Eastern Subterranean Termite Reticulitermes Flavipes Research Articles

Differential Selection on Caste-Associated Genes in a Subterranean Termite.

Simple SummarySocial insects can sometimes boldly invade new habitats, including areas of human habitation where they can become unwanted domestic or agricultural pests. In this study, we use molecular sequence analysis to study genetic patterns associated with the invasion and division of labour in the Eastern subterranean termite Reticulitermes flavipes. By studying how genes vary by caste and by population, we show that even termites invasive to a metropolitan city can still harbour plenty of genetic variation, as much or more as native termite populations. We suggest therefore that invasive termites do not necessarily suffer long term loss-of-variation upon invasion. Second, we show that genes associated in their expression with the soldier caste evolve approximately twice as fast as genes expressed by other castes of this species, regardless of from what population the castes were sampled. Why termite soldier genes evolve quickly is not known, but it seems unrelated to invasion or the invaded habitat. Given that soldiers are sterile and thus have no direct fitness, the evidence for gene-level selection in the soldier caste is an intriguing example of kin selection.Analyzing the information-rich content of RNA can help uncover genetic events associated with social insect castes or other social polymorphisms. Here, we exploit a series of cDNA libraries previously derived from whole-body tissue of different castes as well as from three behaviourally distinct populations of the Eastern subterranean termite Reticulitermes flavipes. We found that the number (~0.5 M) of single nucleotide variants (SNVs) was roughly equal between nymph, worker and soldier caste libraries, but dN/dS (ratio of nonsynonymous to synonymous substitutions) analysis suggested that some of these variants confer a caste-specific advantage. Specifically, the dN/dS ratio was high (~4.3) for genes expressed in the defensively specialized soldier caste, relative to genes expressed by other castes (~1.7–1.8) and regardless of the North American population (Toronto, Raleigh, Boston) from which the castes were sampled. The populations, meanwhile, did show a large difference in SNV count but not in the manner expected from known demographic and behavioural differences; the highly invasive unicolonial population from Toronto was not the least diverse and did not show any other unique substitution patterns, suggesting any past bottleneck associated with invasion or with current unicoloniality has become obscured at the RNA level. Our study raises two important hypotheses relevant to termite sociobiology. First, the positive selection (dN/dS > 1) inferred for soldier-biased genes is presumably indirect and of the type mediated through kin selection, and second, the behavioural changes that accompany some social insect urban invasions (i.e., ‘unicoloniality’) may be detached from the loss-of-diversity expected from invasion bottlenecks.

Breeding Patterns and Population Genetics of Eastern Subterranean Termites Reticulitermes flavipes in Urban Environment of Nebraska, United States

Reticulitermes flavipes (Kollar) has become the most destructive subterranean termite pest, on urban structures in Nebraska. In this study, we used seven microsatellite loci to infer the colony breeding system and population genetic structure among 20 infested urban structures in Nebraska. Our data revealed that 17 structures were infested by simple family colonies of R. flavipes, while, the remaining three were infested with mixed family colonies. The measure of population differentiation, FCT value (0.459) indicated that all the 20 urban colonies (10 - 410 km apart) represented pronounced levels of genetic differentiation. The Mantel test disclosed a weak and significantly-positive correlation between genetic and geographic distance (slope = 0.0009, P = 0.001). The urban populations of R. flavipes in Nebraska possessed a breeding system characterized by monogamous pairs of outbred reproductives with excessive heterozygosity.

A Lab-Based Study of Temperate Forest Termite Impacts on Two Common Wood-Rot Fungi.

Termites and fungi are the primary decomposers of dead wood. Interactions between wood-feeding termites and wood-rot fungi are inevitable given their shared food source. Termites have developed multiple defense strategies against infectious fungi, such as Metarhizium spp., that include antifungal proteins in their saliva and fungal inhibition properties in their gut. The antifungal properties of termite salivary secretions depend on β-1,3-glucanases that are likely to be effective against a broad spectrum of filamentous fungi. Given the overlap in niches, there is opportunity for interference competition between termites and wood-rot fungi to occur. Here we demonstrate that β-1,3-glucanases in the saliva and the antifungal properties of the gut of the eastern subterranean termite Reticulitermes flavipes (Kollar) (Blattodea: Rhinotermitidae) affects the growth of two common wood-rot fungi, Gloeophyllum trabeum Persoon (Murrill) (Gloeophyllales: Gloeophyllaceae) and Phanerochaete chrysosporium (Burdsall) (Polyporales: Phanerochaetaceae).

