The Role of the ADAMTS18 Gene-Induced Immune Microenvironment in Mouse Kidney Development.

The aim of this study is to investigate the role of the ADAMTS18 gene in regulating the renal development of mice. PAS staining was used to observe the kidney development of E12.5-E17.5 mice, while immunofluorescence staining and RT-PCR were used to observe the expression of ADAMTS18. Ureteric bud (UB) branches were observed using immunofluorescence staining using the UB marker E-cadherin, and the apoptosis and proliferation of posterior renal mesenchymal cells were analyzed using TUNEL and PH3 fluorescence staining. Flow cytometry was used to analyze the immune cell infiltration, and western blotting (WB) was used to analyze the expression of PD-1/PD-L1 and CTLA-4. As a result, the ADAMTS18 gene expression gradually increased as the kidney continued to mature during embryonic development. Compared with that in the control and vector groups, UB branching was significantly reduced in the ADAMTS18 deletion group (p < 0.05), but that deletion of ADAMTS18 did not affect posterior renal mesenchymal cell proliferation or apoptosis (p > 0.05). Compared with those in the control and vector groups, the proportion of embryonic kidney B cells and the proportion of CD8+ cells were significantly greater after ADAMTS18 was knocked down (p < 0.05), but the difference in neutrophil counts was not significant (p > 0.05). The WB analysis revealed that the PD-1/PD-L1 and CTLA-4 expression was significantly increased after ADAMTS18 was knocked down (p < 0.05). In conclusion, the ADAMTS18 gene may be involved in mice kidney development by regulating the immune microenvironment and activating immune checkpoints. Deletion of the ADAMTS18 gene may be unfavorable for kidney development.

The progression of obstructive renal fibrosis in rats is regulated by ADAMTS18 gene methylation in the embryonic kidney through the AKT/Notch pathway.

This study aimed to explore the mechanism by which postembryonic renal ADAMTS18 methylation influences obstructive renal fibrosis in rats. After exposure to transforming growth factor (TGF)-ß1 during the embryonic period, analysis of postembryonic renal ADAMTS18 methylation and expression levels was conducted. Histological analysis was performed to assess embryonic kidney lesions and damage. Western blot analysis was used to determine the expression of renal fibrosis markers. Rats with ureteral obstruction and a healthy control group were selected. The methylation levels of ADAMTS18 in the different groups were analyzed. Western blot analysis and immunohistochemistry were performed to analyze the expression of renal fibrosis markers, and kidney-related indicators were measured. Treatment with TGF-ß1 resulted in abnormal development of the postembryonic kidney, which was characterized by rough kidney surfaces with mild depressions and irregularities on the outer surface. TGF-ß1 treatment significantly promoted ADAMTS18 methylation and activated the protein kinase B (AKT)/Notch pathway. Ureteral obstruction was induced to establish a renal hydronephrosis model, which led to renal fibrotic injury in newborn rats. Overexpression of the ADAMTS18 gene alleviated renal fibrosis. The western blot results showed that compared to that in the control group, the expression of renal fibrosis markers was significantly decreased after ADAMTS18 overexpression, and there was a thicker renal parenchymal tissue layer and significantly reduced p-AKT/AKT and Notch1 levels. TGF-ß1 can induce ADAMTS18 gene methylation in the postembryonic kidney, and the resulting downregulation of ADAMTS18 expression has long-term effects on kidney development, potentially leading to increased susceptibility to obstructive renal fibrosis. This mechanism may involve activation of the AKT/Notch pathway. Reversing ADAMTS18 gene methylation may reverse this process.

Curcumin inhibits lipopolysaccharide-induced proximal renal tubular epithelial cell death by regulating ADAMTS18 methylation and AKT/Notch pathways.

