Resistance of the termite, <Emphasis Type="Italic">Coptotermes formosanus</Emphasis> Shiraki to <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> due to grooming

Termites, Coptotermes formosanus, reared individually, were highly susceptible to the entomopathogenic fungus, Metarhizium anisopliae, while termites reared in␣groups were highly resistant. When reared in groups, the termites treated with M.␣anisopliae conidia on the body surface were groomed by their nestmates and more than 80% of the conidia were removed from the cuticle within 3 h. However, there was not a significant reduction in the numbers of conidia on the body surfaces of termites reared individually. For the termites maintained in groups, conidia were found in foreguts, midguts and hindguts, but very few conidia were detected in the guts of termites reared individually. Conidia in the alimentary tracts did not germinate, but some of were alive. As a result, it seems that the removal of foreign bodies, such as fungal conidia, from the␣cuticle is one function of termite mutual grooming behavior and that conidia removed from the cuticle are eliminated through alimentary tracts. This study indicates that mutual grooming behavior is very effective in protecting these termites from M.␣anisopliae infection.     

Mechanism,induction factors,and adaptive significance of dealation in the subterranean termite <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Isoptera,Rhinotermitidae)

Summary. We studied the dealation of a subterranean termite Reticulitermes speratus Kolbe from three different angles; mechanism, induction factors, and adaptive significance. During nuptial flight alates prevent the wings from breaking off by holding the wings in a 3-dimensional configuration. In the process of dealation, the alates hold the wings in a 2-dimensional configuration, half spread the wings in a lowered position, and bend the abdomen back toward the wings, thereby exerting high pressure at the basal sutures. This separates the wings from the body. The primary factor determining the timing of dealation appears to be isolation from other alates: It induces alates to shed their wings, whereas pairing has no effect on dealation in this species. Alates of termites are especially vulnerable to ant predation during the period between flight and colony foundation. An experiment designed to compare the risk of alates and dealates to predation by a termite-hunting ant, Brachyponera chinensis Emery, showed that dealation reduced the predation risk during the period spent running on the ground.     

DNA methylation in the termite <Emphasis Type="Italic">Coptotermes lacteus</Emphasis>

Social insects are key examples of organisms that display polyphenism. Their genomes encode instructions for the development of multiple phenotypes, known as castes, which typically have highly divergent morphology, physiology and behaviour. DNA methylation, an epigenetic mechanism associated with modulation of gene expression in various eukaryotes, has recently been shown to provide a key link between environmental cues and caste-specific gene expression in honey bees (Hymenoptera). In termites—a major social insect group phylogenetically distant from Hymenoptera—the existence of DNA methylation has not, to our knowledge, been reported to date. Since genes encoding key DNA methylation enzymes are known to be absent in the genomes of a number of insect species, we sought to test whether termites are able to methylate their DNA, and, if so, whether caste-specific patterns of DNA methylation exist. We performed methylation-specific amplified fragment length polymorphism on the termite Coptotermes lacteus, and found evidence for DNA methylation. However, a comparison of methylation levels in different castes did not reveal any significant differences in methylation levels. The demonstration of DNA methylation in termites sets the stage for future epigenetic studies in these important social insects.     

Comparative nutrient economy,stable isotopes,and related adaptive traits in <Emphasis Type="Italic">Picea rubens</Emphasis>, <Emphasis Type="Italic">Picea mariana</Emphasis>, and their hybrids

Nutrient- and water economy-related traits in plants have significant implications for growth and fitness. We explored, examined, and compared nutrient concentrations, use efficiencies, assimilation, and informative isotopic elements in a seedling provenance experiment, and in seedling and mature tree controlled-cross hybrid experiments of red spruce (RS) (Picea rubens Sarg.) and black spruce (BS) (P. mariana (Mill.) BSP). Provenance experiment results showed RS had consistently lower carbon (C) and nitrogen (N) concentrations, and N assimilation ratio (NAR), but higher N-use efficiency (NUE), C:N ratio, water-use efficiency (WUE), and needle calcium (Ca) and magnesium (Mg) concentrations than BS. The hybrid seedling experiment showed similar results and additive inheritance for needle N, C:N ratio, NAR, Ca, and Mg, evident by a near-linear progression from one species to the other. Within both species, seedling height showed a negative relationship with needle N and a positive relationship with NUE. However, across hybrid indices, seedling height showed a positive relationship with needle N and a negative relationship with NUE. Also across hybrid indices, seedling height showed a negative relationship to Ca and C:N, and a positive relationship with NAR and ¹³C discrimination (without hybrid 25). Mature tree hybrid experiment results were similar to those of the seedling experiment, but with a dampening of differences caused by low nutrient availability and possibly age effects. The similarity was not true for ¹³C discrimination as mature tree height showed a strong negative relationship to ¹³C discrimination, indicating that BS had greater WUE. The reversal is most probably caused by the large difference in water availability.     

