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1.
J Econ Entomol ; 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32484504

RESUMO

The whitefly, Bemisia tabaci Gennadius, is a major phloem-feeding pest of agricultural crops that is also an important vector of many plant diseases. The B. tabaci Mediterranean ('MED') biotype is a particularly effective vector of Tomato yellow leaf curl virus (TYLCV), a devastating plant pathogen. Although insecticides play an important role in the control of MED and TYLCV, little is known about how TYLCV infection affects MED susceptibility to insecticides. We conducted research addressing how MED susceptibility to flupyradifurone, the first commercially available systemic control agent derived from the butenolide class of insecticides, was affected by TYLCV infection. We first conducted bioassays determining the LC15 and LC50 for control and viruliferous MED feeding on either water- or insecticide-treated plants. We next measured several demographic parameters of control and viruliferous MED exposed to either insecticide- or water-treated plants. TYLCV infection increased MED tolerance of flupyradifurone: the LC15 and LC50 of viruliferous MED were double that of uninfected MED. Viral infection also altered MED demographic responses to flupyradifurone, but in an inconsistent manner. Although the ability of TYLCV and other persistently transmitted viruses to benefit Bemisia via manipulation of host plant defense is well known, this appears to be the first example of virally mediated changes in vector susceptibility to an insecticide.

2.
J Econ Entomol ; 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32484541

RESUMO

The production of Chinese chives is reduced throughout China due to a root-feeding dipteran pest Bradysia odoriphaga Yang et Zhang (Diptera: Sciaridae), therefore deciphering the conditions influencing its growth and development are important in developing ecological control strategies. A study was conducted from 2014 to 2017 to determine the relationship between the abundance of B. odoriphaga and temperature (atmospheric and soil), soil water content, and atmospheric humidity in a Chinese chive field in Beijing City, China. Numbers of adults peaked in March and October to November and were lowest in July to August and December to next February; numbers of larvae were highest in December to next February and lowest in July to August. From 2014 to 2017, the numbers of adults and larvae were significantly correlated with monthly mean atmospheric temperatures and soil temperatures, but were not significantly correlated with monthly mean atmospheric relative humidity and soil water content. However, for both adults and larvae, numbers were significantly greater with high soil water contents compared with drought treatment. The results of this study suggest that the very low soil water contents, high atmospheric temperatures, and high soil temperatures were critical for regulating field populations of B. odoriphaga.

3.
Nat Commun ; 11(1): 3003, 2020 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-32532972

RESUMO

The arms race between entomopathogenic bacteria and their insect hosts is an excellent model for decoding the intricate coevolutionary processes of host-pathogen interaction. Here, we demonstrate that the MAPK signaling pathway is a general switch to trans-regulate differential expression of aminopeptidase N and other midgut genes in an insect host, diamondback moth (Plutella xylostella), thereby countering the virulence effect of Bacillus thuringiensis (Bt) toxins. Moreover, the MAPK cascade is activated and fine-tuned by the crosstalk between two major insect hormones, 20-hydroxyecdysone (20E) and juvenile hormone (JH) to elicit an important physiological response (i.e. Bt resistance) without incurring the significant fitness costs often associated with pathogen resistance. Hormones are well known to orchestrate physiological trade-offs in a wide variety of organisms, and our work decodes a hitherto undescribed function of these classic hormones and suggests that hormonal signaling plasticity is a general cross-kingdom strategy to fend off pathogens.

4.
Pestic Biochem Physiol ; 167: 104602, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32527436

RESUMO

The ecdysteroid hormone 20-hydroxyecdysone (20E), a critical hormone in arthropods, plays an essential role in insect growth, molting and reproduction. A previous study showed that 20E is actually regulated by six P450 genes (five P450 genes belonging to the Halloween family and a CYP18A1 gene) in model insects. However, the role of the six P450 genes in Bemisia tabaci Q (also call Mediterranean, MED), an important pest of field crops, remains unclear. Here, six P450 genes were cloned by RT-PCR, and the phylogenetic tree indicated a close orthologous relationship of these P450 genes between MED and other insects. Spatiotemporal expression profiling revealed that five P450 genes (CYP18A1, CYP306A1, CYP307A2, CYP314A1 and CYP315A1) were expressed at significantly higher levels in the head than in the abdomen and thorax. Four P450 genes (CYP302A1, CYP307A2, CYP314A1 and CYP315A1) were expressed at the highest levels in males, and CYP18A1 was expressed at the highest levels in the 4th nymph stage. The molting process was delayed by approximately 1-3 days after knockdown of these genes at the 4th nymph stage, and the mean proportion of shriveled or dead insects reached 8.3% (CYP18A1), 20.8% (CYP302A1), 7.0% (CYP307A2), 31.8% (CYP306A1), 28.6% (CYP314A1) and 24.1% (CYP315A1). In addition, 20E rescued the negative effect of ds-CYP306A1, ds-CYP314A1 and ds-CYP315A1 on the eclosion rate. We concluded that these Halloween genes and CYP18A1 likely participate in the development of MED, and in particular, CYP306A1 could be used as a putative insecticide target for controlling this piercing-sucking insect.