Introduction: This study aimed to explore the effect of curcumin on the activity of lipopolysaccharide (LPS)-induced proximal renal tubular epithelial cells (PRTECs) and to analyze the molecular mechanism by which curcumin regulates their occurrence. Material and methods: LPS-induced PRTECs were used to construct an inflammatory cell model. RT-qPCR and western blot (WB) were used to detect ADAMTS18 expression. Methylation-specific PCR was used to detect ADAMTS18 methylation levels. After curcumin treatment, MTT assay was used to analyze cell viability, flow cytometry was used to analyze apoptosis, and ADAMTS18 expression and methylation levels were detected again. After transfection with siADAMTS18, cell viability and apoptosis were analyzed again. The levels of inflammatory factors were measured by enzyme-linked immunosorbent assay, and the expression levels of AKT, Notch-1 and Notch-2 were analyzed by WB. Results: Curcumin strongly inhibited LPS-induced PRTEC inflammatory lesions, restored normal cell proliferation, and reduced the apoptosis rate by downregulating ADAMTS18 methylation and restoring ADAMTS18 expression. After siADAMTS18, the ability of curcumin to improve cell viability was reduced, and the ability of curcumin to downregulate inflammatory factors was significantly reduced. Curcumin could also inhibit the expression of AKT, Notch-1 and Notch-2 simultaneously. siADAMTS18 attenuated the abovementioned effects of curcumin. Conclusions: Curcumin inhibits LPS-induced PRTEC death by regulating ADAMTS18 methylation and AKT/Notch pathways.

Curcumin Interferes with TGF- ß 1-Induced Fibrosis in NRK-49F Cells by Reversing ADAMTS18 Gene Methylation.

OBJECTIVE: To explore the molecular mechanism by which curcumin affects renal interstitial fibrosis (RIF) progression by regulating ADAM metallopeptidase with thrombospondin type 1 motif 18 (ADAMTS18) methylation. METHODS: NRK-49F cells RIF model were induced with transforming growth factor ß 1 (TGF- ß 1). Effects of different concentrations of curcumin (0, 10, 20, and 30 µmol/L) on cell proliferation, cell cycle, cell apoptosis as well as cyclin D1 expression were analyzed by cell counting kit-8, flow cytometry and Western blot, respectively. ADAMTS18 methylation levels were determined by methylation-specific polymerase chain reaction. ADAMTS18, fibronectin (FN), type I collagen (Col- I) and alpha-smooth muscle actin (α -SMA) mRNA and protein expressions were analyzed by real-time PCR (RT-PCR) and Western blot, respectively. Meanwhile, cells were treated with 50 mmol/L 5-aza-2'-deoxycytidine (5-aza-dC, demethylation agent) for 72 h. Effect of curcumin on extracellular matrix (ECM) deposition was evaluated by immunochemical staining and Western blot. NRK-49F cells were transfected with ADAMTS18 small interfering RNA and grouped into a normal control, ADAMTS18-knock-out (KO), and ADAMTS18-KO+ 30 µmol/L curcumin groups, and whether curcumin can reverse the effect of ADAMTS18 knockdown on RIF was evaluated. RESULTS: Compared with the control group, TGF-ß 1 significantly inhibited the proliferation of NRK-49F cells, blocked the G1/G0 phase, promoted cell apoptosis and inhibited cyclin D1 expression (P<0.01). Among the different concentrations of curcumin, 30 µmol/L curcumin significantly reversed these processes (P<0.01). Immunochemical staining and Western blot results showed that curcumin significantly inhibited the deposition of FN, Col- I and α-SMA (P<0.01). Curcumin and 5-zaz-dC had synergistic effects, promoting ADAMTS18 expression, removing ADAMTS18 methylation, and reducing ECM deposition. ADAMTS18 knockdown promoted ECM accumulation, and curcumin reversed this process (P<0.01). CONCLUSION: TGF-ß 1-induced fibrosis in NRK-49F cells. Curcumin promoted ADAMTS18 expression, reduced ECM accumulation, and alleviated RIF progression by inhibiting ADAMTS18 methylation.

