Functional anatomy of five endangered tropical timber wood species of the family Dipterocarpaceae

Wood anatomy of five dipterocarp species endemic to the Philippines was studied with the goal to explore functional wood traits of ecological significance. Stem wood of 6-year-old trees grown under similar environmental conditions in a plantation (Leyte, Philippines) was used. Wood densities decreased in the following order Hopea plagata > Dipterocarpus kerrii > Parashorea malaanoman > Shorea almon ≈ Shorea contorta. This was mainly caused by significantly thicker fiber cell walls of H. plagata and D. kerrii than those of the other three species. Wood density was negatively correlated with the abundance of axial parenchyma cells. Predicted conductance was independent from wood density and lowest in H. plataga and highest in D. kerrii and S. contorta. These results indicate that H. plagata and D. kerrii is woods have higher constructions costs in term of carbon per unit of biomass, and that H. plagata is probably better acclimated to varying soil moisture than the other species.     

Molecular database for classifying Shorea species (Dipterocarpaceae) and techniques for checking the legitimacy of timber and wood products

The extent of tropical forest has been declining, due to over-exploitation and illegal logging activities. Large quantities of unlawfully extracted timber and other wood products have been exported, mainly to developed countries. As part of the export monitoring effort, we have developed methods for extracting and analyzing DNA from wood products, such as veneers and sawn timbers made from dipterocarps, in order to identify the species from which they originated. We have also developed a chloroplast DNA database for classifying Shorea species, which are both ecologically and commercially important canopy tree species in the forests of Southeast Asia. We are able to determine the candidate species of wood samples, based on DNA sequences and anatomical data. The methods for analyzing DNA from dipterocarp wood products may have strong deterrent effects on international trade of illegitimate dipterocarp products. However, the method for analyzing DNA from wood is not perfect for all wood products and need for more improvement, especially for plywood sample. Consequently, there may be benefits for the conservation of tropical forests in Southeast Asia.     

Population Genetic Structure and Reproductive Strategy of the Introduced Grass Centotheca lappacea in Tropical Land-Use Systems in Sumatra

Intensive transformation of lowland rainforest into oil palm and rubber monocultures is the most common land-use practice in Sumatra (Indonesia), accompanied by invasion of weeds. In the Jambi province, Centotheca lappacea is one of the most abundant alien grass species in plantations and in jungle rubber (an extensively used agroforest), but largely missing in natural rainforests. Here, we investigated putative genetic differentiation and signatures for adaptation in the introduced area. We studied reproductive mode and ploidy level as putative factors for invasiveness of the species. We sampled 19 populations in oil palm and rubber monocultures and in jungle rubber in two regions (Bukit Duabelas and Harapan). Amplified fragment length polymorphisms (AFLP) revealed a high diversity of individual genotypes and only a weak differentiation among populations (F_ST = 0.173) and between the two regions (F_ST = 0.065). There was no significant genetic differentiation between the three land-use systems. The metapopulation of C. lappacea consists of five genetic partitions with high levels of admixture; all partitions appeared in both regions, but with different proportions. Within the Bukit Duabelas region we observed significant isolation-by-distance. Nine AFLP loci (5.3% of all loci) were under natural diversifying selection. All studied populations of C. lappacea were diploid, outcrossing and self-incompatible, without any hints of apomixis. The estimated residence time of c. 100 years coincides with the onset of rubber and oil palm planting in Sumatra. In the colonization process, the species is already in a phase of establishment, which may be enhanced by efficient selection acting on a highly diverse gene pool. In the land-use systems, seed dispersal might be enhanced by adhesive spikelets. At present, the abundance of established populations in intensively managed land-use systems might provide opportunities for rapid dispersal of C. lappacea across rural landscapes in Sumatra, while the invasion potential in rainforest ecosystems appears to be moderate as long as they remain undisturbed.     

