Late Larval Stages Research Articles

MiR-3017b contributes to metamorphosis by targeting sarco/endoplasmic reticulum Ca2+ ATPase in Tribolium castaneum.

In recent years, increasing numbers of microRNAs (miRNAs) have been reported to regulate insect metamorphosis. One thousand, one hundred fifty-four miRNAs have been previously identified from Tribolium castaneum by high-throughput sequencing; however, little is known about which miRNAs can participate in metamorphosis, leaving the role of miRNAs in regulating the underlying mechanism elusive. Here, we report the participation of miR-3017b in the metamorphosis of T. castaneum. Temporal profiles revealed that miR-3017b was highly expressed at the late larval stage, but significantly decreased at the early pupal stage. Overexpression of miR-3017b caused larval to pupal to adult metamorphosis arrested. Dual-luciferase reporter assay and miRNA-mRNA interaction assay illustrated that miR-3017b interacts with the coding sequence of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) and suppresses its expression. Knockdown of SERCA caused metamorphosis arrested, similar to that observed in miR-3017b overexpression beetles. Further functional mechanism analyses revealed that 20-hydroxyecdysone application downregulates miR-3017b and up-regulates SERCA expression. The expression level of downstream genes in the 20E pathway was disrupted after overexpressing miR-3017 and the knockdown of SERCA. These results provided evidence miR-3017b-SERCA contributes to metamorphosis by regulating the 20E pathway in T. castaneum. It could advance our understanding of the coordination of 20E and miRNA regulation in insect metamorphosis.

Field life tables and key mortality factors of Helicoverpa armigera infesting tomato (Solanum lycopersicon)

Field life table studies of Helicoverpa armigera (Hübner) infesting tomato (Solanum lycopersicon L.) wereundertaken during 2018 and 2019 at research farm of Punjab Agricultural University, Ludhiana, Punjab. Key mortality factors observed in early and late larval instar groups were Campoletis chlorideae (Uchida), Beauveria bassiana (Bals.), HaNPV, Bacillus thuringiensis (Berliner), along with other unknown factors. The egg stage was the most vulnerable exhibiting the highest loss (32.0–91.0%) during main and spring season tomato crop followed by early larval stage (40.27–44.86%), late larval stage (35.84–40.95%), and pupal (24.45–29.23%) stage. Trend index was positive (4.54) on main season crop which showed that total mortality factors during this season were not effective in causing the pest decline and it was negative (0.22) on spring season crop which depicted that total mortality factors were more effective and the chances of increase in population during next season would be less. Generation survival was highest on main season crop (0.29) as compared to spring season (0.23) and indicated that maximum generation survival was in main season whereas, in spring season the generation survival was lower and contributed maximum towards mortality

Drosophila septin interacting protein 1 regulates neurogenesis in the early developing larval brain

Neurogenesis in the Drosophila central brain progresses dynamically in order to generate appropriate numbers of neurons during different stages of development. Thus, a central challenge in neurobiology is to reveal the molecular and genetic mechanisms of neurogenesis timing. Here, we found that neurogenesis is significantly impaired when a novel mutation, Nuwa, is induced at early but not late larval stages. Intriguingly, when the Nuwa mutation is induced in neuroblasts of olfactory projection neurons (PNs) at the embryonic stage, embryonic-born PNs are generated, but larval-born PNs of the same origin fail to be produced. Through molecular characterization and transgenic rescue experiments, we determined that Nuwa is a loss-of-function mutation in Drosophila septin interacting protein 1 (sip1). Furthermore, we found that SIP1 expression is enriched in neuroblasts, and RNAi knockdown of sip1 using a neuroblast driver results in formation of small and aberrant brains. Finally, full-length SIP1 protein and truncated SIP1 proteins lacking either the N- or C-terminus display different subcellular localization patterns, and only full-length SIP1 can rescue the Nuwa-associated neurogenesis defect. Taken together, these results suggest that SIP1 acts as a crucial factor for specific neurogenesis programs in the early developing larval brain.

