The phylogenetic positions of the conifer genera Amentotaxus,Phyllocladus, and Nageia inferred from 18s rRNA sequences

To determine the evolutionary positions of the conifer genera Amentotaxus, Phyllocladus, and Nageia, we obtained 18S rRNA sequences from 11 new taxa representing the major living orders and families of gymnosperms. With the published Chlamydomonas as an outgroup, phylogenetic analyses of our new data and available sequences indicate that (1) the Gnetales form a monophyletic group, which is an outgroup to the conifers, (2) the conifers are monophyletic, (3) Taxaceae, Cephalotaxaceae, Cupressaceae, and Taxodiaceae form a monophyletic group, (4) Amentotaxus is closer to Torreya than to Cephalotaxus, suggesting that Amentotaxus is better to be classified as a member of Taxaceae, (5) Phyllocladus, Dacrycarpus, Podocarpus, and Nageia form a monophyletic group, and (6) Pinaceae is an outgroup to the other families of conifers. Our finding that Phyllocladus is a sister group of the Podocarpaceae disagrees with the suggestion that the phylloclade of the genus is an ancient structure and that the genus is a terminal taxon within the Podocarpaceae. The genus Nageia is more closely related to Podocarpus than to Dacrycarpus and was derived from within the Podocarpaceae. In conclusion, our data indicate that in conifers, the uniovulate cone occurred independently in Taxacaeae and Cephalotaxaceae, and in Podocarpaceae after the three families separated from Pinaceae, and support the hypothesis that the uniovulate cone is derived from reduction of a multiovulate cone.Correspondence to: S.-M. Chaw     

Implications of fossil conifers for the phylogenetic relationships of living families

Fossils have played a central role in our understanding of the evolution of conifers. Interpretation of the seed cone as a compound strobilus and the homologies of the ovuliferous scales of modern conifers with the axillary dwarf shoot of Pennsylvanian forms are based on fossils. Similarly, early evolutionary trends involving the reduction, fusion, and planation of the fertile and sterile elements of the axillary dwarf shoot, leading to structures characteristic of modern families, are documented in Late Permian and Triassic conifers. However, a phylogeny elucidating the derivation of modern families from fossil forms based on shared derived features has been elusive. The present cladistic treatment using 11 characters of ovulate cones and one of pollen grains suggests three phylogenetic groups of Late Paleozoic conifers, represented loosely by the Emporicaceae, Utrechtiaceae, and Majonicaceae of Mapes and Rothwell. The Taxaceae appears to have diverged from ancestors within the Utrechtiaceae, whereas the other modern families owe their origins to the Majonicaceae. The origin of the Taxodiaceae appears to have been biphyletic.Taxodium, Cupressus andSciadopitys are strongly linked toDolmitia of the Majonicaceae, butCryptomeria, Cunninghamia andAraucaria are grouped together and diverge basal to the former taxa.Pinus branches from a position basal to the known genera of the Majonicaceae and all modern families except the Taxaceae.Podocarpus also diverges basal toMajonica but may share an ancestor with this genus;Cepahalotaxus diverges basal to theDolmitiaPseudovoltzia subclade but distal toMajonica. Similarly, the Cheirolepidiaceae originated from basal members of the Majonicaceae and shows no close phylogenetic relationship with any modern family. Except for a strong linkage betweenCycadocarpidium and theAraucariaCunninghamia subclade, genera of the Voltziaceae appear to have branched more or less independently from within the Majonicaceae and show no strong affinity with modern conifers. Thus differences between modern conifer families are due mainly to their divergence from different Paleozoic ancestors.     

