Roland Schubert

Genetic response of forest systems to changing environmental conditions

Contributors. Part 1: Verification of Response to Stress. Stress responses in Scots pine (Pinus sylvestris L.). Cloning and characterisation of an ozone-inducible pinysylvin methyltransferase H. Chiron, et al. Screening of Sitka spruce genotypes for resistances to the White Pine Weevil in British Columbia J.N. King, R.I. Alfaro. Genetic variation in two heavily polluted stands of Norway spruce (Picea abies [L Karst.) as indicated by nuclear and organelle DNA markers R. Riegel, et al. Effects of extreme SO2-air pollution in winter 1995/96 on vitality and growth of SO2-tolerant Norway spruce (Picea abies [L. Karst.) clones in the Ore mountains H. Wolf. Variation in adaptation and growth as indicated by provenance trial Platycladus orientalis (L.) Franco X. Shen, X. Chen. Influence of nursery environment and pollution on alders L. Mejnartowicz. Part 2: Genetic Variation under Diverse Environmental Conditions. Small scale spatial genetic structure of six tropical tree species in French Guiana B. Degen, et al. Genetic variation in natural populations of Araucaria angustifolia (Bert.) O. Kuntze in Brazil V.A. Sousa, H.H. Hattemer. Microsatellite DNA markers and their usefulness in poplars, and conservation of microsatellite DNA loci in Salicaceae O.P. Rajora, M.H. Rahman. PCR-RFLP analysis of introns of nuclear genes in Populus and Prunus B. Heinze. Genetic types in white oak populations north of the Alps and in the Danube valley U.M. Csaikl, A.O. Konig. Highly polymorphic uniparentally inherited DNA markers for spatial genetic analysis of silver fir (Abies alba Mill.) populations B. Ziegenhagen, et al. Levels of genetic differentiation inPinus halepensis Mill. in Spain using quantitative traits, isozymes, RAPDs and cp-microsatellites R. Alia, et al. Geographical variation of gene diversity of Pinus pinaster Ait. in the Iberian Peninsula S.C. Gonzalez-Martinez, et al. Is autochthony an operational concept? F.N. Schoppa, H.-R. Gregorius. Part 3: Genetic Resources, Reproduction, Management. Molecular markers in sustainable management, conservation, and restoration of forest genetic resources O.P. Rajora, A. Mosseler. Sustainable treatment of resources: The genetic basis H.-R. Gregorius. Genetic diversity and differentiation of individual effective pollen clouds in trees H.H. Hattemer, et al. Microsatellite analysis of small anonymous seedlot samples from pedunculate oak (Quercus robur): a promising approach to monitor the number of different seed parents and pollen donors C. Lexer, et al. Fructification and genetic structures of Fagus sylvatica mixed stands in upper regions of the Harz mountains D. Krabel, et al. Dispersal of seed and effective pollen in small stands of European beech (Fagus sylvatica L.) K. Wang, H.H. Hattemer. Patterns of seed dispersal in a scattered forest tree species (Sorbus torminalis) based on multi-scale investigation of population genetic structure for chloroplast DNA S. Oddou-Muratorio, et al. Gene flow and mating system in a seedling seed orchard and a natural stand of Pinus merkusii Jungh. et de Vriese in Indonesia I.Z. Siregar, H.H. Hattemer. The pattern of genetic variation in Pinus nigra subspecies pallasiana natural populations from the Kazdag and Bolkar mountains, Turkey: Implications for in situ gene conservation Z. Kaya, et al. Genetic

Quantitative detection of agar-cultivated and rhizotron-grown Piloderma croceum Erikss. & Hjortst. by ITS1-based fluorescent PCR

A real-time quantitative TaqMan-PCR was established for the absolute quantification of extramatrical hyphal biomass of the ectomycorrhizal fungus Piloderma croceum in pure cultures as well as in rhizotron samples with non-sterile peat substrate. After cloning and sequencing of internal transcribed spacer (ITS) sequences ITS1/ITS2 and the 5.8S rRNA gene from several fungi, including Tomentellopsis submollis, Paxillus involutus, and Cortinarius obtusus, species-specific primers and a dual-labelled fluorogenic probe were designed for Piloderma croceum. The dynamic range of the TaqMan assay spans seven orders of magnitude, producing an online-detectable fluorescence signal during the cycling run that is directly related to the starting number of ITS copies present. To test the confidence of the PCR-based quantification results, the hyphal length of Piloderma croceum was counted under the microscope to determine the recovery from two defined but different amounts of agar-cultivated mycelia. Inspection of the registered Ct values (defined as that cycle number at which a statistically significant increase in the reporter fluorescence can first be detected) in a 10-fold dilution series of template DNA represents a suitable and stringent quality control standard for exclusion of false PCR-based quantification results. The fast real-time PCR approach enables high throughput of samples, making this method well suited for quantitative analysis of ectomycorrhizal fungi in communities of natural and artificial ecosystems, so long as applicable DNA extraction protocols exist for different types of soil.

