assurance and adulteration screening programs employed by regulatory agencies could be significantly strengthened.

Our study highlights the utilization of a genetic database for wood-origin identification in Intsia palembanica, a valuable heavy hardwood from the tropical forests. This forensic tool is essential for strengthening the verification of legality in the wood supply chain from the forest to the end-users. An increasing number of rules and regulations are being put in place to promote sustainable practice in the timber trade, one of which involves ensuring that importers declare the correct species name and source of geographic origin of the timber. We aimed to determine the origin of the I. palembanica seed source used in the early establishment on the Forest Research Institute Malaysia (FRIM) campus. DNA samples of I. palembanica individuals from the FRIM campus were obtained and analyzed using four chloroplast (cp) DNA markers to characterize the haplotype variants for population identification. In addition, the DNA samples were also genotyped at 14 short tandem repeat (STR) loci for individual identification. Individual assignment to the possible geographic origin was done through an assignment test. On the basis of our recently developed I. palembanica genetic databases, the I. palembanica seed source for the early establishment was inferred to be originated from a mixture of several sources, with a large portion from the southern region (89%) and a relatively small portion from the northern region (11%) of Peninsular Malaysia. The I. palembanica seed source used for the early establishment on the century old FRIM campus was inferred to be originated from several forest reserves located not far from the planting sites. This study proves the applicability of the DNA method in supply-chain verification, where an unknown I. palembanica tree can be traced to its geographic origin using genetic databases.

Aquilaria malaccensis (Thymelaeaceae) is the main source of high-grade agarwood in Southeast Asia. Aggressive collections and trade activities over the past decades have put great pressure on the natural stands and raised concerns over the long-term survival potential of A. malaccensis. Tracking and authentication of agarwood require method with a high degree of accuracy. Therefore, this study aimed to develop DNA databases of A. malaccensis as the tracking tools at species, population and individual levels for forensic identification and chain of custody certification. Using two cpDNA (rbcL and matK) and an rDNA (ITS2) markers, species identification database of Aquilaria was developed to distinguish A. malaccensis from A. hirta, A. microcarpa, A. beccariana, A. crassna, A. sinensis and A. rostrata. In addition, based on 35 populations of A. malaccensis throughout Peninsular Malaysia, cpDNA haplotype and STR allele frequency databases were developed for population and individual identification. A haplotype distribution map based on 29 haplotypes derived from seven cpDNA showed that the A. malaccensis in Peninsular Malaysia can be associated to Kedah-Perak and Kelantan-Johor regions. Similarly, genetic relatedness and Bayesian clustering analyses based on 10 STR markers also divided the 35 populations into two main genetic clusters, corresponding to Kedah-Perak and Kelantan-Johor regions. The STR allele frequency databases were established and characterized according to these two regions. To determine the performance of the STR allele frequency databases for population identification, independent self-assignment tests showed that the percentage of individuals correctly assigned into the origin population was 93.88% in Kedah-Perak and 90.29% in Kelantan-Johor. For the STR allele frequency databases to be used for individual identification, conservativeness tests showed that the θ should be adjusted to 0.250 and 0.200 in the Kedah-Perak and Kelantan-Johor databases, respectively. To ensure consistency in allele calling for the dinucleotide repeat loci across different electrophoretic platforms or laboratories, allelic ladders have been developed for the 10 STR loci. Two case studies are presented of how these databases were used to track A. malaccensis to the origin population and stump. These databases are ready to be used to provide admissible forensic evidence for legal proceedings against the illegal harvesters of agarwood and for agarwood certification to meet the consumer country regulations.

Background: Genetic divergence is one of the key processes in speciation. In the Begoniaceae, genetic divergence caused by limited gene flow may explain its high species diversity and endemicity. T...