Recovery of Haemal Lordosis in European Seabass Dicentrarchus labrax (Linnaeus 1758)

Lordosis of the haemal part of the vertebral column is a frequent abnormality in reared fish. Haemal lordosis develops during the late larval and early juvenile life stages of fish, mainly due to the high swimming activity of the fish in the rearing tanks. In the present study, we have examined whether haemal lordosis recovers during the growth of European seabass. Furthermore, we aimed to develop simple morphometric indices (PrAn1 and PrAn2) that might link the severity of lordosis at the juvenile stage with fish morphological quality at harvesting. At 111 days post-hatching (dph, 53 ± 4 mm standard length, SL), 600 seabass juveniles with lordotic (L, 200 fish) or normal (N, 400 fish) external morphology were selected and introduced in a common tank. At 150 dph (75 ± 7 mm SL), 350 fish were randomly selected, pit-tagged and transferred to a sea cage for on-growing up to 502 dph (234 ± 16 mm SL). The morphological examination of the fish at 150 and 502 dph revealed that 60% (46 out of 77) of L juveniles turned into normal phenotype by the end of on-growing period. Interestingly, 56% of the fish with recovered external morphology (N-Rec) presented either a completely normal vertebral column (31%) or minor abnormalities of individual vertebrae (25%). Following the results of geometric morphometric analysis, the differences in the body shape between N-Rec and N fish were not statistically significant (p > 0.05, canonical variate analysis). The examined morphometric indices were effective in discriminating the normal fish from 58% (PrAn1) to 65% (PrAn2) of lordotic juveniles. Results are discussed with respect to the mechanism of lordosis recovery, and spotlights on their application for quality control and cull out of the abnormal fish in commercial hatcheries.

Eueides lybia (Fabricius, 1775) and Eueides olympia (Fabricius, 1793) are Distinct Species: Evidence from Barcodes and Early Stages of E. L. Lybia in Suriname (Nymphalidae: Heliconiinae)

Larval and pupal stages are described for Eueides lybia lybia (Nymphalidae: Heliconiinae) in Suriname. Its hostplants are Passiflora nitida and P. glandulosa (Passifloraceae). Based on characters of the habitus of the imago, the male genitalia, the late larval stages and barcodes, it is argued that Eueides lybia consists of two species, Eueides lybia (Fabricius, 1775) and Eueides olympia (Fabricius, 1793) revised status. The most straightforward way to distinguish between the two is the costal streak on the ventral forewing of the imago: red in E. lybia and yellow in E. olympia. E. olympia is considered to have three subspecies, E. olympia olympia (Fabricius, 1793) new status, E. olympia lybioides Staudinger 1876 new status and E. olympia salcedoi K. Brown & F. Fernández Yépez, 1985 new status. E. olympia lybioides may turn out to be a distinct species, as may populations of E. lybia from Peru.

Sex-Specific Stress Tolerance in Embryos of Lake Char (Salvelinus umbla)

Salmonid fish have become important models in evolution and ecology, but possible effects of embryo or larval sex are mostly ignored, probably because morphological gonad formation starts only months after hatching and sexual maturation years later. However, recent gene expression studies and first observations in domestic strains suggest that sex-specific life histories could already start at an embryonic stage. Here we test this hypothesis in embryos and larvae of lake char (Salvelinus umbla). We sampled wild char and used their gametes to produce embryos of 40 different families. Embryos were raised singly in a stress or a non-stress environment until a late larval stage (stress was induced by allowing remainders of ovarian fluids to support microbial growth). Genetic markers were then used to sex the fish and reconstruct paternity (N= 1,463, including dead embryos). Primary sex ratio did not differ among families and was about 1:1. Female embryos hatched on average later and showed lower stress tolerance than male embryos. There were significant parental effects on offspring growth and mortality, but the sex differences in embryo performance were not family specific. We conclude that the sexes differ in their life history and susceptibilities to environmental stress already at embryonic stages. Environmental stress during incubation can therefore affect population sex ratio and hence population growth and genetics.

Maternal diet affects utilization of endogenous lipids by red drum Sciaenops ocellatus embryos and early larvae.

