Antennal excision reveals disparate olfactory expression patterns within castes in Reticulitermes aculabialis (Isoptera: Rhinotermitidae).

In lower termites, which exhibit a high degree of compound eye degradation or absence, antennae play a pivotal role in information acquisition. This comprehensive study investigates the olfactory system of Reticulitermes aculabialis, spanning five developmental stages and three castes. Initially, we characterize the structures and distribution of antennal sensilla across different developmental stages. Results demonstrate variations in sensilla types and distributions among stages, aligning with caste-specific division of labor and suggesting their involvement in environmental sensitivity detection, signal differentiation, and nestmate recognition. Subsequently, we explore the impact of antennal excision on olfactory gene expression in various caste categories through transcriptomics, homology analysis, and expression profiling. Findings reveal that olfactory genes expression is influenced by antennal excision, with outcomes varying according to caste and the extent of excision. Finally, utilizing fluorescence in situ hybridization, we precisely localize the expression sites of olfactory genes within the antennae. This research reveals the intricate and adaptable nature of the termite olfactory system, highlighting its significance in adapting to diverse ecological roles and demands of social living.

Embryonic Development of Parthenogenetic and Sexual Eggs in Lower Termites.

Worldwide, termites are one of few social insects. In this research, the stages of embryonic development in the parthenogenetic and sexual eggs of Reticulitermes aculabialis and R. flaviceps were observed and described. In R. flaviceps, the egg development of the FF and FM groups happened during the early phases of development, whereas in R. aculabialis, this appeared mainly during the late phase of development. The variance in the number of micropyles between the R. flaviceps FF colony type and the R. aculabialis FF colony type was statistically significant. Five stages of egg development were found in both types of R. aculabialis but only the sexual eggs of R. flaviceps. In R. flaviceps, 86% of the parthenogenetic eggs stopped growing during the blastoderm development, with the yolk cell assembling frequently in the center of the egg. According to the results of the single-cell transcriptome sequencing, we investigated the egg-to-larval expression level of genes (pka, map2k1, mapk1/3, hgk, mkp, and pax6) and indicated that the levels of essential gene expression in RaFF were considerably higher than in RfFF (p < 0.05). We also discovered that the oocyte cleavage rate in the FF colony type was considerably lower in R. flaviceps compared to R. aculabialis, which gave rise to a smaller number of mature oocytes in R. flaviceps. During ovulation in both species, oocytes underwent activation and one or two cleavage events, but the development of unfertilized eggs ceased in R. flaviceps. It was shown that termite oocyte and embryonic development were heavily influenced by genes with significant expressions. Results from the databases KEGG, COG, and GO unigenes revealed the control of numerous biological processes. This study is the first to complete a database of parthenogenetic and sexual eggs of R. flaviceps and R. aculabialis.

Elevated expression of immune and DNA repair genes in mated queens and kings of the Reticulitermes chinensis termites.

Studies have identified that mating induces a series of physiological changes in animals. In this period, males tending to invest more energy, immune peptides, and other substances to reduce the cost of living for females. This results in lower survival rates in later life than females. Meanwhile, both males and females shorten lifespans due to reproduction. However, the reasons why termites' queens and kings are both extremely long-lived and highly fecund are unclear. Therefore, this study aimed to examine the effects of mating on the expression of immune and DNA repair genes for lifespan extension in termite queens and kings. Here, we reported that mated queens show relatively higher expression of immune genes (phenoloxidase, denfensin, termicin, transferrin), antioxidant genes (CAT, SOD), detoxification genes (GST, CYP450) than virgin queens in the Reticulitermes chinensis. In addition, mated kings also highly expressed these genes, except for termicin, transferrin, GST, and CYP450. After mating, both queens and kings significantly upregulated the expression of DNA repair genes (MLH1, BRCA1, XRCC3, RAD54-like). Mismatch repair genes (MMR) MSH2, MSH4, MSH6 were considerably increased in mated queens, while MSH4, MSH5, MSH6 were upregulated in mated kings. Our results suggest that mating increases the expression of immune and DNA repair genes in the termite queens and kings, and thus possibly improving their survival during reproductive span due to the omnipresent pathogens.