To inform product users about the origin of timber, the implementation of a traceability system is necessary for the forestry industry. In this study, we developed a comprehensive genetic database for the important tropical timber species Merbau, Intsia palembanica, to trace its geographic origin within peninsular Malaysia. A total of 1373 individual trees representing 39 geographically distinct populations of I. palembanica were sampled throughout peninsular Malaysia. We analyzed the samples using a combination of four chloroplast DNA (cpDNA) markers and 14 short tandem repeat (STR) markers to establish both cpDNA haplotype and STR allele frequency databases. A haplotype map was generated through cpDNA sequencing for population identification, resulting in six unique haplotypes based on 10 informative intraspecifically variable sites. Subsequently, an STR allele frequency database was developed from 14 STRs allowing individual identification. Bayesian cluster analysis divided the individuals into two genetic clusters corresponding to the northern and southern regions of peninsular Malaysia. Tests of conservativeness showed that the databases were conservative after the adjustment of the θ values to 0.2000 and 0.2900 for the northern (f = 0.0163) and southern (f = 0.0285) regions, respectively. Using self-assignment tests, we observed that individuals were correctly assigned to populations at rates of 40.54−94.12% and to the identified regions at rates of 79.80−80.62%. Both the cpDNA and STR markers appear to be useful for tracking Merbau timber originating from peninsular Malaysia. The use of these forensic tools in addition to the existing paper-based timber tracking system will help to verify the legality of the origin of I. palembanica and to combat illegal logging issues associated with the species.

Baeckea frutescens or Cucur Atap, is a medicinal plant that has been traditionally used as herbal medicine and has great potential for pharmaceutical application due to its biological properties. The increasing demand for B. frutescens herbal medicinal products has led to its over-exploitation. Therefore, it is crucial to generate baseline genetic information in order to formulate conservation and breeding programmes for this species. In this study, a total of 26 polymorphic simple sequence repeat (SSR) markers have been developed in B. frutescens based on the leaf transcriptome sequence data obtained via next generation sequencing (NGS) technology. These markers were evaluated using 32 individuals from Forest Research Institute Malaysia Research Station in Setiu, Terengganu. The mean number of alleles was two and the average expected (HE) and observed heterozygosity (HO) were 0.484 and 0.900, respectively. Cluster analysis revealed high genetic similarity among the individuals, whereby many of them were of the same clones. It is essential to assess more individuals from other populations for better understanding on the genetic diversity of this useful medicinal plant.

Background: Dipterocarpaceae is a pantropical tree family that plays an important role in our understanding of the ecology of Asian tropical rain forests. However, genome sizes for members of the Dipterocarpaceae are still poorly known.Aims: To report the genome size of 115 dipterocarp species and examine the variation and evolution of genome size in this family.Methods: Genome size was estimated using flow cytometry. Both the rpoB and trnL intron were sequenced to uncover the evolution of genome size within a phylogenetic framework.Results: The 1C genome size varied between 0.267 and 0.705 pg in Shorea hemsleyana and Shorea ovalis, respectively, a 2.64-fold variation across the family. Most dipterocarps are characterised by very small genomes with a mean 1C value of 0.416 pg (sd = 0.075) and five polyploids are recorded. The ancestral genome size for dipterocarps was reconstructed as 1Cx = 0.481 pg (95% CI = 0.433–0.534).Conclusions: Genome size variation in dipterocarps was characterised by very small v...

Taxonomic classification of the important medicinal plant, Ficus deltoidea (Moraceae), is challenging because of the variability of its leaves and fig forms that occur within the species. We developed 16 nuclear Simple Sequence Repeat (nSSR) markers, and characterized them using 24 individuals from a natural population. We then studied the intraspecific variation of F . deltoidea subsp. deltoidea in Peninsular Malaysia using morphological and molecular approaches. Based on the morphological variations, we further determined the varieties that occur regionally under the above subspecies based on the leaf characteristics. As for molecular data sets, we used both chloroplast DNA (cpDNA) and nSSR markers to elucidate the phylogenetic relationship among the varieties. The cpDNA dendrogram yielded poorer resolution where most of the clades were forming paraphyletic complex. The cluster analysis based on nSSR is largely congruent with the morphological classification, with F . deltoidea subsp. deltoidea classified into four main varieties, namely var. deltoidea , var. angustifolia , var. kunstleri and var. lutescens . Our study demonstrates the applicability of molecular approach in complementing the conventional taxonomic classification.

