Phylogeny of the Altingiaceae based on cpDNA matK, PY-IGS and nrDNA ITS sequences

 Phylogenetic relationships of the three genera of the family Altingiaceae, i.e., Altingia, Liquidambar and Semiliquidambar, based on matK sequences and the intergenic spacer between the psaA and ycf3 genes (PY-IGS) of cpDNA, and on the internal transcribed spacer (ITS) of nrDNA were studied. Phylogenetic trees based on the three data sets (matK, PY-IGS and ITS) were generated using Hamamelis japonica and Mytilaria laosensis (Hamamelidaceae), Cercidiphyllum japonicum (Cercidiphyllaceae), and Daphniphyllum calycinum (Daphniphyllaceae) as outgroups. The partition-homogeneity tests indicated that the three data sets and the combined data are homogeneous. A combined analysis also generated a strongly supported phylogeny. The phylogenetic trees show that the North American and western Asian species, L. styraciflua and L. orientalis, respectively, form a monophyletic group which is sister to the clade including all Asian species in the family. The genus Liquidambar is paraphyletic with Altingia and Semiliquidambar nested within. Phylogenetic analyses of the molecular data indicate that taxonomic reexamination of the generic delimitation in the Altingiaceae is needed. Received December 20, 2000 Accepted June 25, 2001     

Molecular phylogeny of relict-endemic <Emphasis Type="Italic">Liquidambar orientalis</Emphasis> Mill based on sequence diversity of the chloroplast-encoded <Emphasis Type="Italic">mat</Emphasis>K gene

The genetic diversity and evolutionary divergence in Liquidambar species and Liquidambar orientalis varieties were compared with respect to the matK gene. A total of 66 genotypes from 18 different populations were sampled in southwestern Turkey. The matK region, which is about 1,512 bp in length, was sequenced and studied. L. orientalis, L. styraciflua, and L. formosana had similar magnitude of nucleotide diversity, while L. styraciflua and L. acalycina possessed higher evolutionary divergence. The highest evolutionary divergence was found between L. styraciflua and eastern Asian Liquidambar species (0.0102). However, the evolutionary divergence between L. orientalis and other species was of a similar magnitude. The maximum-parsimony phylogenetic tree showed that L. styraciflua and L. orientalis formed a closer clade while East Asian species were in a separate clade. This suggests that the North Atlantic Land Bridge through southern Greenland may have facilitated continuous distribution of Liquidambar species from southeastern Europe to eastern North America in early Tertiary period. The maximum-parsimony tree with only 18 Oriental sweetgum populations indicated that there were two main clusters: one with mainly L. orientalis var. integriloba and the other with var. orientalis and undetermined populations. High nucleotide diversity (0.0028) and divergence (0.00072) were found in L. orientalis var. integriloba populations and Muğla-1 geographical region. This region could be considered as the major refugium and genetic diversity center for the species. The low genetic diversity and divergence at intraspecies level suggest that L. orientalis populations in Turkey share an ancestral polymorphism from which two varieties may have evolved.     

ISOZYME DIVERGENCE BETWEEN EASTERN ASIAN,NORTH AMERICAN,AND TURKISH SPECIES OF LIQUIDAMBAR (HAMAMELIDACEAE)

The deciduous woody genus Liquidambar has four morphologically similar species in eastern and western Asia, eastern North America, and Central America. Liquidambar styraciflua is found in the eastern United States and Central America, L. orientalis is native only to southwest Turkey, and L. formosana and L. acalycina occur in eastern Asia. This genus is one of many that contributes to the floristic similarities observed between these different regions. Allelic variation was scored at 22 isozyme loci from 41 populations. The level of genetic divergence between species on different continents is high. Nei's genetic identity was 0.431 between L. formosana and L. styraciflua, 0.485 between L. acalycina and L. styraciflua, 0.512 between L. orientalis and L. styraciflua, 0.256 between L. formosana and L. orientalis, and 0.305 between L. acalycina and L. orientalis. Estimates of time of divergence from the isozyme data suggest that the current species diverged before or during the Miocene. The pattern of relationships portrayed by the isozyme data suggest a longer period of separation between the eastern and western Asian forms of this genus. In addition, the eastern North American and Turkish species appear to be the most closely related intercontinental pair of species providing evidence for a North Atlantic land bridge as late as the Miocene. It would appear, therefore, that the North American populations were in contact with the Asian populations over the North Pacific and North Atlantic possibly as late as the Miocene, but that the separation between the two Asian populations occurred much earlier. The time of divergence as measured from the isozyme data correlates with an independent assessment of the origin of these disjuncts as determined from the fossil record.     