Phylogenetic relationships of conifers inferred from partial 28S rRNA gene sequences

The conifers, which traditionally comprise seven families, are the largest and most diverse group of living gymnosperms. Efforts to systematize this diversity without a cladistic phylogenetic framework have often resulted in the segregation of certain genera and/or families from the conifers. In order to understand better the relationships between the families, we performed cladistic analyses using a new data set obtained from 28S rRNA gene sequences. These analyses strongly support the monophyly of conifers including Taxaceae. Within the conifers, the Pinaceae are the first to diverge, being the sister group of the rest of conifers. A recently discovered Australian genus Wollemia is confirmed to be a natural member of the Araucariaceae. The Taxaceae are nested within the conifer clade, being the most closely related to the Cephalotaxaceae. The Taxodiaceae and Cupressaceae together form a monophyletic group. Sciadopitys should be considered as constituting a separate family. These relationships are consistent with previous cladistic analyses of morphological and molecular (18S rRNA, rbcL) data. Furthermore, the well-supported clade linking the Araucariaceae and Podocarpaceae, which has not been previously reported, suggests that the common ancestor of these families, both having the greatest diversity in the Southern Hemisphere, inhabited Gondwanaland.     

Mapped DNA probes from loblolly pine can be used for restriction fragment length polymorphism mapping in other conifers

A high-density genetic map based on restriction fragment length polymorphisms (RFLPs) is being constructed for loblolly pine (Pinus taeda L.). Consequently, a large number of DNA probes from loblolly pine are potentially available for use in other species. We have used some of these DNA probes to detect RFLPs in 12 conifers and an angiosperm. Thirty complementary DNA and two genomic DNA probes from loblolly pine were hybridized to Southern blots containing DNA from five species of Pinus (P. elliottii, P. lambertiana, P. radiata, P. sylvestris, and P. taeda), one species from each of four other genera of Pinaceae (Abies concolor, Larix laricina, Picea abies, and Pseudotsuga menziesii), one species from each of three other families of Coniferales [Sequoia sempervirens (Taxodiaceae), Torreya californica (Taxaceae) and Calocedrus decurrens (Cupressaceae)], and to one angiosperm species (Populus nigra). Results showed that mapped DNA probes from lobolly pine will cross-hybridize to genomic DNA of other species of Pinus and some other genera of the Pinaceae. Only a small proportion of the probes hybridized to genomic DNA from three other families of the Coniferales and the one angiosperm examined. This study demonstrates that mapped DNA probes from loblolly pine can be used to construct RFLP maps for related species, thus enabling the opportunity for comparative genome mapping in conifers.     

The evolution of gymnosperms redrawn by phytochrome genes: the Gnetatae appear at the base of the gymnosperms

Gymnosperms possess two to four phytochrome types which apparently are the result of successive gene duplications in the genomes of their common ancestors. Phytochromes are nuclear-encoded proteins whose genes, contrary to chloroplast, mitochondrion, and rRNA genes, have hitherto rarely been used to examine gymnosperm phylogenies. Since the individual phytochrome gene types implied phylogenies that were not completely congruent to one another, conflicting branching orders were sorted by the number of gene lineages present in a taxon. The Gnetatae (two gene types) branched at the base of all gymnosperms, a position supported by bootstrap sampling (distance and character state trees, maximum likelihood). The Gnetatae were followed by Ginkgo, Cycadatae, and Pinaceae (three gene types) and the remaining conifers (four gene types). Therefore, in phytochrome trees, the most ancient branch of the conifers (Pinatae) seems to be the Pinaceae. The next split appears to have separated Araucariaceae plus Podocarpaceae from the Taxaceae/Taxodiaceae/Cupressaceae group. Structural arrangements in the plastid genomes (Raubeson and Jansen 1992) corroborate the finding that there is no close connection between Pinaceae and Gnetatae as suggested by some publications. The analyses are based on 60 phytochrome genes (579 positions in an alignment of PCR fragments) from 28 species. According to rough divergence time estimates, the last common ancestor of gymnosperms and angiosperms is likely to have existed in the Carboniferous.     

