A revision of the <Emphasis Type="Italic">Heterospathe elegans</Emphasis> (<Emphasis Type="Italic">Arecaceae</Emphasis>) complex in New Guinea

Summary  Three closely affiliated species of Heterospathe Scheff. (H. elegans (Becc.) Becc., H. humilis Becc. and H. versteegiana Becc.) from New Guinea are revised. They are reduced to a single species which is divided into two subspecies, and the new combination H. elegans subsp. humilis (Becc.) M. S. Trudgen & W. J. Baker is made. The subspecies can be readily distinguished by their growth habit. Epitypes are designated for the three previously published names, as informative material on the habit is not included in the existing type specimens. A new, potentially related species is described as H. pullenii M. S. Trudgen & W. J. Baker.     

<Emphasis Type="Italic">Athyrium crassicaule</Emphasis> (<Emphasis Type="Italic">Woodsiaceae</Emphasis>), a new fern species from the Drakensberg,southern Africa

Summary  A new Athyrium species, A. crassicaule J. P. Roux (Woodsiaceae), is described from the Drakensberg in Lesotho and South Africa. A key to the Athyrium species in Lesotho, South Africa and Swaziland is provided.     

New names,combinations and synonyms in <Emphasis Type="Italic">Justicia</Emphasis> and <Emphasis Type="Italic">Stenostephanus</Emphasis> (Acanthaceae)

Summary  Preparation of a catalogue of the plants of Colombia requires 30 new combinations in Stenostephanus Nees and five in Justicia L. to bring nomenclature up to date. Three new names Justicia littoralis J. R. I. Wood, J. albicans J. R. I. Wood and J. magdalenensis J. R. I. Wood are coined for Jacobinia lindaviana Rusby, Chaetothylax leucantha Leonard and Justicia disparifolia J. R. I. Wood respectively. The genus Pelecostemon Leonard is considered to belong to sect. Chaetothylax (Nees) V. A. W. Graham of Justicia L. Several species of Stenostephanus from Colombia and Venezuela are treated as synonyms of earlier species.     

A new species of <Emphasis Type="Italic">Macroptilium</Emphasis> (Benth.) Urb. (<Emphasis Type="Italic">Leguminosae</Emphasis>: <Emphasis Type="Italic">Papilionoideae</Emphasis>: <Emphasis Type="Italic">Phaseolinae</Emphasis>) from North-Eastern Brazil

Summary  A new species of Macroptilium sect. Microcochle (Benth.) J. A. Lackey is herein described from the states of Piauí and Bahia, Brazil. Macroptilium cochleatum is characterized by few-flowered inflorescences, calyx teeth longer than the tube, a tightly four-coiled keel, and linear, patent fruits. The discovery of this unique Macroptilium species, with its distally coiled keel, expands the diagnostic features of the genus. A key to the Brazilian species of sect. Microcochle is provided.     

High susceptibility to respiratory Acinetobacter baumannii infection in A/J mice is associated with a delay in early pulmonary recruitment of neutrophils

Acinetobacter baumannii is an important cause of both community-associated and nosocomial pneumonia, which have become increasingly difficult to treat because of the rapid development of resistance to multiple antibiotics. Despite its clinical importance, the pathogenesis of and host defense against respiratory A. baumannii infection remains largely unknown. To examine host factors that could contribute to the defense, we compared the susceptibilities of A/J and C57BL/6 mice to intranasal (i.n.) inoculation with A. baumannii. We found that A/J mice were significantly more susceptible to infection with higher mortality (P < 0.05) and tissue bacterial burdens (P < 0.01) as well as greater histopathology in the lung and spleen than C57BL/6 mice. More importantly, the high susceptibility of A/J mice was associated with a reduced local proinflammatory cytokine/chemokine (particularly IL-1β, MIP-2 and TNF-α) responses and a significant delay and reduction in the early influx of neutrophils in the lung (P < 0.05). Intranasal administration of neutrophil-inducing chemokine MIP-2 to A/J mice enhanced pulmonary neutrophil influx and partially restored host resistance to A. baumannii to a level comparable to the more resistant C57BL/6 mice. Our results imply that the early recruitment of neutrophils into the lung is critical for initiating an efficient host defense against respiratory A. baumannii infection.     