The Distribution Pattern and Species Richness of Scorpionflies (Mecoptera: Panorpidae).

The uneven distribution of species diversity on earth, with mountainous regions housing half of the high species diversity areas, makes mountain ecosystems vital to biodiversity conservation. The Panorpidae are ecological indicators, ideal for studying the impact of climate change on potential insect distribution. This study examines the impact of environmental factors on the distribution of the Panorpidae and analyzes how their distribution has changed over three historical periods, the Last Interglacial (LIG), the Last Glacial Maximum (LGM), and Current. The MaxEnt model is used to predict the potential distribution area of Panorpidae based on global distribution data. The results show that precipitation and elevation are the primary factors affecting species richness, and the suitable areas for Panorpidae are distributed in southeastern North America, Europe, and southeastern Asia. Throughout the three historical periods, there was an initial increase followed by a decrease in the area of suitable habitats. During the LGM period, there was a maximum range of suitable habitats for cool-adapted insects, such as scorpionflies. Under the scenarios of global warming, the suitable habitats for Panorpidae would shrink, posing a challenge to the conservation of biodiversity. The study provides insights into the potential geographic range of Panorpidae and helps understand the impact of climate change on their distribution.

Morphology, morphogenesis and molecular phylogeny of Lamtostyla granulifera sinensis subsp. nov. (Ciliophora, Hypotrichia) from a wetland in China.

The morphology, morphogenesis and molecular phylogeny of a hypotrichous ciliate, Lamtostyla granulifera sinensis subsp. nov., isolated from northern China, were investigated. This population appeared highly similar in morphology to L. granulifera Foissner, 1997. However, on detailed investigation some non-overlapping features were identified, i.e., the body shape and the arrangement of the cortical granules. These differences suggested the separation at subspecies level. Furthermore, the morphogenesis of the new subspecies is described, which is characterized by: (1) the posterior part of the parental adoral zone of membranelles is renewed; (2) the amphisiellid median cirral row is formed from two anlagen; and (3) the frontoventral-transverse cirral anlagen II to VI generate one transverse cirrus each. Phylogenetic analyses based on SSU rDNA sequence data show that Lamtostyla species are scattered in different clades. The monophyly of the genus Lamtostyla is also rejected by the AU test in this study.

De Novo Transcriptome Assembly and Analysis of Longevity Genes Using Subterranean Termite (Reticulitermes chinensis) Castes.

The longevity phenomenon is entirely controlled by the insulin signaling pathway (IIS-pathway). Both vertebrates and invertebrates have IIS-pathways that are comparable to one another, though no one has previously described de novo transcriptome assembly of IIS-pathway-associated genes in termites. In this research, we analyzed the transcriptomes of both reproductive (primary kings "PK" and queens "PQ", secondary worker reproductive kings "SWRK" and queens "SWRQ") and non-reproductive (male "WM" and female "WF" workers) castes of the subterranean termite Reticulitermes chinensis. The goal was to identify the genes responsible for longevity in the reproductive and non-reproductive castes. Through transcriptome analysis, we annotated 103,589,264 sequence reads and 184,436 (7G) unigenes were assembled, GC performance was measured at 43.02%, and 64,046 sequences were reported as CDs sequences. Of which 35 IIS-pathway-associated genes were identified, among 35 genes, we focused on the phosphoinositide-dependent kinase-1 (Pdk1), protein kinase B2 (akt2-a), tuberous sclerosis-2 (Tsc2), mammalian target of rapamycin (mTOR), eukaryotic translation initiation factor 4E (EIF4E) and ribosomal protein S6 (RPS6) genes. Previously these genes (Pdk1, akt2-a, mTOR, EIF4E, and RPS6) were investigated in various organisms, that regulate physiological effects, growth factors, protein translation, cell survival, proliferation, protein synthesis, cell metabolism and survival, autophagy, fecundity rate, egg size, and follicle number, although the critical reason for longevity is still unclear in the termite castes. However, based on transcriptome profiling, the IIS-pathway-associated genes could prolong the reproductive caste lifespan and health span. Therefore, the transcriptomic shreds of evidence related to IIS-pathway genes provide new insights into the maintenance and relationships between biomolecular homeostasis and remarkable longevity. Finally, we propose a strategy for future research to decrypt the hidden costs associated with termite aging in reproductive and non-reproductive castes.

