Seaweed-herbivore-predator interactions: host-plant specialization reduces predation on small herbivores

Summary Because feeding specialization among marine herbivores is rare, marine communities provide a simplified system for identifying factors selecting for specialization. On Australia's Great Barrier Reef, we investigated interactions among the chemically-defended seaweed Chlorodesmis fastigiata, herbivores specialized on this alga, and potential predators of these herbivores. Chlorodesmis is a low preference food for reef fishes but appears to be the only food of the crab Caphyra rotundifrons and the ascoglossan gastropods Elysia sp. and Cyerce nigricans. The crab is found only in patches of Chlorodesmis, feeds solely on the alga, and selectively shelters in it in laboratory choice experiments. Crab grazing on the red seaweed Acanthophora spicifera was stimulated when this alga was coated with increasing concentrations of the cytotoxic diterpenoid chlorodesmin, the major secondary metabolite of Chlorodesmis. Crabs did not sequester Chlorodesmis metabolites but avoided predators by sheltering in the unpalatable alga. All crabs tethered on the reef without access to Chlorodesmis patches were rapidly eaten; those with access to Chlorodesmis patches were much less susceptible to predation. The cryptic ascoglossan Elysia sp. was found exclusively in patches of Chlorodesmis and sequestered metabolites from the alga. Living Elysia were unpalatable to the common wrasse Thalassoma lunare in laboratory assays, but the crude organic extract of Elysia did not significantly deter feeding by Thalassoma. Elysia sequestered chlorodesmin, which deterred feeding by reef fishes in field assays but was ineffective against Thalassoma in laboratory assays at 5% food dry mass. Unlike Elysia, the aposematically colored ascoglossan Cyerce nigricans sequestered Chlorodesmis metabolites in relatively small amounts, but produced larger amounts of unrelated polypropionate compounds. Cyerce were never attacked by fishes and the crude organic extract of this slug strongly deterred feeding by wrasses in laboratory assays. The dorid nudibranch Gymnodoris sp. was found only in Chlorodesmis patches and appeared to be a specialized predator on Elysia; it would not prey on Cyerce. Data from this and other recent investigations demonstrate that some small marine herbivores feed selectively or exclusively on seaweeds that are chemically defended from fishes. This association reduces predation on the herbivores and suggests that escape from and deterrence of predation may be a dominant factor selecting for specialization among these herbivores.     

Anin vitro rearing system for the propagation of the ectoparasitoidCatolaccus grandis

An experimental ‘closed’ rearing system, where egg and larval manipulations were eliminated, was developed for the in vitro rearing of Catolaccus grandis (Hymenoptera: Pteromalidae) Burks, an important ectoparasitoid of the cotton boll weevil, Anthonomus grandis Boheman. In this rearing model, n-hexane (a synthetic ovipositional stimulant for this parasitoid), was smeared on the Parafilm® cover of a modified rearing chamber (a Multiwell®) tissue culture plate) to induce the deposition of uncontaminated eggs, on the inner side of this waxy membrane, and on or around an agar retained diet that had been dispensed into the individual chamber wells. When the efficiency of the in vitro rearing system was compared to the current in vivo rearing method for this species, the duration of the life cycle was significantly shorter in parasitoids reared in vivo, but this difference was less than one day (17.8 vs 17.1 days, respectively). On the other hand, the number of eggs laid in the in vitro rearing chamber during a 4 h period was c. 2.5 times greater than in the conventional in vivo rearing apparatus, and adult yields were c. 25% greater when using the in vitro closed rearing method. Male to female ratios were c. 1:9 when reared in vitro as compared to 1.0:1.5 for those reared in vivo. There were no apparent adverse effects of this in vitro rearing system on the parasitoid's general behavior and reproduction after two consecutive generations.     

