scholarly journals Regulation of RNA Interference Pathways in the Insect Vector Laodelphax striatellus by Viral Proteins of Rice Stripe Virus

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1591
Author(s):  
Yan Xiao ◽  
Qiong Li ◽  
Wei Wang ◽  
Yumei Fu ◽  
Feng Cui
Keyword(s):  
Rna Interference ◽  
Viral Replication ◽  
Protein Binding ◽  
Brown Planthopper ◽  
Rice Stripe Virus ◽  
Transcript Level ◽  
Infection Process ◽  
Insect Vector ◽  
Binding Assays ◽  
Protein Levels

RNA interference (RNAi), especially the small interfering RNA (siRNA) and microRNA (miRNA) pathways, plays an important role in defending against viruses in plants and insects. However, how insect-transmitted phytoviruses regulate the RNAi-mediated antiviral response in vector insects has barely been uncovered. In this study, we explored the interaction between rice stripe virus (RSV) and the miRNA and siRNA pathways of the small brown planthopper, which is a vector insect. The transcript and protein levels of key genes in the two RNAi pathways did not change during the RSV infection process. When the expression of insect Ago1, Ago2, or Translin was silenced by the injection of double-stranded RNAs targeting these genes, viral replication was promoted with Ago2 silencing but inhibited with Translin silencing. Protein-protein binding assays showed that viral NS2 and RNA-dependent RNA polymerase interacted with insect Ago2 and Translin, respectively. When NS2 was knocked down, the transcript level of Ago2 increased and viral replication was inhibited. Therefore, viral NS2 behaved like an siRNA suppressor in vector insects. This protein-binding regulation of insect RNAi systems reflects a complicated and diverse coevolution of viruses with their vector insects.

scholarly journals Coordination between terminal variation of the viral genome and insect microRNAs regulates rice stripe virus replication in insect vectors

2021 ◽  
Vol 17 (3) ◽  
pp. e1009424
Author(s):  
Wan Zhao ◽  
Jinting Yu ◽  
Feng Jiang ◽  
Wei Wang ◽  
Le Kang ◽  
...  
Keyword(s):  
Viral Replication ◽  
Viral Genome ◽  
Promoter Activity ◽  
Brown Planthopper ◽  
Rice Stripe Virus ◽  
Genomic Variation ◽  
Reverse Genetic System ◽  
Insect Vector ◽  
Insect Vectors ◽  
Extension Sequence

Maintenance of a balance between the levels of viral replication and selective pressure from the immune systems of insect vectors is one of the prerequisites for efficient transmission of insect-borne propagative phytoviruses. The mechanism regulating the adaptation of RNA viruses to insect vectors by genomic variation remains unknown. Our previous study demonstrated an extension of the 3’-untranslated terminal region (UTR) of two genomic segments of rice stripe virus (RSV). In the present study, a reverse genetic system for RSV in human cells and an insect vector, the small brown planthopper Laodelphax striatellus, was used to demonstrate that the 3’-terminal extensions suppressed viral replication in vector insects by inhibiting promoter activity due to structural interference with the panhandle structure formed by viral 3’- and 5’-UTRs. The extension sequence in the viral RNA1 segment was targeted by an endogenous insect microRNA, miR-263a, which decreased the inhibitory effect of the extension sequence on viral promoter activity. Surprisingly, the expression of miR-263a was negatively regulated by RSV infection. This elaborate coordination between terminal variation of the viral genome and endogenous insect microRNAs controls RSV replication in planthopper, thus reflecting a distinct strategy of adaptation of phytoviruses to insect vectors.

scholarly journals The nucleocapsid protein of rice stripe virus in cell nuclei of vector insect regulates viral replication

2021 ◽  
Author(s):  
Wan Zhao ◽  
Junjie Zhu ◽  
Hong Lu ◽  
Jiaming Zhu ◽  
Fei Jiang ◽  
...  
Keyword(s):  
Viral Replication ◽  
Nucleocapsid Protein ◽  
Brown Planthopper ◽  
Rice Stripe Virus ◽  
Host Cells ◽  
Cell Nuclei ◽  
Nuclear Entry ◽  
Genomic Rnas ◽  
Importin Α ◽  
Vector Insect

