An enigmatic Ephedra-like fossil lacking micropylar tube from the Lower Cretaceous Yixian Formation of Liaoning,China

Angiosperms and gymnosperms are two well-separated groups in seed plants according to the current understanding. The huge gap between these two groups constitutes a serious threat against the Darwinism, which expects a continuous transitional series between them. The Lower Cretaceous Yixian Formation of Liaoning, China is famous for its megafossil angiosperms, including some early angiosperms and putative gnetalean plants. Here we document another Ephedra-like fossil plant, Pseudoephedra n. gen. n. sp., from the Yixian Formation on the basis of light microscopic (LM) and scanning electron microscopic (SEM) observations. Although its general morphology demonstrates a great resemblance to Ephedra, the expected micropylar tube characteristic of Ephedra is missing in Pseudoephedra. Instead a solid projection is seen on the top of the female parts. Such a puzzling character combination makes Pseudoephedra perplexing in seed plant phylogeny. If put in Ephedraceae (Gnetales), Pseudoephedra would destroy the only synapomorphy (micropylar tube) of the BEG clade. If put in angiosperms, Pseudoephedra would bridge the formerly huge gap between gymnosperms and angiosperms. Apparently, further investigation is needed to clarify the uncertain position of Pseudoephedra.     

Variable wood formation and adaptation to the alpine environment of Ephedra pachyclada (Gnetales: Ephedraceae) in the Mustang district, western Nepal

BACKGROUND AND AIMS: Plants of Ephedra normally have vessels, but are known to become nearly vessel-less in some alpine localities. Previous studies implied that wood formation in Ephedra differs fundamentally from that in dicotyledons in which vessel-bearing and vessel-less taxa are systematically distinct. Using E. pachyclada in the Mustang district of Nepal, growing in an altitudinal range of over 2000 m, variation in wood formation and adaptation to alpine environment was studied in this normally vessel-bearing species. METHODS: Variation in wood anatomy and wood formation was observed with conventional optical microscopy. The lengths of three kinds of tracheary elements were measured and statistically analysed against habitat altitude and plant size of the individuals studied. KEY RESULTS: In E. pachyclada three kinds of tracheary elements, vessel elements, tracheids and fibre-tracheids, were nearly equal in length within individuals showing no elongation after differentiation from cambial initials. Tracheary element lengths among individuals had a negative correlation with altitude and a positive correlation with plant size. Multivariate analyses showed that altitude has a stronger correlation with tracheary element lengths than plant height or stem diameter. Moreover, several individuals from high elevations completely lacked vessels, and vessel formation fluctuated even in individuals from lower elevations. CONCLUSIONS: Wood anatomical trends in E. pachyclada are considered as an adaptation to extremely dry conditions in high mountains. Fluctuation in vessel formation in individuals from low elevations indicated that vessels differentiate only when their lateral expansion is allowed. These results showed that E. pachyclada has a different system of wood formation from dicotyledons and supported the opinion that the wood structure of Gnetales is fundamentally different from that of angiosperms.     

A new Permian gnetalean cone as fossil evidence for supporting current molecular phylogeny

• Background and aims The order Gnetales has been the central focus of controversy in seed plant phylogeny. Traditional treatment of morphology supports the anthophyte hypothesis with Gnetales sister to angiosperms but current molecular data reject this hypothesis. A new fossil gnetalean cone, Palaeognetaleana auspicia gen. et sp. nov., is reported from the Upper Permian in North China, and its phylogenic implications are considered.• Methods Samples of cones from the upper part of the Upper Permian redbeds of Baode section, northwestern Shanxi Province, China, were examined.• Key results The cone is characterized by its unusual nature of reproduction that combines features of post-Triassic gnetaleans and some of the Palaeozoic conifers. It is made up of a number of imbricate axillary units, each simply formed by an ovule and a subtending bract, which may be comparable with the axillary seed-scale complex of some of the Palaeozoic conifer cones. The cone exhibits at least a partially bisexual character that appears to have pollen sacs with monosulcate ribbed pollen grains and sessile, asymmetric, and radiospermic ovules. The ovule has an integument of three envelopes: an outer one of pointed scales; a middle sclerified one; and an inner cuticle that extends upward into a micropyle with an oblique tip.• Conclusions The new Permian cone has unequivocal affinity with the Gnetales. The fossil has considerably extended the divergence time of the Gnetales from 140 (210?) back to 270 myr ago and, therefore, provides the first significant fossil evidence to support the current conclusion based on molecular data of seed plants, i.e. monophyletic gymnosperms, comprising the Gnetales are closely related to conifers.Key words: Palaeognetaleana auspicia (gen. et sp. nov.), Gnetales, bisexual fossil cone, phylogeny of seed plants, radiation of gymnosperms, molecular data, Late Permian, North China     

Double fertilization in Gnetum gnemon (Gnetaceae): its bearing on the evolution of sexual reproduction within the Gnetales and the anthophyte clade

