RESUMEN
RNA viruses are extremely diverse and rapidly evolving in various organisms. Our knowledge on viral evolution with interacted hosts in the manner of ecology is still limited. In the agricultural ecosystem, invasive insect species are posing a great threat to sustainable crop production. Among them, fruit flies (Diptera: Tephritidae Bactrocera and Zeugodacus) are destructive to fruits and vegetables, which are also closely related and often share similar ecological niches. Thus, they are ideal models for investigating RNA virome dynamics in host species. Using meta-transcriptomics, we found 39 viral sequences in samples from 12 fly species. These viral species represented the diversity of the viromes including Dicistroviridae, negev-like virus clades, Thika virus clades, Solemoviridae, Narnaviridae, Nodaviridae, Iflaviridae, Orthomyxoviridae, Bunyavirales, Partitiviridae, and Reoviridae. In particular, dicistrovirus, negev-like virus, orthomyxovirus, and orbivirus were common in over four of the fly species, which suggests a positive interaction between fly viromes that exist under the same ecological conditions. For most of the viruses, the virus-derived small RNAs displayed significantly high peaks in 21 nt and were symmetrically distributed throughout the viral genome. These results suggest that infection by these viruses can activate the host's RNAi immunity. Our study provides RNA virome diversity and evidence on their infection activity in ecologically associated invasive fruit fly species, which could help our understanding of interactions between complex species and viruses.
Asunto(s)
Virus ARN , Tephritidae , Animales , Ecosistema , Kenia , Virus ARN/genética , TranscriptomaRESUMEN
Ecdysteroids regulate molting in arthropods by binding to heterodimers of the ecdysone receptor and retinoid-X-receptor, homologous to the ultraspiracle protein, to induce the expression of downstream signal response genes including the nuclear receptor HR3. However, the detailed expression dynamics of HR3 during molting in spider mites are not yet clear. In this study, the full length of PcHR3 was retrieved based on the genome of citrus red mite, Panonychus citri. The open reading frame is 1707 bp encoding 568 amino acids, which contains a DNA binding domain and a ligand binding domain. Then, the expression pattern of PcHR3 was analyzed throughout the development of the deutonymph by RT-qPCR. The result showed that PcHR3 was mainly transcribed in the late deutonymph stage, when the deutonymph was at least 24 h old and motionless, the critical point at which the mites started molting. Transcription reached the highest level in 32-h-old deutonymphs and decreased by 36 h, where the mites remained in a quiescent state. Further silencing of PcHR3 by leaf-disc-based delivery of dsRNA to 8-h-old deutonymph mites, resulted in retarded development and death of 58% of deutonymphs. In summary, we suggest that PcHR3 regulates the latter stages of molting in P. citri.
Asunto(s)
Muda , Interferencia de ARN , Receptores Citoplasmáticos y Nucleares/fisiología , Tetranychidae , Animales , Proteínas de Artrópodos/antagonistas & inhibidores , Proteínas de Artrópodos/fisiología , Ecdisteroides , Receptores Citoplasmáticos y Nucleares/antagonistas & inhibidoresRESUMEN
In insects, the ecdysteroid 20-hydroxyecdysone coordinates with juvenile hormone (JH) to regulate the process of molting, development and metamorphosis; however, this interaction is still unclear in the mites. In this study, we investigated the gene related to ecdysteroid and JH biosynthesis pathways, including four ecdysteroid and 11 JH biosynthesis genes. We examined their expression patterns during molting of different developmental stages of the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), an important agricultural pest that feeds on more than 1100 plant species. The expression of ecdysteroid biosynthesis Halloween genes exhibited a positive zigzag-like pattern, with a peak after 8 h of molting and a drop 8 h after entering each quiescent stage. In contrast, JH biosynthesis genes expression displayed a negative zigzag-like pattern, with a peak at 8 h after entering each quiescent stage and a drop after 8 h of each molting. These opposite patterns imply that ecdysteroid and JH expression is coordinated during the developmental transition. Our data provide an initial perspective on the co-expression of ecdysteroid and JH biosynthesis genes to regulate this important developmental process in the two-spotted spider mite.