Phylogenetic relationships in the Hamamelidaceae: Evidence from the nucleotide sequences of the plastid genematK

The Hamamelidaceae is a family that bridges the basal elements of the Rosidae and the lower Hamamelidae, thus a better understanding of the phylogeny of the family is important for clarifying evolutionary patterns in the diversification of eudicots. However, subfamilial as well as tribal relationships in the Hamamelidaceae have been controversial. Nucleotide sequences of the chloroplast genematK were used to study the intergeneric relationships of the family. In the phylogenetic trees, constructed using parsimony analysis, the clade containingAltingia andLiquidambar (Altingioideae) is sister to a clade that includes all other Hamamelidaceae.Exbucklandia andRhodoleia form a clade, suggesting a close relationship between the two genera.Disanthus is sister to the monophyletic Hamamelidoideae. The paraphyletic arrangement ofDisanthus, Mytilaria andExbucklandia with respect to the Hamamelidoideae does not support the combination of these genera in one subfamily. In the Hamamelidoideae, thematK phylogeny supports the monophyly of several previously recognized groups with modifications, including the tribes Eustigmateae (incl.Molinadendron), Fothergilleae (excl.Molinadendron andMatudaea), and the subtribe Dicoryphinae. However, the Hamamelideae as traditionally circumscribed is polyphyletic. Apetaly has evolved three times independently in the Hamamelidoideae.     

New insights into DNA barcoding of seagrasses

Taxonomists find some plant genera challenging because of the few morphological differences or unclear characters among closely related species, which leads to the misidentification of taxa. DNA barcoding is an approach to identify species by using short orthologous DNA sequences, known as ‘DNA barcodes’. Concatenated rbcL and matK sequences are considered DNA barcodes for seagrasses. However, these markers are not applicable to all members of seagrasses at the species level, especially within the genus Halophila. Our previous studies indicated that the internal transcribed spacer (ITS) showed higher species resolution than the concatenated rbcL and matK sequences in the case of Halophila ovalis and closely related species. In this study, 26 ITS, two rbcL and two matK consensus sequences from 18 seagrass taxa belonging to four families collected in India, Vietnam, Germany, Croatia and Egypt were processed. Molecular ITS analysis resolved five clades. The results also indicate that the Cymodoceaceae family might be a non-monophyletic group. In conclusion, ITS could be applied as a DNA barcode for seagrasses instead of the rbcL/matK system previously proposed.     

<Emphasis Type="Italic">Altingioxylon hainanensis</Emphasis> sp. nov.: earliest fossil wood record of the family Altingiaceae in Eastern Asia and its implications for historical biogeography

A new species, Altingioxylon hainanensis, is described from the Eocene Changchang Formation of the Changchang Basin on Hainan Island, South China. It is the first record of a fossil wood assigned to Altingiaceae found in China, and the most ancient evidence of wood for this family in eastern Asia. The new species is similar to A. rhodoleioides, known since the Miocene in India and Java Island, and to Altingia hisauchii from the Miocene to Pliocene of Japan. The close resemblance between these species and Liquidambar sp., known from the Middle Miocene of western North America, provides additional evidence for the migration of their ancestors from Asia to North America across the Bering land bridge during the Miocene. Distinctions in ray sizes between the eastern Asian specimens and their contemporaries from Europe to Kazakhstan is suggested as a result of the divergence between the large eastern Asian clade and the North American–west Asian clade within Altingiaceae during the Eocene–Oligocene. The presence of crystals in ray cells may be considered an ancestral condition that persists in the eastern Asian lineages up to the extant Altingia and Semiliquidambar, but which was lost in other Altingiaceae in the course of evolution.     

