• The Plant Pathology Journal
• /
• v.17 no.3
• /
• pp.164-168
• /
• 2001

Rice black-streaked dwarf virus (RBSDV) and Maize rough dwarf virus (MRDV) are closely related viruses. Since the two viruses produce identical symptoms on maize, barley, and wheat, diagnosis of infected plants is difficult. Previously, we reported that partial cDNA clones of RBSDV S5 and S6 from the Japanese isolate (RBSDV-H) have lower sequence homology to MRDV than do cDNA clones from other genomic segments. In order to test whether cDNA clones of RBSDV-H S5 and S6 can be used for molecular diagnosis, RBSDV field isolates from Korea and from Hokkaido, Japan were tested in dot blot hybridizations probed with RBSDV-H S5 and S6 cDNA colnes. Hybridization with these probes was more intense against the RBSDV genome than against the MRDV genome. Therefore, RBSDV-H S5 and S6 cDNA clones can be used to differentiate between the two viruses. Furthermore, RBSDV-H S5 and S6 clones reacted more strongly against the viruses from stunted maize plants from Korean fields than to MRDV, indicating that RBSDV may be the causal disease agent in maize plants in Korea.

• Research in Plant Disease
• /
• v.22 no.1
• /
• pp.32-37
• /
• 2016

In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice black-streaked dwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted from RBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBank JX994211), a pair of S10 specific primers were designed and used to amplify the fragment encoding the N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR. Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). Recombinant RBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified protein was used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV from infected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, we provide purified RBSDV P10 (1-278 aa), which would be good material for the serological study of RBSDV-miryang isolates.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.7 no.3
• /
• pp.140-145
• /
• 1987

In 1985 and 1986, silage productivity of 8 Korean improved and 7 US introduced corn hybrids and their resistance to rice black-streaked dwarf virus (RBSDV) were tested in the southern part of Korea where RBSDV is prevalent. The results obtained are summarized as followes: 1. Percentage of RBSDV diseased plantes differed depending on the year and hybrid. Suweon 19, Kwangok, Hoengseungok, Jecheonok, Pioneer 3424, Pioneer 3H001, NC6 13 1 were most susceptible, Pioneer 3 160 and Pioneer 3358 were moderately susceptible, and Nampyungok, Suweon 90, Pioneer XCF38, PB 2, and Jinjuok were least suceptible. No hybrid was completely resistant to RBSDV. 2. There were negative correlations between percentage of RBSDV diseased plants and culm length, percentage of ear bearing plants, ear yield, silage yield, or digestible dry matter, but percentage of RBSDV diseased plants was not correlated with stover yield. 3. Silage yield of Jinjuok, Pioneer 3160, Pioneer 3358, and Pioneer XCF38 were higher than that of other hybrids in both years. However, digestible dry matter of Nampyungok, Suweon 89, and Suweon 90 were higher than that of other hybrids due to a higher proportion of ear in 1986 when RBSDV infection was severer.

• Research in Plant Disease
• /
• v.15 no.2
• /
• pp.57-62
• /
• 2009

Genetic diagnosis method of Virion Captured (VC)/RT-PCR for Rice stripe virus (RSV) and Rice black-streaked dwarf virus (RBSDV), Korean major rice viruses transmitted by small brown plant hopper, Laodelphax striatellus, was developed. Virion extraction buffer for rice plant was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. However, the extraction buffer for L. striatellus was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite and 2% polyvinylpyrrolidone wt 40,000 (PVP-40). Specific primers for detection of RSV and RBSDV were selected for VC/RT-PCR method. The specific primers were used as a duplex primer to detect viruliferous small brown plant hopper collected from Gimpo, Pyeongtaek and Siheung areas in Gyeonggi province. The genetic diagnosis methods of single and duplex VC/RT-PCR for RSV and RBSDV could be used easily and economically, especially on the diagnosis of L. striatellus. The rate of viruliferous insect (RVI) for RSV was compared with ELISA and VC/RT-PCR for L. striatellus collected from fields. RVI by ELISA was same as 9.2% with RVI by VC/RT-PCR. However, there were some different detection results between the methods. It could be suggested that there is a possibility of serological and/or genomic differences among RSV isolates. The portion of RVI detected simultaneously by ELISA and VC/RT-PCR was 71.0%, and the detection rate from VC/RT-PCR was higher as 3.2% than that from ELISA, which had a reason of simultaneous detection ability both RSV and RBSDV of VC/RT-PCR.

• The Plant Pathology Journal
• /
• v.21 no.2
• /
• pp.172-173
• /
• 2005

Until now, occurrence of Rice black-streaked dwarf virus (RBSDV) is observed in Gyeongsang provinces, southeastern part of Korea. However, recently, the occurrence of RBSDV is increasing and spreading in Jeonra provinces including Gochang-gun, southwestern part of Korea. RBSDV infected plants showed typical symptoms including stunted, deformed leaves with white waxy or black-streaked swelling along the veins. We extracted viral genomic dsRNA from infected leaves and analyzed dsRNA pattern by polyacrylamide gel electrophoresis. Ten genomic segments with similar sized dsRNAs were observed. We also detected RBSDV by reverse transcription (RT)-PCR using specific primers for S10 from genomic dsRNA and observed amplified DNA fragment specific for RBSDV S10.