Assuntos
Hemípteros , Inseticidas , Animais , Sistema Enzimático do Citocromo P-450 , Proteínas de Insetos , Insetos , Filogenia
5.
Pestic Biochem Physiol ; 165: 104555, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32359544

RESUMO

The development of genetic based techniques, specifically RNA interference (RNAi), has emerged as a powerful tool in novel pest management strategies for pestiferous coleoptera. The 28-spotted ladybird beetle, Henosepilachna vigintioctopunctata, is a dynamic foliar pest of solenaceous plants, primarily potato plants, and has quickly become one of the most important pests attacking many crops in Asian countries. In this study, we demonstrate the efficacy of dietary RNAi targeting vATPase B, which led to significant gene silencing. Downstream effects of vATPase B silencing appeared to be both time- and partial dose-dependent. Our results indicate that silencing of vATPase B caused a significant decrease in survival rate, as well as reduced the food stuffs consumption and inhibited the overall development of H. vigintioctopunctata. Furthermore, results demonstrate expression of insect melanism related genes, TH and DDC, was significantly up regulated under the dsvATPase B (RNAi molecule designed against vATPase B) treatment. The impact of oral dsvATPase B delivery on the survival of 1st, 3rd instars, and adults was investigated through bacterially expressed dsRNA. The effectiveness of RNAi-based gene silencing in H. vigintioctopunctata provides a powerful reverse genetic tool for the functional annotation of its genes. This study demonstrates that vATPase B may represent a candidate gene for RNAi-based control of H. vigintioctopunctata.


Assuntos
Besouros , RNA de Cadeia Dupla , Animais , Dieta , Controle de Pragas , Interferência de RNA
6.
Trends Plant Sci ; 25(7): 627-630, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32349926

RESUMO

Unlike the situation for humans and microbes, the active multiprotein assemblies of plants have not been systematically defined. A recent report by McWhite et al. remedies this by analyzing the protein complexes of 13 plant species, thereby defining core assemblies and providing an essential resource for interpreting the genotype-phenotype space of plants.

7.
Genomics ; 2020 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-32353477

RESUMO

The gnat, Bradysia odoriphaga Yang et Zhang, is an important underground pest in Asia. B. odoriphaga differ in heat and cold tolerance and exhibit quite different developmental strategies. To understand the underlying mechanisms, we sequenced and compared the transcriptome of B. odoriphaga under 40 °C (a stressful high temperature), 25 °C, and 4 °C (a stressful low temperature) for 1 h. We found that metabolism- and ribosome-related genes were modulated. In high temperature (40 °C), heat shock protein (HSP) genes, detoxication genes, metabolism genes, protein turnover genes, and stress signal transduction genes were differentially expressed. In low temperature (4 °C), genes related with heat shock protein (HSP) and detoxication were differentially expressed. Our study increases our understanding of the complex molecular mechanisms involved in the responses of B. odoriphaga to acute temperature stress and provides a potential strategy for pest management.

8.
Artigo em Inglês | MEDLINE | ID: mdl-32368925

RESUMO

SIGNIFICANCE: Alongside well-characterized permanent multimeric enzymes and multi-enzyme complexes relatively unstable transient enzyme-enzyme assemblies, including metabolons, provide an important mechanism for the regulation of energy and redox metabolism. CRITICAL ISSUES: Despite the fact that enzyme-enzyme assemblies have been proposed for many decades and experimentally analyzed for at least 40 years there are very few pathways for which unequivocal evidence for the presence of metabolite channeling, the most frequently evoked reason for their formation, has been provided. Furthermore, in contrast to the stronger, permanent interactions for which a deep understanding of the subunit interface exists the mechanism(s) underlying transient enzyme-enzyme interactions remain poorly studied. Recent advances: The widespread adoption of proteomic and cell biological approaches to characterize protein-protein interaction is defining an ever-increasing number of enzyme-enzyme assemblies as well as enzyme protein interactions that likely identify factors which stabilize such complexes. Moreover, the use of microfluidic technologies provided compelling support of a role for substrate-specific chemotaxis in complex assemblies. FUTURE DIRECTIONS: Embracing current and developing technologies should render the delineation of metabolons from other enzyme-enzyme complexes more facile. In parallel, attempts to confirm that the findings reported in microfluidic systems are indeed representative of the cellular situation will be critical to understanding the physiological circumstances requiring and evoking dynamic changes in the levels of the various transient enzyme-enzyme assemblies of the cell.