The fertilization process in Gnetum is critical to our understanding of the evolution of sexual reproduction within the Gnetales, a monophyletic group of nonfiowering seed plants that are the closest living relatives to flowering plants. Although much is known about the fertilization process in Ephedra, which is basal within the Gnetales, little is known about sexual reproduction in the derived sister groups Gnetum and Welwitschia. Ovules of Gnetum gnemon were collected at various stages after hand pollination and processed for light, fluorescence, and electron microscopy. Approximately 5 d after pollination, pollen tubes reach sexually mature female gametophytes, which are coenocytic. At that time, a binucleate sperm cell is found within each pollen tube. Within 7 d of pollination, double fertilization events occur when each of two sperm nuclei released from a pollen tube fuses with a separate, undifferentiated female nucleus within the free nuclear female gametophyte, which lacks differentiated egg cells. The products of double fertilization are two viable zygotes; endosperm is not formed. The lack of differentiated egg cells in Gnetum gnemon is unparalleled among land plants and the documentation of a regularly occurring process of double fertilization is congruent with the hypothesis that a rudimentary process of double fertilization evolved in a common ancestor of angiosperms and Gnetales.     

Wood and bark anatomy of the African species of Gnetum

Quantitative and qualitative data are given for the two African species of Gnetum ( Gnetum section Gnetum subsection Micrognemones ). These species are lianoid and lack the fibre-tracheids of G. gnemon but have about the same vessel element and tracheid length as in that species. Vessel diameter is related to stem age and organography. Tori are clearly present in tracheary elements of the African Gnetum species, a first report for the genus. In these two species, origin of the lateral meristem, which produces vascular tissue and cambia, can be traced directly and indirecdy to cortical parenchyma. A second kind of meristematic action, newly reported for Gnetum, is produced by proliferation of axial parenchyma, fragmenting secondary xylem. Both presence of tori and site of origin of lateral meristematic activity in Gnetum contrast with corresponding conditions in Welwitschia.     

Perspective: the origin of flowering plants and their reproductive biology--a tale of two phylogenies

Recently, two areas of plant phylogeny have developed in ways that could not have been anticipated, even a few years ago. Among extant seed plants, new phylogenetic hypotheses suggest that Gnetales, a group of nonflowering seed plants widely hypothesized to be the closest extant relatives of angiosperms, may be less closely related to angiosperms than was believed. In addition, recent phylogenetic analyses of angiosperms have, for the first time, clearly identified the earliest lineages of flowering plants: Amborella, Nymphaeales, and a clade that includes Illiciales/ Trimeniaceae/Austrobaileyaceae. Together, the new seed plant and angiosperm phylogenetic hypotheses have major implications for interpretation of homology and character evolution associated with the origin and early history of flowering plants. As an example of the complex and often unpredictable interplay of phylogenetic and comparative biology, we analyze the evolution of double fertilization, a process that forms a diploid embryo and a triploid endosperm, the embryo-nourishing tissue unique to flowering plants. We demonstrate how the new phylogenetic hypotheses for seed plants and angiosperms can significantly alter previous interpretations of evolutionary homology and firmly entrenched assumptions about what is synapomorphic of flowering plants. In the case of endosperm, a solution to the century-old question of its potential homology with an embryo or a female gametophyte (the haploid egg-producing generation within the life cycle of a seed plant) remains complex and elusive. Too little is known of the comparative reproductive biology of extant nonflowering seed plants (Gnetales, conifers, cycads, and Ginkgo) to analyze definitively the potential homology of endosperm with antecedent structures. Remarkably, the new angiosperm phylogenies reveal that a second fertilization event to yield a biparental endosperm, long assumed to be an important synapomorphy of flowering plants, cannot be conclusively resolved as ancestral for flowering plants. Although substantive progress has been made in the analysis of phylogenetic relationships of seed plants and angiosperms, these efforts have not been matched by comparable levels of activity in comparative biology. The consequence of inadequate comparative biological information in an age of phylogenetic biology is a severe limitation on the potential to reconstruct key evolutionary historical events.     

Higher substitution rates and lower dN/dS for the plastid genes in Gnetales than other gymnosperms

Previous studies have shown that Gnetales exhibit an increase in the rates of substitution in plastid genes. However, the relative roles of selection and drift in driving the observed increase in substitution rate remain unclear. Here we estimated the nonsynonymous (dN) and synonymous (dS) substitution rates and the dN/dS ratios for 59 plastid protein-coding genes conserved across Gnetales and other gymnosperms with maximum likelihood methods. Our results show that: (1) values of dN and dS for 48 and 50 genes, respectively, are significantly higher in Gnetales relative to the other sampled gymnosperms; (2) the acceleration of dN is of lower magnitude than that of dS; (3) dN/dS is significantly reduced for 22 of the 59 plastid genes in Gnetales; and (4) dN/dS is extremely heterogeneous among the Gnetalean genes, varying by up to a factor of 40. We propose that while biological characteristics such as generation time and plant height may contribute to the rate increase in Gnetales, the special habitats that they occupy are linked to the gene-specific reduction of dN/dS.