Evaluation of the diagnostic efficiency of voided urine fluorescence in situ hybridization for predicting the pathology of preoperative "low-risk" upper tract urothelial carcinoma.

Objectives: To evaluate the clinical utility of voided urine fluorescence in situ hybridization (FISH) for predicting the pathology of preoperative "low-risk" upper tract urothelial carcinoma (UTUC). Methods: Information of patients preoperatively diagnosed with "low-risk" UTUC receiving radical nephroureterectomy (RNU) between May 2014 and October 2019 were retrospectively collected. All of the patients accepted the FISH test and then were divided into two groups according to the results of FISH. The diagnostic value of FISH was assessed through the receiver operating characteristics (ROC) curve and area under the curve. Logistic regression analysis was applied to examine FISH as a predictive factor of tumor final stage and grade of preoperative "low-risk" UTUC. Results: In total, 129 patients were included. Of them, 70 (54.2%) were marked with positive FISH result. The difference at final pathology in tumor stage and tumor grade between these two groups of FISH (-) and FISH (+) had significantly statistical significance (p<0.001). Regarding to the tumor stage at final pathology, the sensitivity, specificity, positive predictive value and negative predictive value of FISH were 70.7 (58.9-80.3)68.5 (54.3-80.1)75.7 (63.7-84.8) and 62.7 (49.1-74.7), respectively. Regarding to the tumor grade at final pathology, the sensitivity, specificity, positive predictive value and negative predictive value of FISH were 64.7 (53.5-74.6), 65.9 (50.0-79.1), 78.6 (66.8-87.1) and 49.1 (36.5-62.3), respectively. The results of logistic regression analysis indicated that FISH could predict the pathologic characteristics of preoperative "low-risk" UTUC independently. Conclusions: FISH was qualified with relatively high diagnostic estimates for predicting tumor stage and grade of preoperative "low-risk" UTUC, and could be an independent predictive factor in clinical practice. For preoperative "low-risk" UTUC patients but with positive FISH result, choosing nephron-sparing surgery may require special caution.

Suitable stocking density of fish in paddy field contributes positively to 2-acetyl-1-pyrroline synthesis in grain and improves rice quality.

BACKGROUND: Fragrant rice is increasingly popular with the public owing to its fresh aroma, and 2-acetyl-1-pyrroline (2-AP) is the main characteristic component of the aroma in fragrant rice. Rice-fish co-culture is an environmentally friendly practice in sustainable agriculture. However, the effect of rice-fish co-culture on 2-AP in grains has received little study. A conventional fragrant rice (Meixiangzhan 2) was used, and a related field experiment during three rice growing seasons was conducted to investigate the effects of rice-fish co-culture on 2-AP, as well as the rice quality, yield, plant nutrients, and precursors and enzyme activities of 2-AP biosynthesis in leaves. This study involved three fish stocking density treatments (i.e. 9000 (D1), 15 000 (D2), and 21 000 (D3) fish fries per hectare) and rice monocropping. RESULTS: Rice-fish co-culture increased the 2-AP content in grains by 2.5-49.4% over that of the monocropping, with significant increases in the early and late rice seasons of 2020. Rice-fish co-culture treatments significantly promoted seed-setting rates by 3.39-7.65%, and improved leaf nutrients and rice quality. Notably, the D2 treatment significantly increased leaf total nitrogen (TN), total phosphorus (TP), and total potassium (TK) contents and the head rice rate at maturity stage, while significantly decreased chalkiness degree. There was no significant difference in rice yield. CONCLUSION: Rice-fish co-culture had positive effects on 2-AP synthesis, rice quality, seed-setting rates, and plant nutrient contents. The better stocking density of field fish for rice-fish co-culture in this study was 15 000 fish ha-1 . © 2023 Society of Chemical Industry.

Curcumin Inhibits Proliferation of Renal Cell Carcinoma in vitro and in vivo by Regulating miR-148/ADAMTS18 through Suppressing Autophagy.