Molecular phylogeny of the genus Triticum L

The genus Triticum L. includes the major cereal crop, common or bread wheat (hexaploid Triticum aestivum L.), and other important cultivated species. Here, we conducted a phylogenetic analysis of all known wheat species and the closely related Aegilops species. This analysis was based on chloroplast matK gene comparison along with trnL intron sequences of some species. Polyploid wheat species are successfully divided only into two groups – Emmer (sections Dicoccoides and Triticum) and Timopheevii (section Timopheevii). Results reveal strictly maternal plastid inheritance of synthetic wheat amphiploids included in the study. A concordance of chloroplast origin with the definite nuclear genomes of polyploid species that were inherited at the last hybridization events was found. Our analysis suggests that there were two ancestral representatives of Aegilops speltoides Tausch that participated in the speciation of polyploid wheats with B and G genome in their genome composition. However, G genome species are younger in evolution than ones with B genome. B genome-specific PCR primers were developed for amplification of Acc-1 gene.     

大果马蹄荷(Exbucklandia tonkinensis)群落的纬度地带性

大果马蹄荷主产南亚热带至中亚热带南缘,南至海南岛,北至罗霄山脉中段的井冈山、桃源洞地区,是常绿阔叶林的特征种。以纬度地带性(海南霸王岭、广东黑石顶、广东南岭、江西金盆山、江西井冈山、湖南桃源洞)为依托,选定6个具代表性的大果马蹄荷群落开展群落生态学研究,结果表明:(1)各样地物种多样性较高,尤以金盆山蕨类植物9科10属11种、种子植物42科78属128种和桃源洞蕨类植物9科11属12种、种子植物41科79属134种最为丰富。群落组成的优势科主要为金缕梅科、壳斗科、樟科、山茶科、杜鹃花科、山矾科等。(2)从区系特征和环境梯度看,大果马蹄荷群落以南亚热带为分布中心,向南或向北其物种多样性Simpson指数、Shannon-Wiener指数及相应的均匀度指数均呈下降趋势,其中霸王岭、黑石顶、南岭、金盆山、井冈山、桃源洞的Shannon-Wiener指数分别为3.453、4.021、4.130、3.790、3.415、3.712。(3)群落相似性聚类分析显示群落随纬度和随海拔高度形成两个梯度系列,一是以黑石顶、金盆山、井冈山、桃源洞的纬度地带性为一支,相似性系数0.51;二是以南岭和霸王岭聚成海拔梯度较高的另一支,但其相似性系数0.50,为0.33—0.48。(4)大果马蹄荷群落种类组成在区系性质上很相似,具有明显的南亚热带特征;同时,受海拔、地形、气温、降雨条件等因素的影响,种子植物属的热带成分随纬度增加而呈波动性下降趋势。(5)大果马蹄荷种群在各群落中的重要值水平和径级结构表现出一致性,在纬度地带性差异上无明显的相关性。霸王岭大果马蹄荷的径级结构为增长型,但重要值排名为32,说明向南分布该种群优势度明显下降;在南岭、黑石顶、金盆山、桃源洞该种群优势度较大,且为稳定型种群;在井冈山该群落受到人为干忧,大果马蹄荷的重要值排名第1,但为衰退型种群。     

DNA barcoding of tuberous Orchidoideae: a resource for identification of orchids used in Salep

Tubers of terrestrial orchids are harvested and traded from the eastern Mediterranean to the Caspian Sea for the traditional product Salep. Overexploitation of wild populations and increased middle‐class prosperity have escalated prices for Salep, causing overharvesting, depletion of native populations and providing an incentive to expand harvesting to untapped areas in Iran. Limited morphological distinctiveness among traded Salep tubers renders species identification impossible, making it difficult to establish which species are targeted and affected the most. In this study, a reference database of 490 nrITS, trnL‐F spacer and matK sequences of 133 taxa was used to identify 150 individual tubers from 31 batches purchased in 12 cities in Iran to assess species diversity in commerce. The sequence reference database consisted of 211 nrITS, 158 trnL‐F and 121 matK sequences, including 238 new sequences from collections made for this study. The markers enabled unambiguous species identification with tree‐based methods for nrITS in 67% of the tested tubers, 58% for trnL‐F and 59% for matK. Species in the genera Orchis (34%), Anacamptis (27%) and Dactylorhiza (19%) were the most common in Salep. Our study shows that all tuberous orchid species in this area are threatened by this trade, and further stresses the urgency of controlling illegal harvesting and cross‐border trade of Salep tubers.     