Biological changes in the Eastern subterranean termite, <Emphasis Type="Italic">Reticulitermes flavipes</Emphasis> (Isoptera,Rhinotermitidae) and its protozoa profile following starvation

The effects of starvation on survival, body mass, movement, other behavior, and the symbiotic protozoan community in Reticulitermes flavipes (Kollar) were investigated in a 40-day assay. Groups of 100 termites in their natural worker/soldier ratio (98/2) were tested. Starvation resulted in significantly greater mortality and induced more cannibalism of workers than in the control. Worker survival rate gradually declined to 58% along with an increasing rate of cannibalism during the first 30 days, and then quickly decreased to 5% along with an increasing rate of necrophagia after 30 days. In contrast, starvation had no significant effect on the survival of soldiers (60%) as compared to the control (90%) and starved workers did not cannibalize soldiers. Ten protozoan species residing in the hindgut were identified. When compared with field termites, the 40-day starvation eliminated 5 species (Trichonympha agilis, Pyrsonympha vertens, and P. major, D. gracilis, Holomastigotes elongatum), significantly reduced the numbers of two species (Dinenympha fimbriata, Spironympha kofoidi), had no effect on two species (Trichomitus trypanoides and Spirotrichonympha flagellata), and led to proliferation of one species (Monocercomonas sp.), whereas feeding on filter paper reduced the populations of five species (T. agilis, D. fimbriata, P. vertens, P. major, and S. flagellata) to different degrees. Workers surviving starvation had similar body mass and short-range movement speed to workers fed on filter paper and workers freshly collected from natural setting. The energy-demanding survival and walking indicated that they might obtain sufficient nutrients from cannibalizing other workers. The observed behaviors are important in helping to understand termite survival strategies and the mechanisms by which termites maintain their social structure under stressful conditions.     

Taxonomy and phylogeny of north mediterranean <Emphasis Type="Italic">Reticulitermes</Emphasis> termites (Isoptera,Rhinotermitidae): a new insight

Summary The molecular characterisation of 18 new populations of Reticulitermes is here presented for COII and 16S genes; results are elaborated and compared to all available ones, at distances and gene trees (Maximum Parsimony, Maximum likelihood, Bayesian analysis) levels. Within the R. lucifugus complex, a subspecific rank of differentiation appears tenable for Italian (R. lucifugus lucifugus, R. lucifugus corsicus) and European (R. lucifugus banyulensis, R. lucifugus grassei) taxa; a subspecific differentiation emerges also for the Sicilian samples. The existence of two different entities in the area formerly defined as inhabited by R. balkanensis is demonstrated; The north-eastern Italian Reticulitermes sp. is found to be more widely distributed in northern and south-eastern Italy and shows a close relationship to the sample from Peloponnese; the GenBank sample from continental Greece, on the contrary, appears more related to other eastern taxa such as R. lucifugus from Turkey and R. clypeatus from Israel; the distribution and differentiation of eastern Reticulitermes taxa are explained through the role that southern Balkans should have played as glacial refuge. Present data also evidences instances of anthropic involvement in taxa distribution; for one of these, the importation of at least a family group is taken into account.Received 8 May 2003; revised 1 July and 28 August 2003; accepted 24 September 2003.     

Alate differentiation and compound-eye development in the dry-wood termite <Emphasis Type="Italic">Neotermes koshunensis</Emphasis> (Isoptera,Kalotermitidae)

In social insects, caste-specific characters develop in the postembryonic differentiation processes. However, the mechanisms of caste-specific organ development have yet to be elucidated. In order to obtain insights into the relationship between caste differentiation and the regulation of organ development, we determined the caste-developmental pathway and observed compound-eye development accompanying alate differentiation in the dry-wood termite, Neotermes koshunensis. As previously reported in other Neotermes, this species has a linear caste-developmental pathway, comprising six larval- and two nymphal-instar stages. Although the apparent eye formation occurs during the last nymphal stages, just prior to the imaginal molt, individuals possess eye primordia from the first larval-instar stage. The outer morphological structure of the eye was observed from the third larval-instar stage. The detailed differentiation of cells constituting ommatidia appeared to occur in relatively young larval instars (fourth stage), although the pigmentation of pigment cells and detailed structural formation of ommatidia occurred during the final stage of alate development, i.e., during the late second nymphal-instar stage. This suggests that eye development is arrested in the larval stages, and then resumed during the late nymphal stage to complete functional eye formation, which is required for nuptial flight. In comparison to major hemimetabolous insects, which possess functional compound eyes even at the first instar larva, this termite species shows the heterochronic shift in terms of compound-eye development. Received 20 March 2006; revised 24 September 2006; accepted 4 October 2006.     

