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Background: Rice stripe virus (RSV) caused a serious disease pandemic in rice in East China between 2001 and 2010. The continuous integrated managements reduced virus epidemic year by year until it was non-epidemic. As an RNA virus, its genetic variability after undergoing a long-term non-epidemic period was meaningful to study. While in 2019, the sudden occurrence of RSV in Jiangsu provided an opportunity for the study. Methods and results: The complete genome of JY2019, an RSV isolate from Jiangyan, was determined. A genotype profile of 22 isolates from China, Japan and Korea indicated that the isolates from Yunnan formed the subtype II, and other isolates clustered the subtype I. RNA 1–3 of JY2019 isolate well-clustered in the subtype I clade, and RNA 4 was also in subtype I, but it had a slight separation from other intra-group isolates. After phylogenetic analyses, it was considered NSvc4 gene contributed to the tendency, because it exhibited an obvious trend towards the subtype II (Yunnan) group. High sequence identity (100%) of NSvc4 between JY2019 and barnyardgrass isolate from different regions demonstrated genetic variation of NSvc4 was consistent in RSV natural populations in Jiangsu in the non-epidemic period. In the phylogenetic tree of all 74 NSvc4 genes, JY2019 belonged to a minor subtype Ib, suggesting the subtype Ib isolates might have existed in natural populations before the non-epidemic period, but not a dominant population. Conclusions: Our results suggested that NSvc4 gene was susceptible to selection pressure, and the subtype Ib might be more adaptable for the interaction between RSV and hosts in the non-epidemic ecological conditions. [ABSTRACT FROM AUTHOR]

Biotic stresses represent a serious threat to rice production to meet global food demand and thus pose a major challenge for scientists, who need to understand the intricate defense mechanisms. Proteomics and metabolomics studies have found global changes in proteins and metabolites during defense responses of rice exposed to biotic stressors, and also reported the production of specific secondary metabolites (SMs) in some cultivars that may vary depending on the type of biotic stress and the time at which the stress is imposed. The most common changes were seen in photosynthesis which is modified differently by rice plants to conserve energy, disrupt food supply for biotic stress agent, and initiate defense mechanisms or by biotic stressors to facilitate invasion and acquire nutrients, depending on their feeding style. Studies also provide evidence for the correlation between reactive oxygen species (ROS) and photorespiration and photosynthesis which can broaden our understanding on the balance of ROS production and scavenging in rice-pathogen interaction. Variation in the generation of phytohormones is also a key response exploited by rice and pathogens for their own benefit. Proteomics and metabolomics studies in resistant and susceptible rice cultivars upon pathogen attack have helped to identify the proteins and metabolites related to specific defense mechanisms, where choosing of an appropriate method to identify characterized or novel proteins and metabolites is essential, considering the outcomes of host-pathogen interactions. Despites the limitation in identifying the whole repertoire of responsive metabolites, some studies have shed light on functions of resistant-specific SMs. Lastly, we illustrate the potent metabolites responsible for resistance to different biotic stressors to provide valuable targets for further investigation and application. [ABSTRACT FROM AUTHOR]

Rice stripe virus (RSV) causes viral disease and yield losses in rice. Leaves of rice plants showing yellowing, stripes and curling were collected in Sichuan Province, China and tested for rice strip virus (RSV) by RT-PCR using specific primers. Nine of the nineteen symptomatic samples were positive for RSV. The complete sequence of RSV RNA1 to RNA4 was determined by assembling cDNA fragments obtained from isolate Huid04 in Huidong County, Sichuan Province (GenBank accession Nos. KX611337 to KX611340). Phylogenetic analyses of RNA-dependent RNA polymerase sequences showed a close relationship of RSV isolate Hui0d4 and isolates DaL08 and KunM08 from Yunnan Province, China. This is the first complete genomic sequence of an RSV isolate from Sichuan Province, China. [ABSTRACT FROM AUTHOR]