New chromosome reports in the subtribes Diocleinae and Glycininae (Phaseoleae: Papilionoideae: Fabaceae)

The base chromosome number of x = 11 is the most probable in all the subtribes included in tribe Phaseoleae, although some aneuploid reduction is evident in Collaea and Galactia (Diocleinae) and chromosome duplications are seen in Amphicarpaea, Cologania and Glycine (Glycininae). The aims of this study were to improve the cytological knowledge of some species of Collaea and Galactia and to examine the anomalous counts reported for Calopogonium (Glycininae) and verify its taxonomic position. In addition, a molecular phylogeny was constructed using nuclear ribosomal DNA sequences (internal transcribed spacer region), and the chromosome number was optimized on the topology. In this work, the chromosome counts for Galactia lindenii, Galactia decumbens and Collaea cipoensis (all 2n = 20), and Calopogonium sericeum (2n = 22) are reported for the first time. The new reports for Galactia and Collaea species are in agreement with the chromosome number proposed for subtribe Diocleinae. The study rejects the concept of a cytologically anomalous Calopogonium and, based on the phylogenetic analysis, corroborates the position of this genus within subtribe Glycininae. The ancestral basic chromosome number of x = 11 proposed for Phaseoleae is in agreement with the evolutionary pathway of chromosome numbers analysed in this work. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 336–341.     

Can members of the south‐western Gila robusta species complex be distinguished by morphological features?

The goal for this project was to re‐examine key morphological characters hypothesized to differentiate Gila intermedia, Gila robusta and Gila nigra and outline methods better suited for making species designations based on morphology. Using a combination of meristic counts, morphological measurements and geometric morphometrics, morphological dissimilarities were quantified among these three putative species. Traditional meristic counts and morphological measurements (i.e. distances between landmarks) were not useful for species identification. Geometric morphometrics, however, identified differences among species, while also suggesting an effect of geographic location on morphological variation. Using canonical variate analysis for the 441 fish sampled in this study, geometric morphometrics accurately predicted true group membership 100% of the time for G. nigra, 97% of the time for G. intermedia and 91% of the time for G. robusta. These results suggest that geometric morphometric analysis is necessary to identify morphological differences among the three species. Geometric morphometric analysis used in this study can be adopted by management officials as a tool to classify unidentified individuals.     

THE CYTOTAXONOMY OF FILIPENDULA (ROSACEAE) AND ITS IMPLICATIONS

The genus Filipendula Mill. is generally separated from Spiraea L. in systematic keys on the basis of a single fruit character. In some taxonomic treatments of the Rosaceae, where subfamilies are used, this places the genera in separate subfamilies. Karyological studies can be useful in assaying the justifiability of such treatment and are needed because of serious discrepancies between previous reports of chromosome numbers and the recent textbook designation, on dubious grounds, of F. vulgaris as an example of a “permanent chromosome hybrid.” The results given in this paper show that x = 7 in this genus (compared with x = 9 in Spiraea) and the reasons for rejecting previous counts of 2n = 15 for F. vulgaris are presented. “Permanent chromosome hybridity” for this species is also rejected. The possibility that a cytotype with 2n = 16 may exist in the northern part of the range of F. ulmaria cannot be completely discounted, but positive evidence is presented for 2n = 14 in this species (even though 2n = 16 has been reported most frequently recently). The basic number 7 for Filipendula is in agreement with the placing of this genus in the subfamily Rosoideae even though the hereditary peculiarities (apomixis and permanent chromosome hybridity) shown by some other members of this subfamily are apparently not now needed to explain the cytological situation in Filipendula.     

Microsynteny between pea and Medicago truncatula in the SYM2 region

The crop legume pea (Pisum sativum) is genetically well characterized. However, due to its large genome it is not amenable to efficient positional cloning strategies. The purpose of this study was to determine if the model legume Medicago truncatula, which is a close relative of pea, could be used as a reference genome to facilitate the cloning of genes identified based on phenotypic and genetic criteria in pea. To this end, we studied the level of microsynteny between the SYM2 region of pea and the orthologous region in M. truncatula. Initially, a marker tightly linked to SYM2 was isolated by performing differential RNA display on near-isogenic pea lines. This marker served as the starting point for construction of a BAC physical map in M. truncatula. A fine-structure genetic map, based on eight markers from the M. truncatula physical map, indicates that the two genomes in this region share a conserved gene content. Importantly, this fine structure genetic map clearly delimits the SYM2-containing region in pea and the SYM2-orthologous region in M. truncatula, and should provide the basis for cloning SYM2. The utility of the physical and genetic tools in M. truncatula to dissect the SYM2 region of pea should have important implications for other gene cloning experiments in pea, in particular where the two genomes are highly syntenic within the region of interest.     

