Chromosome evolution in the genus Cicindela: physical mapping and activity of rDNA loci in the tiger beetle species Cicindela littoralis and C. flexuosa

Cicindela littoralis and Cicindela flexuosa were analysed at population level to determine the localization and activity of ribosomal genes. Fluorescence in situ hybridization (FISH), using a PCR‐amplified 18S rDNA fragment as a probe, revealed the presence of polymorphism regarding the number of chromosomes with ribosomal genes as well as their localization within the genome. Nine populations of C. littoralis showed a consistent pattern of two loci located in an autosomal pair (active during spermatogenesis as shown by silver staining) and one locus located in one of the multiple X chromosomes (silent during spermatogenesis), whereas individuals from the population of Punta Entinas showed only signals in the autosomal pair, lacking the heterosomal locus. In C. flexuosa, two patterns were also observed. Nine populations showed two loci in an autosomal pair whereas individuals from the population of San Pedro del Pinatar showed the two loci located in the heterosomes (one of the multiple Xs and in the Y). The hypothesis that these two different populations may reflect a status of well‐differentiated phylogenetic entities is not supported for C. littoralis after the phylogenetic analysis of a fragment of the cytochrome oxidase I gene.     

Microhabitat segregation and physiological differences in co-occurring tiger beetle species,Cicindela oregona and Cicindela tranquebarica

Summary The daily movements of two co-occurring tiger beetle species were monitored in conjunction with changes in microclimate along streams in Northeast Arizona. Cicindela oregona and C. tranquebarica temporarily segregated across areas of beach exhibiting different microclimates. C. oregona progressively moved from the dry upper beach to the wet stream edge as beach temperatures increased and humidity decreased. The actively foraged throughout the day in this moist habitat at air temperatures between 25 and 38°C. C. tranquebarica remained on the dry, upper portions of the beach and shuttled between sun and shade at air temperatures above 35°C. Only when stream edge temperatures exceeded 30°C was tranquebarica found in this subhabitat. Both species exhibited physiological tolerances in the laboratory that were consistent with their microhabitat preferences in the field. Although both species had similar high lethal temperatures (47–48°C) in saturated air, oregona died at lower temperatures (39–43°C) than tranquebarica (46–47°C) under dry (0% RH) conditions. C. oregona was considerably more active than tranquebarica at body temperatures below 30°C and exhibited higher levels of active metabolism between 25 and 40°C. In addition, C. tranquebarica exhibited significantly lower water loss rates than oregona at 30, 35 and 40°C.     

Characterization of auditory afferents in the tiger beetle,Cicindela marutha Dow

We have identified a nerve carrying auditory afferents and characterized their physiological responses in the tiger beetle,Cicindela marutha.

Evolution and phylogenetic information content of the ITS-1 region in the tiger beetle Cicindela dorsalis

Sequence divergence in the internal transcribed spacer region 1 (ITS-1) of the ribosomal DNA locus was assessed in subspecies of the coastal North American tiger beetle, Cicindela dorsalis. The spacer region was amplified using the polymerase chain reaction and cloned for sequencing. Of a total of 50 clones obtained from 12 specimens, 42 clones were different in at least one nucleotide position. In a parsimony analysis of these sequences, the main phylogenetic distinction was found to separate sequences from the Gulf of Mexico and the Atlantic Ocean. Within these two assemblages phylogenetic resolution was low, and the variation within individuals was almost as high as the variation within the entire lineage. The pattern of sequence variation suggests the existence of two forms of the ITS-1 that are maintained on different chromosomes. Polymorphisms of limited geographical distribution could be detected, and 41 additional clones were partly sequenced, to assess the geographic distribution of these polymorphisms in more detail. In a population aggregation analysis, the geographic pattern of ITS-1 distribution was basically congruent with that obtained in earlier studies from mitochondrial DNA in the same C. dorsalis populations.      