OBJECTIVE: To explore the effect of curcumin on the proliferation of renal cell carcinoma and analyze its regulation mechanism. METHODS: In RCC cell lines of A498 and 786-O, the effects of curcumin (2.5, 5, 10 µ mo/L) on the proliferation were analyzed by Annexin V+PI staining. Besides, A498 was inoculated into nude mice to establish tumorigenic models, and the model mice were treated with different concentrations of curcumin (100, 200, and 400 mg/kg), once daily for 30 days. Then the tumor diameter was measured, the tumor cells were observed by hematoxylin-eosin staining, and the protein expressions of miR-148 and ADAMTS18 were detected by immunohistochemistry. In vitro, after transfection of miR-148 mimics, miR-148 inhibitor or si-ADAMTS18 in cell lines, the expression of ADAMTS18 was examined by Western blotting and the cell survival rate was analyzed using MTT. Subsequently, Western blot analysis was again used to examine the autophagy phenomenon by measuring the relative expression level of LC3-II/LC3-I; autophagy-associated genes, including those of Beclin-1 and ATG5, were also examined when miR-148 was silenced in both cell lines with curcumin treatment. RESULTS: Curcumin could inhibit the proliferation of RCC in cell lines and nude mice. The expression of miR-148 and ADAMTS18 was upregulated after curcumin treatment both in vitro and in vivo (P<0.05). The cell survival rate was dramatically declined upon miR-148 or ADAMTS18 upregulated. However, si-ADAMTS18 treatment or miR-148 inhibitor reversed these results, that is, both of them promoted the cell survival rate. CONCLUSION: Curcumin can inhibit the proliferation of renal cell carcinoma by regulating the miR-148/ ADAMTS18 axis through the suppression of autophagy in vitro and in vivo. There may exist a positive feedback loop between miR-148 and ADAMTS18 gene in RCC.

Trophic transfer of methylmercury and brominated flame retardants in adjacent riparian and aquatic food webs: 13C indicates biotransport of contaminants through food webs.

Biomagnification of persistent toxic substances (PTSs) in food chains is of environmental concern, but studies on biotransport of PTSs across aquatic and riparian food chains are still incomplete. In this study, biomagnification of several PTSs including methylmercury (MeHg), polybrominated diphenyl ethers (PBDEs), and 1,2-bis (2,4,6-tribromophenoxy) ethane (BTBPE) was investigated in adjacent aquatic and riparian food webs. Concentrations of MeHg and PBDEs ranged from 2.37 to 353 ng/g dry weight (dw) and not detected (Nd) to 65.1 ng/g lipid weight (lw) in riparian samples, respectively, and ranged from Nd to 705 ng/g dw and Nd to 187 ng/g lw in aquatic samples, respectively. Concentrations of MeHg were significantly correlated with δ13C (p < 0.01) rather than δ15N (p > 0.05) values in riparian organisms, while a significant correlation was observed between concentrations of MeHg and δ15N (p < 0.01) in aquatic organisms. Biomagnification factors (BMFs) and trophic magnification factors (TMFs) of PBDE congeners were similar in riparian and aquatic food webs, while BMFs and TMFs of MeHg were much higher in aquatic food web than those in riparian food web. The results indicate the biotransport of MeHg from aquatic insects to terrestrial birds, and δ13C can be a promising ecological indicator for biotransport of pollutants across ecosystems.

Acid Rain Increases Impact of Rice Blast on Crop Health via Inhibition of Resistance Enzymes.