Asunto(s)
Proteínas de Artrópodos/genética , Ecdisteroides/biosíntesis , Expresión Génica , Hormonas Juveniles/biosíntesis , Muda/genética , Tetranychidae/genética , Animales , Proteínas de Artrópodos/metabolismo , Ecdisteroides/genética , Hormonas Juveniles/genética , Larva/genética , Larva/crecimiento & desarrollo , Ninfa/genética , Ninfa/crecimiento & desarrollo , Óvulo/crecimiento & desarrollo , Tetranychidae/crecimiento & desarrolloRESUMEN
Insulin receptors play key roles in growth, development, and polymorphism in insects. Here, we report two insulin receptor genes (AcInR1 and AcInR2) from the brown citrus aphid, Aphis (Toxoptera) citricidus. Transcriptional analyses showed that AcInR1 increased during the nymph-adult transition in alate aphids, while AcInR2 had the highest expression level in second instar nymphs. AcInR1 is important in aphid development from fourth instar nymphs to adults as verified by dsRNA feeding mediated RNAi. The silencing of AcInR1 or/and AcInR2 produced a variety of phenotypes including adults with normal wings, malformed wings, under-developed wings, and aphids failing to develop beyond the nymphal stages. Silencing of AcInR1 or AcInR2 alone, and co-silencing of both genes, resulted in 73% or 60%, and 87% of aphids with problems in the transition from nymph to normal adult. The co-silencing of AcInR1 and AcInR2 resulted in 62% dead nymphs, but no mortality occurred by silencing of AcInR1 or AcInR2 alone. Phenotypes of adults in the dsInR1 and dsInR2 were similar. The results demonstrate that AcInR1 and AcInR2 are essential for successful nymph-adult transition in alate aphids and show that RNAi methods may be useful for the management of this pest.
Asunto(s)
Áfidos/crecimiento & desarrollo , Áfidos/genética , Silenciador del Gen , Genes de Insecto , Estadios del Ciclo de Vida , Ninfa/genética , Receptor de Insulina/genética , Animales , Áfidos/clasificación , Fenotipo , Filogenia , Interferencia de ARNRESUMEN
The citrus red mite, Panonychus citri, is one of the most economically and globally destructive mite pests of citrus. Acaricide resistance has been a growing problem in controlling this pest. As the main inhibitory neurotransmitter in organisms, γ-aminobutyric acid (GABA) is synthesized from the amino acid glutamate by the action of glutamate decarboxylases (GADs). In the present study, one novel GAD gene, PcGAD, was identified and characterized from P. citri. The opening reading frame of PcGAD contained 1548 nucleotides that encode 515 amino acids. The subsequent spatiotemporal expression pattern by RT-qPCR revealed that the expression levels of PcGAD were significantly higher in larvae than in adults. Challenging with various concentrations of abamectin resulted in the upregulation of PcGAD transcript levels. Furthermore, biochemical characterization indicated that changes in GAD activity coincided with its mRNA levels. High-performance liquid chromatography confirmed that the GABA contents of P. citri increased upon abamectin treatment. The application of abamectin induces PcGAD expression and activates GAD activity, thereby resulting in an increase in GABA content in P. citri, which contributes to the adaptability of the mite to abamectin challenge.