The Antennal Sensilla and Expression Patterns of Olfactory Genes in the Lower Termite Reticulitermes aculabialis (Isoptera: Rhinotermitidae).

The insect olfactory system plays pivotal roles in insect survival and reproduction through odor detection. Morphological and physiological adaptations are caste-specific and evolved independently in workers, soldiers, and reproductives in termites. However, it is unclear whether the olfactory system is involved in the division of labor in termite colonies. In the present study, the antennal sensilla of alates, workers, soldiers, nymphs, and larvae of the termite Reticulitermes aculabialis Tsai et Hwang ( Isoptera: Rhinotermitidae) were investigated. Transcriptomes were used to detect olfactory genes, and differential expression levels of olfactory genes were confirmed in various castes by qRT-PCR analysis. Nine types of sensilla were identified on the antennae of R. aculabialis, and soldiers possessed all 9 types. In 89,475 assembled unigenes, we found 16 olfactory genes, including 6 chemosensory protein (CSP) and 10 odorant-binding protein (OBP) genes. These OBP genes included 8 general odorant-binding protein genes (GOBPs) and 2 pheromone-binding protein-related protein (PBP) genes. Five CSP genes were more highly expressed in alates than in workers, soldiers, larvae, and nymphs, and the expression levels of CSP6 were significantly higher in nymphs. Seven GOBP and two PBP genes exhibited significantly higher expression levels in alates, and there were no significant differences in the expression levels of GOBP2 among workers, soldiers, alates, and larvae. These results suggest that alates, as primary reproductives, have unique expression patterns of olfactory genes, which play key roles in nuptial flight, mate seeking, and new colony foundation.

Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici.

ROPs (Rho-like GTPases from plants) are a unique family of small GTP-binding proteins in plants and play vital roles in numerous cellular processes, including growth and development, abiotic stress signaling, and plant defense. In the case of the latter, the role of ROPs as response regulators to obligate parasitism remains largely enigmatic. Herein, we isolated and identified ShROP7 and show that it plays a critical role in plant immune response to pathogen infection. Real-time quantitative PCR analysis revealed that the expression of ShROP7 was significantly increased during incompatible interactions. To establish its requirement for resistance, we demonstrate that virus-induced gene silencing (VIGS) of ShROP7 resulted in increased susceptibility of tomato to Oidium neolycopersici (On) Lanzhou strain (On-Lz). Downstream resistance signaling through H2O2 and the induction of the hypersensitive response (HR) in ShROP7-silenced plants were significantly reduced after inoculating with On-Lz. Taken together, with the identification of ShROP7-interacting candidates, including ShSOBIR1, we demonstrate that ShROP7 plays a positive regulatory role in tomato powdery mildew resistance.

Endocuticle is involved in caste differentiation of the lower termite.

