Relish involved in immunity and larval survival in the willow leaf beetle Plagiodera versicolora.

BACKGROUND: Insects mainly rely on innate immunity against pathogen infection. Plagiodera versicolora (Coleoptera: Chrysomelidae), is a worldwide leaf-eating forest pest in salicaceous trees. However, the mechanisms behind the immunodeficiency pathway (IMD) remain poorly understood. RESULTS: In this study, we obtained a Relish gene from transcriptome analysis. Tissue and instar expression profiles were subsequently obtained using quantitative real-time polymerase chain reaction analysis. The results showed that Relish has high expression levels in eggs, larvae and adults, and especially in fat bodies. Transcripts of the tested antimicrobial peptides (AMPs), defensin1, defensin2 and attacin2 were downregulated by dsRelish. Knockdown of Relish led to greater mortality in larvae after Staphylococcus aureus infection. In addition, we performed bacterial 16S ribosomal RNA-based high-throughput sequencing. The results showed that the relative abundance of some gut bacteria was significantly altered after dsRelish ingestion. CONCLUSION: This study provides a greater understanding of the IMD signaling pathway, facilitating functional studies of Relish in P. versicolora. Moreover, a genetic pest management technique might be developed using Relish as a lethal gene to control the pest P. versicolora. © 2024 Society of Chemical Industry.

Constructing the Polymer Molecules to Regulate the Electrode/Electrolyte Interface to Enhance Lithium-Metal Battery Performance.

Lithium-ion batteries, with high energy density and long cycle life, have become the battery of choice for most vehicles and portable electronic devices; however, energy density, safety and cycle life require further improvements. Single-functional group electrolyte additives are very limited in practical applications, a ternary polymer bifunctional electrolyte additive copolymer (acrylonitrile-butyl hexafluoro methacrylate- poly (ethylene glycol) methacrylate- methyl ether) (PMANHF) was synthesized by free radical polymerization of acrylonitrile, 2, 2, 3, 4, 4, 4-hexafluorobutyl methacrylate and poly (ethylene glycol) methyl ether methacrylate. A series of characterizations show that in Li metal anodes, the preferential reduction of PMANHF is conducive to the formation of a uniform and stable solid electrolyte interphase layer, and Li deposition is uniform and dense. At the NCM811 cathode, a film composed of LiF- and Li3N-rich is formed at the cathode-electrolyte interface, mitigating the side reaction at the interface. At 1.0 mA cm-2, the Li/Li cell can be stabilized for 1000 cycles. In addition, the Li/NCM811 cell can stabilize 200 cycles with a cathode capacity of 153.7 mAh g-1, with the capacity retention of 89.93 %, at a negative/positive capacity ratio of 2.5. This study brings to light essential ideas for the fabrication of additives for lithium-metal batteries.

Programmable synthesis of difluorinated hydrocarbons from alkenes through a photocatalytic linchpin strategy.

The introduction of difluoromethylene moieties into organic molecules has garnered significant attention due to their profound influence on the physicochemical and biological properties of compounds. Nonetheless, the existing approaches for accessing difluoroalkanes from readily available feedstock chemicals remain limited. In this study, we present an efficient and modular protocol for the synthesis of difluorinated compounds from alkenes, employing the readily accessible reagent, ClCF2SO2Na, as a versatile "difluoromethylene" linchpin. By means of an organophotoredox-catalysed hydrochlorodifluoromethylation of alkenes, followed by a ligated boryl radical-facilitated halogen atom transfer (XAT) process, we have successfully obtained various difluorinated compounds, including gem-difluoroalkanes, gem-difluoroalkenes, difluoromethyl alkanes, and difluoromethyl alkenes, with satisfactory yields. The practical utility of this linchpin strategy has been demonstrated through the successful preparation of CF2-linked derivatives of complex drugs and natural products. This method opens up new avenues for the synthesis of structurally diverse difluorinated hydrocarbons and highlights the utility of ligated boryl radicals in organofluorine chemistry.