Molecular and biochemical characterization of an extracellular serine-protease from <Emphasis Type="Italic">Vibrio metschnikovii</Emphasis> J1

A protease-producing bacterium was isolated from an alkaline wastewater of the soap industry and identified as Vibrio metschnikovii J1 on the basis of the 16S rRNA gene sequencing and biochemical properties. The strain was found to over-produce proteases when it was grown at 30°C in media containing casein as carbon source (14,000 U ml⁻¹). J1 enzyme, the major protease produced by V. metschnikovii J1, was purified by a three-step procedure, with a 2.1-fold increase in specific activity and 33.3% recovery. The molecular weight of the purified protease was estimated to be 30 kDa by SDS-PAGE and gel filtration. The N-terminal amino acid sequence of the first 20 amino acids of the purified J1 protease was AQQTPYGIRMVQADQLSDVY. The enzyme was highly active over a wide range of pH from 9.0 to 12.0, with an optimum at pH 11.0. The optimum temperature for the purified enzyme was 60°C. The activity of the enzyme was totally lost in the presence of PMSF, suggesting that the purified enzyme is a serine protease. The kinetic constants K _m and K _cat of the purified enzyme using N-succinyl-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide were 0.158 mM and 1.14 × 10⁵ min⁻¹, respectively. The catalytic efficiency (K _cat /K _m) was 7.23 × 10⁸ min⁻¹ M⁻¹. The enzyme showed extreme stability toward non-ionic surfactants and oxidizing agents. In addition, it showed high stability and compatibility with some commercial liquid and solid detergents. The aprJ1 gene, which encodes the alkaline protease from V. metschnikovii J1, was isolated, and its DNA sequence was determined. The deduced amino acid sequence of the preproenzyme differs from that of V. metschnikovii RH530 detergent-stable protease by 12 amino acids, 7 located in the propeptide and 5 in the mature enzyme.     

Two new Brazilian species and new combinations in <Emphasis Type="Italic">Euploca</Emphasis> (Heliotropiaceae)

Summary  Two new species of Euploca (Heliotropiaceae): E. krapovickasii J. I. M. Melo & Semir and E. pottii J. I. M. Melo & Semir are described for Brazil, and 12 new combinations are proposed in neotropical Euploca. Discussions of taxonomic relationships based on morphology, with illustrations and information on geographical distribution, flowering and fruiting phenology are presented.     

A modified assay to determine hemolytic toxin variability among Karenia clones isolated from the Gulf of Mexico

Karenia brevis (formerly Gymnodinium breve) is a toxic marine dinoflagellate generally restricted to the Gulf of Mexico and is the main causative organism in fish kills, shellfish intoxications and respiratory distress in humans following bloom events. K. mikimotoi is a morphologically similar co-occurring species which is toxic in other parts of the world oceans, but has not been recognized as a major contributor in toxicity of blooms within the Gulf of Mexico. Recently there has been increasing evidence of the simultaneous production of a variety of bioactive compounds in addition to potent neurotoxins (brevetoxin) in Karenia brevis isolates. These compounds are potentially ichthyotoxic and have been shown to cause hemolysis in several bioassays [Eshbach, E., Scharsack, J., John, U., Medlin, L., 2001. Improved erythrocyte lysis assay in microtitre plates for the sensitive detection and efficient measurement of haemolytic compounds from ichthyotoxic algae. J. Appl. Toxicol. 21, 513–519; Kirkpatrick, B., Fleming, L.E., Squicciarini, D., Backer, L.C., Clark, R., Abraham, W., Benson, J., Cheng, Y.S., Johnson, D., Pierce, R., Zaias, J., Bossart, G.D., Baden, D.G., 2004. Literature review of Florida red tide: implications for human health effects. Harmful Algae 3, 99–115]. Presence of hemolytic compounds may therefore add to the overall toxicity levels of bloom events. Current monitoring methods include assays which are highly sensitive in brevetoxin detection and yet may not target other harmful compounds.By adapting protocols developed by Eshbach et al. [Eshbach, E., Scharsack, J., John, U., Medlin, L., 2001. Improved erythrocyte lysis assay in microtitre plates for the sensitive detection and efficient measurement of haemolytic compounds from ichthyotoxic algae. J. Appl. Toxicol. 21, 513–519], Red drum (Sciaenops ocellatus) erythrocytes were used to create a modified bioassay to detect hemolytic activity of crude algal extracts. Red drum was selected because it is endemic to coastal areas throughout the Gulf of Mexico and is sensitive to Karenia blooms, and thus makes this species a valid ecological target. Preliminary data has shown this method is sensitive for use in assessing hemolysis induced by laboratory cultures down to levels of 1 × 10³ cells mL⁻¹. Results showed an unexpectedly high level of hemolytic activity among K. mikimotoi clones, with one Texas strain inducing significantly higher hemolysis compared to Florida K. brevis isolates. Using this approach, future research efforts will examine the difference in production of hemolytic compounds among various Karenia clones.     

