The article presents the results of the search for microsatellite or simple sequence repeat (SSR) loci with tri-, tetra-, penta- and hexanucleotide tandem repeat motifs in the draft de novo assembly of the Siberian fir (Abies sibirica Ledeb.) genome and the development of convenient relatively highly and moderately polymorphic markers that can be easily genotyped even by simple gel electrophoresis. In total, 64 pairs of oligonucleotide polymerase chain reaction (PCR) primers for 32 detected microsatellite loci were designed and tested. Based on the testing results, 10 most promising polymorphic loci were selected and genotyped in eight natural populations of Siberian fir. Homologous microsatellite loci in the genome of European silver fir (Abies alba Mill.) were also identified by mapping Siberian fir contigs, containing SSR loci to the European silver fir genome assembly. A multiplex panel of 14 universal microsatellite loci was developed and genotyped in samples from four natural populations of A. alba and a small sample of eight Nordmann fir (Abies nordmanniana (Steven) Spach) trees.

We report the first Hettangian theropod tracksite (~200 Ma) yielding a rich accumulation of plant remains from the Bleymard Strait (southern France). It constitutes an excellent opportunity to reconstruct lowermost Jurassic ecosystems hosting dinosaurs and which are still poorly documented in this area. Two morphotypes of tridactyl tracks are distinguished. They share similarities with Grallator and Kayentapus. Plant-bearing beds yield abundant leafy axes (Pagiophyllum peregrinum), male cones (Classostrobus sp.), wood (Brachyoxylon sp.) and pollen of conifers (Classopollis classoides). Sedimentological, petrological and mineralogical analyses demonstrated that, in the Dolomitic Formation from Bleymard, the palaeoenvironment progressively evolved from (1) a shoreface to a foreshore domain; to (2) a shallow environment that is restricted or occasionally open to the sea; then to (3) an intertidal to supratidal zone. The Hettangian theropod ecosystem of the Bleymard Strait was composed of tidal flats that were periodically emerged and bordered paralic environments inhabited by a littoral conifer-dominated forest in which Cheirolepidiaceae were the main component. The paucity of the palaeobotanical assemblage, as well as the xerophytic characteristics of Pagiophyllum, show that flora from Bleymard was adapted to withstand intense sunlight and coastal environments exposed to desiccant conditions coupled with salty sea spray, and dry conditions. These features are those of a conifer-dominated flora under a tropical to subtropical climate. The flora as well as the clay mineral analyses suggest contrasting seasons (cyclically dry then humid). This study supports that theropods were abundant and particularly adapted to this type of littoral environment bordering Cheirolepidiaceae-dominated forests.

Although there are reports of permineralised woods in Mesozoic and Cenozoic strata in Mexico, there are few palaeobotanical studies based on coniferous stems. Four taxa of fossil wood from three localities in Mexico are described at the anatomical level and identified taxonomically based on detailed comparisons with woods of extant and fossil taxa. Agathoxylon gilii sp. nov. and A. jericonse sp. nov. are determined from the Todos Santos Formation (Upper Jurassic), in Chiapas; A. parrensis sp. nov. is described from the Las Encinas Formation (Palaeocene) in Coahuila; and Taxodioxylon cabullensis sp. nov. corresponds with material from the Packard Shale Formation in Sonora. Dominance of Araucariaceae and Cupressaceae in this region during the Mesozoic and early Cenozoic is further supported by leaf records, but these materials may also represent other taxa, like Cheirolepidaceae, which may have araucarioid secondary xylem. Records of Brachyphyllum and/or Pagiophyllum, which may represent araucarioid or cheirolepidaceous taxa, and different cupressaceous leaves, have been collected in rocks of the same period in nearby formations. At present, the amalgamation of isolated organs to form one plant is uncertain and, thus, there is an open challenge to reconstruct the plants and better understand these extinct communities. The new fossil taxa will add to the diversity of extinct plants and conifer communities during the Mesozoic and early Cenozoic in low-latitude North America.

Pleistocene biogeography of conifer species is poorly known in much of western North America. We conducted morphological studies on 201 conifer cones and cone fragments recovered from Pleistocene sediments at the Ziegler Reservoir fossil site (2705 m) near Snowmass Village, Colorado. The basin, formed ~155–130 ka, contains fossil-bearing lacustrine, palustrine, and colluvial sediments spanning approximately 85 ka. Using a suite of morphological characters, particularly cone-scale bracts, we differentiated species of Abies, Picea, and Pseudotsuga. All fossil Abies specimens were assignable based on bract morphology to Abies concolor, which is currently absent from central Colorado (nearest populations are 160 km southwest of the site). A. concolor occurs only in sediments of MIS 5d and 5c. Pseudotsuga menziesii and Picea engelmannii cones occurred in sediments corresponding to MIS 5e, 5d, 5c, and 5a. A fourth conifer species, occurring in sediments of MIS 5e, 5d, 5c, and 5a, is difficult to assign to any extant species. Bract morphology is similar to Picea pungens, which grows near the site today, but scale morphology is unlike P. pungens. These fossils may represent ancestral P. pungens, an extinct variant, or an extinct sister species.