Assessing detectability for monitoring of rare species: a case study of the cobblestone tiger beetle (Cicindela marginipennis Dejean)

Interpreting data on distribution or population trends may be difficult unless detection probability is accounted for. We wished to determine the detectability of the rare and patchily distributed cobblestone tiger beetle (Cicindela marginipennis) along the upper Genesee River in western New York for development of a monitoring strategy. We used occupancy surveys and distance sampling to examine two types of detectability. The first type was site-level detectability: the probability of detecting a single cobblestone tiger beetle on an occupied cobble bar, calculated using program PRESENCE. The second type was individual-level detectability: the probability of detecting an individual cobblestone tiger beetle in a population on a single cobble bar, calculated using program DISTANCE. Our occupancy surveys consisted of collecting presence and absence data on cobble bars along the Genesee River; these showed a relatively narrow range of site-level detection probabilities (0.60?C0.68) for cobblestone tiger beetles in 2008 and 2009. Three visits were necessary to detect cobblestone tiger beetles on 90% of occupied cobble bars. Individual cobblestone tiger beetles were detectable one-half of the time (0.50) in our surveys. It is important for ecologists to distinguish between the two kinds of detectability, as monitoring implications could differ substantially depending on which is calculated. Our monitoring recommendations include (1) continuing occupancy surveys with at least three visits to each cobble bar; (2) conducting occupancy surveys between 10:00 and 17:00 on warm sunny days in mid-July and mid-August; and (3) conducting surveys at three- to five-year intervals depending on the study objective.     

Using molecular and ecological data to diagnose endangered populations of the puritan tiger beetle Cicindela puritana

Populations of the puritan tiger beetle Cicindela puritana in the eastern United States were found to be highly threatened at the Connecticut River, whereas several large populations on the western shore and newly discovered populations on the eastern shore of the Chesapeake Bay appeared to be less endangered. We assessed if the disjunct C. puritana subgroups are genetically distinct and therefore should be treated as separate units for conservation purposes. A total of 13 individuals from the Connecticut River and 27 individuals from the Chesapeake Bay were each analysed by sequencing of up to 837 base pairs of mitochondrial DNA per individual. Five different haplotypes could be distinguished. In a phylogenetic analysis of these DNA sequences that included four related Cicindela species as out-groups, haplotypes from the Chesapeake Bay represent a distinct clade. The conservation status of these populations was evaluated using a phylogenetic approach based on cladistic analysis and the framework of the phylogenetic species concept. According to this analysis, beetles from the Connecticut River and the Chesapeake Bay have to be considered as independent units. Populations from the eastern and western shore of Chesapeake Bay are not split in more than one unit using the same criteria, although they exhibited some degree of genetic subdivision. The results from the mtDNA analysis were corroborated by ecological parameters in that the Chesapeake Bay populations can be distinguished from all congeners by their different tat association.     

Structure of the visual system of the larva of the tiger beetle (Cicindela chinensis)

The visual system of the larval tiger beetle (Cicindela chinensis) consists of six (two large, two mediumsized, and two small) stemmata on either side of the head, and an underlying neuropil mass. Each stemma exhibits a corneal lens and an underlying rhabdom layer. Retinular cells extend single proximal axons into the neuropil mass. The neuropil mass has a flattened heart-shape, and consists of two juxtaposed identical structures, each being a neuropil complex of each of the two large stemmata. The complex consists of lamina and medulla neuropils. Most retinular axons terminate in the lamina neuropil. Axons of two types of lamina monopolar neurons descend parallel to each other into the lamina neuropil. Moreover, each lamina neuropil contains a single giant monopolar neuron. Possible centrifugal processes and tangential neurons also occur. Lamina monopolar axons descend straight into the medulla neuropil. Medulla neurons spread fan-shaped dendrites distally in the medulla neuropil and send single axons toward the protocerebrum. These data are discussed with respecct to the unique visual behavior of this larva and in comparison with other insect visual systems.     