The use and limits of AFLP data in the taxonomy of polyploid wild potato species in Solanum series Conicibaccata

Solanum sect. Petota (tuber-bearing wild and cultivated potatoes) are a group of approximately 190 wild species distributed throughout the Americas from the southwestern United States south to Argentina, Chile, and Uruguay. Solanum series Conicibaccata are a group of approximately 40 species within sect. Petota, distributed from central Mexico to central Bolivia, composed of diploids (2n = 2x = 24), tetraploids (2n = 4x = 48) and hexaploids (2n = 6x = 64); the polyploids are thought to be polysomic polyploids. This study initially was designed to address species boundaries of the four Mexican and Central American species of series Conicibaccata with AFLP data with the addition of first germplasm collections of one of these four species, Solanum woodsonii, as a follow-up to prior morphological, chloroplast DNA, and RAPD studies; and additional species of series Conicibaccata from South America. AFLP data from 12 primer combinations (1722 polymorphic bands) are unable to distinguish polyploid species long thought to be distinct. The data suggest a complex reticulate history of the tetraploids or the need for a broad downward reevaluation of the number of species in series Conicibaccata, a trend seen in other series of sect. Petota. Separately, through flow cytometry, we report the first ploidy level of S. woodsonii, as tetraploid (2n = 48). The U.S. Government’s right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Colonization of cucumber plants by the biocontrol fungus Clonostachys rosea f. catenulata

Clonostachys rosea f. catenulata (Gliocladium catenulatum) strain J1446 (formulated as Prestop WP) suppressed Fusarium root and stem rot caused by Fusarium oxysporum f. sp. radicis-cucumerinum (Forc) on cucumber plants grown hydroponically in rockwool medium. Sixty days following application at seeding, the biocontrol agent had proliferated through the rockwool blocks and was present on cucumber roots and the crown region of the stem at populations >1 × 10⁵ CFU/g fresh weight. Scanning electron micrographs showed that C. rosea had rapidly colonized the root surface and was associated with root hairs and epidermal cell junctions. Following transformation of the fungus with Agrobacterium tumefaciens strain AGL-1 containing the hygromycin resistance (hph) and β-glucuronidase (uidA) genes, blue-stained mycelia could be seen growing on the surface and within epidermal and cortical cells of roots, stems and shoots 3 weeks after treatment. Quantification of GUS activity by fluorometric assays showed that fungal biomass was highest in the roots and crown area, while the extent of colonization of upper stems and true leaves was variable. Higher population levels resulted following application to rockwool blocks compared to seed treatment. Application of C. rosea preceding inoculation with Forc significantly reduced pathogen populations on roots compared to plants inoculated with Forc alone. Colonization of infection sites in the root zone reduced pathogen development and disease incidence. Densities of the biocontrol agent appeared to increase in the presence of the pathogen.     

Onychomycosis in São Paulo,Brazil

Onychomycosis is a fungal infection of the nails with broad aetiological scope, and it represents 18–40% of all onychopathies and 39% of all superficial mycotic infections. From July 1996 to December 1999, samples of nails were collected from 588 patients with presumptive diagnosis of onychomycosis at the Dermatology and Mycology Divisions EPM\UNIFESP, Brazil, and the diagnosis was confirmed in 247 of these cases. The most common pathogens isolated in this study were yeasts in 52% of positive cultures (Candida albicans 18.3%, Candida parapsilosis 13.8%, other species of Candida 15.4% and other yeasts 4.6%), followed by dermatophytes in 40.6% of positive cultures (the most commonly isolated organisms were Trichophyton rubrum in 33.2%, followed by Trichophyton mentagrophytes in 6.3% and others 1.2%). Non-dermatophyte moulds were isolated in 7.4% of positive cultures (Fusarium spp. 4.5%, Nattrassia mangiferae 2.3% and Aspergillus spp. 0.6%). Distal and lateral subungual onychomycosis (DLSO) was the commonest clinical pattern 44.6% followed by free edge onycholysis (FEO) 38.8% and others. In conclusion, this study demonstrated that T. rubrum is the main agent causing onychomycosis in toenails, and species of genus Candida were the main agents isolated in fingernail onychomycosis in our region.     