Embryonic and early larval development and metabolism of most teleost fishes are fueled entirely by maternally derived nutritional resources (yolk and oil) before the onset of exogenous feeding. The composition of those resources depends, in part, on maternal diet. To examine how diet-induced variations in egg composition affect embryonic and larval utilization of endogenous lipids and fatty acids, we fed red drum (Sciaenops ocellatus) broodstock four different diets to produce distinctive fatty acid compositions in eggs but with no significant difference in total lipid content. We sampled embryos and unfed larvae every 12h from 12h post-fertilization (hpf) until starvation (96-120 hpf; n=5 spawns per diet group). Rates of utilization of the oil globule were significantly different among diet groups, resulting in significant differences in mean oil globule size at the first feeding stage (84 hpf). Utilization rates for 15 fatty acids were significantly different among diet groups and were proportional to their initial concentration. As a result, differences in larval fatty acid compositions among maternal diet groups diminished over time but remained different even at later stages. In addition, larval standard length at 84 hpf was positively correlated with egg total lipid content. This study suggests that the fatty acid composition of yolk and oil affects energy metabolism and tissue composition in red drum larvae. These effects could have consequences for the survival and essential physiological functions of fish larvae during the critical period of transition to exogenous feeding.

Odorant binding protein C12 is involved in the defense against eugenol in Tribolium castaneum

Tribolium castaneum (T. castaneum) is a worldwide pest of stored grain that mainly harms flour, and not only causes serious loss of flour quality but also leads to deterioration of flour quality. Chemical detection plays a key role in insect behavior, and the role of odorant-binding proteins (OBPs) in insect chemical detection has been widely studied. However, the mechanism of OBPs in insect defense against exogenous toxic substances is still unclear. In this study, biochemical analysis showed that eugenol, the active component of A. vulgaris essential oil, significantly induced the expression of the OBP gene OBPC12 from T. castaneum (TcOBPC12). The mortality of late larvae treated with eugenol was higher than that of the control group after RNA interference (RNAi) against TcOBPC12, which indicates that the OBP gene is involved in the eugenol defense mechanism and leads to a decrease in sensitivity to eugenol. Tissue expression profiling showed that the expression of TcOBPC12 in the epidermis, hemolymph, and intestine was higher than in other larval tissues, and TcOBPC12 was expressed mainly in the epidermis, head, and fat body of adults. The developmental expression profile showed that the expression of TcOBPC12 in late eggs, early and late larval stages, and late adult stages was higher than in other developmental stages. These data suggest that TcOBPC12 may be involved in the absorption of exogenous toxic substances by the larvae from T. castaneum. Our results provide a theoretical basis for the metabolism and degradation mechanism of exogenous toxic substances and help explore more potential target genes of insect pests.

Myogenesis of Siboglinum fiordicum sheds light on body regionalisation in beard worms (Siboglinidae, Annelida)

BackgroundMany annelids, including well-studied species such as Platynereis, show similar structured segments along their body axis (homonomous segmentation). However, numerous annelid species diverge from this pattern and exhibit specialised segments or body regions (heteronomous segmentation). Recent phylogenomic studies and paleontological findings suggest that a heteronomous body architecture may represent an ancestral condition in Annelida. To better understand the segmentation within heteronomous species we describe the myogenesis and mesodermal delineation of segments in Siboglinum fiordicum during development.ResultsEmploying confocal and transmission electron microscopy we show that the somatic longitudinal musculature consists of four separate strands, among which ventrolateral one is the most prominent and is proposed to drive the search movements of the head of the late metatrochophore. The somatic circular musculature lies inside the longitudinal musculature and is predominantly developed at the anterior end of the competent larva to support the burrowing behaviour. Our application of transmission electron microscopy allows us to describe the developmental order of the non-muscular septa. The first septum to form is supported by thick bundles of longitudinal muscles and separates the body into an anterior and a posterior region. The second group of septa to develop further divides the posterior body region (opisthosoma) and is supported by developing circular muscles. At the late larval stage, a septum reinforced by circular muscles divides the anterior body region into a forepart and a trunk segment. The remaining septa and their circular muscles form one by one at the very posterior end of the opisthosoma.ConclusionsThe heteronomous Siboglinum lacks the strict anterior to posterior sequence of segment formation as it is found in the most studied annelid species. Instead, the first septum divides the body into two body regions before segments are laid down in first the posterior opisthosoma and then in the anterior body, respectively. Similar patterns of segment formation are described for the heteronomous chaetopterid Chaetopterus variopedatus and serpulid Hydroides elegans and may represent an adaptation of these annelids to the settlement and transition to the sedentarian-tubiculous mode of life.