Worldwide, rice blast (Pyricularia oryzae) causes more rice crop loss than other diseases. Acid rain has reduced crop yields globally for nearly a century. However, the effects of acid rain on rice-Pyricularia oryzae systems are still far from fully understood. In this study, we conducted a lab cultivation experiment of P. oryzae under a series of acidity conditions as well as a glasshouse cultivation experiment of rice that was inoculated with P. oryzae either before (P. + SAR) or after (SAR + P.) simulated acid rain (SAR) at pH 5.0, 4.0, 3.0 and 2.0. Our results showed that the growth and pathogenicity of P. oryzae was significantly inhibited with decreasing pH treatments in vitro culture. The SAR + P. treatment with a pH of 4.0 was associated with the highest inhibition of P. oryzae expansion. However, regardless of the inoculation time, higher-acidity rain treatments showed a decreased inhibition of P. oryzae via disease-resistance related enzymes and metabolites in rice leaves, thus increasing disease index. The combined effects of high acidity and fungal inoculation were more serious than that of either alone. This study provides novel insights into the effects of acid rain on the plant-pathogen interaction and may also serve as a guide for evaluating disease control and crop health in the context of acid rain.

Invasive Chromolaena odorata species specifically affects growth of its co-occurring weeds.

Plant-plant interaction is essential to weed invasion success and is related to impacts on the environment. To understand interactions of the well-known invasive plant siamweed (Chromolaena odorata) and its neighbors (exotic Praxelis clematidea and native cadillo) in South China, and their competitive mechanisms above- and belowground, a multicultivation experiment was conducted. Competitive indices, plant morphological traits, soil nutrient contents, enzyme activities, and microbial biomass were measured. Competitive balance index and morphological traits revealed balanced competition between P. clematidea and siamweed, and suppressive effect of siamweed on cadillo. In coculture of siamweed and P. clematidea, the branch length of siamweed slightly lengthened, while the branch number of P. clematidea increased compared with their respective monocultures accordingly. Overall impacts of the two invaders on soil properties were near averages of their single impacts. In coculture of siamweed and cadillo, siamweed was more competitive in both light and nutrient capture; soil urease activity and acid phosphatase activity were magnified and mitigated compared with the averages of those in their respective monocultures, respectively. The species-specific results of siamweed competing with its co-occurring weeds would contribute to a better understanding of mechanism in synergistic effect of siamweed with the other invasive plants.

Tobacco Cutworm (Spodoptera Litura) Larvae Silenced in the NADPH-Cytochrome P450 Reductase Gene Show Increased Susceptibility to Phoxim.

Nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome P450 reductases (CPRs) function as redox partners of cytochrome P450 monooxygenases (P450s). CPRs and P450s in insects have been found to participate in insecticide resistance. However, the CPR of the moth Spodoptera litura has not been well characterized yet. Based on previously obtained transcriptome information, a full-length CPR cDNA of S. litura (SlCPR) was PCR-cloned. The deduced amino acid sequence contains domains and residues predicted to be essential for CPR function. Phylogenetic analysis with insect CPR amino acid sequences showed that SlCPR is closely related to CPRs of Lepidoptera. Quantitative reverse transcriptase PCR (RT-qPCR) was used to determine expression levels of SlCPR in different developmental stages and tissues of S. litura. SlCPR expression was strongest at the sixth-instar larvae stage and fifth-instar larvae showed highest expression in the midgut. Expression of SlCPR in the midgut and fat body was strongly upregulated when fifth-instar larvae were exposed to phoxim at LC15 (4 µg/mL) and LC50 (20 µg/mL) doses. RNA interference (RNAi) mediated silencing of SlCPR increased larval mortality by 34.6% (LC15 dose) and 53.5% (LC50 dose). Our results provide key information on the SlCPR gene and indicate that SlCPR expression levels in S. litura larvae influence their susceptibility to phoxim and possibly other insecticides.

Mikania Micrantha Wilt Virus Alters Insect Vector's Host Preference to Enhance Its Own Spread.