Comparative gene mapping in cattle,Indian muntjac,and Chinese muntjac by fluorescence in situ hybridization

The Indian muntjac (Muntiacus muntjak vaginalis) has a karyotype of 2n = 6 in the female and 2n = 7 in the male. The karyotypic evolution of Indian muntjac via extensive tandem fusions and several centric fusions are well documented by molecular cytogenetic studies mainly utilizing chromosome paints. To achieve higher resolution mapping, a set of 42 different genomic clones coding for 37 genes and the nucleolar organizer region were used to examine homologies between the cattle (2n = 60), human (2n = 46), Indian muntjac (2n = 6/7) and Chinese muntjac (2n = 46) karyotypes. These genomic clones were mapped by fluorescence in situ hybridization (FISH). Localization of genes on all three pairs of M. m. vaginalis chromosomes and on the acrocentric chromosomes of M. reevesi allowed not only the analysis of the evolution of syntenic regions within the muntjac genus but also allowed a broader comparison of synteny with more distantly related species, such as cattle and human, to shed more light onto the evolving genome organization. For convenience and to avoid confusion we added for each species a three letter abbreviation prior to the chromosomal band location discussed in this paper: BTA, Cattle chromosome; HSA, Human chromosome; MMV, M. m. vaginalis chromosome; MRE, M. reevesi chromosome.     

A novel extracellular phospholipase C purified from a marine bacterium, <Emphasis Type="Italic">Pseudoalteromonas</Emphasis> sp. J937

Marine bacterial isolates were screened for phospholipase C (PLC) activity on PCY agar plates containing phosphatidylcholine (PC) as substrate. The strain that showed the highest activity on a PCY screening agar plate and a thin-layer chromatography was identified as a strain of Pseudoalteromonas and subsequently designated Pseudoalteromonas sp. J937. The extracellular PLC of the strain J937 was purified to a specific activity of 33 U mg⁻¹ protein by serial ion exchange and gel filtration column chromatography. It had a molecular mass of 32 kDa estimated by SDS–PAGE. The optimal pH and temperature of the enzyme were about pH 8 and 45°C, respectively. The PLC hydrolyzed phosphatidylethanolamine as well as PC but not other glycerophospholipids. Its activity was enhanced by 150% with Ca²⁺ (200 mM) and by 180% with Na⁺ (500 mM), suggesting that the purified PLC is a marine-type enzyme.     

Effects of carbon concentration and carbon to nitrogen ratio on the growth and sporulation of several biocontrol fungi

Effects of carbon concentration and carbon to nitrogen (C:N) ratio on six biocontrol fungal strains are reported in this paper. All fungal strains had extensive growth on the media supplemented with 6–12 g l⁻¹ carbon and C:N ratios from 10:1 to 80:1, and differed in nutrient requirements for sporulation. Except for the two strains of Paecilomyces lilacinus, all selected fungi attained the highest spore yields at a C:N ratio of 160:1 when the carbon concentration was 12 g l⁻¹ for Metarhizium anisopliae SQZ-1-21, 6 g l⁻¹ for M. anisopliae RS-4-1 and Trichoderma viride TV-1, and 8 g l⁻¹ for Lecanicillium lecanii CA-1-G. The optimal conditions for P. lilacinus sporulation were 8 g l⁻¹ carbon with a C:N ratio of 10:1 for M-14 and 12 g l⁻¹ carbon with a C:N ratio of 20:1 for IPC-P, respectively. The results indicated that the influence of carbon concentration and C:N ratio on fungal growth and sporulation is strain dependent; therefore, consideration for the complexity of nutrient requirements is essential for improving yields of fungal biocontrol agents.     

Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance

The bar gene was introduced into the cork oak genome. Cork oak embryogenic masses were transformed using the Agrobacterium strain AGL1 which carried the plasmid pBINUbiBar. This vector harbours the genes, nptII and bar, the latter under control of the maize ubiquitin promoter. The transgenic embryogenic lines were cryopreserved. Varying activities of phosphinothricin acetyl transferase were detected among the lines, which carried 1–4 copies of the insert. Molecular and biochemical assays confirmed the stability and expression of the transgenes 3 months after thawing the cultures. These results demonstrate genetic engineering of herbicide tolerance in Quercus spp. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Rubén álvarez, Ricardo J. Ordás are contributed equally.     