Asunto(s)
Glutamato Descarboxilasa/metabolismo , Ivermectina/análogos & derivados , Tetranychidae , Ácido gamma-Aminobutírico/metabolismo , Animales , Glutamato Descarboxilasa/efectos de los fármacos , Ivermectina/farmacologíaRESUMEN
To better characterize the resistance development and therefore establish effective pest management strategies, this study was undertaken to investigate the inheritance mode and biochemical mechanisms of malathion resistance in the oriental fruit fly, Bactrocera dorsalis (Hendel), which is one of the most notorious pests in the world. After 22 generations of selection with malathion, the malathion-resistant (MR) strain of B. dorsalis developed a 34-fold resistance compared with a laboratory susceptible strain [malathion-susceptible (MS)]. Bioassay results showed that there was no significant difference between the LD50 values of malathion against the progenies from both reciprocal crosses (F(1)-SR and F(1)-RS). The degree of dominance values (D) was calculated as 0.39 and 0.32 for F(1)-RS and F(1)-SR, respectively. The logarithm dosage-probit mortality lines of the F(2) generation and progeny from the backcross showed no clear plateaus of mortality across a range of doses. In addition, Chi-square analysis revealed significant differences between the mortality data and the theoretical expectations. The realized heritability (h(2)) value was 0.16 in the laboratory-selected resistant strain of B. dorsalis. Enzymatic activities identified significant changes of carboxylesterases, cytochrome P450 (general oxidases), and glutathione S-transferases in MR compared with the MS strain of B. dorsalis. Taken together, this study revealed for the first time that malathion resistance in B. dorsalis follows an autosomal, incompletely dominant, and polygenic mode of inheritance and is closely associated with significantly elevated activities of three major detoxification enzymes.
Asunto(s)
Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Herencia , Insecticidas/farmacología , Malatión/farmacología , Tephritidae/genética , Animales , Femenino , Ligamiento Genético , Inactivación Metabólica , Resistencia a los Insecticidas , Masculino , Tephritidae/efectos de los fármacosRESUMEN
Chitinases are hydrolytic enzymes that are required for chitin degradation and reconstruction in arthropods. In this study, we report a cDNA sequence encoding a putative chitinase (PcCht1) from the citrus red mite, Panonychus citri. The PcCht1 (564 aa) possessed a signal peptide, a conserver domain, and a chitin-binding domain. Structural and phylogenetic analyses found that PcCht1 had high sequence similarity to chitinases in Tetranychus urticae. Real-time quantitative PCR analyses showed that the transcript levels of PcCht1 peaked periodically in larval and nymph stages. Moreover, significant increase of PcCht1 transcript level in the larvae was observed upon the exposure of diflubenzuron. In contrast, exposures of the larvae to diflubenzuron resulted in the decreased chitin content. Furthermore, through a feeding-based RNA interference approach, we were able to reduce the PcCht1 transcript level by 59.7 % in the larvae, and consequently the treated larvae showed a very low molting rate compared with the control. Our results expanded the understanding of the important role of PcCht1 in the growth and development of P. citri.
Asunto(s)
Proteínas de Artrópodos/genética , Quitinasas/genética , Metamorfosis Biológica , Interferencia de ARN , Tetranychidae/crecimiento & desarrollo , Tetranychidae/genética , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/metabolismo , Secuencia de Bases , Quitinasas/metabolismo , Clonación Molecular , ADN Complementario/genética , Larva/genética , Larva/crecimiento & desarrollo , Larva/metabolismo , Ninfa/genética , Ninfa/crecimiento & desarrollo , Ninfa/metabolismo , Filogenia , ARN Mensajero/genética , Tetranychidae/enzimologíaRESUMEN
The vitellogenin receptor (VgR) functions as an essential component in uptaking and transporting vitellogenin (Vg) in female adults, which is involved in ovary development and oviposition. This study aimed to clarify the molecular characteristics and function of VgR in the oriental fruit fly Bactrocera dorsalis (Hendel). Here, we identified the full-length of BdVgR (GenBank Accession No. JX469118), encoding a 1925 residue (aa) protein with a 214.72 kDa molecular mass and several typical motifs of low-density lipoprotein receptor superfamily (LDLR). Phylogenic analysis suggested that BdVgR was evolutionary conserved with other Dipteran VgRs. The expression of BdVgR was exclusively detected in the ovaries rather than head, thorax or other tissues. The developmental expression patterns showed that the signal of BdVgR was detectable in very beginning of adult stage, and positively correlated with the growth rate of ovaries and the expression levels of its ligands. In addition, we also demonstrated that the expression level of BdVgR, and ovary development were significantly suppressed after being injected with BdVgR-targeted dsRNA. Together, all of these results indicated that BdVgR was critical for yolk protein absorption and ovary maturation in B. dorsalis, playing a vital role in female reproduction.