Molecular phylogeny of the genus Pieris (Ericaceae) with special reference to phylogenetic relationships of insular plants on the Ryukyu Islands

Phylogenetic relationships within the genus Pieris (Ericaceae) were investigated based on the rbcL and matK genes along with five spacer sequences of chloroplast DNA to address questions regarding the phylogeography of the genus in association with insular plants on the Ryukyu Islands. The most parsimonious trees indicated that P. floribunda from eastern North America is a sister taxon to the remaining taxa examined, and suggested that the East Asian taxa examined are monophyletic. A morphologically cohesive group, section Pieris, was revealed to be paraphyletic. Within the East Asian clade, insular endemics from the Ryukyu Islands, Taiwan, and mainland Japan formed a sister group to P. formosa from the Himalayas and southern China. Our data suggest that the insular endemics of the Ryukyu Islands and Taiwan arose via allopatric divergence as a result of a paleogeographical land configuration of a landbridge during the early–middle Pleistocene in the Quaternary Period.     

What is a bona fide mating-type gene? Internuclear complementation of mat mutants in Podospora anserina

In the heterothallic ascomycete Podospora anserina, the mating-type locus is occupied by two mutually exclusive sequences termed mat+ and mat–. The mat+ sequence contains only one gene, FPR1, while the mat– sequence contains three genes: FMR1, SMR1 and SMR2. Previous studies have demonstrated that FPR1 and FMR1 are required for fertilization. Further analyses have led to the hypothesis that mat+ and mat– genes establish a mat+ and mat– nuclear identity, allowing recognition between nuclei of opposite mating type within the syncytial cells formed after fertilization. This hypothesis was based on the phenotypes of strains bearing mutations in ectopic mat genes. Here we present an analysis of mutations in resident mat– genes which suggests that, unlike FMR1 and SMR2, SMR1 is not involved in establishing nuclear identity. In fact, mutations in these two genes impair nuclear recognition, leading to uniparental progeny, while mutations in SMR1 block the sexual process, probably at a step after nuclear recognition. The nuclear identity hypothesis has also been tested through internuclear complementation tests. In these experiments, the mat– mutants were crossed with a mat+ strain carrying the wild-type mat– genes. Our rationale was that internuclear complementation should not be possible for nuclear identity genes: the relevant genes should show nucleus-restricted expression, and diffusion of their products to other nuclei should not occur. This test confirmed that SMR1 is not a bona fide mat gene since it can fulfill its function whatever its location, in either a mat− or a mat+ nucleus, and even when present in both nuclei. SMR2, but not FMR1, behaves like a nuclear identity gene with respect to internuclear complementation tests. A model is proposed that tentatively explains the ambiguous behaviour of the FMR1 gene and clarifies the respective functions of the three mat– proteins. Received: 15 October 1996 / Accepted: 25 April 1997     

Phytoremediation potential of transplanted bare-root seedlings of trees for lead/zinc and copper mine tailings

Selecting plant species that can overcome unfavorable conditions and increase the recovery of degraded mined lands remains a challenge. A pot experiment was conducted to evaluate the feasibility of using transplanted tree seedlings for the phytoremediation of lead/zinc and copper mine tailings. One-year-old bare-root of woody species (Rhus chinensis Mill, Quercus acutissima Carruth, Liquidambar formosana Hance, Vitex trifolia Linn. var. simplicifolia Cham, Lespedeza cuneata and Amorpha fruticosa Linn) were transplanted into pots with mine tailings and tested as potential metal-tolerant plants. Seedling survival, plant growth, root trait, nutrient uptake, and metal accumulation and translocation were assessed. The six species grew in both tailings and showed different tolerance level. A. fruticosa was highly tolerant of Zn, Pb and Cu, and grew normally in both tailings. Metal concentrations were higher in the roots than in the shoots of the six species. All of the species had low bioconcentration and translocation factor values. However, R. chinensis and L. formosana had significantly higher translocation factor values for Pb (0.88) and Zn (1.78) than the other species. The nitrogen-fixing species, A. fruticosa, had the highest tolerance and biomass production, implying that it has great potential in the phytoremediation of tailing areas in southern China.     

