Evidence of oligogenic sex determination in the apple snail <Emphasis Type="Italic">Pomacea canaliculata</Emphasis>

A small number of genes may interact to determine sex, but few such examples have been demonstrated in animals, especially through comprehensive mating experiments. The highly invasive apple snail Pomacea canaliculata is gonochoristic and shows a large variation in brood sex ratio, and the involvement of multiple genes has been suggested for this phenomenon. We conducted mating experiments to determine whether their sex determination involves a few or many genes (i.e., oligogenic or polygenic sex determination, respectively). Full-sib females or males that were born from the same parents were mated to an adult of the opposite sex, and the brood sex ratios of the parents and their offspring were investigated. Analysis of a total of 4288 offspring showed that the sex ratios of offspring from the full-sib females were variable but clustered into only a few values. Similar patterns were observed for the full-sib males, although the effect was less clear because fewer offspring were used (n?=?747). Notably, the offspring sex ratios of all full-sib females in some families were nearly 0.5 (proportion of males) with little variation. These results indicate that the number of genotypes of the full-sibs, and hence genes involved in sex determination, is small in this snail. Such oligogenic systems may be a major sex-determining system among animals, especially those with variable sex ratios.     

Thermal limits for the establishment and growth of populations of the invasive apple snail <Emphasis Type="Italic">Pomacea canaliculata</Emphasis>

Pomacea canaliculata is a South American freshwater snail considered as one of the world’s worst invasive alien species. A temperature of around 25 °C has usually been considered to be optimal for rearing P. canaliculata. Nevertheless, snails have not been reared under a wide range of temperatures to reveal the optimum for performance in terms of population increase. We investigated the effect of temperature on growth, survival and reproduction, estimating demographic parameters for P. canaliculata in the wide range of temperatures at which these snails are active (15–35 °C). No reproductive activity was evidenced for the snails reared at 15 °C, probably explained by the small sizes attained at this temperature. Temperatures above 25 °C did not promote a significant acceleration in growth so higher temperatures will not result in a reduction in time to reach maturity. In fact, snails from 25 and 30 °C began reproduction at the same age. We report here for the first time a detrimental effect of high temperatures that provoked a significant decrease in the contribution of snails to the next generation: the viability of eggs from the snails reared at 30 °C was very low and the snails exposed to a constant water temperature of 35 °C were unable to produce eggs. Our findings reveal a new environmental constraint that could be a determinant of the range limits of this species in invaded regions, especially during the coming decades, anticipating the scenario predicted from global warming.     

Herbivory on aquatic vascular plants by the introduced golden apple snail (<Emphasis Type="Italic">Pomacea canaliculata</Emphasis>) in Lao PDR

The effect of naturally found densities of the exotic and herbivorous golden apple snail (Pomacea canaliculata) on three dominant aquatic plants – duckweed (Lemna minor), water hyacinth (Eichhornia crassipes) and morning glory (Ipomoea aquatica) – was assessed in a wetland survey and quantified in a field experiment in Laos in southeast Asia. Snail grazing reduced plant biomass, but plant species were differently affected by grazing. Duckweed had almost disappeared after 6 and water hyacinth after 21 days, whereas morning glory remained at 80% of initial biomass after 32 days. Snail growth was lowest on morning glory and, when all plant species were simultaneously presented to snails, this plant was not preferred. We suggest that the negative effect the golden apple snail had on the growth of these plant species in field enclosures is present in the natural environment as well. This new and intense herbivory could have serious negative effects on invaded freshwater ecosystems in this region.     

Eleven microsatellite loci for two invasive freshwater apple snails, <Emphasis Type="Italic">Pomacea canaliculata</Emphasis> and <Emphasis Type="Italic">P. maculata</Emphasis> (Ampullariidae)

Pomacea canaliculata and P. maculata are major invasive species that continue to spread and negatively impact aquatic habitats globally. Herein we report the development of a set of highly polymorphic microsatellite loci that readily amplify in both species, and that will be informative for understanding the pathways of spread and population structure across the native and non-native range. The high polymorphism of these markers should also provide substantial power for refining the estimated origins of the introduced populations. However, they are uninformative for assessing introgressive hybridization between the two species because of shared alleles.     