Linking environmental drivers, juvenile growth, and recruitment for Japanese jack mackerel Trachurus japonicus in the Sea of Japan

AbstractGrowth traits of juvenile Japanese jack mackerel (Trachurus japonicus) were estimated based on otolith daily increments in spring during 2011–2016 in the southwestern part of the Sea of Japan (SWSJ). Spatial variability in growth traits was examined between the western and eastern areas of the study area. Back‐calculated body length at 50 days post hatch varied among areas and years between 26 and 40 mm. Body lengths at age in the eastern area were significantly smaller than those in the western area. Slower growth rate in the eastern area was associated with the lower ambient temperature in the eastern area than in the western area. In the eastern area, interannual variations in the instantaneous growth rate positively correlated with ambient temperature. Furthermore, interannual variations in back‐calculated body length during the late larval and early juvenile stages positively correlated with the juvenile abundance. These results suggest that the thermal conditions in the eastern area are indicative of the growth rate and abundance of juvenile T. japonicus in SWSJ. We further examined relationships between juvenile abundance and thermal conditions during 2003–2019 using archived data of this survey and found that the ambient temperature in the eastern area positively correlated with the juvenile abundance and also the strength of the Tsushima Warm Current. Thus, higher temperatures derived from the Tsushima Warm Current result in a faster growth rate during the late larval and early juvenile stages and subsequently higher abundance of T. japonicus juveniles in SWSJ.

Oxidative Stress Parameters in Goitrogen-Exposed Crested Newt Larvae (Triturus spp.): Arrested Metamorphosis.

Thiourea is an established disruptor of thyroid hormone synthesis and is frequently used as an inhibitor of metamorphosis. The changes caused by thiourea can affect processes associated with the oxidative status of individuals (metabolic rate, the HPI axis, antioxidant system). We investigated the parameters of oxidative stress in crested newt (Triturus spp.) larvae during normal development in late larval stage 62 and newly metamorphosed individuals, and during thiourea-stimulated metamorphosis arrest in individuals exposed to low (0.05%) and high (0.1%) concentrations of thiourea. Both groups of crested newts exposed to thiourea retained their larval characteristics until the end of the experiment. The low activities of antioxidant enzymes and the high lipid peroxidation level pointed to increased oxidative stress in larvae at the beginning of stage 62 as compared to fully metamorphosed individuals. The activities of catalase (CAT) and glutathione-S-transferase (GST) and the concentration of sulfhydryl (SH) groups were significantly lower in larvae reared in aqueous solutions containing thiourea than in newly metamorphosed individuals. The high thiourea concentration (0.1%) affected the antioxidative parameters to the extent that oxidative damage could not be avoided, contrary to a lower concentration. Our results provide a first insight into the physiological adaptations of crested newts during normal development and simulated metamorphosis arrest.

Light colour affect the survival rate, growth performance, cortisol level, body composition, and digestive enzymes activities of different Snubnose pompano (Trachinotus blochii (Lacépède, 1801) larval stages

Snubnose pompano (Trachinotus blochii) are widely cultured in the world. Light colours are among factors affecting fish performance in captivity. Snubnose pompano larvae often reared under white light. However, no study supports its choice or informs the extent of its effect or its constituent colour. The study investigated the influence of green, blue, yellow, purple, and white lights on growth, survival, cortisol, body composition, and digestive enzymes activities of one-day post-hatch Snubnose pompano larvae reared in light magenta tanks under a photoperiod of 24 Light:0 Dark hours for 25 days. Overall, the best growth performance of snubnose pompano larvae was in purple and white, and green lights during early and late larval stages, respectively. White light promoted survival rate while that of green light reduced. Larvae were less stressed and had enhanced body contents under white and purple light environments. Purple and yellow lights influenced low trypsin and pepsin activities. The study reports for the first time the effect of the light colour environment on the digestive enzyme activities and body composition of the finfish larvae. Generally, the study provides novel insights on optimal light colour in the larval rearing protocol of this species. Through this study, the use of white light during the first feeding (zero to four days post-hatch) and purple light in later stages (five days post-hatch until metamorphosis) is encouraged while discourages green and yellow lights in the larval rearing of Snubnose pompano.