Characterization of the S locus genes, SLG and SRK, of the Brassica S3 haplotype: identification of a membrane-localized protein encoded by the S locus receptor kinase gene

The S locus, which controls the self-incompatibility response in Brassica, has been shown to contain at least two genes. SLG encodes a secreted S locus glycoprotein whilst SRK encodes a putative S locus receptor kinase. SRK has been shown potentially to encode a functional kinase and genetic evidence indicates that this gene is essential for the self-incompatibility response. Here the characterization of the SRK and SLG genes of a Brassica line homozygous for the S₃ haplotype is described. A 120 kDa glycoprotein was identified in stigmas and several lines of evidence indicated that this protein is encoded by the SRK₃ gene. First, the 120 kDa glycoprotein was recognized by antibodies raised against peptides based on the SRK₃ gene sequence. Secondly, this protein is polymorphic and, in an F₂ population segregating for the S₃ haplotype, was expressed only in plants possessing the S₃ haplotype. Thirdly, the 120 kDa protein was expressed specifically in stigmas. Finally, the 120 kDa protein was only extracted from stigmas in the presence of detergent indicating that it is anchored in the membrane. SRK has been predicted to encode a transmembrane glycoprotein based on the deduced amino acid sequence. Located on the membrane, SRK is in a position to interface between an extracellular recognition event between pollen and pistil and an intracellular signal transduction pathway which initiates the self-incompatibility response.     

Acclimation of photosynthesis and respiration to elevated atmospheric CO2 in two Scrub Oaks

Abstract For two species of oak, we determined whether increasing atmospheric CO₂ concentration (C_a) would decrease leaf mitochondrial respiration (R) directly, or indirectly owing to their growth in elevated C_a, or both. In particular, we tested whether acclimatory decreases in leaf‐Rubisco content in elevated C_a would decrease R associated with its maintenance. This hypothesis was tested in summer 2000 on sun and shade leaves of Quercus myrtifolia Willd. and Quercus geminata Small. We also measured R on five occasions between summer 1999 and 2000 on leaves of Q. myrtifolia. The oaks were grown in the field for 4 years, in either current ambient or elevated (current ambient + 350 µmol mol^?1) C_a, in open‐top chambers (OTCs). For Q. myrtifolia, an increase in C_a from 360 to 710 µmol mol^?1 had no direct effect on R at any time during the year. In April 1999, R in young Q. myrtifolia leaves was significantly higher in elevated C_a—the only evidence for an indirect effect of growth in elevated C_a. Leaf R was significantly correlated with leaf nitrogen (N) concentration for the sun and shade leaves of both the species of oak. Acclimation of photosynthesis in elevated C_a significantly reduced maximum RuBP‐saturated carboxylation capacity (V_{c max}) for both the sun and shade leaves of only Q. geminata. However, we estimated that only 11–12% of total leaf N was invested in Rubisco; consequently, acclimation in this plant resulted in a small effect on N and an insignificant effect on R. In this study measurements of respiration and photosynthesis were made on material removed from the field; this procedure had no effect on gas exchange properties. The findings of this study were applicable to R expressed either per unit leaf area or unit dry weight, and did not support the hypothesis that elevated C_a decreases R directly, or indirectly owing to acclimatory decreases in Rubisco content.     