Illegal logging and smuggling of Gonystylus bancanus (Thymelaeaceae) poses a serious threat to this fragile valuable peat swamp timber species. Using G. bancanus as a case study, DNA markers were used to develop identification databases at the species, population and individual level. The species level database for Gonystylus comprised of an rDNA (ITS2) and two cpDNA ( trnH-psbA and trnL ) markers based on a 20 Gonystylus species database. When concatenated, taxonomic species recognition was achieved with a resolution of 90% (18 out of the 20 species). In addition, based on 17 natural populations of G. bancanus throughout West (Peninsular Malaysia) and East (Sabah and Sarawak) Malaysia, population and individual identification databases were developed using cpDNA and STR markers respectively. A haplotype distribution map for Malaysia was generated using six cpDNA markers, resulting in 12 unique multilocus haplotypes, from 24 informative intraspecific variable sites. These unique haplotypes suggest a clear genetic structuring of West and East regions. A simulation procedure based on the composition of the samples was used to test whether a suspected sample conformed to a given regional origin. Overall, the observed type I and II errors of the databases showed good concordance with the predicted 5% threshold which indicates that the databases were useful in revealing provenance and establishing conformity of samples from West and East Malaysia. Sixteen STRs were used to develop the DNA profiling databases for individual identification. Bayesian clustering analyses divided the 17 populations into two main genetic clusters, corresponding to the regions of West and East Malaysia. Population substructuring ( K =2) was observed within each region. After removal of bias resulting from sampling effects and population subdivision, conservativeness tests showed that the West and East Malaysia databases were conservative. This suggests that both databases can be used independently for random match probability estimation within respective regions. The reliability of the databases was further determined by independent self-assignment tests based on the likelihood of each individual's multilocus genotype occurring in each identified population, genetic cluster and region with an average percentage of correctly assigned individuals of 54.80%, 99.60% and 100% respectively. Thus, after appropriate validation, the genetic identification databases developed for G. bancanus in this study could support forensic applications and help safeguard this valuable species into the future.

The development of timber tracking methods based on genetic markers can provide scientific evidence to verify the origin of timber products and fulfill the growing requirement for sustainable forestry practices. In this study, the origin of an important Dark Red Meranti wood, Shorea platyclados, was studied by using the combination of seven chloroplast DNA and 15 short tandem repeats (STRs) markers. A total of 27 natural populations of S. platyclados were sampled throughout Malaysia to establish population level and individual level identification databases. A haplotype map was generated from chloroplast DNA sequencing for population identification, resulting in 29 multilocus haplotypes, based on 39 informative intraspecific variable sites. Subsequently, a DNA profiling database was developed from 15 STRs allowing for individual identification in Malaysia. Cluster analysis divided the 27 populations into two genetic clusters, corresponding to the region of Eastern and Western Malaysia. The conservativeness tests showed that the Malaysia database is conservative after removal of bias from population subdivision and sampling effects. Independent self-assignment tests correctly assigned individuals to the database in an overall 60.60−94.95% of cases for identified populations, and in 98.99−99.23% of cases for identified regions. Both the chloroplast DNA database and the STRs appear to be useful for tracking timber originating in Malaysia. Hence, this DNA-based method could serve as an effective addition tool to the existing forensic timber identification system for ensuring the sustainably management of this species into the future.

Begonias are hyper-diverse and important horticultural plants. Six polymorphic microsatellite markers were developed from CT- and GT-enriched libraries of Begonia maxwelliana . The number of alleles per locus ranged from 2 to 12 and the observed heterozygosity ranged from 0.036 to 0.813. Null alleles were detected in one locus ( Bma 161) after Bonferroni correction. All the six markers were amplifiable in 23 selected Begonia species with the success rates of 17–100%. On average, species of the same section as B. maxwelliana (i.e. sect. Platycentrum ) yielded higher transferability (91%). These markers will be useful for population genetic studies of the genus Begonia .

Orthosiphon stamineus (Lamiaceae) is an important medicinal plant in Malaysia that has great potential in the herbal medicine industry. In this study, we developed and characterized ten microsatellites for O. stamineus. The microsatellite loci were checked for variability using 28 individuals of O. stamineus sampled from 10 nurseries. The number of alleles ranged from two to four, and the observed heterozygosity ranged from 0.143 to 0.963. Neighbor-joining analysis revealed two main genetic clusters corresponding to the two varieties (white and purple). The application of these microsatellites will facilitate the cultivar/variety identification and germplasm management in the species.

