Lignans in plant cell and organ cultures: An overview

Lignans are found in a wide variety of plant species. The lignan podophyllotoxin is of special interest, since its derivatives like e.g. etopophos® are used in anticancer therapy. As chemical synthesis of podophyllotoxin is not yet economic, it still has to be isolated from wild growing Podophyllum species, some of which are considered to be endangered species. Therefore plant in vitro cultures may serve as alternative sources for podophyllotoxin and for other types of lignans as well. This review describes the establishment of plant cell and tissue cultures for lignan production and the experiments to improve product yields by changing the cultivation parameters, addition of elicitors and feeding of precursors. It also summarizes the use of plant cell and organ cultures to study the biosynthesis of lignans on enzymological level. Abbreviations: DOP – deoxypodophyllotoxin; LARI – lariciresinol; MATAI – matairesinol; 6MPTOX – 6-methoxypodophyllotoxin; PINO – pinoresinol; PTOX – podophyllotoxin; SECO – secoisolariciresinol     

Knots in trees – A new rich source of lignans

Recent research in our group has revealed that knots, i.e. the branch bases inside tree stems, commonly contain 5–10% (w/w) of lignans. Norway spruce (Picea abies) knots contain as much as 6–24% of lignans, with 7-hydroxymatairesinol (HMR) as the predominant (70–85%) lignan. Some other spruce species also contain HMR as the main lignan, but some spruce species have also other dominating lignans. Most fir (Abies) species contain secoisolariciresinol and lariciresinol as the main lignans. Lignans occur also in knots of pines (Pinus spp.), although in lower amounts than in spruces and firs. Scots pine (Pinus silvestris) knots were found to contain 0.4–3% of lignans with nortrachelogenin as the main lignan. Lignans have been identified also in knots of some hardwoods, although flavonoids are more abundant in hardwoods. Knots are detrimental in the manufacture of pulp and paper and should preferably be removed before pulping. This is possible using a recently developed industrially applicable process called ChipSep. Recent research has also established novel synthetic routes to several lignans, such as matairesinol, secoisolariciresinol, lariciresinol and cyclolariciresinol, starting from hydroxymatairesinol by applying fairly straight-forward chemical transformations. We conclude that wood knots in certain spruce and fir species constitute the richest known source of lignans in nature. The lignans occur in knots in free form and are easily extracted by aqueous ethanol, or even by water. Not only HMR, but also other potentially valuable lignans, could be produced in a scale of hundreds of tons per year by extraction of knots separated from wood chips at pulp and paper mills.     

Discriminating ecological processes affecting different dimensions of α‐ and β‐diversity in desert plant communities

Understanding the spatial distribution of plant diversity and its drivers are major challenges in biogeography and conservation biology. Integrating multiple facets of biodiversity (e.g., taxonomic, phylogenetic, and functional biodiversity) may advance our understanding on how community assembly processes drive the distribution of biodiversity. In this study, plant communities in 60 sampling plots in desert ecosystems were investigated. The effects of local environment and spatial factors on the species, functional, and phylogenetic α‐ and β‐diversity (including turnover and nestedness components) of desert plant communities were investigated. The results showed that functional and phylogenetic α‐diversity were negatively correlated with species richness, and were significantly positively correlated with each other. Environmental filtering mainly influenced species richness and Rao quadratic entropy; phylogenetic α‐diversity was mainly influenced by dispersal limitation. Species and phylogenetic β‐diversity were mainly consisted of turnover component. The functional β‐diversity and its turnover component were mainly influenced by environmental factors, while dispersal limitation dominantly effected species and phylogenetic β‐diversity and their turnover component of species and phylogenetic β‐diversity. Soil organic carbon and soil pH significantly influenced different dimensions of α‐diversity, and soil moisture, salinity, organic carbon, and total nitrogen significantly influenced different dimensions of α‐ and β‐diversity and their components. Overall, it appeared that the relative influence of environmental and spatial factors on taxonomic, functional, and phylogenetic diversity differed at the α and β scales. Quantifying α‐ and β‐diversity at different biodiversity dimensions can help researchers to more accurately assess patterns of diversity and community assembly.     