Consequences of invasion by Pseudosuccinea columella on the dynamics of native lymnaeids living on the acid soils of central France

Eight road ditches located on granite and colonised by the native lymnaeids Galba truncatula or Omphiscola glabra were studied for 2 years to follow the distribution and density of these species in their habitats when the invasive lymnaeid Pseudosuccinea columella was introduced. Six other ditches populated only by a native lymnaeid or by P. columella were used as controls. The same experiment was also carried out for 3 years in seven water tanks placed in semi-natural conditions. The colonisation of P. columella was more rapid in habitats with G. truncatula than in those with O. glabra. Similarly, this colonisation led to a progressive decrease in the number of native lymnaeids in these ditches and this decrease was also faster for G. truncatula than for O. glabra. A similar decrease of G. truncatula and O. glabra was also noted when P. columella was introduced into water tanks placed in semi-natural conditions. The habitats of G. truncatula or O. glabra on the acid soils of central France can be successfully colonised by P. columella, and the establishment of this invasive species may lead to a decrease of the two native lymnaeids or even the local extinction of one of these (G. truncatula).     

Evaluation of phytoseiid predators for control of western flower thrips on greenhouse cucumber

Ten predatory mite species, all phytoseiids, were evaluated for control of western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), on greenhouse cucumber. This study was done to further improve biological control of thrips on this crop. Neoseiulus cucumeris (Oudemans) is at present used for biological control of thrips in greenhouses. Compared to this species, Typhlodromalus limonicus (Garman & McGregor), Typhlodromips swirskii (Athias-Henriot) and Euseius ovalis (Evans) reached much higher population levels resulting in a significantly better control of thrips. T. limonicus was clearly the best predator of WFT. Also Euseius scutalis (Athias-Henriot) increased to higher populations levels than N. cucumeris, but without controlling the thrips, probably because of an unequal distribution of this predator on the plant. Iphiseius degenerans (Berlese), Neoseiulus barkeri (Hughes), Euseius finlandicus (Oudemans) and Typhlodromus pyri (Scheuten) did not establish better than N. cucumeris. A non-diapausing exotic strain of N. cucumeris did not differ from the North European strain. The best performers in this study were all of sub-tropical origin. T. limonicus, T. swirskii and E. ovalis have good potentials for controlling not only thrips but also whiteflies. Factors affecting the efficacy of phytoseiids on greenhouse cucumbers are discussed.     

Life history and host range of the leaf blotcher Eucosmophora schinusivora: a candidate for biological control of Schinus terebinthifolius in the USA

The host range of Eucosmophora schinusivora Davis and Wheeler (Lepidoptera: Gracillariidae) was studied to assess its suitability as a biological control agent of Schinus terebinthifolius Raddi (Anacardiaceae), a serious environmental and agricultural weed in the USA and elsewhere in the world. The life history of this insect species and its host range were determined in the laboratory with adult no-choice oviposition and larval development tests. This species has five instars, the first three are sap-feeding miners and the last two are tissue feeding. Total development time was 31.7 days. To examine specificity of this species, 10 plant species in Anacardiaceae were selected based on taxonomic relatedness to S. terebinthifolius, economic importance, and availability. In the laboratory, except for Anacardium occidentale and Cotinus obovatus, all of the tested species were accepted for oviposition with a marked preference for the weed S. terebinthifolius, Schinus molle, Rhus copallinum, Rhus sandwicensis and Pistacia chinensis. Complete development, from egg to adult, was achieved only on S. terebinthifolius, S. molle, R. copallinum, P. chinensis and Metopium toxiferum. In conclusion, E. schinusivora will not be considered for the biological control of S. terebinthifolius in the continental USA. However, the utilisation of this species in other infested areas such as Hawai'i and Australia should be considered.     

The second phantom aquatic leaf beetle in Japan: Macroplea mutica rediscovery in the wetlands (Coleoptera: Chrysomelidae)

Wetland biodiversity is currently declining on a global scale. Wetland biodiversity understanding is critical for determining the wetlands' conservation value. In this study, Macroplea Samouelle, 1819 (Coleoptera: Chrysomelidae) was discovered in Aomori Prefecture, Honshu Island, Japan. Only two Macroplea species have been recorded in Japan, M. japana (Jacoby, 1885) and M. mutica (Fabricius, 1792). Macroplea japana had been unrecorded for 60 years before being rediscovered in Honshu Island in 2022, and a single adult M. mutica female was discovered in Hokkaido Prefecture in 2003. The discovered individuals were concluded to be M. mutica based on morphological and molecular analyses. Although morphological differences were observed with the Eurasian M. mutica individuals, the male genitalia was nearly identical to M. mutica. For the molecular phylogenetic analysis based on COI and 28S sequences, Macroplea individuals in Japan were clustered with M. mutica on the Eurasian Continent. This is the first record of this species on Honshu Island (and the second in Japan), as well as the first record of adult males. This species would require conservation policies and additional distributional surveys.     