Evaluation of the DNA barcoding approach to develop a reference data-set for the threatened flora of Sicily

The Mediterranean Basin is one of the most significantly altered World Biodiversity Hotspots with extensive habitat loss and fast genetic population erosion, for which urgent biodiversity reconnaissance and preservation actions are required. In particular, Sicily has about 600 taxa classified as threatened or near-threatened. The correct recognition and identification of such biodiversity is required for supporting further activities. The objective of this work is to assess the ability of the DNA barcoding approach to identify different taxonomic groups from a collection of the most threatened plant taxa, throughout natural Sicilian populations. The evaluation of the DNA barcoding core markers, rbcL and matK, was carried out on 30 taxa belonging to 13 families. DNA barcode fragments were recovered from all taxa (100%). The rbcL gene was recovered from 97% of the taxa and matK gene from 73%. In this test, 19 taxa overall (63%) were totally resolved at the specific or subspecific level, by at least one of the core markers. Fourteen of the 17 most threatened taxa (EN, CR) included in this work were totally discriminated. The matK and rbcL locus, respectively, resolved 64% and 48% of the taxa successfully sequenced. The matK gene expressed the highest genetic distance (K2P value), from 0.4% to 8.6%, against a range of 0.1–2% of rbcL gene. However, the rbcL gene appeared a good compromise between PCR, sequencing success and species-level resolution. Cryptic groups suggest the implementation of additional barcoding markers or different primer combinations, particularly for matK, in order to increase the performances. However, this preliminary result confirms the potential of the barcoding approach for quick identification of unknown and heterogeneous plant groups to generate a dedicated reference data-set of the threatened Sicilian flora for a wide range of applications.     

Astragalus (Fabaceae): A molecular phylogenetic perspective

Nucleotide sequences of the plastidmatK gene and nuclear rDNA internal transcribed spacer region were sampled fromAstragalus L. (Fabaceae), and its closest relatives within tribe Galegeae, to infer phylogenetic relationships and estimate ages of diversification. Consistent with previous studies that emphasized sampling for nrDNA ITS primarily within either New World or Old World species groups,Astragalus, with the exception of a few morphologically distinct species, is strongly supported as monophyletic based on maximum parsimony and Bayesian analyses ofmatK sequences as well as a combined sequence dataset. ThematK data provides better resolution and stronger clade support for relationships amongAstragalus and traditionally related genera than nrDNA ITS.Astragalus sensu stricto plus the genusOxytropis are strongly supported as sister to a clade composed of strictly Old World (African, Australasian) genera such asColutea. Sutherlandia, Lessertia, Swainsona, andCarmichaelia, plus several morphologically distinct segregates of EurasianAstragalus. Ages of these clades and rates of nucleotide substitution estimated from a fossil-constrained, rate-smoothed, Bayesian analysis ofmatK sequences sampled from Hologalegina indicateAstragalus diverged from its sister group,Oxtropis, 12–16 Ma, with divergence of Neo-Astragalus beginning ca 4.4. Ma. Estimates of absolute rates of nucleotide substitution forAstragalus and sister groups, which range from 8.9 to 10.2×10⁻¹⁰ substitutions per site per year, are not unusual when compared to those estimated for other, mainly temperate groups of papilionoid legumes. The results of previously published work and other recent developments on the phylogenetic relationships and diversification ofAstragalus are reviewed.     

Intrapopulational genetic diversity of Araucaria angustifolia (Bertol.) Kuntze is different when assessed on the basis of chloroplast or nuclear markers

A natural population of Araucaria angustifolia (Bertol.) Kuntze was evaluated to quantify genetic diversity, using nucleotide sequences of chloroplast maturase K genes (matK) and six specific nuclear polymorphic microsatellite loci (nuSSR). Estimates of nucleotide diversity assessed from matK sequences were low (π = 0.001), but they indicated the potential of this gene for evaluating divergence among widespread populations. The matK sequence (1,565 bp) was used for analysis of phylogenetic relationships, enabling clustering of the South American species (A. angustifolia and A. araucana). For analysis of intraspecific genetic diversity, nuSSR were more informative than matK, resulting in a polymorphic loci percentage of 69, a number average of alleles per locus of 11.6, expected genetic diversity 0.741, and significant spatial genetic structure up to 25 m. Our data can help in the selection of genetic sources for conservation of this species, which is at risk of extinction.     