Feeding of the leech <Emphasis Type="Italic">Glossiphonia weberi</Emphasis> on the introduced snail <Emphasis Type="Italic">Pomacea bridgesii</Emphasis> in India

The predation potential of the indigenous leech Glossiphonia weberi on the snail Pomacea bridgesii, introduced in India, was evaluated in the laboratory. Snails used belonged to the size-classes ≤‰3.0, 3.1–5.0, 5.1–7.0 and 7.1–9.0 mm in shell height, using them both separately and together (mixed) in combinations. In each experiment lasting 24 h a single leech belonging to the size-classes 2.0–3.9, 4.0–5.9, 6.0–7.9, 8.0–9.9 and 10.0–11.9 mm in length was used. Except the 4.0–5.9 mm size-class, leeches were able to capture and kill P. bridgesii irrespective of latter’s size; the predation, however, was confined to snails ≤3.0 mm. The rate of predation varied with the size of the predator and the prey, and a leech was able to kill a maximum of three snails per day. In India, in nature G. weberi feeds mostly on the pulmonate snail, Lymnaea (Radix) luteola. Experimental studies, however, revealed that G. weberi prefers the snails P. bridgesii and L. (R) luteola at the same rate from amongst the many other either less or not-preferred native operculate and non-operculate snails.     

Alarm response of hatchlings of the apple snail, <Emphasis Type="Italic">Pomacea canaliculata</Emphasis> (Gastropoda: Ampullariidae), to aqueous extracts of other individuals

We examined how hatchlings of the freshwater snail, Pomacea canaliculata, responded to aqueous extracts of conspecific hatchlings. Three, 3-day-old hatchlings were macerated in deionized water (1mg hatchling per 1ml water). When 0.5ml of the aqueous extract was added to a test tube containing 10 hatchlings of the same age and 50ml of water, the hatchlings in the water began to crawl out of the water within 5min. The proportion of hatchlings that crawled out of the water approached 0.6–0.9 after 1h, but gradually decreased to 0.4 after 24h. The relatedness between the live and the macerated hatchlings had no significant influence on the response. Hatchlings of egg masses obtained either in the laboratory or in the wild responded similarly to aqueous extracts of hatchlings from either egg mass. This suggests that the conditions under which the egg masses were incubated or the conditions that their parents had experienced had no effect on the hatchlings response. When compared with experiments reported on other aquatic animals, we consider the behavior of the hatchlings to be an alarm response of escaping from predators.     

Feeding behaviour,efficiency and food preference in yabbies <Emphasis Type="Italic">Cherax</Emphasis><Emphasis Type="Italic"> destructor</Emphasis>

The influence of body size on the consumption of live zooplankton (Daphnia spp.) by freshwater crayfish was examined using yabbies (Cherax destructor) ranging from 5 to 45 g. Food preference between live zooplankton and inert pellets was also assessed under experimental conditions. In experimental tanks, yabbies of four size classes (<15, 15–24.9, 25–34.9 and 35–45 g) were presented with live Daphnia. All yabbies were held in separate tanks with five animals per size class. In yabbies less than 15 g, the feeding mode on zooplankton involved rapid searching and probing with the first two pairs of walking legs. Once a prey was located, the chelae on the end of these walking legs would grasp the zooplankton and then rapidly move it towards the mouthparts. Yabbies larger than 25 g tended to use their walking legs to push the Daphnia nearer to their third maxilliped which would then force or scoop the zooplankton towards the mouthparts. A short-term feeding trial showed that there was no significant difference between size classes in regards to zooplankton consumption (P > 0.05). Capture efficiency of live Daphnia by yabbies less than 15 g was significantly lower (76%, P = 0.008) than the three larger size classes (93.6%). Yabbies less than 15 g consumed a significantly (P < 0.001) higher percentage (5.2%) of their body weight than the other size classes (1.1%, 0.8%, and 0.6%, respectively). In the presence of both live zooplankton and a pellet diet, yabbies spent significantly (P = 0.005) more time feeding on zooplankton (85%) than on inert pellets (15%). This was the first study to quantify zooplankton consumption by yabbies and the results provide insights into understanding the trophic role of freshwater crayfish in structuring zooplankton communities and the husbandry management of crayfish farming. Handling editor: S. I. Dodson     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.     

<Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-mediated transformation of callus cells of <Emphasis Type="Italic">Crataegus</Emphasis><Emphasis Type="Italic">aronia</Emphasis>

A genetic transformation system has been developed for callus cells of Crataegus aronia using Agrobacterium tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ Indole-3-butyric acid (IBA) and 0.5 mg l⁻¹ 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red colored callus was obtained on MS medium supplemented with 2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l⁻¹ kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l⁻¹ hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this is the first time to report an Agrobacterium-mediated transformation system in Crataegus aronia.     