9.
Mol Plant ; 2020 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-32305499

RESUMO

While the structures of plant primary metabolic pathways are generally well defined and highly conserved across species, those defining specialized metabolism are less well characterized and more highly variable across species. In this study, we investigated polyphenolic metabolism in the lycopersicum complex by characterizing the underlying biosynthetic and decorative reactions that constitute the metabolic network of polyphenols across eight different species of tomato. For this purpose, GC-MS- and LC-MS-based metabolomics of different tissues of Solanum lycopersicum and wild tomato species were carried out, in concert with the evaluation of cross-hybridized microarray data for MapMan-based transcriptomic analysis, and publicly available RNA-sequencing data for annotation of biosynthetic genes. The combined data were used to compile species-specific metabolic networks of polyphenolic metabolism, allowing the establishment of an entire pan-species biosynthetic framework as well as annotation of the functions of decoration enzymes involved in the formation of metabolic diversity of the flavonoid pathway. The combined results are discussed in the context of the current understanding of tomato flavonol biosynthesis as well as a global view of metabolic shifts during fruit ripening. Our results provide an example as to how large-scale biology approaches can be used for the definition and refinement of large specialized metabolism pathways.

10.
Insect Sci ; 2020 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-32240577

RESUMO

RNA interference (RNAi) techniques have emerged as powerful tools that facilitate development of novel management strategies for insect pests, such as Henosepilachna vigintioctopunctata (Coleoptera: Coccinellidae), which is a major pest of solanaceous plants in Asia. In this study, the potential of oral delivery of in vitro-synthesized and bacterially expressed dsHvlwr to manage of H. vigintioctopunctata was investigated. Our results showed that the H. vigintioctopunctata lesswright (lwr) gene (Hvlwr) had a 480-bp open reading frame and encoded a 160-amino acid protein. Hvlwr expression levels were greater in the fat body than other tissue types. Hvlwr silencing led to greater H. vigintioctopunctata mortality rates and appeared to be time- and partially dose-dependent, likely as a result of the number of hemocytes increasing with dsRNA concentration, but decreasing with time. Bacterially expressed dsHvlwr that was applied to leaf discs caused 88%, 66%, and 36% mortality in 1st instars, 3rd instars, and adults after 10, 10, and 14 days, respectively; when applied to living plants, there was greater mortality in 1st and 3rd instars, but there was no effect on adults. Furthermore, dsHvlwr led to improved plant protection against H. vigintioctopunctata. Our study shows an effective dietary RNAi response in H. vigintioctopunctata and that Hvlwr is a promising RNAi target gene for control of this pest species. This article is protected by copyright. All rights reserved.

11.
Proc Natl Acad Sci U S A ; 117(19): 10246-10253, 2020 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-32327610

RESUMO

The evolution of insect resistance to pesticides poses a continuing threat to agriculture and human health. While much is known about the proximate molecular and biochemical mechanisms that confer resistance, far less is known about the regulation of the specific genes/gene families involved, particularly by trans-acting factors such as signal-regulated transcription factors. Here we resolve in fine detail the trans-regulation of CYP6CM1, a cytochrome P450 that confers resistance to neonicotinoid insecticides in the whitefly Bemisia tabaci, by the mitogen-activated protein kinase (MAPK)-directed activation of the transcription factor cAMP-response element binding protein (CREB). Reporter gene assays were used to identify the putative promoter of CYP6CM1, but no consistent polymorphisms were observed in the promoter of a resistant strain of B. tabaci (imidacloprid-resistant, IMR), which overexpresses this gene, compared to a susceptible strain (imidacloprid-susceptible, IMS). Investigation of potential trans-acting factors using in vitro and in vivo assays demonstrated that the bZIP transcription factor CREB directly regulates CYP6CM1 expression by binding to a cAMP-response element (CRE)-like site in the promoter of this gene. CREB is overexpressed in the IMR strain, and inhibitor, luciferase, and RNA interference assays revealed that a signaling pathway of MAPKs mediates the activation of CREB, and thus the increased expression of CYP6CM1, by phosphorylation-mediated signal transduction. Collectively, these results provide mechanistic insights into the regulation of xenobiotic responses in insects and implicate both the MAPK-signaling pathway and a transcription factor in the development of pesticide resistance.