Hopea bilitonensis is an extremely rare and predominantly selfing dipterocarp in Peninsular Malaysia. A comprehensive research was initiated to assess the ecological genetics of H. bilitonensis to elucidate specific ecological and genetic requirements and subsequently to recommend conservation strategies. The objective for conservation of a rare plant such as H. bilitonensis differs from that of a common plant. For common plants, the conservation strategies are to prevent the species from becoming endangered. In contrast, for rare plants, the final race against extinction is being fought. Tropical forests are rich in plant species diversity and obtaining adequate knowledge to set conservation strategies for the majority of these species might be difficult. Thus, it is suggested that for the conservation of tree species, the species can be grouped according to their life history traits. The information generated for a species can then be adapted to species that have similar types of life history traits. We have recently generated the ecological genetics information for a rare and predominantly outcrossed dipterocarp (Shorea lumutensis). This study on H. bilitonensis will provide ecological genetics information for the conservation of rare and predominantly selfing dipterocarps.

Termites from the genus Odontotermes are known to contain numerous species complexes that are difficult to tell apart morphologically or with mitochondrial DNA sequences. We developed markers for one such cryptic species complex, that is, Odontotermes srinakarinensis sp. nov. from Maxwell Hill Forest Reserve (Perak, Malaysia), and characterised them using a sample of 41 termite workers from three voucher samples from the same area. We then genotyped 150 termite individuals from 23 voucher samples/colonies of this species complex from several sites in Peninsular Malaysia. We analysed their population by constructing dendograms from the proportion of shared-alleles between individuals and genetic distances between colonies; additionally, we examined the Bayesian clustering pattern of their genotype data. All methods of analysis indicated that there were two distinct clusters within our data set. After the morphologies of specimens from each cluster were reexamined, we were able to separate the two species morphologically and found that a single diagnostic character found on the mandibles of its soldiers could be used to separate the two species quite accurately. The additional species in the clade was identified as Odontotermes denticulatus after it was matched to type specimens at the NHM London and Cambridge Museum of Zoology.

Full-sib families are produced from artificially controlled pollination in tree breeding. This mating design is, however, very costly and logistically difficult as most tree species are large in size. With the development of molecular markers, known relationships among the offspring could be established from the maximum likelihood in paternity analysis. In this study, we successfully established full-sib families on the basis of paternity analysis of offspring derived from two Shorea platyclados seed parents from the natural forest. A total of 2,049 and 970 offspring were collected from two mother trees M01 and M06, respectively. Potential pollen donors surrounding the mother trees were sampled, specifically a total of 77 adult trees within the 10-ha plot for M01 and 28 adult trees within the 4-ha plot for M06. For mother tree M01, parentage was assigned to 45.4% of the offspring at the 95% confidence level with 59 full-sib families established. For mother tree M06, parentage was assigned to 19.9% of the offspring at the 95% confidence level with 19 full-sib families established. The establishment of full-sib families in S. platyclados enables breeding in dipterocarp tree species, once a decade-long proposition, to become more efficient in a cost-saving manner.

Tectonic movements, climatic oscillations, and marine transgressions during the Cenozoic have had a dramatic effect on the biota of the tropical rain forest. This study aims to reveal the phylogeography and evolutionary history of a Peninsular Malaysian endemic tropical timber species, Neobalanocarpus heimii (Dipterocarpaceae). A total of 32 natural populations of N. heimii, with 8 samples from each population were investigated. Fifteen haplotypes were identified from five noncoding chloroplast DNA (cpDNA) regions. Overall, two major genealogical cpDNA lineages of N. heimii were elucidated: a widespread southern and a northern region. The species is predicted to have survived in multiple refugia during climatic oscillations: the northwestern region (R1), the northeastern region (R2), and the southern region (R3). These putative glacial refugia exhibited higher levels of genetic diversity, population differentiation, and the presence of unique haplotypes. Recolonization of refugia R1 and R2 could have first expanded into the northern region and migrated both northeastwards and northwestwards. Meanwhile, recolonization of N. heimii throughout the southern region could have commenced from refugia R3 and migrated toward the northeast and northwest, respectively. The populations of Tersang, Pasir Raja, and Rotan Tunggal exhibited remarkably high haplotype diversity, which could have been the contact zones that have received an admixture of gene pools from the northerly and also southerly regions. As a whole, the populations of N. heimii derived from glacial refugia and contact zones should be considered in the conservation strategies in order to safeguard the long-term survival of the species.