• Research in Plant Disease
• /
• v.18 no.4
• /
• pp.387-390
• /
• 2012

Rice black-streaked dwarf virus (RBSDV) was reported to occur on maize plants in Gochang-gun of Jeonllabuk-do region in 2011. The symptoms typically include stunted and deformed leaves. Virus infected plants usually produce poor or no head. RT-PCR analysis of genomic dsRNA extracted from the plant confirmed the infection. Specific primers for full length genome of segments 8 and 10 were used for RNA amplification. Full-length genomes of S8 and S10 were cloned and sequenced. Sequence analysis revealed that the S8 and S10 sequences of the maize isolate were same with rice isolate in size, 1,936 nt and 1,801 nt, respectively. In comparison with rice RBSDV, S8 and S10 showed 94.9-99.6% and 94.1-98.4% sequence identity, respectively. Phylogenetic analysis showed that RBSDV S8 and S10 of maize plants are categorized into the same group as RBSDV of rice plant.

• Applied Biological Chemistry
• /
• v.37 no.3
• /
• pp.148-153
• /
• 1994

Rice black-streaked dwarf virus (RBSDV), a member of the plant reoviridae fijivirus group, causes a serious damage for rice production in Korea. To characterize the RBSDV genome, virus particles were produced by feeding of planthopper (Laodelphax striatellus F.) carring RBSDV to maize plants for 2 days. In $30{\sim}40$ days after feeding, the viral particles were purified from the infected maize roots by using $10{\sim}40%$ sucrose gradient centrifugation. After treatment of 10% SDS to remove the viral coat proteins, ten viral double-stranded RNAs were resolved in agrose gel electrophoresis. Total dsRNA was then used to synthesize cDNA by reverse transcriptase and a cDNA library was constructed in the ${\lambda}gt11$ vector. The phages that contain RBSDV cDNA fragments were selected by hybridizing with the random-primed probe prepared from RBSDV dsRNAs. After subcloning of several cDNA fragments into the pUC19 plasmid vector, one clone (pRV3) was chosen for sequencing. The pRV3 clone was shown to be located on the RBSDV genome fragment No.3 by RNA gel-blot analysis. Sequence analysis of the clone revealed that the pRV3 contains two partial open reading frames.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.33 no.2
• /
• pp.111-116
• /
• 2013

This experiment was carried out to select a resistant corn variety for rice black-streaked dwarf virus (RBSDV) disease in a RBSDV prevalent area (Gochang of Jeollabukdo) from 2006 to 2008. Ten corn varieties for silage preparation were cultivated with first cropping and second cropping system in an RBSDV field, and were tested outbreak rates of RBSDV and dry matter (DM) yield of forage. The outbreak rates of RBSDV were significantly different between corn varieties. Therefore, the resistance degree of corn varieties for RBSDV were divided into 4 groups; very resistant ('Kwangpeyongok' and 'Kwanganok'), resistant ('P3156' and 'P3394'), mildly resistant ('Cheonganok' and 'P32P75') and susceptible ('Suwon19', 'DK697' and 'GW6959') groups respectively. DM yield of forage in RBSDV fields exhibited comparatively significant differences between varieties (p<0.05). DM yield of resistant varieties ('Kwangpeyongok', 'Kwanganok', 'P3156' and 'P3394'), were higher (14~26%) than those of susceptible varieties ('Suwon 19', 'DK697' and 'GW6959'). Therefore, resistant varieties were recommended for increasing forage productivity in field of RBSDV prevalent areas.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.31 no.2
• /
• pp.156-161
• /
• 1986

Silage productivity and resistance to rice black streaked dwarf virus (RSDV) of six Korean-improved and six US introduced corn genotypes were tested in the southern part of Korea. There was a negative correlation between culm length retarded by RBSDV and coefficients of variance of culm length. Frequency distribution of culm length could be classified as three genotypic groups according to the type of distribution and percentage of RBSDV diseased plants. There were negative correlations between percent RBSDV diseased plants at harvest and culm length, percent ear bearing plants, silage yield, or ear yield, but percent RBSDV diseased plants did not related to the ear/silage ratio and stover yield. Silage yield of Pioneer XCF38 was highest, but that of Suweon 89 and NC 6131 was lowest. However, there was not signi-ficant difference in silage yield among the remaining genotypes. Pioneer XCF38, Suweon 89, and Jinjuok were quite resistant to RBSDV, but Suweon 19, Kwangok, Hoengseongok, Jecheonok, and Pioneer 3424 were susceptible and NC 6131 was most susceptible to RBSDV. Although Jinjuok and Suweon 89 were resistant to RBSDV, silage yield was not high because of early senescence of leaves after silkillg.

• The Plant Pathology Journal
• /
• v.40 no.4
• /
• pp.408-413
• /
• 2024

The emergence of rice black-streaked dwarf virus (RBSDV) poses a significant threat to global cereal crop cultivation, necessitating the urgent development of reliable detection and quantification techniques. This study introduces a reliable approach for the precise and sensitive quantification of the RBSDV in cereal crop samples, employing a reverse transcription digital polymerase chain reaction (RT-dPCR) assay. We assessed the specificity and sensitivity of the RT-dPCR assay proposed for precise RBSDV detection and quantification. Our findings demonstrate that RT-dPCR was specific for detection of RBSDV, with no cross-reactivity observed with other viruses infecting cereal crops. The RT-dPCR sensitivity was over 10 times that of RT-quantitative PCR (RT-qPCR). The detection limit of RT-dPCR was 0.096 copies/㎕. In addition, evaluation of RT-dPCR assay with field samples was conducted on 60 different cereal crop samples revealed that RT-dPCR (45/60) exhibited superior accuracy compared with RT-qPCR (23/60). In this study, we present a specific and accurate RT-dPCR assay for the detection and quantification of RBSDV.