Different strategies for the chemical synthesis of lignans

This paper summarises the results of three projects. The first is concerned with developing general routes for the synthesis of lignans. In particular, two routes involving tandem conjugate addition reactions and Diels Alder reactions respectively that have been used to synthesise podophyllotoxin derivatives are described. The second project is concerned with the asymmetric synthesis of lignans and involves the application of these reactions, with the introduction of a menthyloxy group as a chiral auxiliary, to achieve the asymmetric synthesis of podophyllotoxin derivatives. The third project is concerned with the attempted biomimetic syntheses of podophyllotoxin derivatives using oxidative coupling reactions. Attention is focussed primarily on the use of hypervalent iodine reagents, which yield stegane and isostegane derivatives rather than podophyllotoxin derivatives. Other examples of biaryl coupling leading to stegane and isostegane derivatives are included, and other examples of lignan synthesis involving hypervalent iodine reagents are also described. Abbreviations: DDQ – 2,3-dichloro-5,6-dicyano-1,4-benzoquinone; DMAD – dimethyl acetylenedicarboxylate (dimethyl butynedioate); PIDA – phenyliodonium diacetate, iodobenzene diacetate; PIFA – phenyliodonium bis(trifluoroacetate), [bis(trifluoroacetoxy) iodobenzene]; Ra-Ni – Raney nickel; TBAF – tetrabutylammonium fluoride; TBDMS –t-butyldimethylsilyl; TFA – trifluoroacetic acid; TFAA – trifluoroacetic anhydride; TFE – 2,2,2-trifluoroethanol; TTFA – thallium(III) trifluoroacetate.     

Ribosomal DNA non-transcribed spacer polymorphism in subterranean mole rats: genetic differentiation,environmental correlates and phylogenetic relationships

Summary An analysis is presented of genetic differentiation in the non-transcribed spacers of ribosomal DNA (NTS rDNA). Diversity, environmental correlates and the phylogenetic relationships are examined within and between species of the actively speciating subterranean mole rat, superspeciesSpalax ehrenbergi (2n=52, 54, 58, 60) in Israel. This analysis is based on a previous study of the geographic distribution of restriction fragment length polymorphisms of NTS rDNA. Here we present results indicating that NTS rDNA diversity exists mostly (66%) within populations, while 20% is between populations within species, and 14% between species. Multivariate discriminant analysis succeeded in separating 10 of the 13 populations (77%) into their correct chromosomal species, on the basis of the combination of three NTS rDNA repetypes. The phylogenetic relationships suggest that the complex involves two pairs of closely related species (2n=52–54 and 2n=58–60). NTS rDNA diversity, as well as the decrease southward in frequency of repetype C, are correlated with climatic factors of humidity and temperature. These data are discussed in terms of the evolutionary forces of migration and selection which may cause NTS rDNA differentiation. Climatic selection appears to be the major differentiating factor of NTS rDNA.     

Linum persicum: Lignans and placement in Linaceae†

Aryltetralin lignans (podophyllotoxin type) are the main lignan constituents of species belonging to Linum section Syllinum (Linaceae). Linum persicum, a perennial plant native to Iran closely related to L. album, has not yet been studied. To evaluate the lignan profile, fresh plants of L. persicumwere collected and divided into different parts and analyzed by HPLC. The main aryltetralin lignans found inL. persicumplant parts, callus and cell cultures were podophyllotoxin (PTOX), 6-methoxypodophyllotoxin (MPTOX) and - and -peltatin. Furthermore, the systematic relationship between L. persicum and other Linum species are discussed in the light of morphological and phytochemical aspects. Abbreviations: MPTOX – 6-methoxypodophyllotoxin; PTOX – podophyllotoxin; DOP – deoxypodophyllotoxin.     

Low Genetic Differentiation of and Close Evolutionary Relationships between Anas platyrhynchos and Anas poecilorhyncha: RAPD–PCR Evidence