Blooming characteristics and breeding system of an invasive plant Gaura parviflora.

The invasiveness and dissemination of exotic species are strongly influenced by its sexual reproduction characteristics, including blooming characteristics and breeding system. Exploring the association of these sexual reproductive traits with invasiveness would be helpful for revealing the mechanism of its successful invasion. We examined the blooming characteristics and breeding system of Gaura parviflora based on field observations, out-crossing index (OCI) estimation, and hand-pollination experiments. The results showed flowering duration of the G. parviflora population (flowering period) was short (more than 3 months). The life span of single flower (floral longevity) was 40.46 h. Its flower diameter was 3.99 mm. Over seven flowers in bloom per inflorescence and most individuals often bloomed synchronously, which showed a 'mass-flowering pattern'. The changing trend of pollen and stigma vitality was relatively similar, but the duration of stigma vitality was 2 h longer than that of pollen. The stigma and the anthers were close to each other at the initial flowering stage, but the stigma removed from the anthers at the full-blooming stage with the style curving downwards. Many pollinators visited flowers in late full-blooming stage, which were mainly Apis mellifera and Syrphidae spp. Their average visiting frequency was 9.8 times·m-2·h-1. The fruit set in natural pollination after emasculation treatment (insect or wind pollination) was signi-ficantly higher than that in bagged and emasculation treatment, and the treatment of emasculated and bagged with nylon net (excluding insect pollination) could also bear fruits, indicating possible existence of ambophily in G. parviflora. The results of pollen ovule ratio (P/O) mensuration, OCI estimation and hand-pollination experiments showed that its mating system type belonged to additive mixed mating system. So, its characteristics, such as smaller flower size, shorter floral longevity and flowering period, were conducive to allocating more resources to plant growth and seed development, which would help improve its total fitness. The changes of spatial position of male and female organs not only avoided interference between male and female functions, but also created opportunities for stigmas to receive outcross pollen. In addition, the 'mass-flowering pattern' was conducive to attracting pollinators. The pollination mechanism of ambophily was helpful to ensure cross-pollination. The additive mixed mating system could provide double reproductive assurance for this species. These reproductive characteristics were significant for the successful invasion and expansion of G. parviflora.

d-Orbital Electron Delocalization Realized by Axial Fe4 C Atomic Clusters Delivers High-Performance Fe-N-C Catalysts for Oxygen Reduction Reaction.

Fe-N-C catalyst for oxygen reduction reaction (ORR) has been considered as the most promising nonprecious metal catalyst due to its comparable catalytic performance to Pt in proton exchange membrane fuel cells (PEMFCs). The active centers of Fe-pyrrolic N4 have been proven to be extremely active for ORR. However, forming a stable Fe-pyrrolic N4 structure is a huge challenge. Here, a Cyan-Fe-N-C catalyst with Fe-pyrrolic N4 as the intrinsic active center is constructed with the help of axial Fe4 C atomic clusters, which shows a half-wave potential of up to 0.836 V (vs. RHE) in the acid environment. More remarkably, it delivers a high power density of 870 and 478 mW cm-2 at 1.0 bar in H2 -O2 and H2 -Air fuel cells, respectively. According to theoretical calculation and in situ spectroscopy, the axial Fe4 C can provide strong electronic perturbation to Fe-N4 active centers, leading to the d-orbital electron delocalization of Fe and forming the Fe-pyrrolic N4 bond with high charge distribution, which stabilizes the Fe-pyrrolic N4 structure and optimizes the OH* adsorption during the catalytic process. This work proposes a new strategy to adjust the electronic structure of single-atom catalysts based on the strong interaction between single atoms and atomic clusters.

Comparative analysis of anesthesia and clinical efficacy of inhalation anesthesia and intravenous anesthesia in Trigeminal Nerve Balloon Avulsion.