AbstractRice stripe virus (RSV) transmitted by the small brown planthopper causes severe rice yield losses in Asian countries. Although viral nuclear entry promotes viral replication in host cells, whether this phenomenon occurs in vector cells remains unknown. Therefore, in this study, we systematically evaluated the presence and roles of RSV in the nuclei of vector insect cells. We observed that the nucleocapsid protein (NP) and viral genomic RNAs were partially transported into vector cell nuclei by utilizing the importin α nuclear transport system. When blocking NP nuclear localization, cytoplasmic RSV accumulation significantly increased. In the vector cell nuclei, NP bound the transcription factor YY1 and affected its positive regulation to FAIM. Subsequently, decreased FAIM expression triggered an antiviral caspase-dependent apoptotic reaction. Our results reveal that viral nuclear entry induces completely different immune effects in vector and host cells, providing new insights into the balance between viral load and the immunity pressure in vector insects.

scholarly journals Unusual characteristics of dicistrovirus-derived small RNAs in the small brown planthopper, Laodelphax striatellus

2014 ◽  
Vol 95 (3) ◽  
pp. 712-718 ◽  
Author(s):  
Junmin Li ◽  
Ida Bagus Andika ◽  
Yanru Zhou ◽  
Jiangfeng Shen ◽  
Zongtao Sun ◽  
...  
Keyword(s):  
Rna Interference ◽  
Small Rna ◽  
Small Rnas ◽  
Viral Genome ◽  
Size Range ◽  
Brown Planthopper ◽  
Insect Vector ◽  
Laodelphax Striatellus ◽  
Sense Strand ◽  
Wide Size Distribution

In this study, sequences of small RNA (sRNA) libraries derived from the insect vector Laodelphax striatellus were assembled into contigs and used as queries for database searches. A large number of contigs were highly homologous to the genome sequence of an insect dicistrovirus, himetobi P virus (HiPV). Interestingly, HiPV-derived sRNAs had a wide size distribution, and were relatively abundant throughout the 18–30 nt size range with only a slight peak at 22 nt. HiPV sRNAs had a strong bias towards the sense strand, whilst the antisense sRNAs were predominantly 21 and 22 nt. HiPV sRNAs do not have the typical features of PIWI-interacting RNAs, but their 3′ ends were preferentially cleaved at UA-rich sequences. Our data suggest that HiPV sRNAs may be derived both from activities of the RNA interference pathway and from cleavage of the viral genome by other host RNases.

scholarly journals An insect cell line derived from the small brown planthopper supports replication of rice stripe virus, a tenuivirus

2013 ◽  
Vol 94 (6) ◽  
pp. 1421-1425 ◽  
Author(s):  
Yuanyuan Ma ◽  
Wei Wu ◽  
Hongyan Chen ◽  
Qifei Liu ◽  
Dongsheng Jia ◽  
...  
Keyword(s):  
Viral Infection ◽  
Cell Line ◽  
Persistent Infection ◽  
Brown Planthopper ◽  
Insect Cell Line ◽  
Rice Stripe Virus ◽  
Insect Vector ◽  
Structural Protein ◽  
Small Brown Planthopper ◽  
A Cell

A cell line from the small brown planthopper (SBPH; Laodelphax striatellus) was established to study replication of rice stripe virus (RSV), a tenuivirus. The SBPH cell line, which had been subcultured through 30 passages, formed monolayers of epithelial-like cells. Inoculation of cultured vector cells with RSV resulted in a persistent infection. During viral infection in the SBPH cell line, the viral non-structural protein NS3 co-localized with the filamentous ribonucleoprotein particles of RSV, as revealed by electron and confocal microscopy. The knockdown of NS3 expression due to RNA interference induced by synthesized double-stranded RNAs from the NS3 gene significantly inhibited viral infection in the SBPH cell line. These results demonstrated that NS3 of RSV might be involved in viral replication or assembly. The persistent infection of the SBPH cell line by RSV will enable a better understanding of the complex relationship between RSV and its insect vector.

scholarly journals Development of Continuous Cell Culture of Brown Planthopper To Trace the Early Infection Process of Oryzaviruses in Insect Vector Cells