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Infection of rice with Rice stripe virus (RSV) and Rice black streaked dwarf virus (RBSDV) causes a significant loss of grain production. Due to the lack of natural resistance resources against these viruses, it is imperative to discover a biotechnological approach that will provide effective and safe immunity to RSV and RBSDV. In this study, we constructed three dimeric artificial microRNA (amiRNA) precursor expression vectors (pamiR-M, pamiR-3 and pamiR-U) that simultaneously target the CP genes of RSV and RBSDV based on the structure of the rice ( Oryza sativa L.) osa-MIR528 precursor. The transgenic plants were obtained by Agrobacterium tumefaciens-mediated transformation and were shown to express amiRNAs successfully. Viral challenge assays revealed that these transgenic plants demonstrated different resistance (26.66-54.17 %) against RSV and RBSDV infection simultaneously. The amiRNA-targeting 3′-UTR region of CP gene (pamiR-U) induced higher virus resistance: 54.17 % against RSV and 45.83 % against RBSDV. A northern blot assay indicated that there was a good correlation between the resistance level and amiRNAs accumulation. The RNA silencing induced by the original amiRNAs could be bilaterally extended by the siRNA pathway. The amiRNAs, together with the secondary siRNAs, mediated the degradation of viral RNAs. A genetic stability assay showed that transgenes and amiRNA-mediated virus resistance could be stably inherited in the transgenic plants. [ABSTRACT FROM AUTHOR]

The RNA-dependent RNA polymerase (RdRp) of rice stripe virus (RSV) is critical for both the transcription and replication of the viral genome. Despite its importance, little is known about how it functions in cells. In the present study, RSV RdRp was split into three pieces, since expression of the full protein could not be achieved. Then, the intracellular localization of these three RdRp fragments and their interactions with nucleocapsid protein (NP) were investigated, which is another viral protein required for viral RNA synthesis. The data showed that all three RdRp fragments displayed punctuate staining patterns in the cytoplasm, and the C-terminal fragment co-localized with NP in the perinuclear region. Both bimolecular fluorescence complementation and co-immunoprecipitation experiments demonstrated that of the three RdRp fragments, only the C-terminal fragment could interact with NP. Further analysis using a series of truncated NPs identified the N-terminal 50-amino-acid region within NP as the determinant for its interaction with the C-terminus of RdRp. [ABSTRACT FROM AUTHOR]

The nucleocapsid protein (NP) of rice stripe virus (RSV) encapsidates viral genomic RNAs to form virion. The binding of NP with RNA is essential for the formation of virus particle. In this study, the binding specificity of RSV NP to RNA and the domains within the NP that mediate this interaction were investigated by gel electrophoretic mobility shift assays and Northwestern blot analysis. The results demonstrated that RSV NP was able to bind to all synthetic RNAs and DNAs without sequence specificity. Using a series of truncated NPs expressed in E. coli and synthetic peptides, we mapped the RNA-binding domain of NP to the central region from amino acid residues 201-232. Further alanine substitution analysis revealed that Lys, Lys, Lys, and Tyr in the RNA-binding domain were essential for NP to bind with RNA. [ABSTRACT FROM AUTHOR]

Rice stripe virus (RSV) is a major rice-infecting virus in Korea that has caused a severe resurgence in the western provinces in 2007-2008. The outbreak of RSV in rice was caused by the small brown planthopper Laodelphax striatellus (an RSV vector), which overwinters in temperate climates. Mass immigration of its insect vector from China has been proposed as a critical risk factor in development of the disease. A nationwide survey was done in June and July 2012-2013 in 30 Korean provinces to confirm RSV sequence variation. The sequence of the nucleocapsid protein (NCP) gene and the intergenic region (IR) of RSV RNA3 was analyzed. A phylogenetic analysis of the NCP gene and the IR demonstrated that the 33 isolates were closely related to earlier reported isolates from Japan and China, but the secondary IR structure showed strong geographic clustering. This is the first report on a nationwide RSV survey of Korea. The results will provide information on the relationships among RSV isolates to understand the epidemiology of this rice disease and the effect of climate changes and insect migration. [ABSTRACT FROM AUTHOR]

