Genetic differentiation in three Sepia species (Mollusca: Cephalopoda) from Galician waters (north-west Iberian Peninsula)

The systematics of the genus Sepia is not yet clear. Morphological evidence has led to S. officinalis Linnaeus, 1758 being considered as belonging to the subgenus Sepia sensu stricto, and S. orbignyana Férussac, 1826 and S. elegans Blainville, 1827 as belonging to the subgenus Rhombosepion. Samples of 30 individuals of S. officinalis and S. orbignyana from both sides of an oceanographic boundary off the north-west Iberian Peninsula, and a sample of S. elegans from the northern side, were collected in 1993–1994. Allozyme electrophoresis for 32 presumptive loci revealed low levels of genetic variability for the three Sepia species (mean expected heterozygosity estimates were <0.052). No significant differences in allozyme frequencies were detected among populations of either S. officinalis or S. orbignyana. The genetic identities (I) of S. officinalis and S. orbignyana (I=0.12) and of S. elegans (I=0.13) were significantly different from that of S. orbignyana and S. elegans (I=0.49). The former are typical of values for confamilial genera, and a new generic status is proposed for the latter two species, which become Rhombosepion orbignyana (Férussac, 1826) and R. elegans (Blainville, 1827).     

Aesthetascs in Lysmata spp. shrimp: sexual dimorphism and relationship with social environments

Aesthetascs are the olfactory receptor organs responsible for mate recognition in many decapod crustacean species. The number of aesthetascs varies greatly among species in different environments, but the functional significance of this variation is poorly understood. This study examined the relationship between social environment and the number of aesthetascs in six species of caridean shrimp in the genus Lysmata. Generally, the number of aesthetascs was greater in group-living species (L. boggessi and L. wurdemanni) than in low-density (L. ankeri and L. pederseni) and pair-living species (L. amboinensis and L. debelius). Aesthetascs are innervated by olfactory receptor neurons (ORNs). The total number of ORNs is related to the sensitivity of olfactory odor detection, and it was higher in group-living species than in pair-living species. Sexual dimorphism in the number of aesthetascs was present in L. boggessi, L. wurdemanni, and L. ankeri but not in L. pederseni and L. amboinensis.     

Escape responses of four Chilean intertidal limpets to seastars

The Chilean intertidal Lottiidae limpet fauna consists of at least nine species, grouped into the monophyletic taxon Scurria and a single taxon preliminarily designated as `Lottia' orbignyi (Dall). Within the Scurria clade, the most basal species are Scurria variabilis (Sowerby), S. viridula (Lamarck), and S.zebrina (Lesson), and among the most crown groups are S. ceciliana (Orbigny) and S. araucana (Orbigny). This phylogenetic information, obtained by the analysis of molecular characters (16S mtDNA), provides a framework for a comparative study of behavioral characters. In this study, we analyze the escape responses of two basal limpet species (`L.' orbignyi and S. viridula) and two derived limpet species (S. araucana and S. ceciliana) in the presence of the limpet predatory seastar Heliaster helianthus (Lamarck) and the limpet non-predatory seastars Stichaster striatus Muller and Troschel and Patiria chilensis Lutken. Neither P.chilensis nor Stichaster striatus induced escape responses such as those observed with the predatory H. helianthus. Moreover, in the presence of H. helianthus, basal and derived limpet species differed significantly in the percentage of individuals responding, reaction time, and duration of the response. The basal species `L.' orbignyi and Scurriaviridula exhibited instantaneous and vigorous locomotor responses, whereas the derived species S. araucana and S. ceciliana displayed no locomotor responses. The same pattern was found in responses to H. helianthus homogenates. The results strongly suggest a correlation between phylogeny and anti-predator escape responses to the seastar H. helianthus. Alternative escape responses have evolved for basal and derived species within the Chilean clade. Received: 27 April 2000 / Accepted: 16 August 2000     

Sympatry in grapsoid crabs (genera <Emphasis Type="Italic">Planes</Emphasis> and <Emphasis Type="Italic">Plagusia</Emphasis>) from olive ridley sea turtles (<Emphasis Type="Italic">Lepidochelys olivacea</Emphasis>), with descriptions of crab diets and masticatory structures