12.
Inorg Chem ; 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-32223161

RESUMO

Crystal structure prediction (CSP) methods recently proposed a series of new rare-earth (RE) hydrides at high pressures with novel crystal structures, unusual stoichiometries, and intriguing features such as high-Tc superconductivity. RE trihydrides (REH3) generally undergo a phase transition from ambient P63/mmc or P3̅c1 to Fm3̅m at high pressure. This cubic REH3 (Fm3̅m) was considered to be a precursor to further synthesize RE polyhydrides such as YH4, YH6, YH9, and CeH9 with higher hydrogen contents at higher pressures. However, the structural stability and equation of state (EOS) of any of the REH3 have not been fully investigated at sufficiently high pressures. This work presents high-pressure X-ray diffraction (XRD) measurements in a laser-heated diamond anvil cell up to 100 GPa and ab initio evolutionary CSP of stable phases of DyH3 up to 220 GPa. Experiments observed the Fm3̅m phase of DyH3 to be stable at pressures from 17 to 100 GPa and temperatures up to ∼2000 K. After complete decompression, the P3̅c1 and Fm3̅m phases of DyH3 recovered under ambient conditions. Our calculations predicted a series of phases for DyH3 at high pressures with the structural phase transition sequence P3̅c1 → Imm2 → Fm3̅m → Pnma → P63/mmc at 11, 35, 135, and 194 GPa, respectively. The predicted P3̅c1 and Fm3̅m phases are consistent with experimental observations. Furthermore, electronic band structure calculations were carried out for the predicted phases of DyH3, including the 4f states, within the DFT+U approach. The inclusion of 4f states shows significant changes in electronic properties, as more Dy d states cross the Fermi level and overlap with H 1s states. The structural phase transition from P3̅c1 to Fm3̅m observed in DyH3 is systematically compared with other REH3 compounds at high pressures. The phase transition pressure in REH3 shows an inverse relation with the ionic radius of RE atoms.

13.
Trends Biochem Sci ; 2020 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-32345468

RESUMO

Energy-demanding processes, such as cell growth, migration, and differentiation, are tension modulated, begging the question whether metabolism and mechanical tension are tightly linked. A recent report by Park et al. shows that stiffness in the extracellular matrix (ECM) promotes reorganization of actin, resulting in enhanced glycolysis.

14.
Pest Manag Sci ; 2020 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-32112472

RESUMO

BACKGROUND: RNA interference (RNAi) is a potential tool for plant protection against insect pests. The great challenge for effective pest control using RNAi in the field is the development of efficient and reliable methods for the production and delivery of double-stranded RNA (dsRNA). RESULTS: In the present study, we investigated the potential of feeding in vitro synthesized or bacterially expressed dsRNA to populations of the 28-spotted ladybeetle Henosepilachna vigintioctopunctata as a method of biological pest control. Ingestion of in vitro synthesized dsHvRPS18 or dsHvRPL13 led to significant down-regulation of the ribosomal protein-encoding genes HvRPS18 and HvRPL13, respectively, and significantly decreased the survival of H. vigintioctopunctata. Such silencing of HvRPS18 or HvRPL13 expression appeared to be partially dose-dependent and also inhibited the growth of H. vigintioctopunctata and significantly suppressed the expression of digestive enzyme-related genes. Finally, ingestion of bacterially expressed dsHvRPS18 or dsHvRPL13 induced significant mortality in the first and third instars, and in adults. CONCLUSION: The effectiveness of RNAi-based gene silencing in H. vigintioctopunctata provides a powerful reverse genetic tool for the functional annotation of its genes. This study demonstrates that HvRPS18 and HvRPL13 represent candidate genes for RNAi-based biological control of H. vigintioctopunctata. © 2020 Society of Chemical Industry.