Caste differentiation in termites is one of the most conspicuous examples of facultative polyphenism in animals. It is clear that specific cuticular formation occurs in hard exocuticles during caste differentiation. However, the developmental pattern of the soft endocuticle in the differentiation pathways of castes is unknown. To reveal whether the endocuticle is involved in caste differentiation, we compared the exocuticle and endocuticle thickness of individuals in 2 pathways (nymph line and worker line) of caste differentiation in the termite Reticulitermes aculabialis. The endocuticle protein genes were identified by transcriptome analysis and the expression patterns of these genes were confirmed in caste differentiation. We found that the endocuticle structure showed dynamic changes in 2 pathways, and the first difference in endocuticle structure occurred after larvae differentiation bifurcated into workers and nymphs. The thinning of the endocuticle was a significant event from nymphs developing into alates with the thickest exocuticle and thinnest endocuticle. The thickest endocuticle layers were found in the heads of the workers and the ultrastructure of the endocuticle in the heads was more complex than that in the thorax-abdomens. Six endocuticle protein genes were identified and annotated as endocuticle structural glycoproteins SgAbd-2, SgAbd-9, and Abd-5. The expression levels of endocuticle protein genes changed dramatically during caste development and the expression levels in neotenic reproductives (secondary reproductives) were significantly higher than those in alates (primary reproductives). These results reveal the roles of endocuticles in caste differentiation and adaptation to the environment.

Transcriptomic evidence that insulin signalling pathway regulates the ageing of subterranean termite castes.

Insulin is a protein hormone that controls the metabolism of sugar, fat and protein via signal transduction in cells, influencing growth and developmental processes such as reproduction and ageing. From nematodes to fruit flies, rodents and other animals, glucose signalling mechanisms are highly conserved. Reproductive termites (queens and kings) exhibit an extraordinarily long lifespan relative to non-reproductive individuals such as workers, despite being generated from the same genome, thus providing a unique model for the investigation of longevity. The key reason for this molecular mechanism, however, remains unclear. To clarify the molecular mechanism underlying this phenomenon, we sequenced the transcriptomes of the primary kings (PKs), primary queens (PQs), male (WMs) and female (WFs) workers of the lower subterranean termite Reticulitermes chinensis. We performed RNA sequencing and identified 33 insulin signalling pathway-related genes in R. chinensis. RT-qPCR analyses revealed that EIF4E and RPS6 genes were highly expressed in WMs and WFs workers, while mTOR expression was lower in PKs and PQs than in WMs and WFs. PQs and PKs exhibited lower expression of akt2-a than female workers. As the highly conserved insulin signalling pathway can significantly prolong the healthspan and lifespan, so we infer that the insulin signalling pathway regulates ageing in the subterranean termite R. chinensis. Further studies are recommended to reveal the biological function of insulin signalling pathway-related genes in the survival of termites to provide new insights into biomolecular homeostasis maintenance and its relationship to remarkable longevity.

Morphogenesis and molecular phylogeny of two soil ciliates Australocirrus australis (Foissner, 1995) Kumar and Foissner, 2015 and A. aspoecki (Foissner, 2004) Kumar and Foissner, 2015 (Ciliophora, Hypotrichia).

The morphogenesis and molecular phylogeny of two soil hypotrich ciliates, Australocirrus australis and A. aspoecki, collected from Xi'an, Shaanxi Province, China, were investigated using protargol preparation. The main features of morphogenesis of A. australis are as follows: (1) the parental adoral zone of membranelles is retained completely in the proter; (2) parental cirri and the undulating membranes are involved in the formation of six primordial streaks; (3) multiple fragmentation of the third dorsal kinety anlage; (4) more than three dorsomarginal kineties are formed. An early divider of A. aspoecki was also found, and its morphogenetic events resemble those of other Australocirrus species. Furthermore, the SSU rDNA sequence of the Shaanxi populations of A. australis and A. aspoecki was provided. The molecular phylogeny indicates that A. aspoecki clusters together with A. rubrus and two populations of A. australis with full support, which are separated from the clade comprising the two populations of A. shii.

Herba Houttuyniae Extract Benefits Hyperlipidemic Mice via Activation of the AMPK/PGC-1α/Nrf2 Cascade.

