Genomoviruses associated with mountain and western pine beetles.

Genomoviruses are circular single-stranded DNA viruses (∼2 kb in size) classified into nine genera, they are highly diverse and have been identified in a variety of samples ranging from fungi to animal sera. Here we identify five genomoviruses belonging to the Gemycircularvirus genus and one to the Gemykibivirus genus from mountain pine beetle and western pine beetle sampled in Arizona. Collectively these six viral genomes share <77% genome-wide pairwise identity and hence represent six new species of genomoviruses. Four of the gemycircularviruses from the mountain pine beetles are recombinant, with one having a recombinant region that spans the entire capsid protein. Pine beetles have a symbiotic relationship with certain tree pathogenic fungi. Therefore given that Sclerotinia sclerotiorum hypovirulence-associated DNA virus 1, a gemycircularvirus, induces hypovirulence in the plant pathogenic fungus Sclerotinia sclerotiorum and infects the mycophagous insect Lycoriella ingenua, it is possible that the six genomoviruses identified here may be directly associated with the pine beetle fungal symbionts and/or with the insects themselves.

The Role of Multimodal Signals in Species Recognition Between Tree-Killing Bark Beetles in a Narrow Sympatric Zone.

When related species coexist, selection pressure should favor evolution of species recognition mechanisms to prevent interspecific pairing and wasteful reproductive encounters. We investigated the potential role of pheromone and acoustic signals in species recognition between two species of tree-killing bark beetles, the southern pine beetle, Dendroctonus frontalis Zimmermann, and the western pine beetle, Dendroctonus brevicomis LeConte, in a narrow zone of sympatry, using reciprocal pairing experiments. Given the choice of adjacent con- or heterospecific female gallery entrance in a log, at least 85% of walking males chose the entrance of the conspecific, and half the males that initially entered heterospecific galleries re-emerged and entered the conspecific gallery within 15 min. Waveform analysis of female acoustic "chirps" indicated interspecific differences in chirp timing. Males may use information from female acoustic signals to decide whether to enter or remain in the gallery. Individuals in forced heterospecific pairings (produced by confinement of a heterospecific male within the female entrance) did not differ in pheromone production from individuals of conspecific pairs. However, due to the absence of the right species of male, galleries with heterospecific pairs released an abnormal pheromone blend that lacked at least one key component of the aggregation pheromone of either species. The complete aggregation pheromone (i.e., the pheromone blend from entrances with pairs) does not appear to deter interspecific encounters or confer premating reproductive isolation per se; however, it may confer selective pressure for the maintenance of other reproductive isolation mechanisms.

Identification of diverse circular single-stranded DNA viruses in adult dragonflies and damselflies (Insecta: Odonata) of Arizona and Oklahoma, USA.

Next generation sequencing and metagenomic approaches are commonly used for the identification of circular replication associated protein (Rep)-encoding single stranded (CRESS) DNA viruses circulating in various environments. These approaches have enabled the discovery of some CRESS DNA viruses associated with insects. In this study we identified and recovered 31 viral genomes which represent 24 distinct CRESS DNA viruses from seven dragonfly species (Rhionaeschna multicolor, Erythemis simplicicollis, Erythrodiplax fusca, Libellula quadrimaculata, Libellula saturata, Pachydiplax longipennis, and Pantala hymenaea) and two damselfly species (Ischnura posita, Ischnura ramburii) sampled in various locations in the states of Arizona and Oklahoma of the United States of America (USA). We also identified Sclerotinia sclerotiorum hypovirulence-associated DNA virus-1 (SsHADV-1) in P. hymenaea, E. simplicicollis and I. ramburii sampled in Oklahoma, which is the first report of SsHADV-1 in the New World. The genome architectures of the CRESS DNA viruses recovered vary, but they all have at least two major open reading frames (ORFs) that have either a bidirectional or unidirectional arrangement. Four of the viral genomes recovered, in addition to the three isolates of SsHADV-1, show similarities to viruses of the proposed gemycircularvirus group. Analysis of the Rep encoded by the remaining 24 viral genomes reveals that these are highly diverse and allude to the fact that they represent novel CRESS DNA viruses.

Technique for studying arthropod and microbial communities within tree tissues.

Phloem tissues of pine are habitats for many thousands of organisms. Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear young, feed, or hide from natural enemies or harsh environmental conditions outside of the tree. Organisms that persist within the phloem habitat are difficult to observe given their location under bark. We provide a technique to preserve intact phloem and prepare it for experimentation with invertebrates and microorganisms. The apparatus is called a 'phloem sandwich' and allows for the introduction and observation of arthropods, microbes, and other organisms. This technique has resulted in a better understanding of the feeding behaviors, life-history traits, reproduction, development, and interactions of organisms within tree phloem. The strengths of this technique include the use of inexpensive materials, variability in sandwich size, flexibility to re-open the sandwich or introduce multiple organisms through drilled holes, and the preservation and maintenance of phloem integrity. The phloem sandwich is an excellent educational tool for scientific discovery in both K-12 science courses and university research laboratories.