Objective: To observe the anesthesia and clinical efficacy of inhalation anesthesia and intravenous anesthesia in patients with trigeminal neuralgia undergoing surgery. Methods: This is a retrospective study. Eighty patients with trigeminal neuralgia admitted to the Affiliated Hospital of Beihua University from July 2018 to July 2021 were selected and divided into two groups according to different anesthesia methods: inhalation group and intravenous group, with 40 cases in each group. Patients in the inhalation group were given inhalation anesthesia with sevoflurane, while those in the intravenous group were given intravenous anesthesia. Hemodynamics, intubation and extubation time, postoperative consciousness recovery, adverse reactions and clinical effects of surgery were compared between the two groups during anesthesia. Results: During the induction of anesthesia, after induction and after surgery, the levels of hemodynamic parameters in the two groups increased compared with those before induction of anesthesia, and the increase in the inhalation group was smaller (P<0.05). Patients in the inhalation group had a long time from anesthesia to endotracheal intubation but had a short time from completion of surgery to intubation, which was statistically significant compared with the intravenous group (P<0.05). Compared with the intravenous group, the postoperative consciousness recovery time of the inhalation group was significantly shorter and the incidence of adverse reactions was significantly lower (P<0.05). Conclusion: Inhalation anesthesia with sevoflurane is more effective than intravenous anesthesia in trigeminal neuralgia patients treated with trigeminal nerve balloon avulsion, boasting satisfactory safety, less impact on hemodynamics, and shorter recovery time of consciousness.

Electrocatalysis Mechanism and Structure-Activity Relationship of Atomically Dispersed Metal-Nitrogen-Carbon Catalysts for Electrocatalytic Reactions.

Atomically dispersed metal-nitrogen-carbon catalysts (M-N-C) have been widely used in the field of energy conversion, which has already attracted a huge amount of attention. Due to their unsaturated d-band electronic structure of the center atoms, M-N-C catalysts can be applied in different electrocatalytic reactions by adjusting their own microscopic electronic structures to achieve the optimization of the structure-activity relationship. Consequently, it is of great significance for the revelation of electrocatalytic mechanism and structure-activity relationship of M-N-C catalysts. Thus, this review first introduces the relative research methods, including in situ/operando characterization techniques and theoretical calculation methods. Furthermore, clarifying the electrocatalytic mechanism and structure-activity relationship of M-N-C catalysts in different electrochemical energy conversion reactions is focused. Moreover, the future research directions are pointed out based on the discussion. This review will provide good guidance to systematically study the catalytic mechanism of single-atom catalysts and reasonably design the single-atom catalysts.

Disruption of tetrahydrobiopterin (BH4) biosynthesis pathway affects cuticle pigmentation in Henosepilachna vigintioctopunctata.

Tetrahydrobiopterin (BH4) is produced from guanosine triphosphate (GTP) under catalyzation of GTP cyclohydrolase I (GTPCH), 6-pyruvoyltetrahydropterin synthase (PTPS) and sepiapterin reductase (SR), among others. In Drosophila melanogaster, BH4 and other pteridines are required for cuticle tanning and eye pigmentation. In this study, two Hvgtpch (Hvgtpch-a and Hvgtpch-b), an Hvptps and an Hvsr transcripts were identified in a serious defoliator Henosepilachna vigintioctopunctata. Hvgtpch-a and Hvgtpch-b were highly expressed just before and/or right after the molt, in contrast to Hvptps and Hvsr. RNA interference (RNAi) by injection of a dsgtpch targeting the common fragment of Hvgtpch-a and Hvgtpch-b into the third instar larvae caused albino fourth-instar larvae and pupae. Around 80% of the Hvgtpch RNAi larvae failed to pupate. The remaining 20% of Hvgtpch RNAi pupated beetles did not completely remove the larval/pupal exuviae after emerged as adults and eventually died. Depletion of Hvgtpch at the fourth instar stage resulted in under-pigmented pupae and adults, with significantly low pupation and emergence rates. The Hvgtpch RNAi adults rarely moved and fed on plant leaves; they died within a week after emergence. Silence of Hvptps or Hvsr at the third- and fourth-instar stages led to similar but less serious phenotypes, with lowest influence in the Hvsr RNAi ladybirds. Moreover, RNAi of Hvgtpch, Hvptps or Hvsr did not affect coloration of the larval ocelli and pupal/adult compound eyes. Therefore, our results demonstrated that pteridines are involved in melanin formation but not in eye pigmentation in H. vigintioctopunctata. Moreover, our findings will enable the development of a dsgtpch-based pesticide to control H. vigintioctopunctata larvae.