Species designation of the Bruneau Dune tiger beetle (Cicindela waynei) is supported by phylogenetic analysis of mitochondrial DNA sequence data

Beetles comprise nearly one quarter of described species and show high levels of morphological and ecological diversification. Because of their wide distribution, ease of detection, and correlation of species richness patterns with other taxa, tiger beetles have been recommended for use as a global indicator of regional biodiversity, requiring accurate taxonomic designations. The Bruneau Dune tiger beetle (Cicindela waynei), whose habitat consists of an isolated dune field in southern Idaho, was recently described as a distinct species from the St. Anthony Dunes tiger beetle (C. arenicola) based on morphological characteristics. While these characteristics include distinct differences in genital morphology that could indicate intrinsic reproductive isolation, morphological characteristics have sometimes been misleading in tiger beetle taxonomy. To evaluate genetic support for this species designation, we analyzed 1,751 base pairs of mitochondrial DNA sequence from 147 tiger beetles collected throughout the range of both C. arenicola and C. waynei. Maximum-likelihood and Bayesian phylogenetic analyses indicated monophyly for C. waynei on a well-supported, short branch nested within C. arenicola. Bayesian species delimitation analyses strongly supported C. waynei as a distinct species (speciation probability = 1.0) with the estimated time of divergence ca. 14,500–67,000 years ago. This lack of reciprocal monophyly and recency of speciation is consistent with C. waynei as a member of an evolutionary front where speciation has occurred at a rapid rate. Mitochondrial sequence data supports the species designation of C. waynei, emphasizing the need to determine appropriate management for this species and its restricted habitat.     

Behavioral analysis of two distinct visual responses in the larva of the tiger beetle (Cicindela chinensis)

The tiger beetle larva shows two distinct visual responses, a predatory jump and an evasive withdrawal into the burrow (escape). In the present study the visual stimuli controlling these two responses have been behaviorally analyzed in the larva of Cicindela chinensis. The threshold size needed for a target to elicit both responses is a visual angle of 5–7°. The velocities of moving targets needed to elicit the responses are 0.4–33° s⁻¹ for the jump and 0.76–90° s⁻¹ for the escape. Choice between the two responses appears to be controlled by the actual target size rather than by the angular size. It also appears to be controlled by the target height. As the height of the target increases, the probability for the jump decreases, whereas the probability for the escape increases. Response properties of the larva with only a single functional stemma, the other stemmata being occluded, are different from those of the intact larva, which suggests cooperation of at least two stemmata for the release of different visual responses. Visual responses of the one-stemma larva still vary, however, with target size and target height, which suggests the visual responses are partially controlled even by a single stemma. Although our data do not resolve these conflicting results, more than one stemma is necessary for a firm choice between the two responses. Accepted: 13 May 1997     

Neural organization of the lamina neuropil of the larva of the tiger beetle (Cicindela chinensis)

Six neural elements, viz., retinular axons, a giant monopolar axon, straight descending processes (type I), lamina monopolar axons (type II), processes containing clusters of dense-core vesicles (type III), and processes coursing in various directions with varicosities (type IV), have been identified at the ultrastructural level in the lamina neuropil of the larval tiger beetle Cicindela chinensis. Retinular axons make presynaptic contact with all other types of processes. Type I and II processes possess many pre-and postsynaptic loci. Type II processes presumably constitute retinotopic afferent pathways. It remains uncertain whether type I processes are lamina monopolar axons or long retinular axons extending to the medullar neuropil. Type III processes may be efferent neurons or branches of afferent neurons contributing to local circuits. A giant monopolar axon extends many branches throughout the lamina neuropil; these branches are postsynaptic to retinular axons, and may be nonretinotopic and afferent. Type IV processes course obliquely in the neuropil, being postsynaptic to retinular axons, and presynaptic to type I processes.     

Evolutionary dynamics of satellite DNA family PIM357 in species of the genus Pimelia (Tenebrionidae,Coleoptera)

A large number of repeats of a satellite DNA (stDNA) family have been cloned and sequenced from species and populations of the genus Pimelia (Tenebrionidae, Coleoptera). The beetles were collected in the Canary Islands, Morocco, the Iberian Peninsula, and the Balearic Islands in order to analyze the evolutionary forces and processes acting on abundant stDNAs conserved at the genus level. This repetitive family is composed of an abundant A-T-rich stDNA, with basic units of 357 bp. All the sequences obtained showed similarity to the 22 repeat units of the PIM357 stDNA family described previously for six Iberian Pimelia species (Pons et al. 1997 ). An analysis based on similarity shows the presence of three different groups of sequences clearly in accordance with their geographical origin. One is composed of satellite sequences from Iberian and Balearic species, a second group from the Moroccan taxa, whereas the third one is from the Pimelia species endemic to the Canary Islands. The latter group shows higher nucleotide diversities for their stDNA sequences and a lack of relationship between transition stages to fixation and sequence divergence. Phylogeographic data of Canarian Pimelia show that the PIM357 stDNA family has persisted for more than 8 Myr and could probably be traced to the origin of the lineage. The data suggest that distinct demographic and phylogenetic patterns related to the colonization of the volcanic Canarian island chain account for particular evolutionary dynamics of the repeat DNA family in this group.     