Tissue-specific expression of an oat 12S seed glubulin gene in developing tobacco seeds: differential mRNA and protein accumulation

We studied the expression of the oat globulin gene asglo5 in developing transgenic tobacco seeds. The asglo5 gene promoter directed transcription in the endosperm as well as in the provascular tissue, the presumptive root tip and the shoot apical meristem of the embryo as revealed by GUS reporter gene constructs and in situ hybridization. However, immunological tissue printing detected the oat protein exclusively in the tobacco endosperm, suggesting that extensive post-transcriptional regulatory processes influence the expression of the monocot transgene in the dicot host.

Molecular characterization and evolution of the cytosolic class II 17.0 kDa small heat-shock protein gene family from Picea abies (L.) Karst

Abstract In order to investigate the pattern of small heat-shock protein (sHSP) expression in Norway spruce, seedlings were exposed to thermal stress. [35S]methionine in vivo labelling revealed a set of eight predominant sHSPs with molecular weights ranging from 16.0 to 27.0 kDa, comprising a 17.0 kDa cytosolic class II sHSP which is already predicted from a previously cloned cDNA sequence (Forest Genetics 4 (1997) 131). Western blot analysis showed no significant amounts of the 17.0 kDa sHSP in non-stressed vegetative tissues, and upon heat shock it accumulates to levels comparable to those constitutively found in embryo tissues from mature seeds. The Picea abies 17.0 kDa sHSP is encoded by a gene family. Six functional genes and two pseudogenes were obtained from PCR-based cloning, indicating a high degree of sequence conservation with nucleotide identities between 88 and 99%. There is evidence that the gene family is subjected to gene conversion, preferentially homogenizing the 5′ moieties of the genes. The genes code for two distinct polypeptides with molecular weights of 16910 and 16870 Da, the former contains a putative phosphorylation site RXXS. Unlike angiosperm sHSP genes, those from P. abies contain two introns, located in the 5′ un-translated and coding region, respectively. Homologous introns exist in sHSP genes from Picea glauca, and Funaria hygrometrica, suggesting the presence of introns as a retained primitive condition of plant sHSP gene evolution.

Genetic variation in two heavily polluted stands of Norway spruce ( Picea abies [L.] Karst.) as indicated by nuclear and organelle DNA markers

In two polluted neighbouring stands of Norway spruce, “tolerant” and “sensitive” adult trees were selected in pairs and genotyped by recently developed codominant EST markers as well as by chloroplast microsatellites. The results were compared with data previously obtained from isoenzyme gene markers. Larger allelic multiplicities are indicated in the two sensitive subsets as compared to the tolerant ones, i.e. a mean surplus of 11.5% at 6 EST loci and of 27.2% at 19 isoenzyme gene loci. Frequency distributions of genetic types deviate considerably between tolerant and sensitive subsets but statistical significance is indicated by only one of the EST loci tested. Another EST locus is the only one to reveal statistically significant deviations between the observed heterozygosities of the tolerant and the corresponding sensitive subset of the most heavily polluted stand (38% vs. 15%). Based on allelic distances, both categories of nuclear markers reveal a cluster of three subsets in contrast to one and the same sensitive subset which is part of the most heavily polluted stand. A different cluster is indicated by chloroplast markers: the two sensitive and the two tolerant subsets, respectively, reveal greater allelic similarities than the sensitive and tolerant subsets of each stand. Generally, the applied molecular markers appear to be more indicative for the genetic response of Norway spruce to the given environmental stress than isoenzyme gene markers. Different pollution intensities at the two locations are assumed to account for the observed deviations in the genetic response of the two stands.

Ozone and UV-B Responses of Trees and the Question of Forest Sustainability

Forest trees respond to increased ozone levels and UV-B radiation by reduced photosynthesis, allocation and growth, but also by increased expression of plant defence systems. Ozone effects usually are cumulative and affect in particular deciduous trees such as beech and birch. Current concepts of forest succession, climax tree species and timber production are thereby made questionable.