Grapsoid crabs of the genera Planes and Plagusia are commonly referred to as “rafting crabs” due to their propensity to live on flotsam and pelagic marine animals. Planes minutus and Planes major (=Planes cyaneus) are epibionts of sea turtles. Occurrences of grapsoid crabs in the genera Planes and Plagusia were evaluated on a total of 27 olive ridley sea turtles, Lepidochelys olivacea, from the eastern tropic Pacific (1998–2001) and the Hawaiian Islands (2002) captured in July–December each year. This is the first report of Planes marinus and Plagusia squamosa on sea turtles, and of P. major, P. marinus, and P. squamosa in sympatry on a confined substrate. Stomach content analyses showed P. major and P. marinus consumed a variety of neuston and marine vegetation, with the former consuming considerably more animal material. Epibiotic P. squamosa consumed mostly plant material. The three Planes species had distinctive differences in gastric mill tooth morphology. The versatile mouthparts of P. marinus are described and resemble those of their congeners. Most female P. major and P. marinus collected were ovigerous and present in all survey months.     

Highly repetitive DNA sequences provide evidence for a lack of gene flow between two morphological forms of Herdmania momus (Ascidiacea: Stolidobranchia)

The genomic structure of two morphological forms of the pyurid ascidian Herdmania momus — H. momus forma curvata and H. momus forma grandis—are compared using a rapidly evolving highly repetitive element isolated from the genome of H. momus forma curvata. A 663 bp (base pair) Cla I satellite sequence is present only in the genome of H. momus forma curvata, and hence can be used as a form-specific marker. Low-stringency Southern blot analyses, using the Cla I satellite as a probe, revealed that the genomes of H. momus forma grandis and another pyurid ascidian, Pyura stolonifera, do not contain similar sequences to the H. momus forma curvata repetitive element. In addition to this genomic dissimilarity, there are a number of significant reproductive and developmental differences between the two H. momus forms, and interform fertilisation rates are significantly lower than intraform rates. The molecular, reproductive and developmental differences between H. momus forma curvata and H. momus forma grandis indicate the presence of strong barriers to gene flow.     

Effects of dietary fatty acids on the reproductive success of the calanoid copepod<Emphasis Type="Italic"> Temora longicornis</Emphasis>

Egg production and hatching success of the copepod Temora longicornis were measured in laboratory experiments and in the field (North Sea). In the laboratory, ingestion of four algal species (Thalassiosira weissflogii, Phaeocystis globosa, Isochrysis sp. and Dunaliella tertiolecta) was followed and the content of fatty acids in the algae was determined. The two food types (T. weissflogii and Isochrysis sp.) that provided the highest ingestion of carbon and long chain fatty acids also resulted in the highest egg production rate (E_r) and hatching success (H%). In contrast, D. tertiolecta led to both low ingestion of carbon and long chain fatty acids, resulting in low reproductive success. There was a positive relationship between the amount of eicosapentaenoic fatty acid [20:5(n-3), EPA] ingested and E_r and H%, and of the ratio between docosahexaenoic and eicosapentaenoic fatty acid [22:6(n-3)/20:5(n-3), DHA/EPA] in the ingested food and H%. In the field, chlorophyll a and specific fatty acids were measured and protists were enumerated, in order to investigate the link between these factors and the reproductive success of T. longicornis. Hatching time was found to be related to temperature and exceeded 120 h at 6°C. No relationship was found between chlorophyll a and reproductive success (E_r or H%). E_r correlated with the concentration of diatoms and ciliates, which were the dominating protists in early spring, indicating that food quantity was the limiting factor for E_r. As in the laboratory experiments, H% was dependent on the fatty acid DHA and the ratio of DHA/EPA, which indicates that the quality of eggs (H%) is linked to the quality of food.Communicated by M. Kühl, Helsingør     

Feeding in dominant Antarctic copepods—does the morphology of the mandibular gnathobases relate to diet?