The function of turret building behaviour in the larval tiger beetle, Cicindela willistoni (Coleoptera: Cicindelidae)

Abstract. 1. Cicindela willistoni LeConte (Coleoptera: Cicindelidae) adults and larvae occur only on saline and alkali flats with little or no vegetation in southwestern U.S.A. Unlike other larval tiger beetle species, C.willistoni constructs a chimney-like extension (turret) 1–4 cm above its burrow.
2. The function of these unique turrets was tested in relation to several biotic and abiotic factors in the Sulphur Springs Valley, Arizona, U.S.A.
3. The turrets did not prevent flooding of the larval burrows.
4. They did not prevent or reduce the frequency of parasitoids (mainly bombyliid flies of the genus Anthrax ).
5. The turrets did not make it possible for the adult and larval beetles to feed on obviously different types or sizes of prey.
6. However, larvae with turrets were raised sufficiently above the substrate surface and boundary layer to allow them to thermoregulate in the lower temperatures at these heights and remain active throughout the day.
7. The shade produced by the turrets attracted significantly more potential prey items than were available to turretless larvae in the same habitat.     

Quaternary vicariance of tiger beetle,Cicindela chinensis,in Ryukyu,Japan, Taiwan and Korea–China

We show vicariance of Cicindela chinensis in Okinawa, Japan (differentiated within Japan) and Korea–China through construction of Bayesian inference trees by BEAST2. Calibration was done using an assumption of the MRCA expansion of C. chinensis at 1.55 Ma (=geologically obtained formative time of the Ryukyu islands) following the protocol of BEAUti. We derived substitution rates for mitochondrial COI (1.66%/m.y.) and nuclear 28SrRNA (0.109%/m.y.) of analyzed Cicindela. Cicindela ferriei is a sister of C. chinensis, and these two species differentiated from each other at ca. 3 Ma before the expansion of C. chinensis. However, they are not strongly differentiated between Amami‐Oshima and Tokuno‐shima, although they display different color. Vicariance at 1.55 Ma is also recognized between Cicindela batesi in Taiwan and Cicindela aurulenta and virgula in continental China. From the sequence data we obtained, it is also evident that C. c. okinawana recently colonized Ishigaki‐jima from Okinawa‐jima, as did C. batesi in Iriomote‐jima from Taiwan.     

Water loss in three species of tiger beetles (Cicindela): Correlations with epicuticular hydrocarbons

Water loss rates in dry air at 30°C and cuticular lipid/hydrocarbon composition were determined for three species (eight populations) of tiger beetles (Cicindela oregona, C. tranquebarica and C. obsoleta). The highest water loss rates were found in C. oregona (0.049 to 0.052 mg cm⁻²h⁻¹ mmHg⁻¹), a species active in spring and fall along water courses in Arizona, while the lowest rates were exhibited by C. obsoleta (0.022 to 0.028 mg cm⁻²h⁻¹ mmHg⁻¹), a summer-active species that inhabits dry grasslands. Water loss rates for C. tranquebarica were closer to those of C. obsoleta even though C. tranquebarica often coexists with C. oregona. Hydrocarbons were an important constituent of the cuticular lipids of all three species; smaller quantities of wax and cholesterol esters, triacylglycerols, free fatty acids, alcohols, and cholesterol were also detected. C. obsoleta contained the greatest amount of hydrocarbon per surface area. All of its hydrocarbon molecules were saturated, with branched components accounting for about 60% of the total fraction. Saturated (primarily n-alkanes) and unsaturated (n-alkenes) molecules were present in both C. oregona, which contained the lowest hydrocarbon surface density, and C. tranquebarica. The hydrocarbon composition of the three species is discussed in terms of its contribution to the epicuticular waterproofing barrier and its potential use as a chemotaxonomic tool.     