2014 ◽  
Vol 88 (8) ◽  
pp. 4265-4274 ◽  
Author(s):  
H. Chen ◽  
L. Zheng ◽  
Q. Mao ◽  
Q. Liu ◽  
D. Jia ◽  
...  
Keyword(s):  
Cell Culture ◽  
Brown Planthopper ◽  
Infection Process ◽  
Early Infection ◽  
Insect Vector

scholarly journals Ubiquitin-Conjugating Enzyme E2 E Inhibits the Accumulation of Rice Stripe Virus in Laodelphax striatellus (Fallén)

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 908
Author(s):  
Yao Li ◽  
Ze Zhou ◽  
Mi Shen ◽  
Linquan Ge ◽  
Fang Liu
Keyword(s):  
Brown Planthopper ◽  
Rice Stripe Virus ◽  
Ubiquitin Proteasome System ◽  
Plant Viruses ◽  
Pcr Analysis ◽  
Laodelphax Striatellus ◽  
Protein Levels ◽  
Ubiquitin Conjugating Enzyme ◽  
Rapid Amplification ◽  
Conjugating Enzyme

The ubiquitin–proteasome system (UPS) is an essential protagonist in host–pathogen interactions. Among the three classes of enzymes in the UPS, ubiquitin-conjugating enzyme E2 plays a dual role in viral pathogenesis; however, the role of insect E2s in interactions with plant viruses is unclear. Twenty E2-encoding genes in Laodelphax striatellus, the small brown planthopper, were identified and classified into 17 groups by transcriptomic and phylogenetic analysis. Full-length cDNAs of four LstrE2s (LstrE2 A/E/G2/H) were obtained by rapid-amplification of cDNA ends (RACE-PCR) analysis. Expression of the four LstrE2s showed tissue- and development-specific patterns. RT-qPCR analyses revealed that Rice stripe viruse (RSV) infection increased the level of LstrE2 A/E/G2/H. Further study indicated that repression of LstrE2 E via RNAi caused significant increases in the expression of RSV coat protein mRNA and protein levels. These findings suggest that LstrE2 E inhibits RSV accumulation in the planthopper body. Understanding the function of LstrE2 E in RSV accumulation may ultimately result in the development of novel antiviral strategies.

scholarly journals Interplay of Rice Stripe Virus and Rice Black Streaked Dwarf Virus during Their Acquisition and Accumulation in Insect Vector

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1121
Author(s):  
Marcia Beatriz Moya Fernández ◽  
Wenwen Liu ◽  
Lu Zhang ◽  
Jamal-U-Ddin Hajano ◽  
Xifeng Wang
Keyword(s):  
Epithelial Cells ◽  
Crop Production ◽  
Brown Planthopper ◽  
Rice Stripe Virus ◽  
Plant Viruses ◽  
Dwarf Virus ◽  
Insect Vector ◽  
Laodelphax Striatellus ◽  
Persistent Viruses ◽  
Vector Insect

Plant viruses transmitted by hemipteran vectors commonly cause losses to crop production. Rice stripe virus (RSV) and rice black streaked dwarf virus (RBSDV) are transmitted to rice plants by the same vector, the small brown planthopper (SBPH), Laodelphax striatellus Fallén, in a persistent propagative manner. However, rarely do the respective diseases they cause occur simultaneously in a field. Here, we determined the acquisition efficiency of RSV and RBSDV when acquired in succession or simultaneously by SBPH. When RBSDV was acquired first, RSV acquisition efficiency was significantly lower than when only acquiring RSV. However, RBSDV acquisition efficiency from insects that acquired RSV first was not significantly different between the insects only acquiring RBSDV. Immunofluorescence assays showed that the acquisition of RBSDV first might inhibit RSV entry into midgut epithelial cells, but RSV did not affect RBSDV entry. SBPHs were more likely to acquire RBSDV when they were feeding on plants coinfected with the two viruses. When RBSDV was acquired before RSV, RBSDV titer was significantly higher and RSV titer first declined, then increased compared to when only acquiring RBSDV or RSV. Only 5% of the SBPHs acquired both viruses when feeding on plants coinfected with RSV and RBSDV. These results provide a better understanding of the interaction between two persistent viruses when present in the same vector insect and explain why RSV and RBSDV occur in intermittent epidemics.