Effect of tank colour on growth, survival, stress level, digestive enzymatic activities and body composition of snubnose pompano ( Trachinotus blochii Lacépède) larvae

Snubnose pompano (Trachinotus blochii, Lacépède) is a commercially important fish species that shows robust growth in captive environment. Tank colour is an important factor affecting the growth of fish larvae in aquaculture. Although the effect of tank colour has been studied in several fish species, it has not been studied in the snubnose pompano. We aimed to investigate how tank colour affects overall growth, survival, body composition, stress levels and digestive enzyme activities of snubnose pompano larvae at early larval stage and late larval stage. We noted that early–larval–stage larvae showed higher growth, crude protein contents and lower digestive enzyme activities when reared in blue and white tanks. Larvae showed low-stress levels when reared in grey and blue tanks. The same larvae showed low-survival and high-stress levels when reared in red tanks. The late–larval–stage larvae showed high growth and lipids, and low-digestive enzymes activities in white and yellow tanks. The larvae showed high-stress levels and low-ash contents when reared in red tanks. These differences are most likely due to the differences in light intensity inside tanks, which affects the ability of larvae to find and catch prey. Overall, our study sheds new light on cultivation conditions for snubnose pompano larvae. We recommend blue tanks for growing early–larval–stage larvae and white or yellow tanks for late-larval-stage larvae.

Characterization of a member of the CEACAM protein family as a novel marker of proton pump-rich ionocytes on the zebrafish epidermis.

In humans, several members of the CEACAM receptor family have been shown to interact with intestinal pathogens in an inflammatory context. While CEACAMs have long been thought to be only present in mammals, recent studies have identified ceacam genes in other vertebrates, including teleosts. The function of these related genes remains however largely unknown. To gain insight into the function of CEACAM proteins in fish, we undertook the study of a putative member of the family, CEACAMz1, identified in Danio rerio. Sequence analysis of the ceacamz1 gene product predicted a GPI-anchored extracellular protein containing eleven immunoglobulin domains but revealed no evident orthology with human CEACAMs. Using a combination of RT-PCR analyses and in situ hybridization experiments, as well as a fluorescent reporter line, we showed that CEACAMz1 is first expressed in discrete cells on the ventral skin of zebrafish larvae and later on in the developing gills. This distribution remains constant until juvenile stage is reached, at which point CEACAMz1 is almost exclusively expressed in gills. We further observed that at late larval stages, CEACAMz1-expressing cells mostly localize on the afferent side of the branchial filaments and possibly in the inter-lamellar space. Using immunolabelling and 3D-reconstructions, we showed that CEACAMz1 is expressed in cells from the uppermost layer of skin epidermis. These cells are embedded within the keratinocytes pavement and we unambiguously identified them as proton-pump rich ionocytes (HR cells). As the expression of ceacamz1 is turned on concomitantly to that of other known markers of HR cells, we propose that ceacamz1 may serve as a novel marker of mature HR cells from the zebrafish epidermis.

Combination of Modified Atmosphere and Irradiation for the Phytosanitary Disinfestation of Trogoderma granarium Everts (Coleoptera: Dermestidae).