Seed Morphology, Anatomy and Ultrastructure ofPhyllocladusL. C. & A. Rich. ex Mirb. (Phyllocladaceae (Pilg.) Bessey) in Connection with the Generic System and Phylogeny

To analyse the status and phylogeny of the genusPhyllocladus,seedsof all seven species of the genus were studied. The complexof features of thePhyllocladusreproductive system point to theisolated position of the genus within the conifers. Relationswith Podocarpaceae s. l., Taxaceae s. l. and Cephalotaxaceaeappeared to be remote because a complex of features clearlydistinguishesPhyllocladusfrom the afore-mentioned taxa. We findit advisable to circumscribe the family Phyllocladaceae as Bessey(1907) did, and place it into the order Taxales Knobl. in Warming(1890). From investigations of the seeds it appears the genusPhyllocladusconsistsof seven species, forming five groups. There is a significanttendency for transformation of the female reproductive structureswithin the generic boundaries ofPhyllocladus—seeds, originallysolitary, tending to aggregate in various kinds of compact clusters.Copyright1999 Annals of Botany Company PhyllocladusL. C. & A. Rich, ex Mirb., seed anatomy, seed morphology, systematics, phylogenetic relationships, Phyllocladaceae, Podocarpaceae s. str., Acmopyleaceae, Nageiaceae, Austrotaxaceae, Amentotaxaceae, Torreyaceae, Taxaceae, s. str., Cephalotaxaceae, Taxales.     

红豆杉科、三尖杉科和罗汉松科植物叶片结构的比较观察

扫描电镜和光镜下的叶表皮特征以及叶片解剖结构的研究结果,支持在红豆杉科内建立白豆杉属的处理方式;赞同在三尖杉属内建立篦子三尖杉组,叶片解剖结构方面,三尖杉科和红豆杉科的穗花杉属的相似之处颇多。而叶表皮形态特征方面又表现为三尖杉科和罗汉松科更加接近。反映了出了红豆杉科、三尖杉科和罗汉松科之间的密切而又复杂的系统关系。     

Chemotaxonomic differentiation of conifer families and genera based on the seed oil fatty acid compositions: multivariate analyses

 The fatty acid compositions of seed oils from 34 conifer species, mainly Pinaceae and secondarily Cupressaceae, have been determined by gas-liquid chromatography of the methyl esters. As noted in earlier studies, these oils were characterized by the presence of several Δ5-olefinic acids, i.e., 5,9-18:2, 5,9,12-18:3, 5,9,12,15-18:4, 5,11-20:2, 5,11,14-20:3, and 5,11,14,17-20:4 acids, in addition to the more common saturated, oleic, linoleic and α-linolenic acids. Based on these fatty acid compositions, and on those established in earlier systematic studies (totalling 82 species), we established a chemotaxonomic grouping of the main conifer families, i.e., of the Pinaceae, Taxodiaceae, Cupressaceae, and Taxaceae. This was achieved using multivariate analyses (principal component analysis and discriminant analysis). The fatty acids that discriminate best in this classification are the 5,11,14,17-20:4, 9,12,15-18:3 and 5,9,12-18:3 acids. Moreover, it was possible to differentiate between several genera of the Pinaceae: Pinus (including Tsuga and Pseudotsuga), Abies, Cedrus, and Picea plus Larix, represented quite distinct groups. Other fatty acids such as oleic, linoleic, and 5,9-18:2 acids were also important for this purpose. The fatty acid compositions, and particularly the Δ5-olefinic acid contents of conifer seed oils, may thus be applied to the chemosystematic distinction among conifer families as well as genera of the Pinaceae. Received: 3 January 1997 / Accepted: 17 April 1997     

Phylogeny and Divergence Times of Gymnosperms Inferred from Single-Copy Nuclear Genes