Using RAPD–PCR, we examined genetic diversity and phylogenetic relationships in two groups of river ducks: Anas platyrhynchos, A. poecilorhyncha, A. streperaand A. crecca, A. formosa, A. querquedula. Molecular taxon-specific markers were found for teals (A. crecca, A. formosa, A. querquedula) and gadwall (A. strepera). Each of the species examined was shown to exhibit high genetic diversity. The mean levels of intraspecific genetic polymorphism in the groups of mallards (P = 77%) and teals (P = 74.5%) were approximately equal whereas the mean interspecific genetic distances in teals were significantly higher than in mallards (D = 0.432 and D= 0.336, respectively). The levels of interspecific genetic differentiation in the species groups were also different. The genetic distances between the teal species and between gadwall and mallards were equal to 0.668–0.971 while the genetic distance between mallard A. platyrhynchos and spot-billed duck A. poecilorhyncha was 0.401, which slightly exceeds the intraspecific values for mallards (0.356–0.377). The RAPD patterns for this species pair showed high variability and a lack of fixed differences. This was adequately reflected on both intra- and interspecific differences and on phylogenetic constructions in which the morphological species did not form their own clusters but were intermixed. In contrast to mallards, the other species, which showed high genetic variability, were reliably separated in phenogenetic and phylogenetic reconstructions. The possible explanations of the low genetic differentiation of A. platyrhynchos and A. poecilorhynchaare discussed.     

Genetic Diversity in Remnant Mainland and “Pristine” Island Populations of Three Endemic Australian Macropodids (Marsupialia): Macropus Eugenii, Lagorchestes Hirsutus and Petrogale Lateralis

Since European settlement, mainland Australia has experienced a wave of mammal extinctions and population declines. However, some species have persisted on off-shore islands, which are now viewed as important wildlife refuges. In this study, we assessed the level of genetic diversity, at 7–11 microsatellite loci, in island and remnant mainland populations of three endemic species of macropodid marsupial; the tammar wallaby Macropus eugenii(n = 92); rufous hare-wallaby Lagorchestes hirsutus(n = 40) and black-footed rock-wallaby Petrogale lateralis(n = 164). There was a consistent pattern of significantly higher levels of microsatellite diversity in the remnant mainland population (A= 4.9–13.9; He= 0.61–0.86) of each species compared to conspecific pristine island populations (A= 1.2–3.7; He= 0.05–0.44). These marked differences are even apparent where island populations currently have a substantially larger census size. In addition, island populations were substantially inbred (Fe= 0.49–0.91). Although island populations have been insulated from the relatively recent threatening processes operating on the mainland, they have nevertheless been significantly impacted by increased inbreeding and the substantial erosion of genetic diversity. Despite the difficulties of ensuring the survival of remnant mainland populations, they appear to retain substantially more genetic diversity than their island counterparts and therefore are more likely to contribute to the long-term persistence of their species. These data also demonstrate that small remnant mainland populations (n < 10–20) are often capable of rapid recovery and are not necessarily genetically depauperate.     

The western red cedar (Thuja plicata) 8-8′ DIRIGENT family displays diverse expression patterns and conserved monolignol coupling specificity

The isolation and characterization of a multigene family of the first class of dirigent proteins (namely that mainly involved in 8-8 coupling leading to (+)-pinoresinol in this case) is reported, this comprising of nine western red cedar (Thuja plicata) DIRIGENT genes (DIR1–9) of 72–99.5% identity to each other. Their corresponding cDNA clones had coding regions for 180–183 amino acids with each having a predicted molecular mass of ca. 20 kDa including the signal peptide. Real time-PCR established that the DIRIGENT isovariants were differentially expressed during growth and development of T. plicata (P<0.05). The phylogenetic relationships and the rates and patterns of nucleotide substitution suggest that the DIRIGENT gene may have evolved via paralogous expansion at an early stage of vascular plant diversification. Thereafter, western red cedar paralogues have maintained an high homogeneity presumably via a concerted evolutionary mode. This, in turn, is assumed to be the driving force for the differential formation of 8-8-linked pinoresinol derived (poly)lignans in the needles, stems, bark and branches, as well as for massive accumulation of 8-8-linked plicatic acid-derived (poly)lignans in heartwood.     

The effect of<Emphasis Type="Italic"> pfl</Emphasis> gene knockout on the metabolism for optically pure <Emphasis Type="SmallCaps">d</Emphasis>-lactate production by<Emphasis Type="Italic"> Escherichia coli</Emphasis>