This paper reports 10 polymorphic microsatellite markers for a valuable peat swamp timber species, Gonystylus bancanus (Thymelaeaceae). The primers were designed from a genomic library enriched with dinucleotide (CT) repeats and screened on 24 samples from a natural population. The number of alleles detected per locus ranged from two to nine while the observed and expected heterozygosities ranged from 0.455 to 0.864 and from 0.476 to 0.863, respectively. No significant deviation from Hardy–Weinberg equilibrium (P

Simple sequence repeats (SSRs) derived from expressed sequence tags (ESTs) are valuable markers because they represent transcribed regions and often transferable to related taxa. Here, we report the development and characterization of EST-SSRs from Shorea leprosula. Fifty-four sequences containing SSRs were identified in 2003 unigenes assembled from 3159 ESTs. Twenty-four EST-SSRs were developed, of which four gave multiple amplifications, five were found to be monomorphic and 15 showed polymorphism, with allele numbers ranging from two to 17 in a single Pasoh Forest Reserve population of 24 individuals. The observed and expected heterozygosities ranged from 0.05 to 0.91 and from 0.16 to 0.93, respectively. Cross-species transferability of the 15 loci to 36 species within Dipterocarpaceae revealed between four and 14 loci that gave positive amplification and 10 loci were found to be transferable to more than 15 species.

The maintenance of mixed mating was studied in Shorea curtisii, a dominant and widely distributed dipterocarp species in Southeast Asia. Paternity and hierarchical Bayesian analyses were used to estimate the parameters of pollen dispersal kernel, male fecundity and self-pollen affinity. We hypothesized that partial self incompatibility and/or inbreeding depression reduce the number of selfed seeds if the mother trees receive sufficient pollen, whereas reproductive assurance increases the numbers of selfed seeds under low amounts of pollen. Comparison of estimated parameters of self-pollen affinity between high density undisturbed and low density selectively logged forests indicated that self-pollen was selectively excluded from mating in the former, probably due to partial self incompatibility or inbreeding depression until seed maturation. By estimating the self-pollen affinity of each mother tree in both forests, mother trees with higher amount of self-pollen indicated significance of self-pollen affinity with negative estimated value. The exclusion of self-fertilization and/or inbreeding depression during seed maturation occurred in the mother trees with large female fecundity, whereas reproductive assurance increased self-fertilization in the mother trees with lower female fecundity.

This paper reports the isolation and characterization of 24 polymorphic microsatellite markers in an important tropical timber species, Koompassia malaccensis (Leguminosae). The primers were designed from a genomic library enriched for dinucleotide (CT) repeats and screened on 24 samples from a natural population. The number of alleles detected per locus ranged from two to 13 while the observed heterozygosity ranged from 0.042 to 1.000. Significant departure from Hardy–Weinberg equilibrium (P

To conserve a rare plant, conservation programs must be guided by the biological attributes of the species. What constitutes the most critical biological information for plant conservation has been the issue of discussion for the last 30 years. Most scientists promote an approach that is either ecological or genetic in emphasis. Ecological and genetic processes will often interact synergistically to influence the population viability and to determine the persistence of populations in the long run. Consequently, conservation management of plant species, in addition to ecological information, requires a robust understanding of underlying genetic processes as well as the variation within and between populations. Conservation has a cost and the resources available for conservation programs are always limited. Therefore, conservation management strategies should not only be scientifically justified but also practical in terms of resource availability. Shorea lumutensis is a rare and endemic dipterocarp in Peninsular Malaysia. A comprehensive research activity was initiated to assess the population ecology and population genetics of S. lumutensis to elucidate specific ecological and genetic requirements and subsequently to set conservation strategies and priorities. This paper is apparently the first attempt at applying both the ecological and genetic approaches into conservation management of a rare dipterocarp. In addition, this paper also attempts to link the gaps between conservation research and conservation management in a realistic manner. It is our hope that this study will serve as a model for the study of other rare dipterocarps which should be given priority for conservation.