Phylogenetic reconstruction is fundamental to study evolutionary biology and historical biogeography. However, there was not a molecular phylogeny of gymnosperms represented by extensive sampling at the genus level, and most published phylogenies of this group were constructed based on cytoplasmic DNA markers and/or the multi-copy nuclear ribosomal DNA. In this study, we use LFY and NLY, two single-copy nuclear genes that originated from an ancient gene duplication in the ancestor of seed plants, to reconstruct the phylogeny and estimate divergence times of gymnosperms based on a complete sampling of extant genera. The results indicate that the combined LFY and NLY coding sequences can resolve interfamilial relationships of gymnosperms and intergeneric relationships of most families. Moreover, the addition of intron sequences can improve the resolution in Podocarpaceae but not in cycads, although divergence times of the cycad genera are similar to or longer than those of the Podocarpaceae genera. Our study strongly supports cycads as the basal-most lineage of gymnosperms rather than sister to Ginkgoaceae, and a sister relationship between Podocarpaceae and Araucariaceae and between Cephalotaxaceae-Taxaceae and Cupressaceae. In addition, intergeneric relationships of some families that were controversial, and the relationships between Taxaceae and Cephalotaxaceae and between conifers and Gnetales are discussed based on the nuclear gene evidence. The molecular dating analysis suggests that drastic extinctions occurred in the early evolution of gymnosperms, and extant coniferous genera in the Northern Hemisphere are older than those in the Southern Hemisphere on average. This study provides an evolutionary framework for future studies on gymnosperms.     

Asymmetrical development of biovulate cones resulting in uniovulate cones in Ephedra rhytidosperma (Ephedraceae)

Female cone morphology in Ephedra, including the number of initiated ovules and mature seeds per cone, provides important taxonomic characters used in sectional or species delimitation within Ephedra. Recent molecular phylogenies have indicated, however, that seed number per cone has changed repeatedly during the evolution of the genus. This study reports on the development of the female cone of E. rhytidosperma, based on a large sample of dissected cones studied under SEM. All cones were initially biovulate, and in the majority of cases, both female reproductive units (FRUs) developed a micropylar tube and formed mature seeds. In a few cases, the FRU pair developed asymmetrically in a cone, with one of them eventually aborting. There was no evidence of fusion of the FRU pair. Phylogenetically, E. rhytidosperma is in a clade with E. equisetina, which has uniovulate cones, and E. gerardiana and E. minuta, which have biovulate cones that also become unispermous via abortion. The biovulate condition may thus be ancestral in this clade.     

SILICIFIED PINUS REMAINS FROM THE MIOCENE OF WASHINGTON

Silicified leaves, dwarf shoots, pollen cones, and seed cones of Pinus from a Late Miocene chert bed within the Yakima Basalt Formation near Yakima, Washington are interpreted as coming from a single new species, P. foisyi. The needles and dwarf shoots are those of a three-needle pine. The needles contain two to four medial resin canals, a biform hypodermis, and endodermal cells with uniformly thickened walls. The pollen cones are ellipsoidal and about 1 cm long, and many contain bisaccate pollen grains. The seed cones are at least 6 cm long and are slightly asymmetrical. The cone axis has a broad sclerotic outer cortex, and the seed wing extends from a thick parenchymatous base. The scale apex bears a conspicuously swollen projection. The foliage and seed cones are identifiable with the Subgenus Pinus, Section Pinus, Subsection Oocarpae independently of one another, and together indicate a fossil species related to the modem Californian closed cone pines. Pinus foisyi represents one of the earliest occurrences of cone asymmetry associated with this group. However, cone serotiny characteristic of the modem species appears to have evolved after the Late Miocene.     

The Mesozoic gymnosperms

Features needing elucidation in Mesozoic Cycadales and Bennettitales are given and the Ginkgo—Czekanowskia assemblage is surveyed. Florin's interpretation of the conifer cone scale is discussed in relation to Schweizer's work on Pseudovoltzia. Taxus jurassica Florin is revised and shown to differ generically from Taxus but to share features seen among other Taxaceae: it thus helps to unify the family, but it does not relate the Taxaceae to ordinary conifers. A possible way is offered by which early conifers like Walchia could shift their ovules from a lateral to a terminal position and if this were acceptable, then the class Taxopsida would become unnecessary.     