This study should clarify the importance of morphology and stability of the mandibular gnathobases for the diet of Antarctic copepod species. The gnathobase morphology of the dominant copepod species Calanoides acutus, Calanus propinquus, Ctenocalanus citer, Rhincalanus gigas, Metridia gerlachei, Stephos longipes, Microcalanus pygmaeus and Paraeuchaeta antarctica from the Southern Ocean was investigated in detail by means of a scanning electron microscope. The mandibular gnathobases of C. acutus, C. propinquus and C. citer have relatively short and compact teeth. These species feed mainly on diatoms and are able to crack the silicious diatom frustules with their mandibular gnathobases by directed pressure. In contrast the teeth of the mandibular gnathobases of P. antarctica are very long and pointed. The nutrition of this species consists predominantly of other smaller copepod species. The motile prey can be held by skewering, using the gnathobases, and then eventually minced. The mandibular gnathobases of P. antarctica have notably more small bristles than those of the other investigated copepod species. These bristles are probably associated with receptors and could serve to locate the prey. The morphology of the gnathobases of R. gigas and M. gerlachei is between that of P. antarctica on the one side and that of C. acutus, C. propinquus and C. citer on the other. Based on the morphology of its gnathobases the copepod species S. longipes, which has to date been found to feed primarily on phytoplankton, mainly ice algae, must also be considered a zooplankton feeder. The investigation showed that M. pygmaeus has gnathobases with surprisingly long and pointed teeth, indicating that this species very probably feeds both on phyto- and on zooplankton organisms. While the mandibular gnathobases of the males of C. propinquus, R. gigas, M. gerlachei and S. longipes have the same morphology as the females of the respective species, in the other four investigated copepod species the males have reduced (C. acutus, C. citer and M. pygmaeus) or completely missing mandibular gnathobases (P. antarctica). The teeth of the gnathobases of all studied species with the exception of M. gerlachei consist of a different material than the remaining parts of the gnathobases. This material seems to be silicate, which probably enhances the stability of the gnathobase teeth.Communicated by O. Kinne, Oldendorf/Luhe     

Differing distributions of potentially competing amphipods,copepods and gastropods among specimens of the intertidal alga Pelvetia fastigiata

Possible competitive interactions are inferred from the distributions and abundances of species within three invertebrate groups (Hyale spp., Ampithoe spp. and thallusdwelling species) among unmanipulated and experimental specimens of the brown alga Pelvetia fastigiata collected between 1974 and 1976, inclusive, at La Jolla, California, USA. During the fall, when the vagile amphipods Hyale grandicornis and H. frequens were relatively abundant, they were found mostly on specimens of P. fastigiata at upper mid-intertidal and mid-tide levels, respectively. H. grandicornis often dispersed to mid-tide levels, but the two species cooccurred on few plants. Although H. frequens infrequently occupied P. fastigiata in the spring, H. grandicornis still generally sheltered in upper mid-intertidal plants, in part, because the net reproduction rates of H. grandicornis were low when space was available. The tubiculous amphipod A. tea usually was the only Ampithoe species on P. fastigiata; however, A. lindbergii and A. pollex sometimes dispersed to and released broods on this alga, particularly during the early summer when abundances of A. tea were relatively low. Similarly, net immigration rates of other thallus-dwelling copepods were relatively high when abundances of Scutellidium lamellipes were low. Moreover, several copepods were most abundant on different specimens of small isolated plants, and thallus-dwelling gastropods on larger plants at sites with relatively low densities of S. lamellipes on P. fastigiata; such copepods and gastropods generally were not abundant on aggregated plants sheltering large numbers of S. lamellipes. Such differences suggest that competition for food or space is among the factors causing negative correlations in the densities of Ampithoe spp. and thallusdwelling species on P. fastigiata.     