Intsia palembanica is a high-value tropical hardwood species of the family Leguminosae. A total of 16 microsatellite markers were developed for this important timber species using an enrichment approach. These markers were screened on 24 samples from a natural population and subsequently tested for reproducibility, ease of scoring and locus-specificity, across five populations. The observed and expected heterozygosities ranged from 0.026 to 0.976 and 0.026 to 0.928, respectively, while the mean number of alleles per locus per population was 8.73. These microsatellite markers are informative and will be useful for conservation genetic study, individual identification and timber tracking.

Premise of the study Aggressive collections and trade activities in recent decades have resulted in heavy pressure on the natural stands of Aquilaria malaccensis and concerns over its long-term survival potential. To aid DNA profiling and assessment of its genetic diversity, microsatellite markers were developed for the species. Methods and results Seventeen polymorphic microsatellite markers were developed for A. malaccensis using an enrichment protocol. The markers were screened on 24 samples from a natural population. The number of alleles ranged from two to 11, and the observed heterozygosity ranged from 0.042 to 0.957. No significant deviation from Hardy-Weinberg equilibrium was detected after conservative Bonferroni correction. Conclusions This is the first report on the development of microsatellite markers in A. malaccensis. The markers will be used to establish a DNA profiling database and to estimate the genetic diversity and population genetic structure of the species.

We examined the effect of selective logging on the genetic diversity of Scaphium macropodum using RAPD markers via two approaches: (1) to investigate the immediate effect by studying a same population before and after logging, and (2) to determine the long term effect by comparing two regenerated stands with an adjacent unlogged stand, assuming that they were genetically identical before logging. Results showed no negative immediate impact for the first approach, probably due to the high abundance and heterogeneity of S. macropodum in the compartment investigated. However, for the latter approach, substantial genetic erosion (i.e. 31.5% reduction for Shannon diversity, H) was detected in one of the regenerated stands corresponding to its extremely low tree density for S. macropodum. This implies the possible occurrence of genetic drift and increased inbreeding due to population decline as a result of logging. However, the observed genetic differences among the three sub-populations having prevailed before logging cannot be totally discounted in the second approach. This study also demonstrates the use of tree density as a good surrogate measure of genetic diversity. The present harvesting system in Malaysia based on a general cutting limit need to be refined; the basis for determining cutting limit in a forest management unit should consider abundance of commercial species.

To evaluate the effect of flowering tree density on genetic variation in a dominant tree species, genetic diversities and outcrossing rates of Shorea curtisii (Dipterocarpaceae) were compared between undisturbed and selectively logged forests using microsatellite markers. The mean heterozygosity and the effective number of alleles per locus in the selectively logged plot were not significantly different from those of the undisturbed plot. However, the average number of alleles derived from the pollen donor was significantly lower in the selectively logged forests, indicating the possibility of a loss of genetic variation in future generations. The average outcrossing rate for 11 trees in an undisturbed plot was 96.3%, which was similar to other reports from dipterocarp species based on isozyme analysis. However, the outcrossing rate in the selectively logged plot was 52.2% for five trees, which was significantly lower than that in the undisturbed plot. This decline of outcrossing rate in the selectively logged plot may result from the much lower density of conspecific flowering trees, and the high rate of selfing may result in inbreeding depression and/or a decrease in genetic variation of this forest in future generations.

PREMISE OF THE STUDY Microsatellite markers were developed for Johannesteijsmannia lanceolata to assess the genetic diversity and mating system of this alarmingly endangered species. METHODS AND RESULTS A total of 31 polymorphic microsatellite markers were developed for J. lanceolata using the enrichment protocol. These markers were screened on 24 samples from a natural population. The number of alleles ranged from four to 20, while the observed heterozygosity ranged from 0.391 to 1.000. The 31 loci were further tested for transferability on J. altifrons, J. magnifica, and J. perakensis. Generally, all loci showed positive amplifications in these three Johannesteijsmannia species, except Jla124 (J. magnifica) and Jla168b (J. magnifica and J. perakensis). CONCLUSIONS These microsatellite markers could be employed to study the population genetics and mating system of J. lanceolata and other Johannesteijsmannia species.