Simple SummaryThe khapra beetle is defined as one of the most important quarantine pests globally, and fumigating by methyl bromide, one of the ozone-depleting substances under the Montreal Protocol, is a routine measure used for phytosanitary treatment. To protect the Ozone layer, an environmentally friendly measure is needed to be developed. The middle- to late-stage larvae and adults were treated with irradiation, modified atmosphere (MA) alone, and their combinations at room temperature of 24–26 ℃. As a result, late-stage larvae are determined as the most tolerant stage. Ionizing radiation was used to enhance the effects of 1% and 2% O2 MA treatments that the obvious synergistic effects are presented in all combinations, resulted in saving as high as 60% of the estimated exposure times comparing with MA treatment alone. A total of 111,366 late-stage larvae were exposed to a 1% O2 atmosphere for 14 or 15 days after a 200 Gy irradiation, resulted in no survivor in the validating tests. Therefore, the MA-irradiation combination treatment can provide quarantine security at a very high level, it may be combined with international transportation (train or sea container) to disinfest the commodities infested by khapra beetle and other stored products insect pests.The khapra beetle, Trogoderma granarium Everts, is defined as one of the most important quarantine pests globally, and fumigation with methyl bromide, an ozone-depleting substance, is a common phytosanitary measure currently used. The modified atmosphere (MA), irradiation, and their combination treatments of T. granarium larvae and adults were performed at room temperature (24–26 ℃) to develop an ecofriendly phytosanitary disinfestation measure and to shorten the exposure time and overcome treatment disadvantages of irradiation. Late-stage larvae are determined as the most tolerant stage resulted in large LT99.9968 values of 32.6 (29.2–37.5) and 38.0 (35.1–41.7) days treated under 1% and 2% O2 (with N2 balance) atmosphere, respectively. Ionizing radiation was used to enhance the effect of MA and the mortality was highly significantly affected by all the interaction effects, indicating that the synergistic effects present in all the combined treatments. The synergistic ratios, which is defined as the estimated lethal time for MA treatment (LD90, LD99, and LD99.9968), divided by that of combined treatment, were between 1.47 and 2.47. In the confirmatory tests, no individuals recovered from a sum of 111,366 late-stage larvae treated under 1% O2 atmosphere for 14- or 15-d after 200 Gy irradiation, which resulted in validating the probit estimations and achieving an efficacy of 99.9973% mortality at 95% confidence level. Therefore, these treatment schedules are recommended to disinfest T. granarium infecting commodities for phytosanitary purposes under the warehouse, MA packaging, or in combination with international transportation by train or sea container.

Molecular cloning and expression pattern of the juvenile hormone epoxide hydrolase gene from the giant freshwater prawn Macrobrachium rosenbergii during larval development and the moult cycle

Juvenile hormone epoxide hydrolases (JHEHs) are regulators of moulting, development and metamorphosis in insects, but we have limited understanding of the roles of these crucial hormonal regulator in crustaceans. In this study, the giant freshwater prawn Macrobrachium rosenbergii was used as a model organism to investigate the role of JHEH in larval development and moulting. We cloned and characterized a JHEH gene from M. rosenbergii (designated as MrJHEH). The full-length cDNA of MrJHEH was 1,682 bp in length, encoding 460 amino acids, and the deduced MrJHEH protein possesses typical features of JHEH family. The phylogenetic analysis revealed that MrJHEH clustered with homologues from crustaceans. Investigation of tissue distribution revealed the abundant expression of MrJHEH in the hepatopancreas. Developmental expression studies showed that the MrJHEH mRNA was expressed throughout the embryo to post-larval stages, with higher expression at late larval stages. Analysis of the moult cycle showed that the expression of MrJHEH mRNA was significantly elevated at the pre-moult stage as compared with the inter-moult level. Both exogenous 20-hydroxyecdysone (20E) injection and eyestalk ablation (ESA) significantly induced the transcription of MrJHEH in cephalothoraxes. These data suggest that MrJHEH is indispensable for the regulation of larval development and ecdysteroidogenesis of M. rosenbergii.