Hyperlipidemia is associated with metabolic disorders, but the detailed mechanisms and related interventions remain largely unclear. As a functional food in Asian diets, Herba houttuyniae has been reported to have beneficial effects on health. The present research was to investigate the protective effects of Herba houttuyniae aqueous extract (HAE) on hyperlipidemia-induced liver and heart impairments and its potential mechanisms. Male C57BL/6J mice were administered with 200 or 400 mg/kg/day HAE for 9 days, followed by intraperitoneal injection with 0.5 g/kg poloxamer 407 to induce acute hyperlipidemia. HAE treatment significantly attenuated excessive serum lipids and tissue damage markers, prevented hepatic lipid deposition, improved cardiac remodeling, and ameliorated hepatic and cardiac oxidative stress induced by hyperlipidemia. More importantly, NF-E2 related factor (Nrf2)-mediated antioxidant and peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α)-mediated mitochondrial biogenesis pathways as well as mitochondrial complex activities were downregulated in the hyperlipidemic mouse livers and hearts, which may be attributable to the loss of adenosine monophosphate (AMP)-activated protein kinase (AMPK) activity: all of these changes were reversed by HAE supplementation. Our findings link the AMPK/PGC-1α/Nrf2 cascade to hyperlipidemia-induced liver and heart impairments and demonstrate the protective effect of HAE as an AMPK activator in the prevention of hyperlipidemia-related diseases.

Transcriptome changes reveal the genetic mechanisms of the reproductive plasticity of workers in lower termites.

BACKGROUND: The reproductive plasticity of termite workers provides colonies with tremendous flexibility to respond to environmental changes, which is the basis for evolutionary and ecological success. Although it is known that all colony members share the same genetic background and that differences in castes are caused by differences in gene expression, the pattern of the specific expression of genes involved in the differentiation of workers into reproductives remains unclear. In this study, the isolated workers of Reticulitermes labralis developed into reproductives, and then comparative transcriptomes were used for the first time to reveal the molecular mechanisms underlying the reproductive plasticity of workers. RESULTS: We identified 38,070 differentially expressed genes and found a pattern of gene expression involved in the differentiation of the workers into reproductives. 12, 543 genes were specifically upregulated in the isolated workers. Twenty-five signal transduction pathways classified into environmental information processing were related to the differentiation of workers into reproductives. Ras functions as a signalling switch regulates the reproductive plasticity of workers. The catalase gene which is related to longevity was up-regulated in reproductives. CONCLUSION: We demonstrate that workers leaving the natal colony can induce the expression of stage-specific genes in the workers, which leads to the differentiation of workers into reproductives and suggests that the signal transduction along the Ras-MAPK pathway crucially controls the reproductive plasticity of the workers. This study also provides an important model for revealing the molecular mechanism of longevity changes.

The nest fungus of the lower termite Reticulitermes labralis.

Fitness-determining interactions with fungi have often been considered a by-product of social evolution in insects. In higher termites, the mutualistic association between the basidiomycete genus Termitomyces and Macrotermitinae is well known. However, whether and how lower termites use fungi is unclear. Here, we found a large amount of brown sclerotium-forming fungi in egg piles of the lower termite Reticulitermes labralis and identified the sclerotia as Fibulorhizoctonia sp. There was a significant difference in morphology between the sclerotia and the termite eggs. The workers of R. labralis and R. chinensis actively gathered the sclerotia into the egg piles within their nests, whereas the workers of R. aculabialis did not gather sclerotia outside their nests. None of the sclerotia in the egg piles germinated in the presence of workers. However, the sclerotia germinated in the absence of workers, and then the hyphae killed the termite eggs. The data from cellulase activity demonstrated that Fibulorhizoctonia sp. was able to exhaustively digest cellulose into glucose.We confirmed for the first time that the workers carrying the sclerotia into the piles of eggs is not due to mistaking the sclerotia for their eggs and that the workers of R. labralis may be able to select favourite fungi.

The transition path from female workers to neotenic reproductives in the termite Reticulitermes labralis.