Effects of the estuarine dinoflagellate Pfiesteria shumwayae (Dinophyceae) on survival and grazing activity of several shellfish species

A series of experiments was conducted to examine effects of four strains of the estuarine dinoflagellate, Pfiesteria shumwayae, on the behavior and survival of larval and adult shellfish (bay scallop, Argopecten irradians; eastern oyster, Crassostrea virginica; northern quahogs, Mercenaria mercenaria; green mussels, Perna viridis [adults only]). In separate trials with larvae of A. irradians, C. virginica, and M. mercenaria, an aggressive predatory response of three strains of algal- and fish-fed P. shumwayae was observed (exception, algal-fed strain 1024C). Larval mortality resulted primarily from damage inflicted by physical attack of the flagellated cells, and secondarily from Pfiesteria toxin, as demonstrated in larval C. virginica exposed to P. shumwayae with versus without direct physical contact. Survival of adult shellfish and grazing activity depended upon the species and the cell density, strain, and nutritional history of P. shumwayae. No mortality of the four shellfish species was noted after 24 h of exposure to algal- or fish-fed P. shumwayae (strains 1024C, 1048C, and CCMP2089) in separate trials at ≤5 × 10³ cells ml⁻¹, whereas higher densities of fish-fed, but not algal-fed, populations (>7–8 × 10³ cells ml⁻¹) induced low (≤15%) but significant mortality. Adults of all four shellfish species sustained >90% mortality when exposed to fish-fed strain 270A1 (8 × 10³ cells ml⁻¹). In contrast, adult M. mercenaria and P. viridis exposed to a similar density of fish-fed strain 2172C sustained <15% mortality, and there was no mortality of A. irradians and C. virginica exposed to that strain. In mouse bioassays with tissue homogenates (adductor muscle, mantle, and whole animals) of A. irradians and M. mercenaria that had been exposed to P. shumwayae (three strains, separate trials), mice experienced several minutes of disorientation followed by recovery. Mice injected with tissue extracts from control animals fed cryptomonads showed no response. Grazing rates of adult shellfish on P. shumwayae (mean cell length ±1 standard error [S.E.], 9 ± 1 μm) generally were significantly lower when fed fish-fed (toxic) populations than when fed populations that previously had been maintained on algal prey, and grazing rates were highest with the nontoxic cryptomonad, Storeatula major (cell length 7 ± 1 μm). Abundant cysts of P. shumwayae were found in fecal strands of all shellfish species tested, and ≤45% of the feces produced viable flagellated cells when placed into favorable culture conditions. These findings were supported by a field study wherein fecal strands collected from field-collected adult shellfish (C. virginica, M. mercenaria, and ribbed mussels, Geukensia demissa) were confirmed to contain cysts of P. shumwayae, and these cysts produced fish-killing flagellated populations in standardized fish bioassays. Thus, predatory feeding by flagellated cells of P. shumwayae can adversely affect survival of larval bivalve molluscs, and grazing can be depressed when adult shellfish are fed P. shumwayae. The data suggest that P. shumwayae could affect recruitment of larval shellfish in estuaries and aquaculture facilities; shellfish can be adversely affected via reduced filtration rates; and adult shellfish may be vectors of toxic P. shumwayae when shellfish are transported from one geographic location to another.     

Ontogenetic changes in the ecological function of the association behavior between jack mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis> and jellyfish

Commensal behavior of jack mackerel Trachurus japonicus (Temminck & Schlegel) with jellyfishes has been widely observed but its ecological function is still unclear. The goal of the present research is to examine the function of association behavior with jellyfish in the laboratory and in field observations with an emphasis on ontogenetic changes. In the laboratory, jack mackerel juveniles (mean standard length (SL) = 11, 19, 38, and 55 mm) were placed in 500-l polycarbonate tanks with two live moon jellyfish, Aurelia aurita (Linné), and one artificial jellyfish made of silicon. Association behavior with either live or artificial jellyfish was visually observed under the following conditions: control, presence of a predator model, before and after feeding live Artemia, 1 h and 3 h after feeding, and at night. Jack mackerel at 11 mm SL associated with both the moon jellyfish and artificial jellyfish, unrelated to the presence of a predator model or feeding. Juveniles at 19 mm associated with moon jellyfish only in the presence of a predator model. Larger juveniles associated with moon jellyfish at 1 h and 3 h after feeding. Thus the ecological function of association was proposed to develop first from school formation, next as a hiding place from predators, and then as a food source. Underwater observations of jack mackerel associating with giant jellyfish Nemopilema nomurai (Kishinouye) in two different areas in the Sea of Japan supported this hypothesis. High predation pressure from benthic piscivorous fishes in the southern area (Tsushima) may encourage association with jellyfish, whereas pressure from pelagic predators in the northern area (Maizuru) may encourage settlement to rocky reef habitats in temperate waters. Thus the jellyfish may also function as a vehicle for the northward migration of this species. Guest editors: K. A. Pitt & J. E. Purcell Jellyfish Blooms: Causes, Consequences, and Recent Advances     