Wind-powered wheel locomotion, initiated by leaping somersaults, in larvae of the southeastern beach tiger beetle (Cicindela dorsalis media)

Rapid movement is challenging for elongate, soft-bodied animals with short or no legs. Leaping is known for only a few animals with this "worm-like" morphology. Wheel locomotion, in which the animal's entire body rolls forward along a central axis, has been reported for only a handful of animals worldwide. Here we present the first documented case of wind-powered wheel locomotion, in larvae of the coastal tiger beetle Cicindela dorsalis media. When removed from their shallow burrows, larvae easily can be induced to enter a behavioral sequence that starts with leaping; while airborne, larvae loop their body into a rotating wheel and usually either "hit the ground rolling" or leap again. The direction larvae wheel is closely related to the direction in which winds are blowing; thus, all our larvae wheeled up-slope, as winds at our study site consistently blew from sea to land. Stronger winds increased both the proportion of larvae wheeling, and the distance traveled, exceeding 60 m in some cases. In addition, the proportion of larvae that wheel and the distance traveled by wheeling larvae are significantly greater on smooth sandy beaches than on beach surfaces made rough and irregular by pedestrian, equestrian, and vehicular traffic. Like other coastal species of tiger beetles, C. dorsalis media has suffered major declines in recent years that are clearly correlated with increased human impacts. The present study suggests that the negative effects of beach traffic may be indirect, preventing larvae from escaping from predators using wheel locomotion by disrupting the flat, hard surface necessary for efficient wheeling.     

Salinity and shade preferences result in ovipositional differences between sympatric tiger beetle species

1. Adult tiger beetles of the genus Cicindela often co‐occur within a habitat but larvae do not. Larvae are sedentary and form usually permanent burrows at the site of oviposition where they require 1–3 years for development. 2. To test niche partitioning based on ovipositional preference, the behaviour of two sympatric salt marsh tiger beetles, Cicindela circumpicta and C. togata (Coleoptera: Cicindelidae), were examined. 3. In laboratory studies, female C. circumpicta and C. togata distinguished between experimental salinities, with the former preferring 4 parts per thousand (ppt) and the latter preferring 12 ppt. In the field, C. circumpicta larvae were associated with lower salinities (1–3 ppt) and vegetation along the edges of salt flats while C. togata larvae were found on open salt flats often near halophytes (average salinity = 7.8 ppt). 4. In the field, females chose sites for oviposition in response to shade but not vertical landmarks. In a direct test, 53 of 56 new larval burrows occurred in shaded treatments, three in nonshaded controls. 5. Compared with nonshaded controls, shade increased survival of eggs that were collected in the laboratory and placed in the field. For C. circumpicta eggs, 78% placed in shaded treatments hatched, while significantly fewer (22%) hatched in exposed treatments. For C. togata, 43% of eggs placed in the shade hatched, while no eggs placed in exposed treatments hatched. 6. These results support the hypothesis of niche partitioning between C. circumpicta and C. togata based on ovipositional choice and resulting larval habitat.     

Preservation of a complex satellite DNA in two species of echinoderms.

The cloning and sequencing of a tandemly arrayed repetitive DNA sequence from the sea cucumber Holothuria tubulosa has been recently described (Sainz, J., Azorín, F. and Cornudella, L. 1989. Gene 80, 57-64). We have now searched the genomes of several echinoderm species for the presence of homologous repetitive elements. A close but not identical repeated sequence has been identified in a related holothuroid, H. polii. The monomeric repeat unit is 391 bp long and has a base composition of 66.8% A and T residues, lined up in tracts of 4 nt or larger. The monomeric sequence lacks any internal subrepeat organization although it displays a substantial degree of internal redundancy in the form of inverted and direct repeats. The repeated element accounts for 0.34% of the genome which corresponds to a repetition frequency of about 0.5 x 10(5) copies per haploid complement. The intra- and interspecific homologies among monomers of the satellite DNA as derived from sequence analyses are very high, averaging 97%. The results suggest that the homogeneity of the highly reiterated DNA sequence may be attributed to evolutionary conservative trends.