Conifers are evolutionarily more ancient than their angiosperm counterparts, and thus some adaptive mechanisms and features influenced by epigenetic mechanisms appear more highly displayed in these woody gymnosperms. Conifers such as Norway spruce have very long generation times and long life spans, as well as large genome sizes. This seemingly excessive amount of genomic DNA without apparent duplications could be a rich source of sites for epigenetic regulation and modifications. In Norway spruce, an important adaptive mechanism has been identified, called epigenetic memory. This affects the growth cycle of these trees living in environments with mild summers and cold winters, allowing them to adapt rapidly to new and/or changing environments. The temperature during post-meiotic megagametogenesis and seed maturation epigenetically shifts the growth cycle programme of the embryos. This results in significant and long-lasting phenotypic change in the progeny, such as advance or delay of vital phenological processes of high adaptive value, like bud break and bud set. This phenomenon is not only of important evolutionary significance but has clear practical implications for forest seed production and conservation of forest genetic resources. The underlying molecular mechanism that causes the ‘memory’ in long-lived woody species is currently under investigation. Here we summarize the information related to epigenetic memory regulation in gymnosperms, with special emphasis on conifers. The molecular mechanism behind this is still unknown but transcriptional changes are clearly involved. Epigenetic regulation may be realized through several mechanisms, including DNA methylation, histone modification, chromatin remodelling, small non-coding RNAs and transposable element regulation, of which non-coding RNAs might be one of the most important determinants.

The development of techniques for somatic embryogenesis in conifers has led to rapid advances in the ability to culture conifer tissue in vitro. Somatic embryogenesis now provides a means to clonally propagate commercially valuable conifers of several species and offers an in vitro experimental system for studying embryogenesis. As a result of these developments, the conifer zygotic and somatic embryo system has recently attracted the attention not only of the cell biologist but also of biochemists and molecular biologists. In this review I have attempted to consolidate the recent information on ultrastructure, biochemical and molecular aspects of conifer zygotic and somatic embryogenesis. In the first section, salient features of zygotic embryogenesis are highlighted followed by biochemical and ultrastructural characterization of development with reference to storage reserve deposition. In the second section, detailed characterization of seed storage proteins, changes in gene expression with emphasis on storage protein genes and Lea genes during zygotic and somatic embryogenesis are described. Effect of culture treatments such as abscisic acid, osmoticum and desiccation on storage protein gene expression in somatic embryos is discussed. Based on these studies, comparisons are presented between angiosperm and gymnosperm embryogeny regarding the unique morphological and developmental patterns of conifer embryogenesis. Finally, a brief discussion of recent progress in biochemical and molecular characterization of conifer seed germination is presented.

Thirty-three conifer species are currently recorded from Vietnam, Laos and Cambodia. All species are recorded from Vietnam but only six have been recorded from Cambodia and nine from Laos. Recent botanical surveys have recorded one additional species from Cambodia and seven from Laos. New distributions within those countries have also been recorded. This paper discusses those records.

Aspicilia uxoris is proposed as a new combination for a species known from Algeria, Morocco and Spain. It grows on bark and lignum of Cedrus atlantica, Juniperus spp. and Pinus halepensis in the western Mediterranean Region. It is characterized by spore and conidia sizes, granular epihymenium on the surface of paraphyses and a typical apothecial structure and development. The ecology and distribution of the species and a comparison with other epiphytic taxa of the genus are discussed. Notes on the identity of Aspicilia lignicola are provided. Lectotypes for Aspicilia cinerea var. vulgaris f. lignicola, Lecanora lignicola and Lecanora uxoris are designated.

Previous studies have shown that the widely used plant transformation vector Agrobacterium tumefaciens can persist in genetically engineered plants in vitro and in transgenic greenhouse-grown plants, despite the use of counter-selective antibiotics. However, little is known regarding Agrobacterium persistence in tree species. To understand the kinetics of A. tumefaciens decline and persistence in transformation experiments, we assayed for the presence of A. tumefaciens in spruce and pine embryogenic tissue for up to 10 weeks post-transformation. The A. tumefaciens populations declined rapidly in the first five days post-cocultivation but generally declined more slowly in pine, relative to spruce. No bacteria were detected in spruce embryogenic tissue beyond four weeks after cocultivation, however in pine there were ~100 colony forming units per g tissue at 10 weeks post-cocultivation. We present evidence that the detection limit for PCR using virD2 primers to detect A. tumefaciens in a background of pine needle DNA was approximately 109–1010 A. tumefaciens cells per g of tissue. We also assayed for A. tumefaciens in transgenic pine and spruce embryogenic tissue and from needles, branches, stems and roots of transformed plants, up to four years post-inoculation. Occasionally A. tumefaciens was detected in embryogenic tissue up to 12 months post-inoculation. A. tumefaciens was never detected in cultured embryogenic tissue more than twelve months after inoculation, nor in developing somatic embryos or germinating plantlets, nor any of the parts of greenhouse-grown plants. From these data we conclude that if A. tumefaciens persists in transgenic conifers, it does so beneath our ability to detect it.