Profiles of photosynthetic pigments in the ocean using thin-layer chromatography

Phytoplankton pigments at a coastal station off Sydney, Australia, were studied by cellulose thinlayer chromatography. The chromatographic procedure distinguished chlorophylls a, b and c, chlorophyllide a, pheophytin a and pheophorbide a, and the major carotenoids carotene, astaxanthin, fucoxanthin, peridinin, diadinoxanthin and neoxanthin. Chlorophyllide a and pheophorbide a were the most significant chlorophyll-a degradation products in the water column, chlorophyllide a coming from chlorophyllase activity of senescent diatoms, and pheophorbide a from faecal pellets of copepods. Chlorophyll c occurred in every sample, even where there was no trace of chlorophyll a. Because of the greater chemical and biological stability of chlorophyll c compared to chlorophyll a, high chlorophyll c:a ratios result from a large proportion of senescent or detrital material in the samples. Determining the position of patches of copepods, diatoms, green algae and dinoflagellates in the water column was easily done by noting the presence of definitive pigments on the chromatograms.     

Phylogenetic relationships of coral-associated gobies (Teleostei,Gobiidae) from the Red Sea based on mitochondrial DNA data

Bryaninops, Gobiodon, Paragobiodon and Pleurosicya are the most abundant genera of coral-associated gobies. These genera are adapted to live among coral, while other small reef gobies (e.g., the genus Eviota) show no obligate association with this living substrate. Thirteen coral-associated species and two Eviota species were sampled from different regions of the Red Sea, along with four populations/species of Gobiodon from the Indian and western Pacific Oceans. A molecular phylogenetic analysis was performed using partial sequences of 12S rRNA, 16S rRNA and cytochrome b mitochondrial genes, 1,199 base pairs in total. Several clades were consistently resolved in neighbor joining-, maximum parsimony-, maximum likelihood and Bayesian analyses. While each of the four genera Gobiodon, Paragobiodon, Bryaninops and Pleurosicya proved to be monophyletic, their relative position in the phylogeny did not support an emergence of coral-associated gobiids as a monophyletic assemblage. Instead, two separate monophyletic sub-groups were discovered, the first comprising Gobiodon and Paragobiodon, and the second Bryaninops and Pleurosicya. Our molecular phylogenetic examinations also revealed one unassigned species of Gobiodon from the Maldives as a distinct species and confirmed three putative and yet unassigned species from the Red Sea. Moreover, the uniformly black colored species of Gobiodon are not monophyletic but have evolved independently within two distinct species groups. Genetic distances were large in particular within Pleurosicya and Eviota. Estimated divergence times suggest that coral-associated gobies have diversified in parallel to their preferred host corals. In particular, divergence times of Gobiodon species closely match those estimated for their typical host coral genus Acropora.     

Effects of monospecific and mixed-algae diets on survival, development and fatty acid composition of penaeid prawn (Penaeus spp.) larvae

Four species of microalgae (Chaetoceros muelleri, Tetraselmis suecica, Tahitian Isochrysis sp. (T-iso) and Dunaliella tertiolecta) with distinctly different fatty acid profiles were grown in continuous culture and fed to prawn larvae (Penaeus japonicus, P. semisulcatus and P. monodon) as monospecific diets. The best two diets (C. muelleri and T. suecica) were also fed as a mixed diet. Experiments were run until the larvae fed the control diet of C. muelleri metamorphosed to Mysis 1. The survival and development (i.e. performance) of the larvae were affected by algal diet, and the diets were ranked in the order of decreasing nutritional value: C. muelleri ≥ T. suecica > T-iso > D. tertiolecta. Larvae fed a mixed diet of C. muelleri and T. suecica (2:3 by dry weight) performed as well or better than those fed C. muelleri, and the performance of both these groups of larvae was better than those fed T. suecica. The lipid and carbohydrate compositions of the algae had little or no effect on the lipid and carbohydrate compositions of the larvae or their performance. However, the larvae that performed best (i.e. those fed C. muelleri) had significantly more lipid and carbohydrate than those that performed worst (i.e. those fed D. tertiolecta). Larvae fed C. muelleri or the mixed-algae diet had higher proportions of the essential fatty acids eicosapentaenoic acid [EPA, 20:5(n-3)] and arachidonic acid [ARA, 20:4(n-6)] than the larvae fed on other diets. Furthermore, the larvae fed T. suecica, which showed intermediate performance between larvae fed C. muelleri and T-iso or D. tertiolecta, also had higher proportions of EPA and ARA. Both C. muelleri and T. suecica contained EPA and ARA, but T-iso and D. tertiolecta did not, except for trace amounts of EPA in T-iso. The fatty acid ARA appears to be much more important in the diet of larval prawns than has so far been considered. The level of the essential fatty acid docosahexaenoic acid [DHA, 22:6(n-3)] in the algal diet and the larvae was not related to the performance of the larvae; only C. muelleri and T-iso contained DHA. However, the nauplii contained large proportions of DHA, suggesting that these were sufficient to meet the larval requirements for DHA during their development to Mysis 1. Mixed-algae diets could improve the performance of larvae by providing a more comprehensive range of fatty acids. Received: 22 April 1998 / Accepted: 3 December 1998     