As an invasive weed, Mikaniamicrantha Kunth has caused serious damage to natural forest ecosystems in South China in recent years. Mikania micrantha wilt virus (MMWV), an isolate of the Gentian mosaic virus (GeMV), is transmitted by Myzuspersicae (Sulzer) in a non-persistent manner and can effectively inhibit the growth of M. micrantha. To explore the MMWV-M. micrantha-M. persicae interaction and its impact on the invasion of M. micrantha, volatile compounds (VOCs) emitted from healthy, mock-inoculated, and MMWV-infected plants were collected, and effects on host preference of the apterous and alate aphids were assessed with Y-shaped olfactometers. Gas chromatography-mass spectrometry (GC-MS) analysis indicated that MMWV infection changed the VOC profiles, rendering plants more attractive to aphids. Clip-cages were used to document the population growth rate of M.persicae fed on healthy, mock-inoculated, or MMWV-infected plants. Compared to those reared on healthy plants, the population growth of M. persicae drastically decreased on the MMWV-infected plants. Plant host choice tests based on visual and contact cues were also conducted using alate M.persicae. Interestingly, the initial attractiveness of MMWV-infected plants diminished, and more alate M. persicae moved to healthy plants. Taken together, MMWV appeared to be able to manipulate its plant host to first attract insect vectors to infected plants but then repel viruliferous vectors to promote its own dispersal. Its potential application for invasive weed management is discussed.

Complete mitochondrial genome of Rivularia auriculata (Gastropoda, Viviparidae) with phylogenetic consideration.

Complete mitochondrial genome sequence of Rivularia auriculata has a circular genome of 16,552 bp, which is comprised 13 protein-coding genes, 2 rRNA genes, and 22 tRNA genes. The nucleotide composition of the light strand is 43.16% of A, 26.78% of T, 20.18% of C, and 9.88% of G. All genes are encoded on the heavy strand except seven tRNA genes (Met, Tyr, Cys, Trp, Gln, Gly, and Glu) on the light strand. All the protein-coding genes start with ATC initiation codon except ND4 starts with GTG, and two types of inferred termination codons are TAA and TAG. There are 26 intergenic spacers and 4 gene overlaps. It is indicated that R. auriculata has closer genetic relationship with Viviparus chui (88.64% nucleotide sequence identity between them) than the other snail species.

Comparative Characterization of the Complete Mitochondrial Genomes of the Three Apple Snails (Gastropoda: Ampullariidae) and the Phylogenetic Analyses.

The apple snails Pomacea canaliculata, Pomacea diffusa and Pomacea maculate (Gastropoda: Caenogastropoda: Ampullariidae) are invasive pests causing massive economic losses and ecological damage. We sequenced and characterized the complete mitochondrial genomes of these snails to conduct phylogenetic analyses based on comparisons with the mitochondrial protein coding sequences of 47 Caenogastropoda species. The gene arrangements, distribution and content were canonically identical and consistent with typical Mollusca except for the tRNA-Gln absent in P. diffusa. An identifiable control region (d-loop) was absent. Bayesian phylogenetic analysis indicated that all the Ampullariidae species clustered on the same branch. The genus Pomacea clustered together and then with the genus Marisa. The orders Architaenioglossa and Sorbeoconcha clustered together and then with the order Hypsogastropoda. Furthermore, the intergenic and interspecific taxonomic positions were defined. Unexpectedly, Ceraesignum maximum, Dendropoma gregarium, Eualetes tulipa and Thylacodes squamigerus, traditionally classified in order Hypsogastropoda, were isolated from the order Hypsogastropoda in the most external branch of the Bayesian inference tree. The divergence times of the Caenogastropoda indicated that their evolutionary process covered four geological epochs that included the Quaternary, Neogene, Paleogene and Cretaceous periods. This study will facilitate further investigation of species identification to aid in the implementation of effective management and control strategies of these invasive species.

Soil properties and carbon and nitrogen pools in a young hillside longan orchard after the introduction of leguminous plants and residues.