Population structure of food-borne Staphylococcus aureus in China / 中华流行病学杂志

Objective: To understand the population structure of food-borne Staphylococcus (S.) aureus in China. Methods: Whole genome sequencing was used to analyze 763 food-borne S. aureus strains from 16 provinces in China from 2006 to 2020. Multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and staphylococcal chromosome cassettemec (SCCmec) typing were conducted, and minimum spanning tree based on ST types (STs) was constructed by BioNumerics 7.5 software. Thirty-one S. aureus strains isolated from imported food products were also included in constructing the genome phylogenetic tree. Results: A total of 90 STs (20 novel types) and 160 spa types were detected in the 763 S. aureus isolates. The 72 STs (72/90, 80.0%) were related to 22 clone complexes. The predominant clone complexes were CC7, CC1, CC5, CC398, CC188, CC59, CC6, CC88, CC15, and CC25, accounting for 82.44% (629/763) of the total. The STs and spa types in the predominant clone complexes changed over the years. The methicillin-resistant S. aureus (MRSA) detection rate was 7.60%, and 7 SCCmec types were identified. The ST59-t437-Ⅳa (17.24%, 10/58), ST239-t030-Ⅲ (12.07%, 7/58), ST59-t437-Ⅴb (8.62%, 5/58), ST338-t437-Ⅴb (6.90%, 4/58) and ST338-t441-Ⅴb (6.90%, 4/58) were the main types in MRSA strains. The genome phylogenetic tree had two clades, and the strains with the same CC, ST, and spa types clustered together. All CC7 methicillin sensitive S. aureus strains were included in Clade1, while 21 clone complexes and all MRSA strains were in Clade2. The MRSA strains clustered according to the SCCmec and STs. The strains from imported food products in CC398, CC7, CC30, CC12, and CC188 had far distances from Chinese strains in the tree. Conclusions: In this study, the predominant clone complexes of food-borne strains were CC7, CC1, CC5, CC398, CC188, CC59, CC6, CC88, CC15, and CC25, which overlapped with the previously reported clone complexes of hospital and community-associated strains in China, suggesting that close attention needs to be paid to food, a vehicle of pathogen transmission in community and food poisoning.

Association of maternal isolated thyroid peroxidase antibody positive in the first trimester with fetal growth / 北京大学学报(医学版)