Megabenthic decapod crustacean assemblages on the Galician continental shelf and upper slope (north-west Spain)

The structure of megabenthic decapod crustacean assemblages on the Galician (north-west Spain) continental shelf (100 to 200 depth) and upper slope (200 to 500 m) was analyzed based on surveys carried out in autumn and spring, from 1980 to 1987. Forty species belonging to 19 families were caught. The portunid crab Polybius henslowii, a species with pelagic phases, was the most abundant species, but displayed strong spatial and temporal fluctuations. Other dominant species were the Norway lobster Nephrops norvegicus, the portunid Liocarcinus depurator, the galatheids Munida intermedia and M. sarsi, and the shrimps Solenocera membranacea, Plesionika heterocarpus, Pasiphaea sivado and Dichelopandalus bonnieri. Total abundance and biomass (average values excluding Polybius henslowii = 255 individuals and 2.06 kg/30 min tow) and species richness and diversity, H′ (6.85 species and H′ = 1.45 per tow) displayed a significant positive correlation with depth, and strong interannual fluctuations. The factors determining community organization were depth and, to a lesser extent, spatial structure. There was clear evidence of bathymetric zonation, differentiating between species characteristic of the slope (D. bonnieri and Pasiphaea sivado), shelf-slope edge (Macropipus tuberculatus, Pontophilus spinosus, Munida sarsi, S. membranacea, Processa spp.) and shelf (L. depurator, Macropodia tenuirostris, Paguridae and Chlorotocus crassicornis). The spatial zonation was related to changes in oceanography and sediment along the continental margin. Goneplax rhomboides, N. norvegicus, C. crassicornis and Alpheus glaber are benthic species which generally exhibit burrowing behaviour, and they were found mainly in the southern area where there are fine sediments due to the outwelling from the Rías Baixas. Different benthopelagic shrimps (Pontophilus spinosus, Plesionika heterocarpus,Processa spp. and Pasiphaea sivado) were typical of the zone just north of Fisterra, characterized by a convergence of water masses bringing about an increase in productivity due to upwelling. The benthic anomuran and brachyuran crabs Munida intermedia, M.␣sarsi, L.␣depurator and Macropipus tuberculatus were characteristic of the northwestern zone between Fisterra and Estaca, where the infauna reaches high biomass despite coarser sediments with a lower concentration of organic material than in the southern area. Lastly, both the Paguridae and Macropodia tenuirostris were species typically found in the waters in the northern shelf. Based on interannual changes in assemblage structure, two periods could be distinguished: between 1980 and 1984, when Polybius henslowii, D. bonnieri and Pasiphaea sivado had abundance peaks; and another period from 1985 to 1987 when L. depurator, Munida intermedia, M.␣sarsi and Macropipus tuberculatus increased in abundance. Received: 21 May 1996 / Accepted: 9 August 1996     

First genetic validation and diagnosis of the short-finned squid species of the genus <Emphasis Type="Italic">Illex</Emphasis> (Cephalopoda: Ommastrephidae)