15.
Mol Ecol Resour ; 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-32220045

RESUMO

The greenhouse whitefly, Trialeurodes vaporariorum Westwood, is an agricultural pest of global importance. Here we report a 787-Mb high-quality draft genome sequence of T. vaporariorum assembled from PacBio long reads and Hi-C chromatin interaction maps, which has scaffold and contig N50 lengths of 70 Mb and 500 kb, respectively, and contains 18,275 protein-coding genes. About 98.8% of the assembled contigs were placed onto the 11 T. vaporariorum chromosomes. Comparative genomic analysis reveals significantly expanded gene families such as aspartyl proteases in T. vaporariorum compared to Bemisia tabaci Mediterranean (MED) and Middle East-Asia Minor 1 (MEAM1). Furthermore, the cytochrome CYP6 subfamily shows significant expansion in T. vaporariorum and several genes in this subfamily display developmental stage-specific expression patterns. The high-quality T. vaporariorum genome provides a valuable resource for research in a broad range of areas such as fundamental molecular ecology, insect-plant/insect-microorganism or virus interactions and pest resistance management.

16.
Can J Cardiol ; 36(4): 589.e5-589.e7, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32220388

RESUMO

We report an interesting case of coronary arteritis and periaortitis in a 62-year-old man with a history of biopsy-proven IgG4-related pulmonary disease. After 2 years of immune-suppressive therapy, the perivascular tissue surrounding all coronary arteries and the abdominal aorta was significantly attenuated, except that the luminal stenosis was aggravated to 70% in the left anterior descending coronary artery. Treatment with aspirin, atorvastatin, and ezetimibe was added. The patient was discharged under strict lesion surveillance at follow-up.

17.
Pest Manag Sci ; 76(7): 2406-2414, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32030849

RESUMO

BACKGROUND: Tomato spotted wilt orthotospovirus (TSWV), one of the most devastating viruses of ornamental plants and vegetable crops worldwide, is transmitted by the western flower thrips, Frankliniella occidentalis (Pergande), in a persistent-propagative manner. How TSWV influences the reproduction of its vector to enhance transmission and whether infection with TSWV changes the mating behavior of F. occidentalis are not fully understood. RESULTS: TSWV-exposed thrips had a significantly longer developmental time than non-exposed individuals. More importantly, increased developmental time was predominantly associated with adults, a stage critical for dispersal and virus transmission. In addition, TSWV-exposed F. occidentalis produced substantially more progeny than did non-exposed thrips. Interestingly, most of the increase in progeny came from an increase in males, a sex with a greater dispersal and virus transmission capability. Specifically, the female/male ratio of progeny shifted from 1.3-7.0/1 to 0.6-1.1/1. As for mating behavior, copulation time was significantly longer in TSWV-exposed thrips. Finally, females tended to re-mate less when exposed to the virus. Resistance to re-mating may lead to reduced sperm availability in females, which translates to a larger number of male progeny under a haplodiploid system. CONCLUSION: These combined results suggest that TSWV can influence the developmental time, mating behavior, fecundity, and offspring sex allocation of its vector F. occidentalis to facilitate virus transmission. As such, a monitoring program capable of the earlier detection of the virus in host plants and/or its insect vector, thrips, using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), real time quantitative polymerase chain reaction (RT-qPCR) or virus detection strips might be beneficial for long-term, sustainable management. © 2020 Society of Chemical Industry.