The small brown planthopper, Laodelphax striatellus (Fallén) is an important pest of rice in East Asia because it is a vector of rice stripe virus. In this study, we examined the seasonal occurrence of L. striatellus in an agricultural area in central Kyushu in which cropping systems have been changing since the 1970s. The first generation of L. striatellus emerged in the spring, not only on barley, oats, and Italian ryegrass but also on spring crops of forage maize. The second generation of L. striatellus adults emerged in forage maize fields in July, after harvesting of winter crops of barley, oats, and Italian ryegrass. These results suggest that spring crops of forage maize are important hosts of L. striatellus from spring to early summer. The overwintering generation of L. striatellus was collected from rice ratoon, suggesting this is a favorable host for L. striatellus during winter. The appearance of these novel hosts may be a factor in the recent re-emergence of L. striatellus in central Kyushu. [ABSTRACT FROM AUTHOR]

The NS3 protein of rice stripe virus (RSV), encoded by the virion strand of RNA3, is a viral suppressor of RNA silencing (VSR). Rice expressing NS3 had a normal phenotype, was initially sensitive to RSV but recovered at the later stages of infection. RSV accumulated slightly more in transgenic than in wild-type plants at the early stage of infection, but accumulation was similar later. Transgenic rice expressing NS3 also showed enhanced resistance to the fungus Magnaporthe oryzae. Meanwhile, expressional levels of genes related to the salicylic acid (SA) and jasmonic acid (JA) pathways were not significantly altered, indicating that the defense to M. oryzae was independent of the SA and JA pathways. We propose that NS3 may have dual functions, facilitating viral infection as a VSR and inhibiting pathogenic development as an inducer of host defense. [ABSTRACT FROM AUTHOR]

Rice stripe virus (RSV) is the type member of the genus Tenuivirus. RSV is known to have four segmented, single-stranded RNA molecules and causes rice stripe disease in the rice fields of China, Japan, and Korea. Based on the complete genomic sequences of the determined 6 RSV isolates (from Yunnan, Jiangsu, Zhejiang, and Liaoning Provinces, China) and 27 other RSV isolates (from Yunnan, Jiangsu, Anhui, Henan, and Shandong Provinces of China, also Japan and Korea) downloaded from GenBank, we provided a genotyping profile of RSV field isolates and described the population structure of RSV. All RSV isolates, except isolate CX, could be divided into two subtypes, one including 6 isolates from Yunnan Province, and the other including 26 isolates from different parts of China, Japan, and Korea, which were referred to as subtype II and subtype I, respectively. The amino acid distances between subtypes range from 0.053 to 0.085. RSV isolates in Yunnan Province were genetically differentiated from other parts of China, Japan, and Korea and showed infrequent gene flow. The RSV populations collected from other parts of China, Japan, and Korea were only composed of subtype I and showed very low genetic diversity. We speculated that isolate CX may be the result of recombination of isolates from two subtypes. Two potential recombination events were detected in RNA4 of isolate CX. [ABSTRACT FROM AUTHOR]

A twin T-DNA system is a convenient strategy for creating selectable marker-free transgenic plants. The standard transformation plasmid, pCAMBIA 1300, was modified into a binary vector consisting of two separate T-DNAs, one of which contained the hygromycin phosphotransferase ( hpt) marker gene. Using this binary vector, we constructed two vectors that expressed inverted-repeat (IR) structures targeting the rice stripe virus (RSV) coat protein (CP) gene and the special-disease protein (SP) gene. Transgenic rice lines were obtained via Agrobacterium-mediated transformation. Seven independent clones harbouring both the hpt marker gene and the target genes (RSV CP or SP) were obtained in the primary transformants of pDTRSVCP and pDTRSVSP, respectively. The segregation frequencies of the target gene and the marker gene in the T plants were 8.72% for pDTRSVCP and 12.33% for pDTRSVSP. Two of the pDTRSVCP lines and three pDTRSVSP lines harbouring the homozygous target gene, but not the hpt gene, were strongly resistant to RSV. A molecular analysis of the resistant transgenic plants confirmed the stable integration and expression of the target genes. The resistant transgenic plants displayed lower levels of the transgene transcripts and specific small interfering RNAs, suggesting that RNAi induced the viral resistance. [ABSTRACT FROM AUTHOR]

