Phylogenetic assemblage structure of North American trees is more strongly shaped by glacial–interglacial climate variability in gymnosperms than in angiosperms

How fast does biodiversity respond to climate change? The relationship of past and current climate with phylogenetic assemblage structure helps us to understand this question. Studies of angiosperm tree diversity in North America have already suggested effects of current water–energy balance and tropical niche conservatism. However, the role of glacial–interglacial climate variability remains to be determined, and little is known about any of these relationships for gymnosperms. Moreover, phylogenetic endemism, the concentration of unique lineages in restricted ranges, may also be related to glacial–interglacial climate variability and needs more attention. We used a refined phylogeny of both angiosperms and gymnosperms to map phylogenetic diversity, clustering and endemism of North American trees in 100‐km grid cells, and climate change velocity since Last Glacial Maximum together with postglacial accessibility to recolonization to quantify glacial–interglacial climate variability. We found: (1) Current climate is the dominant factor explaining the overall patterns, with more clustered angiosperm assemblages toward lower temperature, consistent with tropical niche conservatism. (2) Long‐term climate stability is associated with higher angiosperm endemism, while higher postglacial accessibility is linked to to more phylogenetic clustering and endemism in gymnosperms. (3) Factors linked to glacial–interglacial climate change have stronger effects on gymnosperms than on angiosperms. These results suggest that paleoclimate legacies supplement current climate in shaping phylogenetic patterns in North American trees, and especially so for gymnosperms.     

The complete nucleotide sequence of the small-subunit ribosomal RNA coding region for the cycadZamia pumila: Phylogenetic implications

Summary The DNA sequence of the small-subunit ribosomal RNA coding region for the cycadZamia pumila L. was determined. TheZamia smallsubunit rRNA was found to be 1813 nucleotides in length and approximately 92% identical to published angiosperm small-subunit rRNA sequences. Conserved regions interspersed with variable regions are observed corresponding to those found in other eukaryotic small-subunit sequences. Using representatives from protist, fungal, plant, and animal groups, a distance matrix was constructed of average nucleotide substitution rates for pairs of organisms. Phylogenetic trees were inferred from similarities between sequences. The sequence ofZamia represents the earliest divergence from the higher plant lineage reported to date for small-subunit rRNA data. Inferred phylogenies also support a monophyletic origin for the angiosperms consistent with studies citing phenotypic characters.     

Changes in growth conditions alter the male strobilus gene expression pattern in<Emphasis Type="Italic"> Cryptomeria japonica</Emphasis>

Two-year old saplings grown from cuttings of Cryptomeria japonica D. Don initiate strobilus development following treatment with gibberellic acid under long-day photoperiods. At 25 °C with a 14-h photoperiod in a phytotron, male strobili initiated normally; however, they remained green and fell from the saplings prematurely. To examine the change in male strobilus development at the molecular level, three genes expressed specifically in male strobili were analyzed. Two were MADS box genes homologous to the B-function genes in angiosperms, CjMADS1 and CjMADS2, and the third was Cry j I, which encodes an allergen protein, and this gene is expressed mainly in microspores. Under phytotron growing conditions, the homeotic genes were expressed constantly, which reflected the extended early developmental stage of male strobili. On the other hand, Cry j I expression was detected after a long delay just before strobilus development ceased. These results indicate that the expression of the genes related to male reproductive development in C. japonica is regulated by a factor(s) that is sensitive to environmental signals.Abbreviation GA₃ gibberellic acid     

Expression analysis of four<Emphasis Type="Italic"> Pinus radiata</Emphasis> male cone promoters in the heterologous host<Emphasis Type="Italic"> Arabidopsis</Emphasis>

Four male cone-specific promoters were isolated from the genome of Pinus radiata D. Don, fused to the -glucuronidase (GUS) reporter gene and analysed in the heterologous host Arabidopsis thaliana (L.) Heynh. The temporal and spatial activities of the promoters PrCHS1, PrLTP2, PrMC2 and PrMALE1 during seven anther developmental stages are described in detail. The two promoters PrMC2 and PrMALE1 confer an identical GUS expression pattern on Arabidopsis anthers. DNA sequence analysis of the PrMC2 and PrMALE1 promoters revealed an 88% sequence identity over 276 bp and divergence further upstream (<40% sequence identity). GUS expression driven by a 276-bp PrMALE1 promoter fragment showed the same pattern in Arabidopsis anthers as observed for the full-length PrMALE1 promoter. Within the 276-bp promoter fragment a region of high homology to a previously described 16-bp anther-box was identified. In gain-of-function experiments the putative PrMALE1 anther-box was fused upstream of a 90-bp CaMV 35S minimal promoter, as a single copy in the sense direction and as an inverted repeat. No GUS expression was conferred to Arabidopsis anthers by either of these two constructs. In a loss-of-function experiment a 226-bp PrMALE1 deletion construct, which did not contain the putative PrMALE1 anther-box, still maintained the originally observed PrMALE1 GUS expression pattern. Hence, gain-of-function as well as loss-of-function experiments consistently showed that the putative anther-box of the PrMALE1 promoter is non-functional in the Arabidopsis genetic background. For the analysis of the four full-length pine promoters PrCHS1, PrLTP2, PrMC2 and PrMALE1, transformation vectors based on pCAMBIA2200 and pCAMBIA1302 were used. It will also be demonstrated in this article that sequences within the T-DNA borders of these vectors caused a characteristic histological background expression in Arabidopsis, with staining observed in vascular tissue of leaves, sepals, roots, filaments of stamens and in stems and pistils.Abbreviation GUS -glucuronidaseGenBank accession numbers for the analysed promoters: AF 337656 (PrCHS1), AF 337655 (PrLTP2), AF 337657 (PrMC2) and AF 337658 (PrMALE1).     