 Premise of the study : Microsatellite markers of an important medicinal plant, Eurycoma longifolia (Simaroubaceae), were developed for DNA profi ling and genetic diversity studies.  Methods and Results : Eighteen polymorphic microsatellite loci were developed for E. longifolia . The primers were designed from a genomic library enriched for dinucleotide (CT) repeats and screened on 32 samples from a natural population. The number of alleles detected per locus ranged from four to 16, while the observed heterozygosity ranged from 0.097 to 0.938. No signifi cant deviation from Hardy – Weinberg equilibrium was detected in all the 18 loci, and no linkage disequilibrium was found between these loci after conservative Bonferroni correction.  Conclusions : The 18 microsatellite markers of E. longifolia are highly polymorphic and informative. These markers would serve as an important tool for DNA profi ling and genetic diversity studies.

Pollen flow and population genetic structure among 30 potentially flowering individuals of Neobalanocarpus heimii , a tropical emergent tree, were investigated in a lowland tropical rainforest of Malaysia using microsatellite polymorphism. The 248 offspring in the vicinity of five reproductive trees of the 30 potentially flowering trees were used in paternity analysis for pollen-flow study. Four primer pairs, developed in different species of dipterocarps, were adopted to detect microsatellite polymorphism. Based upon microsatellite polymorphism, pollen flow and seed migration were detected. Pollen-flow events of more than 400 m were observed directly, based on paternity analysis in the study plot. The estimated average mating distance of the five reproductive trees was 524 m. This result suggests that reproduction of this species is mediated by a long-distance pollinator. The haplotypes of some offspring were not compatible with the nearest reproductive tree. Thus, the results suggest that some seeds are dispersed by a seed dispersal vector. Investigation of genetic structure showed significant and negative correlation of genetic relatedness and spatial distances between the 30 potentially flowering trees, but this correlation was weak. We suggest that long-distance gene flow and seed migration are responsible for the poorly developed genetic structure of this species.

Nine polymorphic microsatellite markers were isolated and characterized in Shorea platyclados (Dipterocarpaceae) using CT-enriched library method. The polymorphism of the microsatellite loci was analyzed by screening 98 samples from a natural population. The number of alleles per locus ranged from 4 to 12, with an average of 8.22 per locus, and the observed heterozygosity ranged from 0.204 to 0.888 (average 0.562). These microsatellite loci will be useful for population genetic studies on S. platyclados and adding genetic information for hill dipterocarps.

Premise of the study: Microsatellite markers were isolated and characterized in Shorea platyclados (Dipterocarpaceae) for DNA profiling and genetic diversity assessment of this tropical timber species. Methods and Results: Fifteen polymorphic microsatellite loci were developed and characterized in S. platyclados using a genomic library enriched for dinucleotide (CT) repeats. The primers amplified dinucleotide repeats with 3–14 alleles per locus across four natural populations. The observed and expected heterozygosities ranged from 0.292 to 1.000 and from 0.301 to 0.894, respectively. No significant deviation from Hardy–Weinberg equilibrium was detected in the 15 loci. Four loci pairs displayed linkage disequilibrium. Conclusions: These highly polymorphic markers are adequate for DNA profiling and studies of population genetics in S. platyclados.

Premise of the study: The exploitation of Labisia pumila for commercial demand is gradually increasing. It is therefore important that conservation is prioritized to ensure sustainable utilization. We developed microsatellites for L. pumila var. alata and evaluated their polymorphism across var. alata, var. pumila, and var. lanceolata. Methods and Results: Ten polymorphic microsatellites of L. pumila were developed using the magnetic bead hybridization selection approach. A total of 84, 48, and 66 alleles were observed in L. pumila var. alata, var. pumila, and var. lanceolata, respectively. The species is likely a tetraploid, with the majority of the loci exhibiting up to four alleles per individual. Conclusions: This is the first report on the development of microsatellites in L. pumila. The microsatellites will provide a good basis for investigating the population genetics of the species and will serve as a useful tool for DNA profiling.

Sustained management of natural forests depends on their ability to regenerate. Successful regeneration of forests accompanied by conservation of genetic diversity of its species is important for both short-term adaptation to environmental change and long-term impact on species and communities (Templeton 1995).