Asunto(s)
Proteínas del Huevo/metabolismo , Proteínas de Insectos/metabolismo , Receptores de Superficie Celular/metabolismo , Tephritidae/crecimiento & desarrollo , Vitelogeninas/metabolismo , Animales , Proteínas del Huevo/genética , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Insectos/genética , Ovario/crecimiento & desarrollo , Ovario/metabolismo , Filogenia , Interferencia de ARN , ARN Interferente Pequeño/genética , Receptores de Superficie Celular/genética , Tephritidae/genética , Tephritidae/metabolismoRESUMEN
The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg) is the precursor of vitellin (Vn), which is the major egg storage protein, and vitellogenin receptor (VgR) is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor). The PcVg1 cDNA is 5748 nucleotides (nt) with a 5553-nt open reading frame (ORF) coding for 1851 amino acids (aa), and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF)-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control.
Asunto(s)
Proteínas del Huevo , Receptores de Superficie Celular , Tetranychidae/genética , Tetranychidae/metabolismo , Vitelogeninas , Secuencias de Aminoácidos , Animales , Clonación Molecular , ADN Complementario/química , ADN Complementario/metabolismo , Proteínas del Huevo/genética , Proteínas del Huevo/metabolismo , Regulación del Desarrollo de la Expresión Génica , Larva/genética , Filogenia , Estructura Terciaria de Proteína , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Tetranychidae/clasificación , Tetranychidae/crecimiento & desarrollo , Vitelogeninas/genética , Vitelogeninas/metabolismoRESUMEN
Chitin synthase synthesizes chitin, which is critical for the arthropod exoskeleton. In this study, we cloned the cDNA sequences of a chitin synthase 1 gene, PcCHS1, in the citrus red mite, Panonychus citri (McGregor), which is one of the most economically important pests of citrus worldwide. The full-length cDNA of PcCHS1 contains an open reading frame of 4605 bp of nucleotides, which encodes a protein of 1535 amino acid residues with a predicted molecular mass of 175.0 kDa. A phylogenetic analysis showed that PcCHS1 was most closely related to CHS1 from Tetranychus urticae. During P. citri development, PcCHS1 was constantly expressed in all stages but highly expressed in the egg stage (114.8-fold higher than in the adult). When larvae were exposed to diflubenzuron (DFB) for 6 h, the mite had a significantly high mortality rate, and the mRNA expression levels of PcCHS1 were significantly enhanced. These results indicate a promising use of DFB to control P. citri, by possibly acting as an inhibitor in chitin synthesis as indicated by the up-regulation of PcCHS1 after exposure to DFB.
Asunto(s)
Proteínas de Artrópodos/genética , Quitina Sintasa/genética , Diflubenzurón/farmacología , Ácaros/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/clasificación , Secuencia de Bases , Quitina Sintasa/clasificación , Citrus/parasitología , Clonación Molecular , ADN Complementario/química , ADN Complementario/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Larva/efectos de los fármacos , Larva/genética , Larva/fisiología , Ácaros/genética , Ácaros/fisiología , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia de AminoácidoRESUMEN
The citrus red mite, Panonychus citri, is known for its ability to rapidly evolve resistance to insecticides/acaricides and to adapt to hosts that produce toxins. To get better insight into the detoxification mechanism of P. citri, two carboxylesterase (CarE) genes, PCE1 and PCE2, were isolated and characterized. PCE1 and PCE2 contained open reading frames of 1,653 and 1,392 nucleotides, encoding proteins of 550 and 463 amino acid residues, respectively. Phylogenetic analyses showed that PCE1 and PCE2 were most closely related to the CarE genes from other phytophagous mites. The transcriptional profiles of two CarE genes among developmental stages (egg, larva, nymph, adult female, and adult male), after exposing to four acaricides (avermectin, azocyclotin, pyridaben, and spirodiclofen) and acid rain were investigated using real-time quantitative PCR (qPCR). The results showed that during development, PCE1 was highly expressed at the egg stage, whereas PCE2 was abundantly expressed at the adult stage of males. The expression levels of PCE1 were highly induced upon exposure to acaricides and acid rain. On the other hand, the expression levels of PCE2 were increased after treatment with avermectin and pyridaben. These results suggest that PCE1 and PCE2 may have distinct roles in different developmental stages and participate in the detoxification of acaricides.