OBJECTIVE@#To investigate the association of isolated thyroid peroxidase antibody (TPOAb) positive in the first trimester with fetal growth.@*METHODS@#A total of 16 446 pregnant women were included in the birth cohort study, whose last menstrual period was between May 2016 and April 2019 and with singleton pregnancy. Maternal serum samples were collected when they firstly came for prenatal care in the first trimester. The pregnant women were consecutively seen and followed in the hospital and the information of pregnant women was extracted from the electronic medical information system. The pregnant women were divided into isolated TPOAb positive group (n=1 654) and euthyroid group (n=14 792). Three fetal ultrasound examinations were scheduled during the routine prenatal visits at the hospital and were performed by trained sonographers. All fetal growth indicators were quantified as gestational age- and gender- adjusted standard deviation score (Z-score) using the generalized additive models for location, scale and shape (GAMLSS). Fetal growth indicators included estimated fetal weight (EFW), abdominal circumference (AC), biparietal diameter (BPD), femur length (FL) and head circumference (HC). Fetal growth restriction (FGR) was defined as AC or EFW Z-score＜3rd centile based on clinical consensus. Generalized estimating equation (GEE) analysis was applied to assess the association of maternal isolated TPOAb positive with fetal growth. The generalized linear model was further used to analyze the association between isolated TPOAb positive and fetal growth indicator at different gestational ages when the fetal growth indicator was significantly associated with isolated TPOAb positive in the GEE mo-del.@*RESULTS@#The median gestational age at three ultrasound measurements was 23.6 (23.3, 24.1), 30.3 (29.7, 30.9), 37.3 (37.0, 37.7) weeks, respectively. The BPD Z-score was higher in isolated TPOAb positive women, compared with the euthyroid pregnant women after adjustment (β=0.057, 95%CI: 0.014-0.100, P=0.009). The generalized linear model showed the BPD Z-score was higher in the isolated TPOAb positive women at the end of 21-25 weeks (β=0.052, 95%CI: 0.001-0.103, P=0.044), 29-32 weeks (β=0.055, 95%CI: 0.004-0.107, P=0.035) and 36-40 weeks (β=0.068, 95%CI: 0.011-0.125, P=0.020), compared with the euthyroid pregnant women. There was no difference in other fetal growth indicators (EFW, AC, FL and HC) and FGR between the isolated TPOAb positive and euthyroid pregnant women.@*CONCLUSION@#The BPD Z-score was slightly increased in the isolated TPOAb positive pregnant women in the first trimester, while other fetal growth indicators were not changed. The reproducibility and practical significance of this result need to be confirmed.

RNA interference of vATPase subunits A and E affects survival of larvae and adults in Plagiodera versicolora (Coleoptera: Chrysomelidae).

Vacuolar-type H+-ATPases (vATPases) are ATP-driven proton pumps and play essential roles in many physiological functions. Plagiodera versicolora (Coleoptera: Chrysomelidae) is a leaf-eating forest pest found in salicaceous trees worldwide. RNA interference (RNAi) is a powerful tool for functional identify and pest control. In this study, we used RNAi as an approach to knock down subunits A and E of the vATPase gene. The phylogenetic analysis showed that vATPase-A and vATPase-E from the same order were clustered together to form Coleoptera subclades, respectively. The expression levels of vATPase-A and vATPase-E were higher in gut, Malpighian tubules and 1st instar larvae. Ingest the dsvATPase-A and dsvATPase-E significantly inhibited the development of 1st to 3th instar larvae, incapacitated of mating and oviposition in adults. In addition, knockdown of vATPase subunit genes caused higher mortality in larvae and adults. The results demonstrate that RNAi efficiencies both vATPase-A and vATPase-E genes at various larvae stages and adults. Moreover, this research suggested that silencing of two vATPase subunits A and E offers a potential strategy to control P. versicolora.

Knockdown of Ecdysone-Induced Protein 93F Causes Abnormal Pupae and Adults in the Eggplant Lady Beetle.

Ecdysone-induced protein 93F (E93) plays triple roles during post-embryonic development in insects whose juvenile instars are more than four. However, it only acts as a specifier of adult structures in Drosophila flies whose larval instars are fixed at three. In this study, we determined the functions of E93 in the eggplant lady beetle (Henosepilachna vigintioctopunctata), which has four larval instars. We uncovered that E93 was abundantly expressed at the prepupal and pupal stages. A precocious inhibition of the juvenile hormone signal by RNA interference (RNAi) of HvKr-h1 or HvHairy, two vital downstream developmental effectors, at the penultimate instar larval stage increased the expression of E93, Conversely, ingestion of JH by the third-instar larvae stimulated the expression of HvKr-h1 but repressed the transcription of either HvE93X1 or HvE93X2. However, disturbance of the JH signal neither drove premature metamorphosis nor caused supernumerary instars. In contrast, depletion of E93 at the third- and fourth-instar larval and prepupal stages severely impaired pupation and caused a larval-pupal mixed phenotype: pupal spines and larval scoli were simultaneously presented on the cuticle. RNAi of E93 at the pupal stage affected adult eclosion. When the beetles had suffered from a dsE93 injection at the fourth-instar larval and pupal stages, a few resultant adults emerged, with separated elytra, abnormally folded hindwings, a small body size and short appendages. Taken together, our results suggest the larval instars are fixed in H. vigintioctopunctata; E93 serves as a repressor of larval characters and a specifier of adult structures during the larval-pupal-adult transition.