Phylogeny of Taxaceae and Cephalotaxaceae Genera Inferred from Chloroplast matK Gene and Nuclear rDNA ITS Region

Phylogeny of the Taxaceae genera and the monotypic family Cephalotaxaceae has been extraordinarily controversial. In this paper chloroplast matK genes and nuclear ITS sequences were determined for all six genera of the two families and representatives of other conifer families. Analysis using either the nonsynonymous sites or the deduced amino acid sequences of matK genes strongly indicates that taxad genera and Cephalotaxaceae are monophyletic, with the Taxodiaceae/Cupressaceae clade as their sister group. Cephalotaxus is basal to the taxad genera, among which two clades, Torreya/Amentotaxus and Taxus/Pseudotaxus/Austrotaxus, are resolved. They correspond to Janchen's two tribes, Torreyeae and Taxeae. In Taxeae, Austrotaxus is the first to branch off. Analyses of the nuclear ITS sequence data corroborated the topology of the matK gene tree. These results refute the views that Cephalotaxaceae has no alliance with Taxaceae and that Austrotaxus and Amentotaxus should be excluded from the Taxaceae. We estimated the divergence time between the Taxodiaceae/Cupressaceae and the Cephalotaxaceae/Taxaceae clades to be 192–230 Myr ago and the divergence time between taxads and Cephalotaxus to be 149–179 Myr ago. Soon after the latter divergence event, within 6–8 Myr, the two taxad tribes originated. In conclusion, our data do not support Florin's claim that taxads could be traced to Devonian psilophytes (359–395 Myr ago).     

Seed cone anatomy of Cheirolepidiaceae (Coniferales): Reinterpreting Pararaucaria patagonica Wieland

? Premise of the study: Seed cone morphology and anatomy reflect some of the most important changes in the phylogeny and evolutionary biology of conifers. Reexamination of the enigmatic Jurassic seed cone Pararaucaria patagonica reveals previously unknown systematically informative characters that demonstrate affinities with the Cheirolepidiaceae. This paper documents, for the first time, internal anatomy for seed cones of this important extinct Mesozoic conifer family, which may represent the ghost lineage leading to modern Pinaceae. ? Methods: Morphology and anatomy of cones from the Jurassic La Matilde Formation in Patagonia are described from a combination of polished wafers and thin section preparations. New photographic techniques are employed to reveal histological details of thin sections in which organic cell wall remains are not preserved. Specific terminology for conifer seed cones is proposed to help clarify hypotheses of homology for the various structures of the cones. ? Key results: Specimens are demonstrated to have trilobed ovuliferous scale tips along with a seed enclosing pocket of ovuliferous scale tissue. Originally thought to represent a seed wing in P. patagonica, this pocket-forming tissue is comparable to the flap of tissue covering seeds of compressed cheirolepidiaceous cones and is probably the most diagnostic character for seed cones of the family. ? Conclusions: Pararaucaria patagonica is assigned to Cheirolepidiaceae, documenting anatomical features for seed cones of the family and providing evidence for the antiquity of pinoid conifers leading to the origin of Pinaceae. A list of key morphological and anatomical characters for seed cones of Cheirolepidiaceae is developed to facilitate assignment of a much broader range of fossil remains to the family. This confirms the presence of Cheirolepidiaceae in the Jurassic of the Southern Hemisphere, which was previously suspected from palynological records.     