Termite workers are characterized by unique flexibility in that a worker can develop in one of three ways: remain a worker, become a soldier within two successive moults, or become a neotenic reproductive (NR) within a single moult. However, is it true that workers can transform into NRs within a single moult? Actually, the developmental pathways of workers turning into NRs remains unclear. In this study, we show for the first time that the female workers of Reticulitermes labralis develop into NRs after a pre-NRs stage. We found that a female worker became a NR after two successive moults, whereas the male workers copulated directly with queens without undergoing any moults. After the first moult led the female workers into the pre-NR stage, the length of their abdomens, seventh sternites and ovaries significantly increased. After the second moult from the pre-NRs stage into NRs, the follicle cells returned to normal, and a few oocytes and follicle cells underwent apoptosis. These results demonstrated that the female pre-NR type was a transitional type during the development of female workers to the NR caste, and the starting point for oogenesis resumption was the NR stage. We confirmed there were fundamental differences in the reproductive pathway of the male and female workers. Therefore, we determined that the transformation process of the female NRs from workers may be a very complex process, and the reproductive biology of the workers has great potential to provide important and spectacular insights into the evolution strategy of termites.

Characterization of the transcriptomes and cuticular protein gene expression of alate adult, brachypterous neotenic and adultoid reproductives of Reticulitermes labralis.

The separation of primary reproductive and secondary reproductive roles based on the differentiation of alate adults and neotenic reproductives is the most prominent characteristic of termites. To clarify the mechanism underlying this differentiation, we sequenced the transcriptomes of alate adults (ARs), brachypterous neotenics (BNs) and adultoid reproductives (ANs) from the last instar nymphs of Reticulitermes labralis. A total of 404,152,188 clean sequencing reads was obtained and 61,953 unigenes were assembled. Of the 54 identified cuticular protein (CP) genes of the reproductives, 22 were classified into the CPR family and 7 were classified into the CPG family. qRT-PCR analyses of the 6 CP genes revealed that the CP genes involved in exocuticle sclerotization were highly expressed in the ARs and RR-1 involved in soft endocuticle was highly expressed in the ARs and ANs. These results suggest that the alate adults might increase cuticular component deposition to adapt to new or changing environments and that the development of reproductive individuals into primary or secondary reproductives is controlled by the expression of cuticular protein genes involved in the hardening of the exocuticle. In addition, the AN caste is a transitional type between the BN and AR castes in the process of evolution.

Complete mitochondrial genome of the "floppy-wing" morph reproductive termite, Reticulitermes labralis (Isoptera: Rhinotermitidae).

The complete mitochondrial genome of Reticulitermes labralis (Isoptera: Rhinotermitidae) was determined for its nucleotide sequence of 16 113 bp. Its gene content and organization were identical with other Reticulitermes species. The 13 protein-coding genes (PCGs) have typical ATN initiation codon. But, the stop codons were TAA, TAG and an incomplete termination codon (T) abutting an adjacent tRNA gene. Twenty-two tRNA genes, in addition to tRNASer(AGN) replaced lacking of the DHU stem with a simple loop, showed the typical clover-leaf secondary structure. The A + T-rich region was 1311 bp in length with 65.98% A + T content. In addition to the A + T-rich region, non-coding sequences of the mtDNA genome harbored 17 intergenic spacers. There were three complete repeats of repeat A in CR, which were not discovered in other termite species. Phylogenetic tree based on the 11 complete mitochondrial genome sequences of closely related termite species accords well with morphological phylogenetic analysis.

The complete mitochondrial genome of the subterranean termite, Reticulitermes chinensis Snyder (Isoptera: Rhinotermitidae).

We have determined the complete mitochondrial genome of Reticulitermes chinensis Snyder. The total length of the R. chinensis is 15,925 bp with 65.27% A + T content. It consists of 13 protein-coding, 22 tRNA, 2 rRNA genes and an A + T-rich control region. All the protein-coding genes used ATN as start codon. But the stop codons were TAA, TAG, and an incomplete termination codon (T) abutting an adjacent tRNA gene. The A + T-rich control region was 1125 bp in length with 67.02% A + T content.