Sequence analyses of the ITS regions and the matK gene for determining phylogenetic relationships of Diospyros kaki (persimmon) with other wild Diospyros (Ebenaceae) species

To elucidate the relationships among Diospyros kaki and species closely related in previous studies, the nuclear ribosomal internal transcribed spacer (ITS) DNA sequence and the chloroplast matK gene were sequenced and compared with those of nine Diospyros species from Thailand, four species from temperate regions, and one species of southern Africa, D. lycioides. Maximum parsimony, maximum likelihood, and neighbor joining analyses of the matK and ITS data sets revealed that D. kaki is closely related to two diploid species, D. oleifera and D. glandulosa. D. kaki, D. glandulosa, and D. oleifera were placed differently in the trees obtained from ITS and matK data sets, suggesting that hybridization and/or introgression may have occurred during the development of these species. D. kaki was not found to be closely related to D. ehretioides, a diploid species from Thailand. These results differed from a prior analysis of this genus performed with chloroplast DNA (cpDNA) restriction site mutations in 3.2- and 2.1-kb amplified sequences. The results supported Ng’s hypothesis that D. glandulosa and D. kaki may share a common ancestor. D. oleifera was also closely associated with D. kaki.     

Phylogeny of Bonatea (Orchidaceae: Habenariinae) based on molecular and morphological data

The genus Bonatea is widely distributed throughout southern and eastern Africa. Considerable debate surrounds the generic status of Bonatea, but there have been neither previous studies of evolutionary relationships among Bonatea species, nor any tests of the monophyly of the genus in relation to its close relative Habenaria. We investigated phylogenetic relationships between Bonatea and selected Habenaria species using morphology, as well as sequences of the chloroplast gene matK and the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA. A fully resolved cladogram was obtained using morphological data, but neither the ITS, matK, nor combined data sets yielded well-resolved and well-supported phylogenetic structure for Bonatea. There is poor congruence between ITS and matK data for interspecific relationships in Bonatea, whilst the morphological results are largely congruent with the ITS analysis. Relative to Habenaria, there is little sequence variation between Bonatea species, which could indicate a recent and rapid radiation of Bonatea. Although the sampled Bonatea species form a distinct clade, more extensive sampling of Habenaria would be required to establish unambiguously whether or not Bonatea is monophyletic.     

基于转录组测序的枫香EST-SSR引物开发及有效性评价

枫香是广西重要的乡土树种之一，具有较高的用材、观赏及药用价值。为验证枫香SSR引物的实际应用效果，该研究基于转录组测序技术，检测枫香SSR位点并设计引物，通过PCR扩增和聚丙烯酰胺凝胶电泳筛选出具有较高多态性的枫香EST-SSR引物，并对1个枫香天然群体进行遗传多样性分析。结果表明：(1)共发掘到23 777个SSR位点，单核苷酸重复类型SSR位点占总位点比例最高(46.54%),在重复次数上5～12次之间的SSR位点占比最高(72.36%)。(2)共开发出262对SSR引物，有效扩增率为53.1%,最终筛选出扩增稳定、条带清晰的引物18对。(3)多态性检测结果显示所有位点均具有多态性，天然群体遗传多样性结果显示该天然群体中等位基因数量(Na)、有效等位基因数量(Ne)、Shannon多样性指数(I)、观测杂合度(Ho)变化范围分别为2～4、1.112 8～2.609 6、0.208 9～1.112 7和0.275 9～1.000 0,平均值分别为2.333 3、1.957 4、0.708 5和0.722 6。综上认为，枫香中占优势的SSR位点重复类型和重复基序与其他物种基本相同，所开...     

Deletion of the mating-type sequences in Podospora anserina abolishes mating without affecting vegetative functions and sexual differentiation

The mating-type locus of Podospora anserina controls fusion of sexual cells as well as subsequent stages of development of the fruiting bodies. The two alleles at the locus are defined by specific DNA regions comprising 3.8 kb for mat+ and 4.7 kb for mat–, which have identical flanking sequences. Here we present the characterization of several mutants that have lost mat+-specific sequences. One mutant was obtained fortuitously and the other two were constructed by gene replacement. The mutants are deficient in mating with strains of either mat genotype but are still able to differentiate sexual reproductive structures. The loss of the mating type does not lead to any discernible phenotype during vegetative growth: in particular it does not change the life span of the strain. The mutants can recover mating ability if they are transformed with DNA containing the complete mat+ or mat– information. The transformants behave in crosses as do the reference mat+ or mat– strains, thus indicating that the transgenic mat+ and mat– are fully functional even when they have integrated at ectopic sites.     

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).