Rice stripe virus (RSV) is one of the most destructive pathogens of rice ( Oryza sativa L.) in East Asia. Development of resistant varieties offers a more economical and efficient way to control this disease. In the present study, tests using four inoculation methods were used on 85 backcross inbred lines of Sasanishiki ( japonica)/Habataki ( indica) to map quantitative trait loci (QTL) conferring resistance to RSV. One QTL on chromosome 3 and two on chromosome 11 were detected, jointly explaining 18-47 % of the trait variance. The QTL ( qSTV11- 1 and qSTV11- 2) on chromosome 11 were closely linked, and mapped in the intervals G257-RM457 and RM457-RM187, respectively. The stabilities of qSTV11- 1 and qSTV11- 2 were validated using a set of 38 established chromosome segmental substitution lines. The two QTL, when combined, showed higher resistance than either of them alone in both field and mass inoculation tests, indicating additivity. Fine mapping of the two genes was carried out using 147 recombined F2:3 lines selected from 2,750 secondary F2 plants of the cross Sasanishiki/SL437. Four SSR (simple sequence repeat) and eight InDel (insertion-deletion) markers newly developed to fine-map the two loci. According to the Nipponbare genomic sequence, qSTV11- 1 was localized to a 333.2-kb interval which was about 230 kb from the well-known Stvb- i. The other locus, qSTV11- 2, which appears to be a new QTL for RSV resistance, was delimited to a 203.9-kb region. Four flanking markers (R15, RM209, R69 and R73) can be used in marker-assisted selection. These results provide an opportunity for map-based cloning of qSTV11- 1 and qSTV11- 2, thereby promoting the breeding program of RSV resistance. [ABSTRACT FROM AUTHOR]

Rice stripe disease (RSD), caused by rice stripe virus (RSV), is a serious disease in temperate rice-growing areas. We have created an RNAi construct containing coat protein gene (CP) and disease specific protein gene (SP) sequences from RSV. The RNAi construct was transformed into two susceptible japonica varieties, Suyunuo and Guanglingxiangjing, to develop resistance against RSD. The homozygous progeny of rice plants in the T and T generations containing RNAi constructs, after self-fertilization were strongly resistant to viral infection. RT-PCR indicated that viral replication of SP and CP in the transgenic plants was significantly inhibited. There were no obvious morphological or developmental differences between the transgenic and wild-type plants from seedling stage to maturity. The excellent agronomic traits of these two varieties, such as high yield and good quality were maintained. Suppression of virus genes using RNAi is therefore a practical and effective strategy for controlling viral infection in crops. [ABSTRACT FROM AUTHOR]

Rice stripe disease, with the pathogen Rice stripe virus (RSV), is one of the most widespread and severe virus diseases. Cultivating a resistant breed is an essential and efficient method in preventing rice stripe disease. Following RNA interference (RNAi) theory, we constructed three RNAi binary vectors based on coat protein (CP), special-disease protein (SP) and chimeric CP/SP gene sequence. Transgenic lines of rice cv. Yujing6 were generated through Agrobacterium-mediated transformation. We inoculated T1 generation plants from each line derived from CP/SP, CP, and SP transgenic rice plants with two RSV isolates from Shandong Province and Jiangsu Province using viruliferous vector insects. In these resistance assays, chimeric CP/SP RNAi lines showed stronger resistance against two isolates than CP or SP single RNAi lines. Stable integration and expression of RNAi transgenes were confirmed by Southern and northern blot analysis of independent transgenic lines. In the resistant transgenic lines, lower levels of transgene transcripts and specific short interference RNAs were observed relative to the susceptible transgenic plant, which showed that virus resistance was increased by RNAi. Genetic analysis demonstrated that transgene and virus resistance was stably inherited in the T2 progeny plants. [ABSTRACT FROM AUTHOR]