Intestinal lactoflora in Estonian and Norwegian patients with antibiotic associated diarrhea

The disruption of intestinal microbiota is an important risk factor for the development of Clostridium difficile caused antibiotic associated diarrhea (AAD). The role of intestinal lactoflora in protection against C. difficile is unclear. Fecal samples (n = 74) from AAD patients were investigated for C. difficile and lactobacilli by culture and real-time PCR. Lactobacilli were identified by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and sequencing of 16S rRNA. In C. difficile negative cases we found somewhat higher counts of intestinal Lactobacilli (5.02 vs. 2.15 CFU log₁₀/g; p = 0.053) by culture and more frequently Lactobacillus plantarum (33.3% vs. 9.4%; p = 0.03) as compared with positive ones. Results of total counts of lactobacilli comparing Estonian and Norwegian samples were conflicting by culture and PCR. We found higher colonization of Norwegian AAD patients with L. plantarum (21% vs. 5%, p = 0.053) and Estonians with Lactobacillus gasseri (19% vs. 2%, p = 0.023). Particular lactobacilli (e.g. L. plantarum) may have a role in protection against C. difficile, whereas the meaning of total counts of lactobacilli remains questionable. In different persons and nations, different lactobacilli species may have a protective role against C. difficile.     

Squamocin mode of action to stimulate biofilm formation of Pseudomonas plecoglossicida J26, a PAHs degrading bacterium

Squamocin, an annonaceous acetogenin (ACG) extracted from Annona cherimolia (Annonaceae), has been shown to increase biofilm production of Pseudomonas plecoglossicida J26 (closely related to P. plecoglossicida), a polycyclic aromatic hydrocarbon degrading bacterium. PAHs have become priority pollutants for bioremediation due to their carcinogenicity and toxicity. The effect of various stressful stimuli (naphthalene, octanol, HCl, and NaCl) on cell growth, biofilm formation and autoinducer production of P. plecoglossicida were evaluated and compared with the effect of squamocin to establish its mode of action on biofilm formation. All stressors that inhibited growth stimulated autoinducer production while squamocin was growth stimulant at concentrations above 2.5 μg ml⁻¹. Despite structural similarities, squamocin is not an autoinducer agonist. It indirectly stimulates autoinducer production and increases P. plecoglossicida J26 cell growth. This is the first report on an ACG mode of action in the formation of biofilm in a naphthalene-degrading strain.     

Generic relationships and dating of lineages in Winteraceae based on nuclear (ITS) and plastid (rpS16 and psbA-trnH) sequence data

Phylogenetic analyses of representative species from the five genera of Winteraceae (Drimys, Pseudowintera, Takhtajania, Tasmannia, and Zygogynum s.l.) were performed using ITS nuclear sequences and a combined data-set of ITS + psbA-trnH + rpS16 sequences (sampling of 30 and 15 species, respectively). Indel informativity using simple gap coding or gaps as a fifth character was examined in both data-sets. Parsimony and Bayesian analyses support the monophyly of Drimys, Tasmannia, and Zygogynum s.l., but do not support the monophyly of Belliolum, Zygogynum s.s., and Bubbia. Within Drimys, the combined data-set recovers two subclades. Divergence time estimates suggest that the splitting between Drimys and its sister clade (Pseudowintera + Zygogynum s.l.) occurred around the end of the Cretaceous; in contrast, the divergence between the two subclades within Drimys is more recent (15.5–18.5 MY) and coincides in time with the Andean uplift. Estimates suggest that the earliest divergences within Winteraceae could have predated the first events of Gondwana fragmentation.