The effect of gene knockout on metabolism in the pflA^–, pflB^–, pflC^–, and pflD^– mutants of Escherichia coli was investigated. Batch cultivations of the pfl ^– mutants and their parent strain were conducted using glucose as a carbon source. It was found that pflA^– and pflB^– mutants, but not pflC^– and pflD^– mutants, produced large amounts of d-lactate from glucose under the microaerobic condition, and the maximum yield was 73%. In order to investigate the metabolic regulation mechanism, we measured enzyme activities for the following eight enzymes: glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), pyruvate kinase, lactate dehydrogenase (LDH), phosphoenolpyruvate carboxylase, acetate kinase, and alcohol dehydrogenase. Intracellular metabolite concentrations of glucose 6-phosphate, fructose 1,6-bisphosphate, phosphoenolpyruvate, pyruvate, acetyl coenzyme A as well as ATP, ADP, AMP, NADH, and NAD⁺ were also measured. It was shown that the GAPDH and LDH activities were considerably higher in pflA^– and pflB^– mutants, which implies coupling between NADH production and consumption between the two corresponding reactions. The urgent energy requirement was shown by the lower ATP/AMP level due to both oxygen limitation and pfl gene knockout, which promoted significant stepping-up of glycolysis when using glucose as a carbon source. It was shown that the demand for energy is more important than intracellular redox balance, thus excess NADH produced through GAPDH resulted in a significantly higher intracellular NADH/NAD⁺ ratio in pfl ^– mutants. Consequently, the homolactate production was achieved to meet the requirements of the redox balance and the energy production through glycolysis. The effect of using different carbon sources such as gluconate, pyruvate, fructose, and glycerol was investigated.     

Selective grazing by the mud snail Ilyanassa obsoleta

Summary Mud snails (Ilyanassa obsoleta) starved for 48 h were allowed to feed on sediments in laboratory microcosms. Sediment cores sliced at 2 mm intervals were compared to snail stomach contents for per cent carbon and nitrogen, plant pigment contents and species composition of benthic diatoms. Concentrations of carbon, nitrogen, phaeopigments, phycocyanin and chlorophyll were enriched in the top 2 mm of the sediments compared to 7–10 mm depth by a factor of 2–10. In turn, these materials were 20–40 times more concentrated in snail guts than in the surface sediments. Snail feces were enriched for carbon and nitrogen by 5–7 times over the surface sediments. Bacterial chlorophyll peaked at about 3–4 mm in the sediments and was not detectable in the snail stomach contents. The C/N ratio of the snail stomach contents was only 6 compared to a ratio of 8.5 for their feces and 12 for the surface sediments.The percentage of migratory diatoms (e.g. Nitzschia and Navicula) decreased with depth where non-migratory species, such as Fragilaria pinnata, dominated. These migratory species were more common in the snails than in the sediments on which they were feeding.A comparison of daily ingestion rates to the animal's energy budget shows that this selective ingestion is sufficient to meet Ilyanassa's energy needs.     

Characterisation of Yeasts Isolated from Deep Igneous Rock Aquifers of the Fennoscandian Shield

The diversity of prokaryotes in the groundwater deep below the surface of the Baltic Sea at the Äspö Hard Rock Laboratory (HRL) in southeast Sweden is well documented. In addition, there is some evidence that eukaryotes, too, are present in the deep groundwater at this site, although their origins are uncertain. To extend the knowledge of eukaryotic life in this environment, five yeast, three yeastlike, and 17 mold strains were isolated from Äspö HRL groundwater between 201 and 444 m below sea level. Phenotypic testing and phylogenetic analysis of 18S rDNA sequences of the five yeast isolates revealed their relationships to Rhodotorula minuta and Cryptococcus spp. Scanning and transmission electron microscopy demonstrated that the strains possessed morphological characteristics typical for yeast, although they were relatively small, with an average length of 3 µm. Enumeration through direct counting and most probable number methods showed low numbers of fungi, between 0.01 and 1 cells mL^–1, at some sites. Five of the strains were characterized physiologically to determine whether they were adapted to life in the deep biosphere. These studies revealed that the strains grew within a pH range of 4–10, between temperatures of 4°C and 25–30°C, and in NaCl concentrations from 0 to 70 g L^–1. These growth parameters suggest a degree of adaptation to the groundwater at Äspö HRL. Despite the fact that these eukaryotic microorganisms may be transient members of the deep biosphere microbial community, many of the observations of this study suggest that they are capable of growing in this extreme environment.     

Influence of a supplementary carbon source on biodegradation of pyridine by freely suspended and immobilizedPimelobacter sp.