Asunto(s)
Carboxilesterasa/genética , Tetranychidae/genética , Acaricidas , Lluvia Ácida , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/química , Femenino , Expresión Génica , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Estrés Fisiológico , Tetranychidae/enzimología , Tetranychidae/crecimiento & desarrolloRESUMEN
The citrus red mite, Panonychus citri (McGregor), is a global citrus pest, and has developed severe resistance to several types of acaricides. However, the molecular mechanisms of resistance in this mite remain unknown. In this study, seven full-length cDNAs encoding glutathione S-transferases (GSTs) genes were identified and characterized in P. citri. The effects of pyridaben and fenpropathrin exposure on the expression of these genes were also investigated. Phylogenetic analysis revealed that the seven GSTs genes in P. citri cloned in this study belong to three different cytosolic classes, including four in mu, two in delta and one in zeta. Among these seven GSTs genes, the relative expression level of PcGSTm1 was significantly higher in adult than in the other life stages (egg, larvae and nymph). Compared with the control, the mRNA levels of the seven GST genes did not change significantly following exposure to pyridaben at LC10. However, RT-qPCR results showed that, when exposed to LC10 of fenpropathrin, six GSTs gene (PcGSTm1, PcGSTm3, PcGSTm4, PcGSTd1, PcGSTd2 and PcGSTz1) transcripts increased in a time-dependent manner. This is the first insight into the molecular characteristics of GSTs gene cDNAs in P. citri. The elevated GSTs gene transcripts following exposure to fenpropathrin might be one of the mechanisms involved in detoxification of this acaricide.
Asunto(s)
Glutatión Transferasa/genética , Ácaros/enzimología , Ácaros/genética , Acaricidas/metabolismo , Acaricidas/toxicidad , Animales , Secuencia de Bases , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Glutatión Transferasa/metabolismo , Ácaros/clasificación , Ácaros/crecimiento & desarrollo , Datos de Secuencia Molecular , Filogenia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Piretrinas/metabolismo , Piretrinas/toxicidad , Piridazinas/metabolismo , Piridazinas/toxicidad , Homología de Secuencia de AminoácidoRESUMEN
Quantitative real time reverse transcriptase polymerase chain reaction (RT-qPCR) is preferred for gene expression analysis in living organisms. Currently, it is a valuable tool for biological and ecological studies as it provides a relatively straightforward way to assess the relevance of transcriptional regulation under developmental and stress tolerance conditions. However, studies have shown that some commonly used reference genes varied among different experimental treatments, thus, systematic evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of arthropods. The aim of this study is to identify the suitable reference genes for RT-qPCR experiments involving various developmental stages and/or under abiotic stresses in citrus red mite Panonychus citri, a key pest in citrus orchards worldwide. GeNorm, NormFinder, and Bestkeeper software analysis indicates that elongation factor-1 alpha (ELF1A), RNA polymerase II largest subunit, alpha tublin, and glyceraldhyde-3-phosphate dehydrogenase (GAPDH) are the most stable reference genes in various developmental stages, meanwhile, ELF1A and GAPDH were the most stable reference genes under various abiotic stresses. Furthermore, this study will serve as a resource to screen reference genes for gene expression studies in any other spider mite species.