Using acoustic technology to reduce bark beetle reproduction.

BACKGROUND: Acoustic signals play a critical role in mate choice, species recognition, communication, territoriality, predator escape and prey selection. Bark beetles, which are significant disturbance agents of forests, produce a variety of acoustic signals. RESULTS: A bioacoustic approach to reducing bark beetle reproduction within wood tissues was explored. Playback of modified biological sounds reduced beetle reproductive output, tunneling distance and adult survival. CONCLUSION: The targeted use of biologically relevant sounds disrupts insect behaviors and could be a species-specific, environmentally friendly method of insect management.

Microclimatic challenges in global change biology.

Despite decades of work on climate change biology, the scientific community remains uncertain about where and when most species distributions will respond to altered climates. A major barrier is the spatial mismatch between the size of organisms and the scale at which climate data are collected and modeled. Using a meta-analysis of published literature, we show that grid lengths in species distribution models are, on average, ca. 10 000-fold larger than the animals they study, and ca. 1000-fold larger than the plants they study. And the gap is even worse than these ratios indicate, as most work has focused on organisms that are significantly biased toward large size. This mismatch is problematic because organisms do not experience climate on coarse scales. Rather, they live in microclimates, which can be highly heterogeneous and strongly divergent from surrounding macroclimates. Bridging the spatial gap should be a high priority for research and will require gathering climate data at finer scales, developing better methods for downscaling environmental data to microclimates, and improving our statistical understanding of variation at finer scales. Interdisciplinary collaborations (including ecologists, engineers, climatologists, meteorologists, statisticians, and geographers) will be key to bridging the gap, and ultimately to providing scientifically grounded data and recommendations to conservation biologists and policy makers.

High global diversity of cycloviruses amongst dragonflies.

Members of the family Circoviridae, specifically the genus Circovirus, were thought to infect only vertebrates; however, members of a sister group under the same family, the proposed genus Cyclovirus, have been detected recently in insects. In an effort to explore the diversity of cycloviruses and better understand the evolution of these novel ssDNA viruses, here we present five cycloviruses isolated from three dragonfly species (Orthetrum sabina, Xanthocnemis zealandica and Rhionaeschna multicolor) collected in Australia, New Zealand and the USA, respectively. The genomes of these five viruses share similar genome structure to other cycloviruses, with a circular ~1.7 kb genome and two major bidirectionally transcribed ORFs. The genomic sequence data gathered during this study were combined with all cyclovirus genomes available in public databases to identify conserved motifs and regulatory elements in the intergenic regions, as well as determine diversity and recombinant regions within their genomes. The genomes reported here represent four different cyclovirus species, three of which are novel. Our results confirm that cycloviruses circulate widely in winged-insect populations; in eight different cyclovirus species identified in dragonflies to date, some of these exhibit a broad geographical distribution. Recombination analysis revealed both intra- and inter-species recombination events amongst cycloviruses, including genomes recovered from disparate sources (e.g. goat meat and human faeces). Similar to other well-characterized circular ssDNA viruses, recombination may play an important role in cyclovirus evolution.

Trichogramma parasitoids alter the metabolic physiology of Manduca eggs.

Egg parasitoids face unique developmental constraints. First, they have exceptionally limited resources to support themselves and their siblings through three life stages. Second, they develop within the physiological system of another species, which they modify to their own ends. We examined how these constraints affect the metabolic physiology of egg parasitism, and whether parasitoids retool their host eggshell to account for their different metabolic demands. Higher-conductance eggshells allow more oxygen to reach the developing parasitoids, but also allow more water to leave the egg. We used Manduca sexta (Lepidoptera: Sphingidae) eggs and Trichogramma (Hymenoptera: Trichogrammatidae) parasitoids from southeastern AZ, USA. Compared with unparasitized Manduca eggs, eggs parasitized by Trichogramma had lower peak metabolic rates and approximately 50 per cent lower metabolic efficiency. However, developing Trichogramma were far more efficient than typical transfer efficiencies between tropic levels (approx. 10%). Even within a few hours of parasitization, eggs containing more Trichogramma had lower per-parasitoid metabolic rates, suggesting that parasitoid larvae have mechanisms for rapidly adjusting their metabolic rates based on number of siblings. Parasitoids also appear to control the conductance of their host eggshell: their different metabolic demands were mirrored by shifts in rates of water loss.

Complex life cycles and the responses of insects to climate change.