Biology, host specificity tests, and risk assessment of the sawfly Heteroperreyia hubrichi, a potential biological control agent of Schinus terebinthifolius in Hawaii

Heteroperreyia hubrichiMalaise (Hymenoptera: Pergidae), a foliagefeeding sawfly of Schinusterebinthifolius Raddi (Sapindales:Anacardiaceae), was studied to assess itssuitability as a classical biological controlagent of this invasive weed in Hawaii. No-choice host-specificity tests were conductedin Hawaiian quarantine on 20 plant species in10 families. Besides the target weed, adultfemales oviposited on four test species. Females accepted the Hawaiian native Rhussandwicensis A. Gray (Sapindales:Anacardiaceae) as an oviposition host equallyas well as the target species. The other threespecies received significantly fewer eggs. Neonate larvae transferred onto test plantssuccessfully developed to pupae on S.terebinthifolius (70% survival) and R.sandwicensis (1% survival). All other 18test plant species failed to support larvaldevelopment. A risk analysis was conducted toquantify the acceptability of non-targetspecies as host plants for H. hubrichi onthe basis of the insect's performance atvarious stages in its life cycle. Risk ofdamage to all plant species tested wasinsignificant except for R. sandwicensis. Risk to this native plant relative to S.terebinthifolius was estimated at 1%. Currently this level of risk is too high torequest introduction of this insect into theHawaiian environment. Detailed impact studiesin the native range of S. terebinthifoliusare needed to identify thepotential benefit that this insect offers. Also, field studies in South America withpotted R. sandwicensis would give a morereliable analysis of the risk this nativeHawaiian plant would face from naturalpopulations of H. hubrichi.     

The use of comparative genome analysis and syntenic relationships allows extrapolating the position of Zn tolerance QTL regions from Arabidopsis halleri into Arabidopsis thaliana

Arabidopsis halleri is a species that has undergone natural selection for zinc (Zn) tolerance. Isolation of the quantitative trait loci (QTL) associated with this trait holds great promise for the identification of the main genes responsible for this adaptation. Using a segregating progeny produced by an interspecific cross, we previously constructed a genetic linkage map of A. halleri × A. lyrata petraea and mapped the three main QTL that confer Zn tolerance in A. halleri (Willems et al.). The goal of the present study is to compare the genetic linkage map of A. halleri × A. l. petraea to the annotated A. thaliana genome sequence to generate a tool for A. halleri genomic approaches. To achieve this aim, we constructed a genetic linkage map with 81 markers anchored on A. thaliana, including 23 genes known to be involved in metal homeostasis. First, this provided an extensive overview of the chromosomal rearrangements that have occurred since the divergence between A. thaliana and its closest relative A. halleri. Second, on the basis of the syntenic relationships assessed experimentally through this work, we transferred the QTL confidence intervals for Zn tolerance to the A. thaliana physical map, allowing access to all the genes localized in the corresponding regions. Third, we validated from the 23 genes involved in metal homeostasis the three ones localized in the QTL regions that can be considered the best candidates for conferring Zn tolerance. Nancy H. C. J. Roosens and Glenda Willems contributed equally to this paper.     

Molecular mapping of gibberellin-responsive dwarfing genes in bread wheat

Opportunities exist for replacing reduced height (Rht) genes Rht-B1b and Rht-D1b with alternative dwarfing genes for bread wheat improvement. In this study, the chromosomal locations of several height-reducing genes were determined by screening populations of recombinant inbred lines or doubled haploid lines varying for plant height with microsatellite markers. Linked markers were found for Rht5 (on chromosome 3BS), Rht12 (5AL) and Rht13 (7BS), which accounted for most of the phenotypic variance in height in the respective populations. Large height differences between genotypes (up to 43 cm) indicated linkage to major height-reducing genes. Rht4 was associated with molecular markers on chromosome 2BL, accounting for up to 30% of the variance in height. Confirming previous studies, Rht8 was linked to markers on chromosome 2DS, whereas a population varying for Rht9 revealed a region with a small but significant height effect on chromosome 5AL. The height-reducing effect of these dwarfing genes was repeatable across a range of environments. The molecular markers developed in this study will be useful for marker-assisted selection of alternative height-reducing genes, and to better understand the effects of different Rht genes on wheat growth and agronomic performance.     