The intensification of young hillside Dimocarpus longan orchard cultivation has led to increase soil erosion and decrease soil fertility in South China. Leguminous crops are often used for improving soil properties. An approximately 2-year-long field experiment in lateritic soil in South China was conducted to evaluate the effects of legume introductions on soil properties and carbon (C) and nitrogen (N) pools. Two leguminous and one non-leguminous plant species, including Arachis hypogaea L. (a leguminous oilseed crop species, DA), Stylosanthes guianensis (a perennial herbaceous leguminous species, DS) and Lolium perenne L. (an annual non-leguminous forage species, DL), were introduced into a D. longan orchard as three treatments and compared to the monoculture of D. longan (the control, D0). And the harvested biomass residues of the three cover plants were returned to their corresponding plots as green manure. Soil samples were collected from depths of 0-10 and 10-20 cm approximately 2 years after treatment application. The results showed that, compared with D0, DA significantly improved the contents of soil available phosphorus, dissolved organic carbon (DOC), total nitrogen, ammonium and the N pool. In addition, DS significantly increased the contents of DOC, microbial biomass carbon and ammonium in the soil. However, DL did not affect any soil properties or the C and N pools. In addition, neither DA nor DS altered the soil bulk density or the contents of available nitrogen, total organic carbon and the C pool. The improvement of soil properties by DS and DA was positively correlated with the plant residues amount, plant N content but negatively correlated with the plant C:N ratios. Besides, the plant growth of longan was significantly improved by DA. In conclusion, compared with that of S. guianensis, the introduction of A. hypogaea L. was more helpful for restoring and improving soil properties, N pool and longan growth within the young hillside orchard in South China.

Characterization of the Complete Mitochondrial Genome Sequences of Three Croakers (Perciformes, Sciaenidae) and Novel Insights into the Phylogenetics.

The three croakers (Nibea coibor, Protonibea diacanthus and Argyrosomus amoyensis, Perciformes, Sciaenidae) are important commercial species inhabiting the Eastern Indian Ocean and Western Pacific. Molecular data employed in previous research on phylogenetic reconstruction have not been adequate and complete, and systematic and comprehensive phylogenetic relationships for these fish are unresolved. We sequenced the complete mitochondrial genomes of the three croakers using next-generation sequencing for the first time. We analyzed the composition and phylogenies between 19 species in the family Sciaenidae using the mitochondrial protein coding sequences of 204 species in the Series Eupercaria. We present the characterization of the complete mitochondrial genome sequences of the three croakers. Gene arrangement and distribution of the three croakers are canonically identical and consistent with other vertebrates. We found that the family Sciaenidae is an independent branch that is isolated from the order Perciformes and does not belong to any extant classification. Therefore, this family is expected to belong to a new classification at the order level and needs further analysis. The evolution of Sciaenidae has lagged far behind the Perciformes differentiation. This study presents a novel insight into the phylogenetics of the family Sciaenidae from the order Perciformes and facilitates additional studies on the evolution and phylogeny of Series Eupercaria.

Identification and Functional Analysis of a Novel Cytochrome P450 Gene CYP9A105 Associated with Pyrethroid Detoxification in Spodoptera exigua Hübner.

In insects, cytochrome P450 monooxygenases (P450s or CYPs) are known to be involved in the detoxification and metabolism of insecticides, leading to increased resistance in insect populations. Spodoptera exigua is a serious polyphagous insect pest worldwide and has developed resistance to various insecticides. In this study, a novel CYP3 clan P450 gene CYP9A105 was identified and characterized from S. exigua. The cDNAs of CYP9A105 encoded 530 amino acid proteins, respectively. Quantitative real-time PCR analyses showed that CYP9A105 was expressed at all developmental stages, with maximal expression observed in fifth instar stage larvae, and in dissected fifth instar larvae the highest transcript levels were found in midguts and fat bodies. The expression of CYP9A105 in midguts was upregulated by treatments with the insecticides α-cypermethrin, deltamethrin and fenvalerate at both LC15 concentrations (0.10, 0.20 and 5.0 mg/L, respectively) and LC50 concentrations (0.25, 0.40 and 10.00 mg/L, respectively). RNA interference (RNAi) mediated silencing of CYP9A105 led to increased mortalities of insecticide-treated 4th instar S. exigua larvae. Our results suggest that CYP9A105 might play an important role in α-cypermethrin, deltamethrin and fenvalerate detoxification in S. exigua.