Squids of the genus Illex are representative of the family Ommastrephidae and account for 65% of the world’s cephalopod captures. Illex is formed by four taxa distributed throughout the Atlantic Ocean (I. argentinus, I. coindetii, I. illecebrosus and I. oxygonius), whose identification and phylogenetic relationships based on morphological characters remain controversial. Thirty-seven enzyme-coding loci were analysed in 230 individuals from seven populations of Illex and ten specimens of Todaropsis eblanae, which were used as the outgroup. Two to four enzyme loci (ALPDH*, IDHP-1*, MEP* and SOD*) were diagnostic among Illex species depending on the species-pair comparison. Individuals morphologically identified as I. oxygonius were also found genetically distinct, which proves the taxonomic validity of this species. No significant intraspecific genetic heterogeneity was detected within Illex argentinus, I. coindetii and I. illecebrosus (Mean G _ST= 0.011, 0.003, 0.017, respectively). I. illecebrosus and I. oxygonius were shown as sister species with a close relationship to I. argentinus, whereas I. coindetii formed a different lineage.     

Mycorrhizal induction of phenolic compounds and antioxidant properties of fungi and seedlings during the early steps of symbiosis

Ectomycorrhizal (ECM) symbiosis plays a major role in nutrient cycling and the functioning of forest ecosystems. Trees with well-developed ectomycorrhizas are more resistant to environmental stresses such as drought and to biotic stresses such as root pathogens. The establishment of ECM symbiosis is triggered by signals produced by both partners. These signals lead to morphological changes and a complex development of specific structures in both the plant root and the fungus. The development of the ECM fungi, Paxillus involutus and Pisolithus arhizus, in the presence and the absence of the symbiont—Pinus pinaster—was evaluated as well as their antioxidant properties and phenolic compounds composition in response to the symbiotic association. ECM fungi grew less in the presence of P. pinaster, with P. arhizus being less affected in growth and thus being more adapted to this association. Protocatechuic acid was found only in P. involutus, while P. pinaster roots, both in association and isolated, proved to have other phenolic acids, such as p-hydroxybenzoic and p-coumaric acids. The symbiosis between P. involutus and P. pinaster had no major effects on the symbionts, while the association between P. arhizus and P. pinaster seems to generally decrease the antioxidant effects of both symbionts, despite the increase in p-coumaric and cinnamic acids in the ECM fungi.     

Molecular phylogeny of the American <Emphasis Type="Italic">Callinectes</Emphasis> Stimpson, 1860 (Brachyura: Portunidae), based on two partial mitochondrial genes

The genus Callinectes encompasses 16 species of commercially important swimming crabs. Most (13) occur on the Pacific and Atlantic coasts of the Americas. We compare mtDNA regions corresponding to 964 basepairs of the large (16S) and small (12S) ribosomal subunits among American Callinectes in order to examine phylogenetic relationships. The status of Callinectes affinis Fausto-Filho and Callinectes maracaiboensis Taissoun is questioned, and C. maracaiboensis is concluded to be a junior synonym of Callinectes bocourti A. Milne-Edwards, from which it cannot be consistently distinguished. We find two major lineages, one of which includes C. affinis, C. bocourti, Callinectes rathbunae Contreras, Callinectes sapidus Rathbun, and Callinectes toxotes Ordway. A second lineage is comprised of Callinectes arcuatus Ordway, Callinectes bellicosus (Stimpson), Callinectes danae Smith, Callinectes exasperatus (Gerstaecker), Callinectes larvatus Ordway, Callinectes ornatus Ordway, and Callinectes similis Williams. Definition of these clades is supported by previously described morphological differences in the length of the gonopods and shared physioecological adaptations. A calibrated molecular clock is used to estimate divergence of the two lineages near 13 mybp. Our analyses suggest that C. ornatus is the closest relative of C. arcuatus, and that C. affinis is closest to C. bocourti.     