Fine structure of Drosophila larval salivary gland ducts as revealed by laser confocal microscopy and SEM

The functions of the larval salivary glands (SGs) of Drosophila are traditionally associated with the production of a massive secretion during puparium formation; it is exocytosed into a centrally located lumen and subsequently expectorated via ducts, the pharynx and mouth. This so-called proteinaceous glue serves as an adhesive to attach the puparial case to a solid substrate. Great attention has been paid to the secretory cells of SGs, which are famous for their giant polytene chromosomes. However, substantially less attention has been devoted to individual or common ducts that form the most proximal portion of the SG organ via which the glue is released into the pharynx. In the present paper, we describe the organization and fine structure of the taenidia, highly specialized circumferential ring-like extracellular (cuticular) components on the internal side of these tubes. Two chitin-specific probes that have previously been used to recognize taenidia in Drosophila tracheae, Calcofluor White M2R (also known as Fluorescent Brightener 28) and the novel vital fluorescent dye SiR-COOH, show positively stained ductal taenidia in late larval SGs. As seen using scanning electron microscopy (SEM), the interior of the ductal tube contains regular and densely-arranged ridge-like circumferential rings which represent local thickenings of the cuticle in various geometries. The microtubular arrays that optically colocalize with taenidia in both the trachea and SG ducts are specifically and strongly recognized by fluorescently-conjugated colchicine as well as anti-tubulin antibody. In contrast to taenidia in the tracheae, the analogous structures in SG ducts cannot be detected by fluorescently-labeled phalloidin or even actin-GFP fusion protein, suggesting that the ducts lack a cortical network made of filamentous actin. We speculate that these taenidia may serve to reinforce the duct during the secretory processes that SGs undergo during late larval and late prepupal stages.

Ionotropic Receptor-dependent cool cells control the transition of temperature preference in Drosophila larvae.

Temperature sensation guides animals to avoid temperature extremes and to seek their optimal temperatures. The larval stage of Drosophila development has a dramatic effect on temperature preference. While early-stage Drosophila larvae pursue a warm temperature, late-stage larvae seek a significantly lower temperature. Previous studies suggest that this transition depends on multiple rhodopsins at the late larval stage. Here, we show that early-stage larvae, in which dorsal organ cool cells (DOCCs) are functionally blocked, exhibit similar cool preference to that of wild type late-stage larvae. The molecular thermoreceptors in DOCCs are formed by three members of the Ionotropic Receptor (IR) family, IR21a, IR93a, and IR25a. Early-stage larvae of each Ir mutant pursue a cool temperature, similar to that of wild type late-stage larvae. At the late larval stage, DOCCs express decreased IR proteins and exhibit reduced cool responses. Importantly, late-stage larvae that overexpress IR21a, IR93a, and IR25a in DOCCs exhibit similar warm preference to that of wild type early-stage larvae. These data suggest that IR21a, IR93a, and IR25a in DOCCs navigate early-stage larvae to avoid cool temperatures and the reduction of these IR proteins in DOCCs results in animals remaining in cool regions during the late larval stage. Together with previous studies, we conclude that multiple temperature-sensing systems are regulated for the transition of temperature preference in fruit fly larvae.

Exogenous administration of dsRNA for the demonstration of RNAi in Maruca vitrata (lepidoptera: crambidae).

The online version contains supplementary material available at 10.1007/s13205-021-02741-8.

Breeding Effort and Hydroperiod Indicate Habitat Quality of Small, Isolated Wetlands for Amphibians Under Climate Extremes

The distribution and function of small, temporarily ponded wetlands such as vernal pools are driven by climate variation, especially precipitation. These wetland ecosystems provide important habitat for amphibians, whose breeding effort (egg deposition) is often used to characterize pool habitat quality. However, whether breeding effort consistently predicts larval survival, juvenile production, and overall reproductive output—key indicators of habitat quality for amphibians in a varying environment—remains unclear. We assessed the occurrence of spotted salamander (Ambystoma maculatum) and wood frog (Lithobates sylvaticus) larvae in 71 vernal pools in New York, USA during the spring and summer of 2016 and 2017—the fifth driest and third wettest years in regional history, respectively—to better understand the relationship between breeding effort and metamorph production under environmental extremes. Breeding effort was an important predictor of whether late-stage larvae occurred in pools for both species. However, occurrence of late-stage larvae was substantially lower for both species during 2016, i.e., the drought year, when pools dried rapidly. These outcomes suggest that, in the face of an increasingly variable climate, prioritizing clusters of pools that encompass a range of hydroperiods and include high-productivity pools would help maintain populations of vernal-pool dependent amphibians.