Temporal variation of antibiotic resistome and pathogens in food waste during short-term storage.

The massive food wastes pose a growing health concern for spreading of antibiotic resistance and pathogens due to food spoilage. However, little is known about these microbial hazards during collection, classification, and transportation before eventual treatment. Here, we profiled the temporal variations of antibiotic resistance genes (ARGs), pathogens, bacterial and fungal communities across four typical food wastes (vegetable, fish, meat, and rice) during storage at room temperature in summer (maximum 28-29 °C) of typical southeast city in China. A total of 171 ARGs and 32 mobile genetic elements were detected, and the absolute abundance of ARGs significantly increased by up to 126-fold with the storage time. Additionally, five bacterial pathogens containing virulence factor genes were detected, and Klebsiella pneumoniae was persistently detected throughout the storage time in all food types except rice. Moreover, fungal pathogens (e.g., Aspergillus, Penicillium, and Fusarium) were also frequently detected. Notably, animal food wastes were demonstrated to harbor higher abundance of ARGs and more types of pathogens, indicating a higher level of hazard. Mobile genetic elements and food types were demonstrated to mainly impact ARG profiles and pathogens, respectively. This work provides a comprehensive understanding of the microbial hazards associated with food waste recycling, and will contribute to optimize the food waste management to ensure biosecurity and benefit human health.

Design of Chopsticks-Shaped Heating Resistors for a Thermal Inkjet: Based on TaN Film.

Efficient printing frequency is critical for thermal bubble inkjet printing, while the difficulty lies in the structural design and material selection of the heating resistors. In this paper, a TaN film was used as the main material of the heating resistors, and two TaN films were placed in parallel to form the chopsticks-shaped structure. The heating time was divided into two sections, in which 0-0.1 µs was the preheating and 1.2-1.8 µs was the primary heating. At 1.8 µs, the maximum temperature of the Si3N4 film could reach about 1100 °C. At the same time, the SiO2 film was added between the TaN film and Si3N4 film as a buffer layer, which effectively avoided the rupture of the Si3N4 film due to excessive thermal stress. Inside the inkjet print head, the maximum temperature of the chamber reached about 680 °C at 2.5 µs. Due to the high power of the heating resistors, the working time was greatly reduced and the frequency of the inkjet printing was effectively increased. At the interface between the back of the chip and the cartridge, the SiO2 film was used to connect to ensure a timely ink supply. Under the condition of 12 V at 40 kHz, the inkjet chip could print efficiently with 10 nozzles at the same time. The inkjet chip proposed in this paper is not limited to only office printing, but also provides a new reference for 3D printing, cell printing, and vegetable and fruit printing.

Silencing tyrosine hydroxylase or dopa decarboxylase gene disrupts cuticle tanning during larva-pupa-adult transformation in Henosepilachna vigintioctopunctata.