The tyrosine <Emphasis Type="Italic">O</Emphasis>-prenyltransferase SirD catalyzes <Emphasis Type="Italic">O</Emphasis>-, <Emphasis Type="Italic">N</Emphasis>-, and <Emphasis Type="Italic">C</Emphasis>-prenylations

Recently, the prenyltransferase SirD was found to be responsible for the O-prenylation of tyrosine in the biosynthesis of sirodesmin PL in Leptosphaeria maculans. In this study, the behavior of SirD towards phenylalanine/tyrosine and tryptophan derivatives was investigated. Product formation has been observed with 12 of 19 phenylalanine/tyrosine derivatives. It was shown that the alanine structure attached to the benzene ring and an electron donor, e.g., OH or NH₂, at its para-position are essential for the enzyme activity. Modifications were possible both at the side chain and the benzene ring. Enzyme products from seven phenylalanine/tyrosine derivatives were isolated and characterized by MS and NMR analyses including HSQC and HMBC and proven to be O- or N-prenylated derivatives at position C4 of the benzene rings. K _M values of six selected derivatives were found in the range of 0.10–0.68 mM. Catalytic efficiencies (K _cat/K _M ) were determined in the range of 430–1,110 s⁻¹·M⁻¹ with l-tyrosine as the best substrate. In addition, 7 of 14 tested tryptophan analogs were also accepted by SirD and converted to C7-prenylated derivatives, which was confirmed by comparison with products obtained from enzyme assays using a 7-dimethylallyltryptophan synthase 7-DMATS from Aspergillus fumigatus.     

Dependence on aerial respiration and its influence on microdistribution in the invasive freshwater snail <Emphasis Type="Italic">Pomacea canaliculata</Emphasis> (Caenogastropoda,Ampullariidae)

The invasive Neotropical snail Pomacea canaliculata is usually regarded as amphibious, although the relative significance of aerial and aquatic respiration is unknown. To investigate the degree of dependence on aerial respiration and its influences on microdistribution, experiments were performed in the laboratory and under seminatural and natural conditions. Restriction of aerial respiration negatively affected survivorship, activity and feeding, its effects worsening with temperature and water fouling; females were more seriously affected than males although the effect depended on reproductive effort. Snails were unevenly distributed relative to the access to air, both in a stream and in an outdoor tank, being concentrated less than 2–4 m from the nearest emergent substratum. Accessibility to air would be an important trait of waterbodies prone to invasions of P. canaliculata, especially in tropical areas. The effectiveness of some control measures could be increased by focusing on areas where snails are concentrated due to their dependence on air.     

Ectopic over-expression of two apple <Emphasis Type="Italic">Flowering Locus T</Emphasis> homologues, <Emphasis Type="Italic">MdFT1</Emphasis> and <Emphasis Type="Italic">MdFT2</Emphasis>, reduces juvenile phase in <Emphasis Type="Italic">Arabidopsis</Emphasis>

To get insight into mechanism by which apple tree (Malus domestica Borkh.) regulates flowering, two apple flowering locus T (FT) homologues, MdFT1 and MdFT2, were isolated from the leaf cDNAs of cultivar Gala. The open reading frames (ORFs) of two MdFTs encoded 174 amino acids. The deduced amino acid sequence of MdFT1 and MdFT2 showed 94.3 % similarity to each other, while 72.6 and 76.0 % to AtFT protein, respectively. Semi-quantitative RT-PCR indicated their specific expression in leaves. Visualization of MdFT2-GFP fusion protein demonstrated its localization on membrane. Ectopic overexpression of either MdFT1 or MdFT2 in Arabidopsis significantly induced early flowering by activating the downstream flowering-related genes.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

A new methanol assimilating yeast, <Emphasis Type="Italic">Ogataea</Emphasis><Emphasis Type="Italic">parapolymorpha</Emphasis>, the ascosporic state of <Emphasis Type="Italic">Candida</Emphasis><Emphasis Type="Italic">parapolymorpha</Emphasis>

Ogataea parapolymorpha sp. n. (NRRL YB-1982, CBS 12304, type strain), the ascosporic state of Candida parapolymorpha, is described. The species appears homothallic, assimilates methanol as is typical of most Ogataea species and forms hat-shaped ascospores in asci that become deliquescent. O. parapolymorpha is closely related to Ogataea angusta and Ogataea polymorpha. The three species can be resolved from gene sequence analyses but are unresolved from fermentation and growth reactions that are typically used for yeast identification. On the basis of multiple isolates, O. angusta is known only from California, USA, in association with Drosophila and Aulacigaster flies, O. parapolymorpha is predominantly associated with insect frass from trees in the eastern USA but O. polymorpha has been isolated from various substrates in the USA, Brazil, Spain and Costa Rica.     