Sexual and asexual colony foundation and the mechanism of facultative parthenogenesis in the termite <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Isoptera,Rhinotermitidae)

Summary. Facultative parthenogenesis has great adaptive significance, especially with regard to low pairing efficiency. In the termite Reticulitermes speratus, females that fail to mate with males reproduce parthenogenetically and found colonies cooperatively with partner females or even alone. Comparison of colony foundation success at 400 days between colonies founded by single females (F), female-female pairs (FF), and male-female pairs (FM) showed that female-female cooperation promoted colony survivorship over monogamous foundation. We report here for the first time the mode of parthenogenesis in Isoptera. Combining chromosome observations and genetic analysis using microsatellites, we show that the mode of parthenogenesis is diploid thelytoky and that the restoration of ploidy is most likely accomplished by terminal fusion. Parthenogens show a higher mortality and a longer egg-development time than sexually produced offspring, probably due to reduced heterozygosity. In addition Wolbachia bacteria were detected in R. speratus. However, since Wolbachia was also detected in non-parthenogenetic R. flavipes, it is unlikely that Wolbachia is the cause of parthenogenesis in R. speratus.Received 30 October 2003; revised 4 March 2004; accepted 17 March 2004.     

<Emphasis Type="Italic">Sporidiobolus johnsonii</Emphasis> and <Emphasis Type="Italic">Sporidiobolus salmonicolor</Emphasis> revisited

The relationship between Sporidiobolus johnsonii and S. salmonicolor was investigated using rDNA sequence data. Two statistically well-supported clades were obtained. One clade included the type strain of S. johnsonii and the other included the type strain of S. salmonicolor. However, some mating strains of S. salmonicolor were found in the S. johnsonii group. These strains belonged to mating type A2 and were sexually compatible with mating type A1 strains from the S. salmonicolor group. DNA–DNA reassociation values were high within each clade and moderate between the two clades. In the re-investigation of teliospore germination, we observed that the basidia of S. salmonicolor were two-celled. In S. johnsonii, basidia were not formed and teliospore germination resulted in direct formation of yeast cells. We hypothesize that the S. johnsonii clade is becoming genetically isolated from the S. salmonicolor group and that a speciation process is presently going on. We suspect that the observed sexual compatibility between strains of the S. johnsonii and S. salmonicolor groups and the possible genetic flow between the two species has little biological relevance because distinct phenotypes have been fixed in the two taxa and intermediate (hybrid) sequences for LSU and ITS rDNAs have not been detected. An erratum to this article can be found at     

Comparative Feeding Ecology of Two Sympatric Greenling Species, <Emphasis Type="Italic">Hexagrammos otakii</Emphasis> and <Emphasis Type="Italic">Hexagrammos agrammus</Emphasis> in Eelgrass <Emphasis Type="Italic">Zostera marina</Emphasis> Beds

Synopsis Feeding ecology was compared between sympatric greenling species, Hexagrammos otakii and H. agrammus in the eelgrass beds in Jindong Bay, Korea, from January to December. These two species had similar diets composed of crustaceans, polychaetes, gastropods and fishes; both species consumed primarily crustaceans throughout study periods. H. otakii, however, fed a greater proportion of polychaetes and fishes than H. agrammus. H. agrammus had a greater proportion of gastropods in their diets. The diet of both species underwent size-related changes; smaller individuals of H. otakii and H. agrammus consumed amphipods (gammarid amphipods and caprellid amphipods), while larger individuals of H. otakii ate polychaetes and fishes and those of H. agrammus fed mainly on gastropods and crabs. The diet of H. otakii underwent seasonal changes; H. otakii consumed mainly polychaetes and fishes during January and February 2002 but amphipods during March and May 2002. H. agrammus, however, ate mainly gastropods and crabs all seasons. H. otakii underwent also a significant diel changes that could be related to difference of prey availability. Thus the nocturnal emergence of gammarid amphipods, polychaetes and fishes explained their greater consumption by H. otakii. Dietary breadth of both species was lower in the smallest individuals (<5 cm SL) and in March and April 2002. This was due to the disproportionate dry mass attributable to the consumption of amphipods by both H. otakii and H. agrammus. Dietary overlap of both species was relatively moderate to high, in particular in <9.9 cm SL (0.62 – 0.71) from May to July 2002 (0.63 – 0.71). This is coincident with higher abundances of crabs, caridean shrimps and polychaetes in the study area, and it was assumed that these prey species were not limited resources. Higher dietary overlap was correlated with an abundance of a shared resource and did not indicate the interspecific competition between H. otakii and H. agrammus.     