The systematic position of some boring sponges (Demospongiae,Hadromerida) studied by molecular analysis

A phylogenetic analysis of some bioeroding sponges of the family Clionaidae (Order Hadromerida) was performed to resolve some taxonomic problems both at the specific and the supraspecific level using the D2 and D3 regions of 28S rDNA. Species belonging to the genera Cliona, Cliothosa, Spheciospongia (fam. Clionaidae) and Diplastrella (fam. Spirastrellidae) from the Mediterranean Sea and Celebes Sea (Indonesia) were analysed. In the phylogenetic tree, the species clustered on two main branches, one comprising Cliona celata, C. rhodensis, C. utricularis, and Cliothosa hancocki, and the other made up of C. viridis, C. nigricans, C. schmidti, C. jullieni, Spheciospongia solida and S. vagabunda. Above the species level, data do not support the separation of the genus Cliothosa from Cliona, while they do support the inclusion of some massive boring species, previously assigned to the genus Spirastrella, in the family Clionaidae. At the species level, data demonstrated the genetic identity of taxa C. viridis and C. nigricans, in spite of their considerable morphological differences. In contrast, the yellow species commonly attributed to C. celata are probably to be considered as a complex of sibling species with a number of distinct taxa present in the Mediterranean. Data also showed the identity of the Mediterranean and Pacific populations of C. schmidti, suggesting the status of a Tethyan relict for this species.     

Rooting response in stem cuttings from five species of mangrove trees: effect of auxins and enzyme activities

An experiment was conducted to optimise rooting in stem cuttings of the mangrove tree species Bruguiera parviflora, Cynometra iripa, Excoecaria agallocha, Heritiera fomes and Thespesia populnea. Cuttings were girdled or non-girdled and treated with the auxins indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and naphthalene-acetic acid (NAA) and amylase, IAA-oxidase, peroxidase and polyphenol oxidase activities were measured. The best rooting performance (in terms of root number) was in stem cuttings of E. agallocha followed by C. iripa and H. fomes when treated with IBA + NAA. IAA and IBA combinations were more effective in T. populnea and B. parvifolia. Variations in rooting response were markedly reflected in amylase, IAA-oxidase, peroxidase and polyphenol oxidase activities. Sharp increases in amylase and polyphenol-oxidase activities were associated with enhanced rooting in E. agallocha, C. iripa and H. fomes. On the contrary, lowered IAA-oxidase and peroxidase activities favoured rooting in T. populnea and B. parviflora. A higher number of roots occurred in pre-girdled cuttings in comparison to non-girdled cuttings. Received: 30 March 1999 / Accepted: 6 October 1999     

Sugar compound products of the periphytic diatom <Emphasis Type="Italic">Navicula ramosissima</Emphasis> induce larval settlement in the barnacle, <Emphasis Type="Italic">Amphibalanus amphitrite</Emphasis>

This study investigated the response of cyprids of the barnacle Amphibalanus amphitrite to 23 strains of laboratory cultured periphytic diatoms isolated from microbial biofilms that formed on glass slides immersed in Tachibana bay, Nagasaki, and those from mass-production tanks in the Fisheries Center of Nagasaki City, Japan. In addition, periphytic diatoms were subjected to various treatments, in order to investigate the nature of the chemical cue in periphytic diatoms. Cyprids of A. amphitrite responded differently to the 23 different periphytic diatom strains and settled in high percentages on Cocconeis sp. and Navicula ramosissima strain A. On the other hand, nine strains of diatoms significantly inhibited settlement. The settlement inducing activity of N. ramosissima strain A increased linearly with diatom density, and its activity was enhanced by culturing the diatom under a bacteria-free condition, suggesting that specific diatom species, i.e., N. ramosissima strain A, may play an important role on larval settlement of the barnacle. Subjecting N. ramosissima strain A biofilm to hydrochloric acid (HCl) and ethanol (EtOH) treatments or heating it at 100°C did not inactivate the film, indicating that the settlement cue was a stable surface bound compound that did not decompose from the above treatments. Moreover, of the various lectins, enzymes, and drugs [H₅IO₆ and sodium dodecyl sulfate or (SDS)] used, only Lentil Agglutinin (LCA) treatment of N. ramosissima strain A biofilm resulted in the reduction its settlement inducing activity. A positive correlation was observed between the settlement inducing activity and the amount of LCA conjugated fluorescein isothiocyanate (FITC-LCA) of N. ramosissima strain A. On the other hand, subjecting biofilms of N. ramosissima strain B, an inactive strain, to various types of treatments resulted in the induction of A. amphitrite larval settlement but LCA treatment also reduced the activity of these treated N. ramosissima strain B biofilms. These findings suggest that a cue containing an LCA-binding sugar chain is present in both A and B strains of N. ramosissima but the large amount of mucous substance covering N. ramosissima strain B biofilm probably makes the sugar chain containing active subunit in strain B unavailable to A. amphitrite cyprids. In conclusion, periphytic diatoms such as N. ramosissima play an important role in larval settlement of the barnacle A. amphitrite. The cue in the diatom was an LCA-binding sugar chain(s) compound that may have similarities to the settlement inducing protein complex (SIPC) from adult shell of the barnacle.     