18.
Pestic Biochem Physiol ; 163: 23-30, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31973862

RESUMO

Insecticidal Cry toxins produced by Bacillus thuringiensis (Bt) have been widely used to control agricultural pests in both foliage sprays and transgenic crops. Nevertheless, rapid evolution of insect resistance to Cry toxins requires elucidation of the molecular mechanisms involved in Cry resistance. Two proposed models have been described to explain the toxicity of Cry proteins, the classic model states that Cry protoxin is activated by midgut proteases resulting in activated toxin that binds to receptors and forms a pore in the midgut cells triggering larval death, and the newly proposed dual model of the mode of action of Bt Cry toxins states that protoxin and activated toxins may have different mechanisms of action since several resistant strains to activated Cry toxins are still susceptible to the same Cry-protoxin. Protoxin activation by midgut proteases is a key step in both models. Herein, we evaluated Cry1Ac protoxin activation in a susceptible Plutella xylostella (L.) strain (DBM1Ac-S) and in the near-isogenic strain (NIL-R) with high field-evolved Cry1Ac resistance. Previous work showed that Cry1Ac resistance in NIL-R correlates with reduced binding to midgut receptors due to enhanced MAPK signaling pathway and down regulation of ABCC2 receptor. However, reduced midgut trypsin levels and altered midgut protease gene transcription were also observed in the Cry1Ac-resistant field isolated strain that is parent of the NIL-R strain. Therefore, we analyzed the midgut protease activities in both DBM1Ac-S and NIL-R strains. Detection of enzymatic activities showed that caseinolytic protease, trypsin and chymotrypsin activities were not significantly different between the susceptible and resistant strains. Furthermore, treatment with different trypsin or chymotrypsin inhibitors, such as Nα-tosyl-l-lysine chloromethyl ketone (TLCK) or Np-tosyl-L-phenylalanine chloromethyl ketone (TPCK) did not affect the susceptibility to Cry1Ac protoxin of the DBM1Ac-S and NIL-R larvae. Bioassay results indicated that the NIL-R larvae showed similar resistant levels to both Cry1Ac protoxin and trypsin-activated toxin. Taken together, our results demonstrated that high-level field-evolved Cry1Ac resistance in the NIL-R strain is independent of Cry1Ac protoxin activation and the specific protoxin mechanism of action. This discovery will strengthen our comprehensive understanding of the complex mechanistic basis of Bt resistance in different insects.


Assuntos
Bacillus thuringiensis , Mariposas , Animais , Proteínas de Bactérias , Endotoxinas , Proteínas Hemolisinas , Proteínas de Insetos , Resistência a Inseticidas , Larva , Peptídeo Hidrolases
19.
Pest Manag Sci ; 76(6): 2040-2048, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31943718

RESUMO

BACKGROUND: Nuclear receptors (NRs) play an essential role in diverse biological processes, such as insect metamorphosis. Here, transcriptome analysis and functional studies were used to determine whether NRs are involved in metamorphosis of whitefly Bemisia tabaci Q, a serious pest to crops, and to find some potential insecticide targets. RESULTS: Twenty NRs were identified in the Bemisia tabaci Q genome and categorized into the NR0-NR6 subfamilies. The phylogenetic tree of NRs from Bemisia tabaci Q and other representative species was constructed, which provided evolutionary insight into their genetic distances. The results of spatiotemporal gene expression indicated that the majority of NR gene expression was higher in the head than the abdomen and higher in eggs than adults. Further functional analysis using RNA interference (RNAi) showed that NR genes play an important role in Bemisia tabaci Q pupation and eclosion. With respect to high mortality and effects on growth, this was reflected in the unable to become pupa when the third-stage nymph treated with double-stranded RNA (dsRNA) and the developmental time delay (4-7 days) when pupae were treated with dsRNA for the 12 NR genes during molting compared with the development time in the control. CONCLUSION: This study provides insight into NR functions during the metamorphosis stages of Bemisia tabaci Q. Several candidate genes could be potential insecticide targets for whitefly pest control due to their important roles in insect development. © 2020 Society of Chemical Industry.

20.
Toxins (Basel) ; 12(2)2020 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-31979385

RESUMO

Bacillus thuringiensis (Bt) produce diverse insecticidal proteins to kill insect pests. Nevertheless, evolution of resistance to Bt toxins hampers the sustainable use of this technology. Previously, we identified down-regulation of a trypsin-like serine protease gene PxTryp_SPc1 in the midgut transcriptome and RNA-Seq data of a laboratory-selected Cry1Ac-resistant Plutella xylostella strain, SZ-R. We show here that reduced PxTryp_SPc1 expression significantly reduced caseinolytic and trypsin protease activities affecting Cry1Ac protoxin activation, thereby conferring higher resistance to Cry1Ac protoxin than activated toxin in SZ-R strain. Herein, the full-length cDNA sequence of PxTryp_SPc1 gene was cloned, and we found that it was mainly expressed in midgut tissue in all larval instars. Subsequently, we confirmed that the PxTryp_SPc1 gene was significantly decreased in SZ-R larval midgut and was further reduced when selected with high dose of Cry1Ac protoxin. Moreover, down-regulation of the PxTryp_SPc1 gene was genetically linked to resistance to Cry1Ac in the SZ-R strain. Finally, RNAi-mediated silencing of PxTryp_SPc1 gene expression decreased larval susceptibility to Cry1Ac protoxin in the susceptible DBM1Ac-S strain, supporting that low expression of PxTryp_SPc1 gene is involved in Cry1Ac resistance in P. xylostella. These findings contribute to understanding the role of midgut proteases in the mechanisms underlying insect resistance to Bt toxins.

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