Asunto(s)
Genes/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Estrés Fisiológico/genética , Tetranychidae/crecimiento & desarrollo , Tetranychidae/genética , Animales , Cartilla de ADN/metabolismo , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Estudios de Asociación Genética , ARN/genética , ARN/normas , Estándares de Referencia , Programas InformáticosRESUMEN
Glutathione S-transferases (GSTs) play central roles in phase II detoxification of both xenobiotics (drugs, insecticides, and herbicides) and endogenous compounds in almost all living organisms. In this study, we successfully purified the GSTs from the citrus red mite, Panonychus citri, by affinity chromatography on Glutathione Sepharose 4B and compared the biochemical characterizations of the purified GSTs from three field populations [beibei (BB), wanzhou (WZ), and zhongxian (ZX)]. SDS-PAGE revealed that the molecular weight of GSTs from three populations consisted of two subunits of 27.3 and 26.1 kDa. The specific activity of the purified GSTs from the WZ and ZX populations was increased 1.5- and 3.8-fold, respectively, compared with the BB population. Accordingly, the pyridaben susceptibility of WZ and ZX populations was less compared with BB population. Kinetic analyses showed that the WZ and ZX populations had higher substrate specificity compared with the BB population based on the values of k (cat) and k (cat) /K (m) to both reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). The in vitro inhibition studies of GSTs indicated that the I (50) values of pyridaben from WZ and ZX populations of P. citri expressed 1.6- and 4.4-fold decreases, respectively, compared to the I (50) value of pyridaben from the BB population. In conclusion, all evidence suggested that the purified GSTs may partially contribute to the susceptibility of acaricide pyridaben in field populations of P. citri.
Asunto(s)
Glutatión Transferasa/metabolismo , Tetranychidae/enzimología , Animales , Electroforesis , Estabilidad de Enzimas , Femenino , Glutatión Transferasa/antagonistas & inhibidores , Glutatión Transferasa/aislamiento & purificación , Concentración de Iones de Hidrógeno , TemperaturaRESUMEN
Carotenoids are involved in many essential physiological functions and are produced from geranylgeranyl pyrophosphate through synthase, desaturase, and cyclase activities. In the pea aphid (Acyrthosiphon pisum), the duplication of carotenoid biosynthetic genes, including carotenoid synthases/cyclases (ApCscA-C) and desaturases (ApCdeA-D), through horizontal gene transfer from fungi has been detected, and ApCdeB has known dehydrogenation functions. However, whether other genes contribute to aphid carotenoid biosynthesis, and its specific regulatory pathway, remains unclear. In the current study, functional analyses of seven genes were performed using heterologous complementation and RNA interference assays. The bifunctional enzymes ApCscA-C were responsible for the synthase of phytoene, and ApCscC may also have a cyclase activity. ApCdeA, ApCdeC, and ApCdeD had diverse dehydrogenation functions. ApCdeA catalyzed the enzymatic conversion of phytoene to neurosporene (three-step product), ApCdeC catalyzed the enzymatic conversion of phytoene to ζ-carotene (two-step product), and ApCdeD catalyzed the enzymatic conversion of phytoene to lycopene (four-step product). Silencing of ApCscs reduced the expression levels of ApCdes, and silencing these carotenoid biosynthetic genes reduced the α-, ß-, and γ-carotene levels, as well as the total carotenoid level. The results suggest that these genes were activated and led to carotenoid biosynthesis in the pea aphid.