Caste-biased genes in a subterranean termite are taxonomically restricted: implications for novel gene recruitment during termite caste evolution

The caste system of social insects presents a classic polyphenism in which widely divergent reproductive and non-reproductive phenotypes are expressed from the same genome. In termites, the sterile soldier caste is particularly divergent in phenotype and presumably evolved under selection for defensiveness. In this study, we use genomic phylostratigraphy to show that genes with soldier- and other caste-biased expression from the Eastern subterranean termite Reticulitermes flavipes are more taxonomically restricted on the tree of life than genes with no caste-biased expression. This pattern suggests that caste-biased genes are relatively young and implies past selection for novel gene recruitment during termite caste evolution. Moreover, a soldier-biased set of 74 genes contains a higher proportion of orphan genes with no known homology than does a nymph-biased set or any null gene sets. This again suggests that the termite caste—and soldiers in particular—makes disproportionate use of evolutionarily novel genes that are potentially recruited from non-coding regions of the genome. Given that Reticulitermes and most termite soldiers are sterile, any past selection for genetic novelty of this caste must have been indirect and mediated through reproducing relatives.

Soldier-biased gene expression in a subterranean termite implies functional specialization of the defensive caste.

In a termite colony, reproduction is typically monopolized by a small number of sexuals that are supported by reproductively altruistic soldiers and workers. We expect caste differentiation to be associated with clear-cut differences in gene expression, and for these differences to reflect caste function and development. Here, we use RNA-Sequencing to compare the gene expression profiles of sexual nymphs and two non-reproductive helper castes (i.e., workers and soldiers) of the Eastern subterranean termite Reticulitermes flavipes. We found that of n = 93 genes that are strictly expressed as a function of caste, a majority (78%) show a soldier-specific pattern. This conspicuous soldier-bias in genome-wide expression suggests that this defensively specialized caste is functionally well-differentiated from both the reproductive and the other non-reproductive caste of this species, despite a shared developmental program with workers. Gene ontology analysis supports the notion of functional specialization by soldiers, as soldier-biased gene sets are enriched for novel biological processes. Whether this pattern reflects ancient or more recent bouts of selection for caste novelty at the gene-regulatory level is not known, but because soldiers are sterile and thus have no direct fitness, any selection for novelty must have been mediated indirectly, through reproducing relatives.

Effect of Temperature and Humidity on Survival of Coptotermes formosanus and Reticulitermes flavipes (Isoptera: Rhinotermitidae)

Two subterranean termite species were subjected to combinations of six temperatures (10°, 15°, 20°, 25°, 30°, or 35°C) and five relative humidities (RH) (55, 65, 75, 85, or 99%) to determine optimum conditions for survival. When small groups of the Formosan subterranean termite Coptotermes formosanus Shiraki or the eastern subterranean termite Reticulitermes flavipes (Kollar) were exposed to all 30 combinations of temperature and RH, survival times were significantly influenced by temperature, RH, and their interaction. For both species, survival times were longest at low temperatures and high RH. Maximum survival of small groups of C. formosanus and R. flavipes workers and soldiers occurred at the combination of 10°C and 99% RH C (LT50= 28.2 d, LT50 = 18.1 d, respectively). Survival of paired C. formosanus dealates was evaluated at combinations of 20°, 25°, or 30°C and 55, 65, 75, 85, or 99% RH. Survival was strongly influenced by temperature and humidity. Longest survival times until 50% mortality occurred at 99% RH and 20° or 25°C (LT50= 2.5 d, LT50 = 3.0 d, respectively). At all temperatures, mortality occurred too quickly for LT50 values to be determined when RH was 55% or 65%.

IDENTIFICATION OF MYCOSIS‐RELATED GENES IN THEEASTERN SUBTERRANEAN TERMITE BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION

The Eastern subterranean termite Reticulitermes flavipes (Isoptera, Rhinotermitidae) is a cosmopolitan, structural pest that is the target of research into termite innate immunity. In this study, we use suppression subtractive hybridization to construct a normalized cDNA library of genes excessively expressed upon fungal infection. At 24 h postinfection with Metarhizium anisopliae, the library revealed 182 expressed sequence tag (EST) clones that potentially represent immune responsive genes. The nucleotide sequence from a majority (97%) of ESTs assembled into a small number (n = 13) of contiguous sequences, with the remainder (n = 6) representing singletons. Our screen therefore captured as many as 19 different mRNAs highly expressed in response to the fungal pathogen at this time. Primary sequencing of all loci revealed that approximately half (n = 10) contained open reading frames with significant similarity to known proteins. These clones represent nuclear and mitochondrial coding genes, as well as putative long noncoding RNA genes. Quantitative polymerase chain reaction analysis of coding genes on independently infected groups of worker termites confirms in each case that the transcripts identified from the library are up-regulated postfungal infection. The genes identified here are relevant to future studies on termite biocontrol and social insect immunity.