Genetic diversity and invasion history of the European subterranean termite <Emphasis Type="Italic">Reticulitermes urbis</Emphasis> (Blattodea,Termitoidae, Rhinotermitidae)

Biological invasions are among key factors of ecological changes, and social insects appear as highly successful invasive animals. Subterranean termites of the holarctic genus Reticulitermes are present in Europe with six native and one invasive (the nearctic R. flavipes) species. The species R. urbis shows a disjunct distribution in the Western Balkans, Eastern Italy and Southern France. Previous molecular and population genetics data suggested that the taxon originated from the Balkans, and that Italian and French populations are invasive, but it is still unknown how many introduction events occurred and from which Balkan source populations. To address these questions, a population genetics analysis was performed on a larger sampling than previous studies, using mitochondrial cytochrome oxidase II and 6 microsatellite markers on 47 colonies collected across the whole distribution area. Mitochondrial analysis confirmed the presence of two major lineages where colonies from Balkans, Italy, and France intermingle. Similarly, microsatellite loci analysis indicated the presence of two genetic clusters, though not corresponding to the two mitochondrial clades, each including colonies from the three sampled areas and with individuals showing mixed cluster membership. Overall, French and Italian populations showed indications of bottleneck (reduced genetic diversity and change of allele frequencies) and do not appear genetically differentiated from the Balkan population. Results presented here support a history of multiple introductions in Italy and France, in a scenario consistent with continuous exchanges between native and invasive areas, as expected along human trades routes.     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

Colony-dependent sex differences in protozoan communities of the lower termite <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Isoptera: Rhinotermitidae)

In many animals, sex differences in hormones, behavior, and immunity lead to differences in their gut microbial communities. One of the best-known examples of mutualistic symbiosis is that between lower termites and their intestinal protozoa. Although differences in the protozoan communities of different castes have been studied in lower termites, nothing is known about the sex differences in protozoan communities in neuter castes. Here, we show that termite workers have different protozoan communities according to sex depending on the colony. We investigated the communities of symbiotic protozoa living in lower termites, Reticulitermes speratus, and how they are affected by sex and caste. Workers had the largest numbers of protozoa, followed by soldiers, whereas reproductives (primary kings and secondary queens) had no protozoa. Workers showed colony-dependent sex differences in the total abundance of protozoa, whereas soldiers showed no such sex differences. There were significant sex effect and/or interaction effect between colony and sex in abundances of five species of protozoa in workers. Workers also showed significant sex differences and/or colony-dependent sex differences in proportion of six species of protozoa. These may result in sex differences in the host–symbiont interaction due to physiological or behavioral sex differences in workers that have not been recognized previously. This study has an important implication: although workers are not engaged in reproduction, their potential sex difference may affect various aspects of social interactions.     

<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.     

Agonistic Behavior of the Termite <Emphasis Type="Italic">Prorhinotermes canalifrons</Emphasis> (Isoptera: Rhinotermitidae)

While intercolonial and interspecific agonism has been broadly documented in termites, detailed studies of individual behavior and its differentiation among castes are scarce. We studied the responses of non-reproductive castes of Prorhinotermes canalifrons (pseudergates, presoldiers, soldiers of different age and artificial intercastes) to conspecific and heterospecific aliens. Each caste displayed a specific behavioral repertoire. Soldiers, but to a lesser extent pseudergates as well, are responsible for colony defense. The behavioral repertoire of the soldier appears on the second day after exuviation and becomes complete on the third day, well before its defensive secretion is present. The behavior of artificial intercastes induced by a JH analog is intermediate between that of pseudergates and soldiers. Reactions to heterospecific aliens were much quicker and more violent than reactions to conspecifics, which suggests that individuals experience difficulties in identifying conspecific aliens.     

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.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation of <Emphasis Type="Italic">Perilla frutescens</Emphasis>

A reproducible plant regeneration and an Agrobacterium tumefaciens-mediated genetic transformation protocol were developed for Perilla frutescens (perilla). The largest number of adventitious shoots were induced directly without an intervening callus phase from hypocotyl explants on MS medium supplemented with 3.0 mg/l 6-benzylaminopurine (BA). The effects of preculture and extent of cocultivation were examined by assaying -glucuronidase (GUS) activity in explants infected with A. tumefaciens strain EHA105 harboring the plasmid pIG121-Hm. The highest number of GUS-positive explants were obtained from hypocotyl explants cocultured for 3 days with Agrobacterium without precultivation. Transgenic perilla plants were regenerated and selected on MS basal medium supplemented with 3.0 mg/l BA, 125 mg/l kanamycin, and 500 mg/l carbenicillin. The transformants were confirmed by PCR of the neomycin phosphotransferase II gene and genomic Southern hybridization analysis of the hygromycin phosphotransferase gene. The frequency of transformation from hypocotyls was about 1.4%, and the transformants showed normal growth and sexual compatibility by producing progenies.     

<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.