Asunto(s)
Áfidos , Vías Biosintéticas , Animales , Áfidos/genética , Vías Biosintéticas/genética , Carotenoides , Pisum sativum , Interferencia de ARNRESUMEN
Spider mites have one ecdysone receptor (EcR) and multiple retinoid X receptors (RXRs). However, the function of these RXRs in spider mite development is unknown. Here, we screened the expression dynamics of two PcRXR isoforms at 4 h intervals in the deutonymphal stage of Panonychus citri. The results showed that PcEcR had an expression pattern similar to that of PcRXR2. For PcRXR1, its expression remained at a certain high level, when there was a decrease of both PcEcR and PcRXR2. In situ hybridization showed that PcRXR2 was detected in the central nervous mass, while the ecdysteroid biosynthesis gene PcSpo was mainly expressed at the edge of the central nervous mass. RNAi-based silencing of PcRXR1 or PcRXR2 showed the same phenotype as in mites with that of silencing PcEcR. Furthermore, RNA-seq was used to mine the genes associated with the expression dynamics of PcRXR1 or PcRXR2, which revealed that the heterodimer of EcR-RXR2 in spider mites might be linked with the cell autophagy and tissue remodeling during apolysis, and RXR1 might be linked with new epicuticle and exocuticle secretion during ecdysis. Taken together, these results increase our understanding of the regulation mechanism of ecdysteroid signal pathway in spider mite development.
Asunto(s)
Ácaros , Tetranychidae , Animales , Ecdisteroides , Muda/genética , Interferencia de ARN , Tetranychidae/genéticaRESUMEN
Pesticide resistance and resurgence are serious problems often occurring simultaneously in the field. In our long-term study of a fenpropathrin-resistant strain of Tetranychus cinnabaribus, enhancement of detoxification and modified fecundity mechanisms were both observed. Here we investigate the network across these two mechanisms and find a key node between resistance and resurgence. We show that the ecdysone pathway is involved in regulating the fecundity of T. cinnabaribus. The concentration change of ecdysone is consistent with the fecundity curve; the concentration of ecdysone is higher in the fenpropathrin-resistant strain which has stronger fecundity. The enhancement of ecdysone is due to overexpression of two P450 genes (CYP314A1 and CYP315A1) in the ecdysone synthesis pathway. Silencing expression of these CYP genes resulted in lower concentration of ecdysone, reduced expression of vitellogenin, and reduced fecundity of T. cinnabaribus. The expression of CYP315A1 is regulated by transcription factors Cap-n-collar isoform C (CncC) and Musculoaponeurotic fibrosarcoma protein (Maf), which are involved in regulating other P450 genes functioning in detoxification of fenpropathrin in T. cinnabaribus. A similar regulation is established in citrus pest mite Panonychus citri showing that the CncC pathway regulates expression of PcCYP315A1, which affects mite fecundity. Transcription factors are activated to upregulate detoxification genes facilitating pesticide resistance, while the "one to multiple" regulation mode of transcription factors simultaneously increases expression of metabolic enzyme genes in hormone pathways and alters the physiology of pests. This is an important response of arthropods to pesticides which leads to resistance and population resurgence.
Asunto(s)
Fibrosarcoma , Ácaros , Plaguicidas , Tetranychidae , Animales , Ecdisona , Ácaros/genética , Tetranychidae/genética , Factores de Transcripción/genéticaRESUMEN
Tomato yellow leaf curl virus (TYLCV) and Tomato chlorosis virus (ToCV) are two of the most devastating cultivated tomato viruses, causing significant crop losses worldwide. As the vector of both TYLCV and ToCV, the whitefly Bemisia tabaci Mediterranean (MED) is mainly responsible for the rapid spread and mixed infection of TYLCV and ToCV in China. However, little is known concerning B. tabaci MED's molecular response to TYLCV and ToCV infection or their co-infection. We determined the transcriptional responses of the whitefly MED to TYLCV infection, ToCV infection, and TYLCV&ToCV co-infection using Illumina sequencing. In all, 78, 221, and 60 differentially expressed genes (DEGs) were identified in TYLCV-infected, ToCV-infected, and TYLCV&ToCV co-infected whiteflies, respectively, compared with non-viruliferous whiteflies. Differentially regulated genes were sorted according to their roles in detoxification, stress response, immune response, transport, primary metabolism, cell function, and total fitness in whiteflies after feeding on virus-infected tomato plants. Alterations in the transcription profiles of genes involved in transport and energy metabolism occurred between TYLCV&ToCV co-infection and single infection with TYLCV or ToCV; this may be associated with the adaptation of the insect vector upon co-infection of the two viruses. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses demonstrated that the single infection with TYLCV or ToCV and the TYLCV&ToCV co-infection could perturb metabolic processes and metabolic pathways. Taken together, our results provide basis for further exploration of the molecular mechanisms of the response to TYLCV, ToCV single infection, and TYLCV&ToCV co-infection in B. tabaci MED, which will add to our knowledge of the interactions between plant viruses and insect vectors.