Effect of Fipronil on Subterranean Termite Colonies (Isoptera: Rhinotermitidae) in the Field

To assess possible colony-level effects of fipronil, a commonly used nonrepellent termiticide, we conducted a field study of eight houses in the Raleigh, NC, area with infestations of the eastern subterranean termite Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae). We installed an extensive grid of in-ground monitoring stations on each property (mean = 68.6 monitors per property) and collected samples from these stations as well as from mud tubes in the structure and wood debris in the yard for approximately 6 mo. We genotyped all samples by using microsatellite markers to identify the number and locations of colonies present on each property. Houses were treated with either a full treatment (n = 5) or exterior/localized interior treatment (n = 3). After treatment, the monitors were checked monthly for 3 mo and then quarterly for 3 yr to track the fate of colonies. Wood debris in natural areas was checked semiannually for 3 yr. All 11 of the treated colonies (those attacking structures) disappeared within 90 d of treatment and were not found again. These colonies were presumed to be eliminated. In contrast, 60% of untreated colonies (those located > 6 m from the foundation wall at the time of treatment) continued to persist throughout the study, as did 25% of the likely treated colonies (those occupying monitors 0.5 m from the foundation wall where the treatment was applied). Our results provide strong evidence for potent colony wide effects of fipronil on subterranean termites leading to colony suppression and likely colony elimination under field conditions.

Effect of Temperature and Termite Starvation on Phagocytosis by Protozoan Symbionts of the Eastern Subterranean Termite Reticulitermes flavipes Kollar

Many termite species rely on intestinal protozoan symbionts to digest their cellulosic foods. We examined cellulose acquisition by the symbionts of the Eastern subterranean termite Reticulitermes flavipes Kollar (Isoptera; Rhinotermitidae) by following their phagocytosis of red paper fed to the termite host. The effects of termite host starvation and environmental temperature on feeding activity were studied in the zooflagellates Trichonympha agilis Leidy (Trichonymphidae), Pyrsonympha vertens Leidy, Dinenympha fimbriata Kirby, and D. gracilis Leidy (Pyrsonymphidae), which are among the largest residents in R. flavipes' hindguts. Protozoans in termites starved for 24 h ingested red paper significantly sooner than protozoans in termites with continuous access to food. Trichonympha, Pyrsonympha, and Dinenympha all ingested red paper particles at approximately the same rate. Red paper appeared significantly sooner in protozoans in termites maintained at 32°C than in those maintained at 22°C or 26°C. At 32°C, numbers of Trichonympha per gut remained constant over 96 h. Pyrsonympha and Dinenympha cells were absent or significantly reduced in number by 72 h at that temperature. These results provide insight into the environmental factors that shape the termite–protozoan symbiosis. They may aid in the development of protozoicides used to control pest termites.

Changes in Morphogenetic Hormone Titers in Isolated Workers of the Termite Reticulitermes flavipes (Kollar)

The daily liters of juvenile hormone and ecdysteroids were determined for workers of the Eastern subterranean termite Reticulitermes flavipes (Kollar) (Isoptera, Rhinotermitidae) following isolation of the workers from soldiers and reproductives. Experiments have demonstrated that isolation leads to biochemical and physiological changes that result in a presoldier molt. Juvenile hormone (JH) and molting hormone (MH) titers were determined in hemolymph samples using radioimmunoassay. The identity of the ecdysteroids as predominantly 20-hydroxyecdysone (90%) and ecdysone (10%) was determined by HPLC using pooled extracts of hemolymph.

Major antitermitic components of the heartwood of southern catalpa

The heartwood of southern catalpa,Catalpa bignonioides Walt., is resistant to attack by the eastern subterranean termiteReticulitermes flavipes (Kollar), but extraction with a ternary solvent mixture of acetone-hexanewater (54∶44∶2) by volume removed the antitermitic characteristics from the heartwood. Four compounds comprised approximately 98% of the antitermitic fraction of the extract: the sesquiterpene alcohol, catalponol (67%); its epimer, epicatalponol (5%); a structurally related ketone, catalponone (1%); and the phthalide, catalpalactone (25%). Pure compounds were isolated by semipreparative scale reversed-phase HPLC and identified by GC-MS and UV spectroscopy. The structure of catalponol was further confirmed by the formation of derivatives. Bioassays indicated that catalponol had the greatest toxicity in cellulose pad tests, but in tests using vacuum impregnation of these compounds into termite-susceptible wood blocks at levels approximating those found in catalpa heartwood, catalpalactone exhibited the highest antitermitic activity.

Behavioral evidence for multicomponent trail pheromone in the termite,Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae)

Evidence is presented for a multicomponent trail pheromone in the eastern subterranean termiteReticulitermes flavipes (Kollar). Choice tests were used to compare strength and persistency of trails made by termites in glass tubes. Tubes connected to termite nests for 24 hr were marked by termites with an extremely long-lasting chemical that persisted for at least one year, and a highly volatile substance that decayed in 15 min. Individual termites varied the trail they deposited under different circumstances. When removed from the nest and placed in a clean tube, they deposited a highly volatile substance. They left stronger and more persistent trails when exploring clean tubes attached to the nest. When they discovered a new food source, they left trails that were more attractive and far more persistent than trails made by exploring or displaced termites. A food trail made by 15 workers was still effective after 24 hr, whereas a trail made by 50 displaced workers, walking one after another through a tube, lasted only 5 min.