On the Systematic Position of Amentotaxus from Its Embryological Investigation

The present paper deals with the embryological study and the systematic position of Amentotaxus argotaenia (Hance) Pilger. The material used was collected during 1980-1981 from Jin-fo Shan, 1400-1600 m, Sichuan Province, China. The species is dioecious. The male cone sheds its pollen during the period from the end of May to the middle of June. The pollen at mature stage is 2-celled. Pollen chamber appears obvious at the end of the nucellus. When pollen grains are dispersed, megaspore mother cell, which is situated deep in the nucellus, is in meiosis. The megaspore divides mitotically after pollination and the free nuclei of female gametophyte divide for the last time at the end of June. The wall formation takes place at the stage of 256 free nuclei. The development of archegonia takes place at the beginning of July and the fertilization occurs about July 20-23. The fertilized egg divides successively four times and results in a 16-nucleate proembryo. The young embryo is developing in August. It is interesting to note that the development of the young embryo is very slow. When the seed reaches the mature stage from June to July in the following year, the multicellular masses of the young embryos resulted from simple polyembryony remain immature within the female gametophyte. No cleavage polyembryony has been found. The subsequent embryogeny takes place after the seed has shed. Keng (1975) considers that Amentotaxus links the Taxaceae with Cephalotaxaceae. Our embryological data support Keng’s conclusion since they share (1) compound microstrobilus, (2) 2-celled pollen grains at shedding stage and (3) the rather long life cycle. Keng (1975) also mentions that Podocarpaceae may connect with Taxaceae through Phyllocladus. According to Keng the Podocarpaceae is related to Taxaceae to certain degree. It is obvious that the primitive spike-like male strobilus like the one in Cordaitales is obviously retained in Podocarpus spicatus and P. andinus of Podocarpaceae and Amentotaxus of Taxaceae. In addition, like in Amentotaxus there are 16 nuclei before wall formation in the proembryo of Podocarpus nivalis. These facts may well indicate that at least the Podocarpaceae and the Taxaceae were derived from a common stock. As far as the Taxaceae is concerned the authors tend to support the view of Koidzumi (1932) that Amentotaxus and Austrotaxus should be put in the same tribe since both possess the spike-like strobilus, the long life cycle and the seed maturation in the following year. They are probably rather primitive genera in the Taxaceae. The proembryogeny of Torreya is more or less specialized. It may be placed in a rather advanced tribe and the tribe Taxeae (including Taxus and Pseudotaxus)may be between the above two tribes. In conclusion, the Taxaceae is related to the Coniferales in certain respects and, as Keng (1975), Harri (1976) and Wang et al. (1979) have pointed out recently, placing of the Taxaceae in Coniferales is rather justifiable.     

Early evolution in the Pinaceae

Nineteen species of structurally preserved ovulate cones of the Pinaceae are known from the Cretaceous. One of these belongs to Pinus, while the others contain anatomical features necessitating their classification in the organ genera Pityostrobus and Pseudoaraucaria. The six species of the latter group are very similar to one another and probably represent a natural, thought extinct, genus. By contrast, there is so much variety in the twelve Cretaceous species of Pityostrobus that when they are considered with respect to the uniformity of cone structure within each of the modern genera, each of the species of this organ genus may well reperesent an natural genus by itself. All expect one of these fossil forms contain features that are today characteristic of Pinus. This, combined with the Early Cretaceous occurrence of a structurally preserved Pinus cone, suggests that Pinus or something very close to it represents the phylogenetic centrum of the ancestral complex. Lack of cones showing distinct affinity with other modern genera supports this idea and further suggests that, while Pinus was in existence in the Early Cretaceous, other Recent genera of the Pinaceae may not have diverged from the complex until the Late Cretaceous or Early Tertiary.     

四种裸子植物叶精油化学成分的研究

采用气相色谱—质谱—计算机联用仪对穗花杉、南方红豆杉、三尖杉和罗汉松叶精油化学成分的研究发现,三尖杉和穗花杉叶精油的组成特点极为相似,相同成分１３个,占各自精油组成的４８．０７％和３３．３２％．三尖杉和南方红豆杉叶精油的相同成分４个,占各自精油组成的１６．１４％和４０．５９％．在一定程度上支持三尖杉科和红豆杉科的亲缘关系接近,红豆杉科可能是通过穗花杉属和三尖杉科相联系的观点．罗汉松和穗花杉叶精油的相同成分４个,占各自叶精油组成的比例为２４．０９％和２０．８２％,比罗汉松和南方红豆杉、三尖杉之间组分的相似性要高．反映出罗汉松科和红豆杉科之间有一定联系,穗花杉属是认识红豆杉科、三尖杉科和罗汉松科之间系统关系的关键属     

Conifer genome sizes of 172 species,covering 64 of 67 genera,range from 8 to 72 picogram