BACKGROUND: The 28-spotted potato ladybird, Henosepilachna vigintioctopunctata, is a notorious defoliator of many solanaceous and cucurbitaceous plants. Tyrosine hydroxylase (TH) and dopa decarboxylase (DDC) are responsible for cuticle tanning pathway in insects. RESULTS: We identified HvTH and HvDDC in H. vigintioctopunctata, and found that high levels of them were accumulated just before or right after molting. Injection of dsHvTH or feeding 3-iodo-tyrosine (3-IT) at the third instar larval stage repressed tanning of the larval cuticle, reduced larval feeding, inhibited larval growth, and consequently caused 100% of larval mortality. Knockdown of HvDDC at the third instar larval stage hardly affected the coloration of larval head, and partially inhibited pigmentation of larval bodies and around 80% of the HvDDC RNAi larvae developed into albino pupae and adults. Moreover, depletion of HvTH or HvDDC at the fourth instar larval stage resulted in albino pupae and adults. The HvTH or HvDDC hypomorph adults fully or partially failed to remove the larval/pupal exuviae, possessed pale and abnormal wings, and poorly tanned heads and bodies, and eventually, struggled for several days without feeding on leaves before death. CONCLUSION: These results show that TH and DDC play key roles in larval and adult cuticle tanning and development in H. vigintioctopunctata. Also, these findings suggest that dopa- and dopamine-originated pigments are essential for larval and adult feeding behavior and the molting process during emergence. © 2022 Society of Chemical Industry.

Dissecting the Isoform-Specific Roles of FTZ-F1 in the Larval-Larval and Larval-Pupal Ecdyses in Henosepilachna vigintioctopunctata.

Fushi Tarazu Factor 1 (FTZ-F1), a member of the nuclear receptor superfamily, is the downstream factor of 20-hydroxyecdysone signaling. In Drosophila melanogaster, alternative transcription start and splicing in the FTZ-F1 gene generate αFTZ-F1 and ßFTZ-F1 isoforms, which are vital for pair-rule segmentation in early embryogenesis and post-embryonic development, respectively. However, whether the same mRNA isoforms are present and exert the conservative roles remains to be clarified in other insects. In the present paper, we first mined the genomic data of representative insect species and unveiled that the same post-transcriptional processing in FTZ-F1 occurred in coleopterans, lepidopterans, dipterans and hymenopterans. Our expression data in Henosepilachna vigintioctopunctata, a serious polyphagous defoliator damaging a wide range of crops in Solanaceae and Cucurbitaceae, showed that both αFTZ-F1 and ßFTZ-F1 were actively transcribed throughout the development, from embryo to adult. The RNA interference-aided knockdown of both isoforms completely arrested larval ecdysis from the third to the fourth instar, in contrast to the depletion of either isoform. In contrast, silencing ßFTZ-F1, rather than αFTZ-F1, severely impaired the larval-pupal transformation. We accordingly propose that both FTZ-F1 isoforms are essential but mutually interchangeable for larval-larval molting, while ßFTZ-F1 is necessary for the larval-pupal transition and sufficient to exert the role of both FTZ-F1s during larval-pupal metamorphosis in H. vigintioctopunctata.

Silencing of Adc and Ebony Causes Abnormal Darkening of Cuticle in Henosepilachna vigintioctopunctata.

N-ß-alanyldopamine (NBAD) is a precursor of N-acylquinone sclerotin utilized for cross-linking between cuticular proteins for cuticle during insect molting. The importance of NBAD in cuticle tanning has not been well compared among different developing stages of insects. Henosepilachna vigintioctopunctata, a typical polyphagous pest feeding on a large number of Solanaceae and Cucurbitaceae plants in Asian countries, displays diverse cuticle pigmentation patterns among developing stages and body regions. Here, we found that the expression of three genes (Hvadc, Hvebony, and Hvtan) involved in NBAD biosynthesis peaked in the 4-day-old pupae or 0-day-old adults of H. vigintioctopunctata. At the first, second, third, and fourth larval instar and pupal stage, their transcript levels were high just before and/or right after the molting. Moreover, they were more abundantly transcribed at the larval heads than in the bodies. RNA interference (RNAi) of either Hvadc or Hvebony at the third instar larvae selectively deepened the color of the larval head capsules, antennae, mouthpart, scoli, strumae, and legs; and depletion of the two genes blackened the pupal head capsules, antennae, mouthpart, and legs. However, the knockdown of either Hvadc or Hvebony darkened the whole bodies of the adults. Conversely, RNAi of Hvtan at the third instar stage had little influence on the pigmentation in the larvae, pupae, and adults. These findings demonstrated that Adc and Ebony are important in cuticle pigmentation of H. vigintioctopunctata and suggested that larger quantities of NBAD were present in adults and play more important roles in pigmentation than larvae/pupae.