The effect of the presence of supplementary glucose or acetate on the growth and pyridine-degrading activity of freely suspended and calcium-alginate-immobilizedPimelobacter sp. was investigated. Although the supplementary carbon sources could be degraded simultaneously with pyridine,Pimelobacter sp. exhibited a preference for pyridine over supplementary carbon sources. Thus, the pyridine-degrading activity of the freely suspended cells was not decreased significantly by the addition of either glucose (1.5–6 mM) or acetate (6–24 mM) to the pyridine (6–24 mM). In the semi-continuous immobilized cell culture, immobilized cells also exhibited a preference for pyridine over supplementary carbon sources and did not switch their substrate preference throughout the culture. Owing to a high cell concentration, the volumetric pyridine degradation rate at 24 mM pyridine in the immobilized cell culture was approximately six times higher than that in the freely suspended cell culture. Furthermore, the immobilized cells could be reused 16 times without losing their pyridine-degrading activity during the culture period tested. Taken together, the use of immobilizedPimelobacter sp. for the degradation of pyridine is quite feasible because of the preference for pyridine over supplementary carbon sources, the high volumetric pyridine degradation rate, and the reusability of immobilized cells.     

Turnover and distribution of root exudates of Zea mays

Decomposition and distribution of root exudates of Zea mays L. were studied by means of ¹⁴CO₂ pulse labeling of shoots on a loamy Haplic Luvisol. Plants were grown in two-compartment pots, where the lower part was separated from the roots by monofilament gauze. Root hairs, but not roots, penetrated through the gauze into the lower part of the soil. The root-free soil in the lower compartment was either sterilized with cycloheximide and streptomycin or remained non-sterile. In order to investigate exudate distribution, 3 days after the ¹⁴C labeling, the lower soil part was frozen and sliced into 15, one-mm thick layers using a microtome. Cumulative ¹⁴CO₂ efflux from the soil during the first 3 days after ¹⁴C pulse labeling did not change during plant growth and amounted to about 13–20% of the total recovered ¹⁴C (41–55% of the carbon translocated below ground). Nighttime rate of total CO₂ efflux was 1.5 times lower than during daytime because of tight coupling of exudation with photosynthesis intensity. The average CO₂ efflux from the soil with Zea mays was about 74 g C g^–1 day^–1 (22 g C m^–2 day^–1), although, the contribution of plant roots to the total CO₂ efflux from the soil was about 78%, and only 22% was respired from the soil organic matter. Zea mays transferred about 4 g m^–2 of carbon under ground during 26 days of growth. Three zones of exudate concentrations were identified from the distribution of the ¹⁴C-activity in rhizosphere profiles after two labeling periods: (1) 1–2 (3) mm (maximal concentration of exudates) 2) 3–5 mm (presence of exudates is caused by their diffusion from the zone 1); (3) 6–10 mm (very insignificant amounts of exudates diffused from the previous zones). At the distance further than 10 mm no exudates were found. The calculated coefficient of exudate diffusion in the soil was 1.9 × 10^–7 cm² s^–1.     

Phenotypic diversity amongst strains of Pleurotus sajor-caju: implications for cultivation in arid environments

In arid regions, biodiversity and biomass are limited by water availability, and this problem has been compounded by desertification associated with global climate change. The saprotrophic macrofungi that are indigenous to hot subtropical and tropical regions, such as Pleurotus spp., can play key roles in water sequestration, nutrient cycling, human nutrition, and bioremediation of waste materials. We studied 15 strains of Pleurotus sajor-caju, a widespread and phenotypically-diverse species, to establish variability in growth response and primordium development over a range of stress parameters: osmotic potential (−0.5 to −5 MPa), temperature (5–40 °C) and pH (2–12). The initiation of primordia precedes basidiome production and therefore represents a key stage in bioremediation strategies and fungi-driven nutrient cycles. Primordia were produced at low pH (4–6), at suboptimal growth temperatures (≤25 °C), and under moderate water stress (−0.5 to −3.5 MPa). Although the growth windows for different strains were similar, their maximum growth rates and the optimum conditions for growth varied. We discuss the phenotypic diversity of Pleurotus strains and discuss their potential for cultivation, bioremediation and ecological regeneration.     