RNA interference-aided knockdown of a putative saccharopine dehydrogenase leads to abnormal ecdysis in the brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae)

2015 ◽  
Vol 105 (4) ◽  
pp. 390-398 ◽  
Author(s):  
P.-J. Wan ◽  
L. Yang ◽  
S.-Y. Yuan ◽  
Y.-H. Tang ◽  
Q. Fu ◽  
...  
Keyword(s):  
Brown Planthopper ◽  
Transcript Level ◽  
Nilaparvata Lugens ◽  
The Body ◽  
Double Stranded Rna ◽  
Saccharopine Dehydrogenase ◽  
Body Weights ◽  
Nilaparvata Lugens Stål ◽  
Phloem Feeding ◽  
Wing Deformation

AbstractThe brown planthopper Nilaparvata lugens is a serious phloem-feeding pest of rice in China. The current study focuses on a saccharopine dehydrogenase (SDH) that catalyzes the penultimate reaction in biosynthesis of the amino acid lysine (Lys), which plays a role in insect growth and carnitine production (as a substrate). The protein, provisionally designated as NlylsSDH [a SDH derived from yeast-like symbiont (YLS) in N. lugens], had a higher transcript level in abdomens, compared with heads, wings, legs and thoraces, which agrees with YLS distribution in N. lugens. Ingestion of Nlylssdh targeted double-stranded RNA (dsNlylssdh) for 5, 10 and 15 days decreased the mRNA abundance in the hoppers by 47, 70 and 31%, respectively, comparing with those ingesting normal or dsegfp diets. Nlylssdh knockdown slightly decreased the body weights, significantly delayed the development of females, and killed approximately 30% of the nymphs. Moreover, some surviving adults showed two apparent phenotypic defects: wing deformation and nymphal cuticles remained on tips of the legs and abdomens. The brachypterours/macropterours and sex ratios (female/male) of the adults on the dsRNA diet were lowered compared with the adults on diets without dsRNA. These results suggest that Nlylssdh encodes a functional SDH protein. The adverse effect of Nlylssdh knockdown on N. lugens implies the importance of Lys in hopper development. This study provides a proof of concept example that Nlylssdh could serve as a possible dsRNA-based pesticide for planthopper control.

F-box protein MdAMR1L1 regulates ascorbate biosynthesis in apple by modulating GDP-mannose pyrophosphorylase

2021 ◽  
Author(s):  
Songya Ma ◽  
Huixia Li ◽  
Lan Wang ◽  
Baiyun Li ◽  
Zhengyang Wang ◽  
...  
Keyword(s):  
Regulatory Mechanism ◽  
Transcript Level ◽  
Ethylene Response Factor ◽  
Protein Abundance ◽  
Protein Levels ◽  
Guanosine Diphosphate ◽  
Physiological Processes ◽  
Ethylene Response ◽  
Ubiquitination Pathway ◽  
F Box Protein

Abstract Ascorbate (Asc) is an important antioxidant in plants and humans that plays key roles in various physiological processes. Understanding the regulation of Asc content in fruit plants is important for improving plant resiliency and optimizing Asc in food. Here, we found that both the transcript level and protein abundance of Asc Mannose pathway Regulator 1 Like 1 (MdAMR1L1) was negatively associated with Asc levels during the development of apple (Malus × domestica) fruit. The overexpression or silencing of MdAMR1L1 in apple indicated that MdAMR1L1 negatively regulated Asc levels. However, in the leaves of MdAMR1L1-overexpressing apple lines, the transcript levels of the Asc synthesis gene Guanosine diphosphate-mannose pyrophosphorylase MdGMP1 were increased, while its protein levels and enzyme activity were reduced. This occurred because the MdAMR1L1 protein interacted with MdGMP1 and promoted its degradation via the ubiquitination pathway to inhibit Asc synthesis at the post-translational level. MdERF98, an apple ethylene response factor, whose transcription was modulated by Asc level, is directly bound to the promoter of MdGMP1 to promote the transcription of MdGMP1. These findings provide insights into the regulatory mechanism of Asc biosynthesis in apples and revealed potential opportunities to improve fruit Asc levels.

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