The indica variety Dular has a high level of resistance to rice stripe virus (RSV). We performed quantitative trait locus (QTL) analysis for RSV resistance using 226 F2 clonal lines at the seedling stage derived from a cross between the susceptible japonica variety Balilla and the resistant indica variety Dular with two evaluation criteria, infection rate (IR) and disease rating index (DRI). The experiments were performed in both 2004 and 2005. Based on IR, three putative QTLs were detected and had consistent locations in the 2 years, one QTL was detected in the RM7324–RM3586 interval on chromosome 3. The other two QTLs were linked and located in the RM287–RM209 and RM209–RM21 intervals on the long arm of chromosome 11, and accounted for 87.8–57.8% of the total phenotypic variation in both years. Based on DRI, three putative QTLs were also detected and had consistent locations in both years. One of them was located in the RM1124–SSR20 interval on the short arm of chromosome 11, while the other two linked QTLs had the same chromosomal locations on chromosome 11 as those detected by IR, and accounted for 55.7–42.9% of total phenotypic variation in both years. In comparison to the mapping results from previous studies, one of the two linked QTLs had a chromosomal location that was similar to Stv-b i , an important RSV resistance gene, while the other appeared to be a newly reported one. [ABSTRACT FROM AUTHOR]

Rice stripe is the most serious virus disease in temperate rice-growing countries. The most economical and environmentally safe practice for controlling this disease is virus-resistant cultivars. 'BL 1' is an elite germplasm line with the blast resistance gene Pib, and has been used as a differential line for testing the pathogenicity of the blast fungus. We found that certain progenies from BL 1 showed resistance to both blast and rice stripe virus (RSV). The objectives of this study were to evaluate the RSV resistance in the field and under artificial conditions, to assess the reaction to the insect vector(small brown plant hopper, SBPH), and to examine its inheritance and its relationship to blast resistance in BL 1.BL 1 was susceptible to SBPH, but resistant to RSV in field and artificial inoculation tests. The inheritance of RSV resistance in F lines from the cross Nipponbare (NPB)/BL 1 was studied using artificial inoculation with a population of viruliferous SBPH. A serological assay for RSV infection using an enzyme-linked immunosorbent assay (ELISA) was used. RSV resistance in BL 1 was controlled by a single major gene with incomplete dominance. The locus responsible for RSV resistance was genetically independent of the blast resistance gene Pib. The resistance gene for RSV infection in BL 1 was also independent of Stvb-i, a gene widely distributed in resistant Japanese cultivars. Resistance to RSV must be diversified in rice cultivars considering the potential for future emergence of new RSV strains. The new resistance gene identified in BL 1, which has improved plant type and blast resistance, is considered useful for breeding RSV-resistant cultivars in japonica rice. [ABSTRACT FROM AUTHOR]

Phages with high affinity to the S protein obtained fromrice stripe virus (RSV) were enriched fromphage-displayed random 12-mer peptide library after threerounds of biopanning. 9 different peptides from theenriched library were selected by ELISA. Circulardichroism (CD) spectra of the GST-S fusion protein withbinding phages and non-binding phages showed thatstructure of the S protein was changed after it bound toeach of these 9 selected 12-mer peptides, which suggestedthat these peptides might disrupt the function of Sprotein. Thus, those peptides might be used to developplant resistance and disrupt virus transmission. 3 of the12-mer peptide genes were fused with the GST gene in pGEX3X. The fusion proteins were also obtained using E.coli expression system and purified. [ABSTRACT FROM AUTHOR]

The sequences were determined of RNAs 3 and 4 of a Chinese isolate (Y) of rice stripe tenuivirus (RStV) and were compared with those of two RStV isolates (M and T) from Japan. Both RNAs of the Y isolate were longer than those of the M and T isolates. There was almost complete conservation in the 5′ and 3′ non-coding regions for each RNA between the isolates. The analogous ambisense coding regions for each isolate were exactly the same size and the sequences were highly conserved. The major differences were in the intergenic regions, the sizes of which accounted for the differences in size of each RNA of the three isolates. There were no obvious patterns of differences in comparisons of the two RNA over the three isolates. The significance of the similarities and differences in sequences of isolates of RStV separated by more than 3500 km is discussed. [ABSTRACT FROM AUTHOR]