Chloroplast DNA haplotype diversity and postglacial recolonization of Hagenia abyssinica (Bruce) J.F. Gmel. in Ethiopia

We investigated genetic variation of 273 individuals from 25 populations of the monotypic species Hagenia abyssinica (Rosaceae) from the highlands of Ethiopia at three chloroplast microsatellite loci. The objectives were to infer the factors that shaped the genetic structure and to reconstruct the recolonization history of the species. Six haplotypes that were phylogenetically grouped into two lineages were identified. Homology of the three loci to the respective regions of the chloroplast genome was confirmed by sequencing. The chloroplast haplotypes found in Hagenia showed a clear pattern of congruence between their geographical distribution and genealogical relationships. A very low haplotype diversity within populations (h _S = 0.079, v _S = 0.058) and a very high population differentiation (G _ST = 0.899, N _ST = 0.926) was observed, reflecting very low mixing between recolonizing lineages. Restricted gene flow through seeds, rare long-distance dispersal, contiguous range expansion and mutation shaped the genetic structure of Hagenia. Fossil pollen records suggested that the trend of postglacial recolonization of Hagenia was first in the south and latter went to the north in Ethiopia.     

Uptake and translocation of manganese in seedlings of two varieties of Douglas fir (Pseudotsuga menziesii var. viridis and glauca)

Douglas fir (Pseudotsuga menziesii) variety glauca (DFG) but not the variety viridis (DFV) showed symptoms of manganese (Mn) toxicity in some field sites. We hypothesized that these two varieties differed in Mn metabolism. To test this hypothesis, biomass partitioning, Mn concentrations, subcellular localization and 54Mn-transport were investigated. Total Mn uptake was three-times higher in DFG than in DFV. DFV retained > 90% of 54Mn in roots, whereas > 60% was transported to the shoot in DFG. The epidermis was probably the most efficient Mn barrier since DFV contained lower Mn concentrations in cortical cells and vacuoles of roots than DFG. In both varieties, xylem loading was restricted and phloem transport was low. However, sieve cells still contained high Mn concentrations. DFV displayed higher biomass production and higher shoot : root ratios than DFG. Our results clearly show that both varieties of Douglas fir differ significantly in Mn-uptake and allocation patterns rendering DFG more vulnerable to Mn toxicity.     

Genetic variation at AFLPs for the Dipterocarpaceae and its relation to molecular phylogenies and taxonomic subdivisions

Genetic differentiation was investigated among 54 Indonesian species of Dipterocarpaceae, a dominant tree family in Asian tropical rainforests, using amplified fragment length polymorphism markers. The tree developed from the resultant unweighted pair group method using arithmetic averages clearly separated all investigated dipterocarps into two major groups that corresponded to tribe Dipterocarpeae and tribe Shoreae, respectively. These results are in accordance with the topology of molecular phylogenetic trees derived from PCR–restriction fragment length polymorphism analysis of chloroplast DNA and generally support the traditional taxonomic assessments. The possibility of interspecific hybridization is also discussed.     

Molecular genetic tools to infer the origin of forest plants and wood

Most forest tree species exhibit high levels of genetic diversity that can be used to trace the origin of living plants or their products such as timber and processed wood. Recent progress to isolate DNA not only from living tissue but also from wood and wood products offers new opportunities to test the declared origin of material such as seedlings for plantation establishment or timber. However, since most forest tree populations are weakly differentiated, the identification of genetic markers to differentiate among spatially isolated populations is often difficult and time consuming. Two important fields of “forensic” applications are described: Molecular tools are applied to test the declared origin of forest reproductive material used for plantation establishment and of internationally traded timber and wood products. These applications are illustrated taking examples from Germany, where mechanisms have been developed to improve the control of the trade with forest seeds and seedlings, and from the trade with wood of the important Southeast Asian tree family Dipterocarpaceae. Prospects and limitations of the use of molecular genetic methods to conclude on the origin of forest plants, wood, and wood products are discussed.     