Sex pheromone communication of tentiform leaf-miners <Emphasis Type="Italic">Phyllonorycter insignitella</Emphasis> and <Emphasis Type="Italic">Ph. nigrescentella</Emphasis> from two related species groups

Summary.  Females of both species start their pheromone-releasing activity on the second day after emergence at the beginning of the photophase. During the present work, a peak of calling activity with close to 100% of active Ph. nigrescentella females was registered 1.5 hour after the light had been put on. The high pheromone release behaviour with 50% active females lasted for 3 hours. The calling activity of the group of females was about 6 h/day. The beginning of a photophase under laboratory conditions or an early morning in nature is a common period for sex pheromone release in the genus Phyllonorycter. (8Z,10E)-tetradecadien-1-yl acetate (8Z,10E-14:Ac), (8Z,10E)-tetradecadien-1-ol (8Z,10E-14:OH) and (8E,10Z)-tetradecadien-1-yl acetate (8E,10Z-14:Ac) in the ratio 96:4:traces as well as 8Z,10E-14:Ac and 8Z,10E-14:OH in the ratio 88:12 collected by Solid Phase Micro Extraction (SPME) were found to be specific for the calling periods of virgin Phyllonorycter insignitella and Ph. nigrescentella females respectively. Field trapping experiments demonstrated that all three compounds are important for the attraction of Ph. insignitella males while only 8Z,10E-14:Ac is the essential sex pheromone component for Ph. nigrescentella. The pheromone activity of all three compounds is reported for the first time. Addition of either 8Z,10E-14:OH or 8E,10Z-14:Ac to 8Z,10E-14:Ac did not have a significant effect on the attraction of Ph. nigrescentella males, while the efficiency of the three component blend was 5 times lower as compared to that of 8Z,10E-14:Ac. Our data demonstrate that 8Z,10E-14:OH and 8E,10Z-14:Ac play a dual function, they are minor sex pheromone components of Ph. insignitella essential for attraction of conspecific males and show an allelochemical, antagonistic effect on Ph. nigrescentella males and, thus, ensuring specificity of the mate location signal in two related Phyllonorycter species.     

Phylogenetic relationships among giant clam species (Mollusca: Tridacnidae) determined by protein electrophoresis

Analyses of allozyme variation at 26 loci in eight species of giant clams, collected from the Western Pacific between 1989 and 1991, have confirmed the major groupings currently recognised within the family Tridacnidae on the basis of morphological characters: the two genera Hippopus and Tridacna, and the subgenera within Tridacna; i.e. Chametrachea (comprising T.squamosa, T.crocea and T.maxima), Tridacna sensu stricto (containing only T. gigas), and Persikima (comprising T. derasa and T. tevoroa). All these groupings were each supported by several shared characters. In contrast to previous views, Persikima was found to be intermediate between Chametrachea and T. gigas. Although statistical support for branch lengths linking Chametrachea and T. gigas was limited in maximum-likelihood analyses, the occurrence of several shared alleles between these groups, and none shared between only Chametrachea and Persikima, suggests a closer relationship between Chametrachea and T. gigas or, at least, an unresolved trichotomy. Morphological data traditionally assumed to support a closer association of Chametrachea and Persikima demonstrated that only one shared character linked subgenera, and this character supported an evolutionary association between Chametrachea and T. gigas. These findings, together with the number of shared characters supporting the subgenus Persikima, suggest that Persikima is the most basal offshoot of the Tridacna line, and that the group may deserve generic status. Received: 21 November 1997 / Accepted: 24 March 1998