Isolation and <Emphasis Type="Italic">S</Emphasis>-genotyping application of <Emphasis Type="Italic">S</Emphasis>-allelic polymorphic <Emphasis Type="Italic">MdSLFB</Emphasis>s in apple (<Emphasis Type="Italic">Malus domestica</Emphasis> Borkh.)

Apple (Malus domestica Borkh.) possesses gametophytic self-incompatibility (GSI) which is controlled by S-RNase in the pistil as well as a pollen S-determinant that has not been well characterized. The identification of S-locus F-box brother (SFBB) genes, which are good candidates for the pollen S-determinant in apple and pear, indicated the presence of multiple S-allelic polymorphic F-box genes at the S-locus. In apple, two SFBB gene groups have been described, while there are at least three groups in pear. In this report, we identified five MdSLFB (S-RNase-linked F-box) genes from four different S-genotypes of apple. These genes showed pollen- and S-allele-specific expression with a high polymorphism among S-alleles. The phylogenetic tree suggested that some of them belong to SFBBα or β groups as described previously, while others appear to be different from SFBBs. In particular, the presence of MdSLFB3 and MdSLFB9 suggested that there are more S-allelic polymorphic F-box gene groups in the S-locus besides α and β. Based on the sequence polymorphism of MdSLFBs, we developed an S-genotyping system for apple cultivars. In addition, we isolated twelve MdSLFB-like genes, which showed pollen-specific expression without S-allelic polymorphism.     

The <Emphasis Type="Italic">Caenorhabditis</Emphasis><Emphasis Type="Italic">briggsae</Emphasis> genome contains active <Emphasis Type="Italic">CbmaT1</Emphasis> and <Emphasis Type="Italic">Tcb1</Emphasis> transposons

The maT clade of transposons is a group of transposable elements intermediate in sequence and predicted protein structure to mariner and Tc transposons, with a distribution thus far limited to a few invertebrate species. We present evidence, based on searches of publicly available databases, that the nematode Caenorhabditis briggsae has several maT-like transposons, which we have designated as CbmaT elements, dispersed throughout its genome. We also describe two additional transposon sequences that probably share their evolutionary history with the CbmaT transposons. One resembles a fold back variant of a CbmaT element, with long (380-bp) inverted terminal repeats (ITRs) that show a high degree (71%) of identity to CbmaT1. The other, which shares only the 26-bp ITR sequences with one of the CbmaT variants, is present in eight nearly identical copies, but does not have a transposase gene and may therefore be cross mobilised by a CbmaT transposase. Using PCR-based mobility assays, we show that CbmaT1 transposons are capable of excising from the C. briggsae genome. CbmaT1 excised approximately 500 times less frequently than Tcb1 in the reference strain AF16, but both CbmaT1 and Tcb1 excised at extremely high frequencies in the HK105 strain. The HK105 strain also exhibited a high frequency of spontaneous induction of unc-22 mutants, suggesting that it may be a mutator strain of C. briggsae.     

Prevalence,intensity and associated factor analysis of <Emphasis Type="Italic">Tropilaelaps</Emphasis><Emphasis Type="Italic">mercedesae</Emphasis> infesting <Emphasis Type="Italic">Apis</Emphasis><Emphasis Type="Italic">mellifera</Emphasis> in China

Tropilaelaps mercedesae is a serious ectoparasite of Apis mellifera in China. The aim of this study was to investigate the infestation rates and intensity of T. mercedesae in A. mellifera in China, and to explore the relative importance of climate, district, management practices and beekeeper characteristics that are assumed to be associated with the intensity of T. mercedesae. Of the 410 participating apiaries, 379 apiaries were included in analyses of seasonal infestation rates and 352 apiaries were included in multivariable regression analysis. The highest infestation rate (86.3%) of T. mercedesae was encountered in autumn, followed by summer (66.5%), spring (17.2%) and winter (14.8%). In autumn, 28.9% (93) of the infested apiaries were in the north (including the northeast and northwest of China), 71.1% (229) were in the central and south (including east, southeast and southwest China), and 306 apiaries (82.9%) were co-infested by both T. mercedesae and Varroa. Multivariable regression analysis showed that geographical location, season, royal jelly collection and Varroa infestation were the factors that influence the intensity of T. mercedesae. The influence of beekeeper’s education, time of beekeeping, operation size, and hive migration on the intensity of T. mercedesa was not statistically significant. This study provided information about the establishment of the linkage of the environment and the parasite and could lead to better timing and methods of control.