Ultrastructure of three Species of Entomoneis (Bacillariophyta) from Lake Qinghai of China, with reference to the external areola occlusions.

Three sympatric Entomoneis species, found at the same specific locality in Lake Qinghai, China, are studied by using light and scanning electron microscope. Two species are proposed as new to science and named as E.sinensis sp. nov. and E.qinghainensis sp. nov. The third species is identified as E.paludosa (W. Smith) Reimer. Entomoneissinensis has a linear-lanceolate valve outline and Ƨ-shaped keel, bears two distinct 8-shaped loops formed by the valvocopula pars media in each cell and each of its stria is composed of either a long hymen strip or a long hymen strip plus one separated areola close to the raphe. Its hymen strip belongs to Type Two, which is a siliceous membrane strip perforated by two rows of linear pores next to transapical costae and two rows of rounded pores between these two rows of linear pores. Entomoneisqinghainensis has large cells, very high keel and evident hymen strip regions like a U-shaped neck pillow at the middle of valve face. Its hymen strip belongs to Type One, which is a siliceous membrane strip perforated by irregularly distributed round pores. Entomoneispaludosa also has the hymen strip regions that are worm-like and close to the raphe canal. Its hymen strip is same as that of E.qinghainensis. The two kinds of the outside areola occlusions in Entomoneis are compared, summarised and discussed.

Yellow-b, -c, -d, and -h are required for normal body coloration of Henosepilachna vigintioctopunctata.

The involvement of yellow genes y-b, y-c, y-e, and y-h in cuticle tanning has poorly been clarified. In the present paper, six putative yellow (y-y, y-b, y-c, y-e y-f, and y-h) genes were identified in Henosepilachna vigintioctopunctata. Hvy-b, Hvy-c, Hvy-e, and Hvy-h were abundantly transcribed at early larval and late pupal stages, especially in the epidermis. Accordingly, RNA interference (RNAi) experiments were performed by an injection of dsy-b, dsy-c, dsy-e, or dsy-h into the second instar larvae and 1-day-old pupae. The head capsule, scoli and strumae, and legs in the fourth-instar larvae became blacker; the blackish spots in the pupae were darkened and widened after RNAi of Hvy-b, compared with those of dsegfp-treated controls. Depletion of Hvy-b at the 1-day-old pupal stage expanded two pair of black markings on the sternum of the metathorax, and darkened the black patched on the sterna of the abdomen segments I-VI in the resultant adults. Depletion of Hvy-e caused darker pigmented adult body and elytral cuticles than those of dsegfp-introduced controls. However, there was no obvious difference in pigmentation of the black markings. Hvy-h-deficient larvae displayed dark yellow body color, whereas the body color of the dsegfp-injected control was pale yellow. There was no obvious difference in coloration of larval specific-black markings or pupal cuticle between dsHvy-h- and dsegfp-treated animals. Moreover, silence of Hvy-c at the second instar larval stage lightened black markings in the resulting larvae and pupae, but had no influence on pale yellow body color. Our results demonstrated their different roles of the four yellow genes during body pigmentation: HvY-b and HvY-c, respectively, inhibit and facilitate the coloration within dark markings, whereas HvY-e and HvY-h, respectively, repress the pigmentation in adult and larval body cuticles outside the black patches in H. vigintioctopunctata.