Understanding the genetic architecture of a quantitative trait in gymnosperms by genotyping haploid megagametophytes

The gymnosperms are a group of plants characterized by a haploid female gametophyte (megagametophyte). With the function of bearing the female gametes and nourishing the developing embryo, the megagametophyte has provided a simple way to understand the genetics of gymnosperm species using biochemical or genetic markers. In this paper, a quantitative genetic approach is proposed to study the genetic architecture of a quantitative trait in gymnosperms by taking advantage of the megagametophyte and the concept of average effect of a gene. Average effect describes the value associated with an allele carried by an individual and transmitted to its offspring. Through the genetic dissection of the average effect and genetic variance associated with a gamete carrying candidate genes, this approach can provide estimates of basic population genetic parameters, such as additive, dominant and epistatic effects, allelic frequencies and linkage disequilibrium. The candidate genes, known through their major mutant phenotype, have been reported in gymnosperms. An example for a candidate gene affecting lignin biosynthesis was applied to demonstrate the statistical procedures of the approach and its advantage. The conditions upon which the approach can be effectively used are discussed. Received: 15 January 1999 / Accepted: 12 March 1999     

Biochemical differences between embryogenic and nonembryogenic calli of conifers

Summary Embryogenic and nonembryogenic calli of loblolly pine (Pinus taeda), Eastern white pine (P. strobus), pond pine (P. serotina), white spruce (Picea glauca), and European larch (Larix decidua) were analyzed for biochemical parameters previously shown to be indicative of an embryogenic state in Norway spruce (Picea abies). Concentrations of glutathione and total reductants as well as rates of ethylene evolution and incorporation of radioactive leucine into protein in the two callus types were consistent with the Norway spruce observations. Embryogenic potential of loblolly pine and pond pine callus was predicted by biochemical analysis in advance of the appearance of somatic embryos. Other parameters such as isozyme patterns and SDS-PAGE of soluble proteins could also be used to distinguish embryogenic from nonembryogenic conifer callus. Among the species investigated, white spruce was the most difficult to sort by these methods.     

Proteomic evaluation of gymnosperm pollination drop proteins indicates highly conserved and complex biological functions

The pollination droplet is a highly conservative pollination mechanism that is observed in all major gymnosperm taxa. Proteomics analysis of the pollination drops was carried out on four gymnosperm species: Juniperus communis (common juniper), Juniperus oxycedrus (prickly juniper), Chamaecyparis lawsoniana (Port Orford cedar), and Welwitschia mirabilis. Pollination drop proteins were purified by SDS-PAGE, and the most abundant proteins were analyzed by mass spectrometry and sequenced. Based on BLAST searching of combined amino acid sequences, the following proteins were identified in the following species: an 83-kDa subtilisin-like proteinase, a 62-kDa glycosyl hydrolase, a 47.5-kDa glucan 1,3-β-glucosidase precursor, a 30-kDa chitinase, and a 25-kDa thaumatin-like protein were identified in J. communis; a 30-kDa chitinase, a 25-kDa thaumatin-like protein, and a 32.5-kDa glucanase-like protein were identified in J. oxycedrus; an 83-kDa subtilisin-like proteinase, a 62-kDa β-d-glucan exohydrolase, a 47.5-kDa glucan 1,3-β-glucosidase, and two 25-kDa thaumatin-like proteins were identified in C. lawsoniana, and a 25-kDa chitinase was identified in W. mirabilis. Based on protein identifications, there is strong evidence that the pollination drop functions in both pathogen defense and pollen development. The discovery of similarities in terms of peptide sequence and protein identifications indicates that ovular secretions are functionally conservative, and that they are essential to reproductive success.     

Adventitious bud development in embryonic tissue ofPinus caribaea andPseudotsuga menziesii

Summary Adventitious bud formation was induced on detached cotyledons and on cotyledons attached to excised embryos ofPinus caribaea andPseudotsuga menziesii. The embryonic tissue was exposed to the cytokinin 6-benzyl amino purine contained within an agar medium. This exposure resulted in the formation of a meristematic zone, involving both epidermal and sub-epidermal cells, and then nodules on the tissue surface. Some of these nodules were induced to differentiate into bud primordia, and thence shoots, following exposure to a combination of auxin (IAA) and cytokinin. Shoots were produced over the entire surface of detached cotyledons ofPs. menziesii but predominantly on the adaxial surfaces of detached cotyledons ofPi. caribaea and the tips and adaxial surfaces of the cotyledons on the entire embryos of both species. Thus, inter-specific differences in the distribution of competent areas for adventitious bud production were detected in embryos.