The complete mitochondrial genome of the apple snail Pomacea maculate (Gastropoda: Ampullariidae).

We present the complete mitochondrial genome of Pomacea maculate in this study. The mitochondrial genome is 15,512 bp in length, containing 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes. Overall nucleotide compositions of the light strand are 41.13% of A, 30.81% of T, 15.25% of C and 12.81% of G. Its gene arrangement and distribution are different from the typical vertebrates. The absence of D-loop is consistent with the Gastropoda, but at least one lengthy non-coding region is essential regulatory element for the initiation of transcription and replication. Phylogenetic tree is constructed by the maximum-likelihood method based on the complete mitochondrial genomes of 15 species of Caenogastropoda, using Helix aspersa as outgroup to assess their actual phylogenetic relationship and evolution. The result provides fundamental data for resolving phylogenetic and genetic problems related to effective management strategies.

Predation risk affects growth and reproduction of an invasive snail and its lethal effect depends on prey size.

The behavior of invasive species under predation risk has been studied extensively, but their growth and reproductive responses have rarely been investigated. We conducted experiments with juveniles and adults of the invasive freshwater snail Pomacea canaliculata, and we observed changes in growth and reproduction in response to predation risk from a caged predator (Trachemys scripta elegans). P. canaliculata produced eggs earlier in the presence of predators and injured conspecifics compared with the control group (no risk), although the total number of egg masses laid by per female was exceeded by that of the controls after 15 days. Egg hatching success noticeably decreased under predation risk, and the incubation period was significantly prolonged; however, the oviposition height of the snails was not affected. A lethal effect of predation risk was detected in juvenile snails but not in adults. The growth of juvenile P. canaliculata was inhibited under predation risk, probably due to a reduction in food intake. Adult females exhibited a greater reduction in growth under predation risk than males, which likely resulted in part from the high reproductive investment of females in egg laying. These results indicate that P. canaliculata snails under predation risk face a trade-off between predator avoidance and growth and reproduction, where the lethal effect of predation risk is linked to the size of the prey.

Identification of Two Cytochrome Monooxygenase P450 Genes, CYP321A7 and CYP321A9, from the Tobacco Cutworm Moth (Spodoptera Litura) and Their Expression in Response to Plant Allelochemicals.

Larvae of the polyphagous tobacco cutworm moth, Spodoptera litura (S. litura), encounter potentially toxic allelochemicals in food. It is therefore important for S. litura to produce detoxification enzymes such as cytochrome P450 monooxygenases (P450s). In this study, we have identified two novel cytochrome P450 genes of S. litura, named CYP321A7 and CYP321A9. Phylogenetic analysis indicated that they belong to the CYP321A subfamily. Expression levels of these genes at different development stages were determined by real-time quantitative polymerase chain reaction (PCR). The highest expression was found in the midgut and the fat body. Larvae fed with a diet supplemented with xanthotoxin or coumarin showed a strongly increased expression of CYP321A7 and CYP321A9 in the midgut and fat body as compared to larvae that consumed a control diet. In contrast, larvae consuming a diet containing aflatoxin B1 or quercetin did not induce the expression of these genes. CYP321A7 and CYP321A9 showed different expression profiles with respect to certain allelochemicals. For example, a diet containing cinnamic acid stimulated the expression of CYP321A9, whereas no changes were observed for CYP321A7. We suggest that the fine tuning of P450 gene expression is an important adaptation mechanism that allows polyphagous S. litura larvae to survive in a changing chemical environment.