Discoveries of new species and new records of species are the order of the day in Vietnam, a country with a high level of biodiversity and many areas still largely unexplored. In recent years many conifers have been discovered for the first time in the country, and this paper focuses on three of these. The Vulnerable Pinus krempfii is an endemic of exceptional scientific interest. The globally Vulnerable Taiwania cryptomerioides, a conifer of ancient lineage in the Cupressaceae previously only known to occur with certainty in Taiwan and the border region between Myanmar and China, is a new discovery for Vietnam, where it is Critically Endangered. The Critically Endangered Xanthocyparis vietnamensis, a new genus and species in the Cupressaceae, was described in 2001 and is restricted to limestone outcrops on the border with China. All three species are of high conservation concern due to their rarity and because of threats to their survival from expanding cultivation, felling and fires. We discuss the suitability of these three conifers as flagship species to promote forest conservation in Vietnam. We also discuss the problems involved in the protection of these species, and the efforts that are currently being undertaken to conserve them.

Coniferous forests are an important component of the Lebanese landscape. Ten species of conifers in five genera are found as wild populations in Lebanon: Cilician fir Abies cilicica, cedar of Lebanon Cedrus libani, Mediterranean Cyprus Cupressus sempervirens, Syrian juniper Juniperus drupacea, Grecian juniper J. excelsa, willdenow J. foetidissima, prickly juniper J. oxycedrus, Calabrian pine Pinus brutia, Aleppo pine P. halepensis and stone pine P. pinea. Although none of these species are seriously threatened in a global context, they exist in Lebanon mostly as fragmented and degraded populations, and their habitats are under great pressure from urbanization and associated development. Conservation efforts in Lebanon have concentrated mainly on the cedar of Lebanon, with two cedar of Lebanon forests protected as Nature Reserves. Large areas of forest, coniferous and otherwise, remain unprotected. In this review, the need for an integrated approach to conservation of the coniferous landscape of Lebanon and the importance of the participation of local communities in this process are emphasized.

Modern cone scales and seeds of Pinus strobus and Sequoia sempervirens, and their fossil (Upper Miocene, c. 6 Mar) counterparts Pinus leitzii and Sequoia langsdorfii have been studied using pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), electron-microprobe and scanning electron microscopy. Microscopic observations revealed only minor microbial activity and high-quality structural preservation of the fossil material. The pyrolysates of both modern genera showed the presence of ligno-cellulose characteristic of conifers. However, the abundance of (alkylated)phenols and 1,2-benzenediols in modern S. sempervirens suggests the presence of non-hydrolysable tannins or abundant polyphenolic moieties not previously reported in modern conifers. The marked differences between the pyrolysis products of both modern genera are suggested to be of chemosystematic significance. The fossil samples also contained ligno-cellulose which exhibited only partial degradation, primarily of the carbohydrate constituents. Comparison between the fossil cone scale and seed pyrolysates indicated that the ligno-cellulose complex present in the seeds is chemically more resistant than that in the cone scales. Principal component analysis (PCA) of the pyrolysis data allowed for the determination of the discriminant functions used to assess the extent of degradation and the chemosystematic differences between both genera and between cone scales and seeds. Elemental composition (C, O, S), obtained using electron-microprobe, corroborated the pyrolysis results. Overall, the combination of chemical, microscopic and statistical methods allowed for a detailed characterization and chemosystematic interpretations of modern and fossil conifer cone scales and seeds.

Lipids were extracted from needles of Norway spruce plants of different provenances. The plants, seeds of which were originally collected from all over Europe, were grown at two sites in England at the same longitude but one 350 km north of the other. Significant correlations were observed between monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG) ratios on the one hand and, on the other, the lowest extreme winter temperature at the original site of seed collection (EWTS).

In the case of lipid changes, however, environmental differences due to site were more important than the inherent genotypic characteristics of the planted trees while provenances with the lowest EWTS values had more MGDG than DGDG. This implies that existing winter hardiness under mild climatic conditions does not come from changes in this ratio although DGDG formation from MGDG is known to be an important feature of winter hardening in Norway spruce elsewhere in Europe. However, closer analysis of the fatty acids in lipids from provenances at individual sites did reveal significant correlations between those provenances with low EWTS values and certain molar amounts or ratios of fatty acids in DGDG, some of which could be traced to genotypic expression. These partly involved the unusual Δ5 desaturations which are mainly confined to the fatty acids of those conifers like Norway spruce that harden to very low temperatures.