Many organisms have complex life cycles with distinct life stages that experience different environmental conditions. How does the complexity of life cycles affect the ecological and evolutionary responses of organisms to climate change? We address this question by exploring several recent case studies and synthetic analyses of insects. First, different life stages may inhabit different microhabitats, and may differ in their thermal sensitivities and other traits that are important for responses to climate. For example, the life stages of Manduca experience different patterns of thermal and hydric variability, and differ in tolerance to high temperatures. Second, life stages may differ in their mechanisms for adaptation to local climatic conditions. For example, in Colias, larvae in different geographic populations and species adapt to local climate via differences in optimal and maximal temperatures for feeding and growth, whereas adults adapt via differences in melanin of the wings and in other morphological traits. Third, we extend a recent analysis of the temperature-dependence of insect population growth to demonstrate how changes in temperature can differently impact juvenile survival and adult reproduction. In both temperate and tropical regions, high rates of adult reproduction in a given environment may not be realized if occasional, high temperatures prevent survival to maturity. This suggests that considering the differing responses of multiple life stages is essential to understand the ecological and evolutionary consequences of climate change.

Insect eggs protected from high temperatures by limited homeothermy of plant leaves.

Virtually all aspects of insect biology are affected by body temperature, and many taxa have evolved sophisticated temperature-control mechanisms. All insects, however, begin life as eggs and lack the ability to thermoregulate. Eggs laid on leaves experience a thermal environment, and thus a body temperature, that is strongly influenced by the leaves themselves. Because plants can maintain leaf temperatures that differ from ambient, e.g. by evapotranspiration, plant hosts may protect eggs from extreme ambient temperatures. We examined the degree to which leaves buffer ambient thermal variation and whether that buffering benefits leaf-associated insect eggs. In particular, we: (1) measured temperature variation at oviposition sites in the field, (2) manipulated temperatures in the laboratory to determine the effect of different thermal conditions on embryo development time and survival, and (3) tested embryonic metabolic rates over increasing temperatures. Our results show that Datura wrightii leaves buffer Manduca sexta eggs from fatally high ambient temperatures in the southwestern USA. Moreover, small differences in temperature profiles among leaves can cause large variation in egg metabolic rate and development time. Specifically, large leaves were hotter than small leaves during the day, reaching temperatures that are stressfully high for eggs. This study provides the first mechanistic demonstration of how this type of leaf-constructed thermal refuge interacts with egg physiology.

Electrical stimulation of the growth plate: a potential approach to an epiphysiodesis.

Epiphysiodesis is an operative procedure that induces bony bridges to form across a growth plate of a bone to stop longitudinal growth. This is a very common orthopedic procedure to correct disproportional long-bone growth discrepancies; however, present techniques require an operation and anesthesia. Our study was designed to develop a minimally invasive method of epiphysiodesis by using electrical stimulation with DC current. In a rabbit model, a thin titanium electrode was inserted into a single location of the distal femoral growth plate in three groups: one without current (control), one group with a constant 10 microA (low current, LC), and one group with a 50 microA (high current, HC). The current was delivered for 2 weeks. The nontreated femur served as a control for each animal. Femur lengths were measured and comparisons were made between operated (left) and nonoperated (right) femurs. Digitized histomorphometric and volumetric analyses were performed on each growth plate, and detailed assessments were made of any morphological changes. Using length measurements, the difference in femur length was significantly larger in the HC group and not in the LC or control groups, showing bone growth inhibition at the higher current. In the HC group, bony bridges and disorganized growth plates were observed. This study shows that delivery of an electrical current of 50 microA for as little as 2 weeks can markedly affect bone growth as evidenced by changes in epiphyseal plate volume and architectural organization, and the study supports the use of this minimally invasive approach as a potential method of achieving an epiphysiodesis.

Androgen-induced vocal transformation in adult female African clawed frogs.

Sex-specific behaviors of some vertebrates are reversible by androgen administered in adulthood. Such behavioral transformations in adulthood provide opportunities to identify how neural systems reconfigure to produce sex-specific behavior. In this study, we focused on the vocalizations of the African clawed frog, Xenopus laevis. Male and female adult Xenopus produce sexually distinct vocalizations; males produce series of rapid clicks, whereas females produce slow trains of clicks. The differences in click rate can be reduced to differences in the firing rate of laryngeal motoneurons in vivo. This behavioral dimorphism is accompanied by various sex-specific characteristics throughout the vocal pathways, including functionally distinct laryngeal muscles and motoneurons in the sexes. In this study, we first determined whether and how testosterone (T) modifies the vocalizations of adult females and then examined changes underlying the behavioral modification at the laryngeal muscle and motoneuron levels. Our results show that, in response to T, the vocalizations of females were transformed within 13 wk. Vocal transformation was preceded by complete masculinization of muscle contractile properties and motoneuron soma size by the fourth week of T treatment, which suggests that the vocal pathways' peripheral components masculinize earlier than the behavior. Therefore the rate of transformation of vocal behavior must reflect a functional transformation of neurons in the central vocal pathways, which leads to the generation of male-like motor rhythms.