RESUMEN
Insect metamorphosis is a complex process involving drastic morphological and physiological changes. microRNAs (miRNAs) are a class of endogenous small non-coding RNAs that play key roles in regulating various biological processes, including metamorphosis, by post-transcriptional repression of mRNAs. The oriental fruit fly, Bactrocera dorsalis, is one of the most destructive insect pests in many Asian countries and the Pacific Islands. The regulatory role of miRNAs in B. dorsalis metamorphosis is unclear. To better understand the molecular regulatory mechanisms of miRNAs in pupariation, Illumina sequencing of the wandering stage (WS), the late WS and the white puparium stage of B. dorsalis were performed. Two hundred forty-nine miRNAs, including 184 known miRNAs and 65 novel miRNAs, were obtained. Among these miRNAs, 19 miRNAs were differentially expressed in pupariation, and eight miRNAs showed relative high expression levels (>50 TPM), of which five differentially expressed miRNAs (DEMs) had target differentially expressed genes (DEGs) predicted by the expected miRNA-mRNA negative regulation pattern using the Illumina HiSeq data. Four sets of DEMs and their predicted target DEGs were confirmed by qPCR. Of the four miRNAs, two miRNAs were down-regulated: miR-981, which may target pdpc, and Bdo-novel-mir-55, which potentially regulates spsX1, psB/C, and chit3. The other two miRNAs were up-regulated: let-7a-3p, which possibly controls lap, and Bdo-novel-mir-24, which may regulate ipc and sp1/2. This study provides a useful resource to elucidate the regulatory role of miRNAs and understand the molecular mechanisms of metamorphosis.
RESUMEN
Ecdysteroids play a crucial role in regulating molting in the phylum of Arthropoda and much is known with members of the subphylum of Hexapoda including the Insecta. However, this is still unclear in key pests as spider mites belonging to the subphylum of Chelicerata that originated earlier in the Cambrian period. In this study, we investigated 14 key genes of ecdysteroid biosynthesis and signaling and their expression over the different developmental stages in the citrus red mite, Panonychus citri (Acari: Stigmaeidae). P. citri is an economically important and widespread pest of citrus crops and it has five developmental stages of egg, larva, protonymph, deutonymph and adult. Typically, the expression of the ecdysteroid-synthesizing Halloween gene Spook (PcSpo) followed a positive zigzag-like pattern with a peak in the first half of each developmental stage and a drop in the second half prior to the molting to the next stage. Similar to PcSpo, PcDib, PcSad, PcRXR2, PcE75 and PcHR38 showed a positive zigzag-like expression pattern, while that of PcE78, PcHR3 and PcFTZ-F1 was opposite that we called a negative zigzag-like pattern. Silencing of the PcSpo gene by RNAi showed that molting was inhibited. Interestingly, we could rescue these RNAi effects by supplementing ponasterone A (PonA) and not by 20E, which is indicative that mites use PonA rather than 20E as ecdysteroid hormone. Modeling of the ecdysteroid receptor (PcEcR) hormone binding cavity also predicted binding of PonA, but showed a steric hindrance for 20E. We believe our data provide insight into the evolution and expression patterns of key ecdysteroid biosynthesis and signaling genes in a distant, non-insect species, and can become a foundation to develop new targets for controlling important agricultural pests such as spider mites.