Gonadal development in a giant threatened reef fish,the humphead wrasse Cheilinus undulatus,and its relationship to international trade

An opportunity arose to obtain humphead wrasse Cheilinus undulatus specimens between 2006 and 2009 from Indonesia, the major source and exporting country of this species, making study on its early gonad development possible for the first time. Protogynous hermaphroditism, previously proposed for this species, was confirmed in this study. Based on histological examination of 178 specimens, mainly <500 mm total length (L_T) and ranging from 208 to 1290 mm L_T (119·1 g to 43·0 kg whole body mass), the minimum body sizes for female and male sexual maturation were determined to be 650 and 845 mm L_T, respectively. Primary male development through juvenile sexual differentiation was not detected. A unique blind pouch, with a possible sperm storage function and associated with the testis, was reported for the first time in the Labridae. In Hong Kong retail markets, the global trading centre for this valuable species, live C. undulatus on sale for food were dominated by body sizes <500 mm L_T between 1995 and 2009, reflecting an international trade largely focused on juveniles. In consideration of these findings, and given the threatened status of this species, management for C. undulatus capture and trade nationally and internationally are discussed with recommendations for ensuring sufficient spawning biomass in exploited populations and for sustainable trade.     

Sawadaea nankinensis comb. nov.: a powdery mildew fungus of Acer buergerianum

The anamorphic state of a powdery mildew on trident maple (Acer buergerianum, Aceraceae), belonging to Sawadaea, has been observed since 1980 in Tokyo and other areas of Japan. Since the autumn of 2003, this fungus has begun to produce chasmothecia in various areas of Japan, which were consistent with Erysiphe nankinensis (= Uncinula nankinensis), but apparently contradictory to the characteristics of the anamorph. Based on a molecular phylogenetic analysis using DNA separately extracted from the anamorph and teleomorph of the fungus on A. buergerianum, it could be demonstrated that sequences of this fungus are sister to Sawadaea. As the anamorph belongs to Oidium subgen. Octagoidium and because of the phylogenetic position within the Sawadaea clade, the new combination Sawadaea nankinensis is proposed for this species. The genus Sawadaea is emended to comprise species with consistently unbranched appendages.     

The Escherichia coli rhamnose promoter rhaP BAD is in Pseudomonas putida KT2440 independent of Crp–cAMP activation

We developed an expression vector system based on the broad host range plasmid pBBR1MCS2 with the Escherichia coli rhamnose-inducible expression system for applications in Pseudomonas. For validation and comparison to E. coli, enhanced green fluorescent protein (eGFP) was used as a reporter. For further characterization, we also constructed plasmids containing different modifications of the rhaP _BAD promoter. Induction experiments after the successful transfer of these plasmids into Pseudomonas putida KT2440 wild-type and different knockout strains revealed significant differences. In Pseudomonas, we observed no catabolite repression of the rhaP _BAD promoter, and in contrast to E. coli, the binding of cyclic adenosine monophosphate (cAMP) receptor protein (Crp)–cAMP to this promoter is not necessary for induction as shown by deletion of the Crp binding site. The crp ⁻ mutant of P. putida KT2440 lacked eGFP expression, but this is likely due to problems in rhamnose uptake, since this defect was complemented by the insertion of the l-rhamnose-specific transporter rhaT into its genome via transposon mutagenesis. Other global regulators like Crc, PtsN, and CyoB had no or minor effects on rhamnose-induced eGFP expression. Therefore, this expression system may also be generally useful for Pseudomonas and other γ-proteobacteria.