Characterization of bacterial diversity in two aerated lagoons of a wastewater treatment plant using PCR-DGGE analysis

Aerated lagoons are commonly used for domestic and industrial wastewater treatment due to their low cost and minimal need of operational requirements. However, little information is known regarding microbial communities that inhabit these ecosystems. In this study, a 16S-DGGE approach was used to estimate bacterial diversity and to monitor community changes in two aerated lagoons from a wastewater treatment plant receiving urban and industrial effluents. Pronounced shifts between bacterial communities collected in winter–spring and summer–autumn months were detected. Temperature, dissolved oxygen (DO) and pH were the variables that most influenced the bacterial communities. Phylogenetic affiliation of predominant members was assessed by the determination of the 16S rDNA sequence of correspondent bands. Affiliations to Cytophaga–Flexibacter–Bacteroides (CFB) group, Firmicutes, and β- and ε-proteobacteria were found.     

天目山物种多样性尺度依赖及其与空间格局关系的多重分形

以浙江省天目山国家级自然保护区为例,采用多尺度分析思想,利用多重分形分析方法,研究了不同尺度下物种多样性的变化、空间分布格局以及多样性与空间格局之间的关系。研究主要得到3方面的结论：（1）物种多样性具有尺度依赖性,随着空间尺度的增大,Shannon-Wiener多样性指数Ⅳ增大,Margalef多样性指数足和均匀度指数E减小;（2）多重分形参数αmin。多重分形谱的变化范围SR等能够定量反映物种的空间分布特征,空间大尺度越大,物种越聚集,空间分布越不均匀;（3）物种多样性与空间格局存在线性或幂函数关系。研究表明多重分形分析定量描述物种空间格局是有效性的,多重分形参数与生物多样性之间的定量关系为研究物种空间格局、生态属性与尺度之间的关系奠定了基础。因此,分形结合传统方法,在生物多样性方面的研究将有很大的潜在价值。     

Colletotrichum species associated with cassava anthracnose in China

Anthracnose caused by Colletotrichum (CAD) is an economically important disease of cassava; however, research on its species diversity and geographical distribution of Colletotrichum in China remains limited. In this study, we investigated the phylogenetic diversity of Colletotrichum isolates associated with symptomatic leaf tissues of cassava from Guangxi and Yunnan provinces in China and aimed to confirm their identification using advanced techniques. Based on multi‐locus phylogenetic analyses and phenotypic characteristics, we identified four species from the isolates, comprising C. plurivorum, C. karstii, C. fructicola and C. siamense, and we found variation in degrees of virulence among the species and between cassava varieties. Our results are the first to report these species from cassava, and they provide a basis for the development of management strategies for CAD in cassava.     

Surface phytoplankton composition and carbon biomass distribution in the Crozet Basin during austral summer of 1999: variability across frontal zones

Surface phytoplankton assemblages were studied in January/February 1999 in the Crozet Basin (43°50S–45°20S; 61°E–64°30E) between the northern Polar Zone and the Agulhas Front. Cell concentrations increased several fold northwards from the SubAntarctic Zone (SAZ) and reached peak numbers (average 2×10⁶ cells l^–l ) in the central and western Subtropical Zone (STZ). The most spectacular increase in cell numbers occurred at the Subtropical Front (STF) and was attributed to dinoflagellates and diatoms. Nanoflagellates and picoplankton were dominant in the entire area (average 2.8×10⁵–1.6×10⁶ cells l^–l). In the SAZ they were followed by coccolithophorids, dinoflagellates and diatoms. In the STZ coccolithophorids were often outnumbered by dinoflagellates. Diatoms were dominated by Pseudonitzschia delicatissima and were generally the least abundant algae, but reached peak densities of 1.2–4×10⁵ cells l^–l at, and north of the STF. Coccolithophorids contained mainly Emiliania huxleyi, but in the SAZ and STF Gephyrocapsa oceanica was a co-dominant species. Dinoflagellates were dominated by nano-sized species of Gymnodinium, Gyrodinium and Prorocentrum. The numbers of dinoflagellate and coccolithophorid species increased considerably in the convergence zone (STZ), which suggests their in-situ development. Heterotrophic dinoflagellates and ciliates were mainly present in the subtropics. Cell carbon biomass was attributed chiefly to auto- and heterotrophic dinoflagellates (av. 23–72 g C l^–l; 68–87%), showing their important contribution to the carbon flow. Variations in cell concentrations across the fronts and water masses, and the distribution of major species were most likely controlled by the combined effect of such factors as nutrient renewal in the convergence zone, availability of iron, increased water-column stability at fronts, and high horizontal gradients in surface-water temperature.