Genetic variation of oaks (<Emphasis Type="Italic">Quercus</Emphasis> spp.) in Switzerland. 3. Lack of impact of postglacial recolonization history on nuclear gene loci

Quercus petraea, Quercus pubescens and Quercus robur are closely related and interfertile white oaks native to Switzerland. The three species are known to share identical cpDNA haplotypes, which are indicative of the postglacial recolonization history of populations. Only two haplotypes are common in Switzerland. We compared variation of cpDNA and of isozymes in 28 oak populations from Switzerland in order to assess the impact of the postglacial population history on current genetic structures of nuclear controlled isozyme gene loci. Species delineation was based on Principal Component Analysis of leaf morphological traits. The species status of populations was reflected at isozyme gene loci, but differentiation between populations with different cpDNA haplotypes and hence different recolonization history was very low at enzyme gene loci for all species. Thus, glacial and postglacial population history was not reflected at nuclear gene loci on the temporal and spatial scale covered by the present study. Extensive gene flow through pollen among populations is likely to have blurred a previously existing genetic differentiation at biparentally inherited gene loci that possibly evolved in the different glacial refugia of the above mentioned cpDNA haplotypes.     

Efficiency of microsatellite enrichment in<Emphasis Type="Italic">Prosopis chilensis</Emphasis> using magnetic capture

Microsatellites (i.e., simple sequence repeats [SSRs]) are highly variable genetic markers that are widely used at an intraspecific level in population genetic studies. Here we employed an enrichment strategy for microsatellite isolation by using microsatellite oligoprobes and magnetic capture of the fragments (Fischer and Bachmann, 1998) inProsopis chilensis (Mol.) Stuntz (Fabaceae). We analyzed the obtained level of enrichment by sequencing 120 enriched genomic fragments. A total of 521 SSR motives were detected. According to specific search criteria (SSR motifs ≥3 repeat units and ≥6 bp length), 95.8% of the clones contained SSR motifs. Of these, 7.8% showed homology to chloroplast sequences and 92.2% to nuclear sequences. When regarding only nuclear SSRs with 5 or more repeat units and a minimum length of 10 bp, the level of enrichment was 30.8%. A FASTA search against the European Molecular Biology Laboratory (EMBL) database univocally revealed 4 clones in transcribed regions, 102 clones in genomic regions with unknown function, and 9 clones in chloroplast regions. Among the loci with longer repeat units (≥10 bp, ≥5 repeat units), 3 were in transcribed regions and 65 were in other genomic regions. We discuss the applicability of these markers for population genetic studies.     

Genetic composition and differentiation of sloe (Prunus spinosa L.) populations in Germany with respect to the tracing of reproductive plant material

Sloe (Prunus spinosa L.) is a shrub native to Europe. In Germany, 50–80 % of all planted sloe is imported. Little is known about the genetic diversity patterns within and between German sloe populations. Thus, a debate arose how to avoid risks for nature and landscape by planting potentially maladapted material. The main objectives of our study are to analyse the genetic differentiation pattern of sloe populations in Germany, to identify geographic/genetic structures and to evaluate their potential for tracing reproductive material. 17 natural populations from Germany and 1 from Italy and Hungary were investigated by Amplified Fragment Length Polymorphisms (AFLP) and PCR–RFLP techniques. The AMOVA analyses based on AFLPs for all populations and for the German populations only result in equally high differentiation values of Φ_PT = 15 % of molecular variance between populations. The analysis of cpDNA PCR–RFLPs resulted in 24 haplotypes with 30 % showing genetic variation between populations. Overall values of genetic variability over all loci and populations are: Na = 0.832, Ne = 1.114 and He = 0.072. Mantel tests for AFLPs and cpDNA haplotypes reveal no association between geographic and genetic distances between populations as a result of a lack of differentiation between German populations and those from southern and southeastern Europe. Weak geographic/genetic patterns were observed on a large scale. However, these concern the German populations only. Our results indicate that vegetative regeneration in combination with founder effects may influence the level of differentiation between populations. Populations with a large amount of vegetative propagation are more differentiated from other populations than those populations which exhibit less vegetative regeneration. The assignment of reproductive material (i.e. plant material) to potential source populations resulted in high values of correct allocations. Hence, such methods can be applied to trace reproductive material of unknown origin.