Nuclear genome size of conifers as measured by flow cytometry with propidium iodide was investigated, striving to collect at least a single species from each genus. 64 out of 67 genera and 172 species were measured. Of the 67 genera, 21 are reported here for the first time and the same is true for 76 species. This nearly doubles the number of measured genera and adds 50% to the number of analyzed species. Conifers have chromosome numbers in the range of n = (7)10–12(19). However, the nuclear DNA content (2C‐value) is shown here to range from 8.3 to 71.6 picogram. The largest genome contains roughly 6 × 10¹⁰ more base pairs than the smallest genome. Genome sizes are evaluated and compared with available taxonomic treatments. For the mainly (sub)tropical Podocarpaceae small genome sizes were found with a 2C‐value of only 8–28 pg, with 13.5 pg on average. For the Taxaceae 2C‐values from 23–60 pg were determined. Not surprisingly, the genus Pinus with 97 species (39 species measured here) has a broad range with 2C = 38–72 pg. A factor of 2 difference is also found in the Cupressaceae (136 species) with nuclear DNA contents in the range 18–35 pg. Apart from the allohexaploid Sequoia, ploidy plays a role only in Juniperus and some new polyploids are found. The data on genome size support conclusions on phylogenetic relationships obtained by DNA sequencing. Flow cytometry is applicable even to young plants or seeds for the monitoring of trade in endangered species.     

Chloroplast matK gene phylogeny of Taxaceae and Cephalotaxaceae,with additional reference to the systematic position of Nageia

Reported in the present paper is a robust chloroplast matK gene phylogeny of Taxaceae, Cephalotaxaceae and Podocarpaceae represented by 10 species of seven genera, with three species of the Pinaceae as outgroups. The matk length of the 13 species ranges from 1488 bp to 1548 bp, which results from indels, in particular, 1-bp(base pair) insertion near the 3’ end of the gene in some groups. A 27 bp deletion was found at the nucleotide position 213 from the 5’ end of the matk gene of Pseudotaxus chienii. The aligned sequences used in PAUP and MEGA analyses were 1568 bp and 1494 bp respectively. In the matK gene, the rates of variation at the first, second and third codon positions are similar although the mean frequency of synonymous substitution is approximately twice as high as that of nonsynonymous substitution. Branch-and-Bound search found only one most parsimonious tree (tree length = 895, CI = 0.850, RI = 0. 876), in which all clades were strongly supported by bootstrap test. According to the tree, Taxaceae and Cephalotaxaceae are monophyletic groups, and the sister group relationship between the two families was confirmed. Taxus is closely related to Pseudotaxus while Torreya is the sister group of Amentotaxus. In addition, the close relationship between Nageia and Podocarpus was resolved. The present study supports the generic status of Pseudotaxus and Amentotaxus in point of cladistic analysis and genetic distance, but contra-dicts the establishment of the family Nageiaceae.     

Morphogenesis of the reproductive shoots of Welwitschia mirabilis and Ephedra distachya (Gnetales), and its evolutionary implications

For decades, Gnetales appeared to be closely related to angiosperms, the two groups together forming the anthophyte clade. At present, molecular studies negate such a relationship and give strong support for a systematic position of Gnetales within or near conifers. However, previous interpretations of the male sporangiophores of Gnetales as pinnate with terminal synangia conflict with a close relationship between Gnetales and conifers. Therefore, we investigated the morphogenesis of the male reproductive structures of Welwitschia mirabilis and Ephedra distachya by SEM and light microscopy. The occurrence of reduced apices to both halves of the antherophores of W. mirabilis gives strong support for the assumption that the male ‘flowers’ of W. mirabilis represent reduced compound cones. We assume that each half of the antherophore represents a lateral male cone that has lost its subtending bract. Although both halves of the antherophores of Ephedra distachya lack apical meristems, the histological pattern of the developing antherophores supports interpreting them as reduced lateral male cones as well. Therefore, the male sporangiophores of Gnetales represent simple organs with terminal synangia. Although extant conifers do not exhibit terminal synangia, similar sporangiophores are reported for some Cordaitales, the hypothetical sister group of conifers. Moreover, several Paleozoic conifers exhibit male cones with terminal sporangia or synangia. Therefore, we propose that conifers, Cordaitales and Gnetales originated from a common ancestor that displayed simple sporangiophores with a terminal cluster of sporangia.