Masatoshi Onuki

Dynamics of Southern rice black-streaked dwarf virus in rice and implication for virus acquisition.

A novel viral disease of rice caused by Southern rice black-streaked dwarf virus (SRBSDV) has spread throughout East and Southeast Asia since the mid-2000s. Outbreaks of this viral disease occur yearly in southern parts of Japan concurrently with overseas migration of the planthopper vector Sogatella furcifera from southern China during the rainy season (from late June to early July). We examined the dynamics (changes in titer and localization) of SRBSDV on rice using reverse-transcription real-time polymerase chain reaction and determined the relationship between virus titer in plants and virus acquisition by S. furcifera. Under a constant temperature of 27°C, a substantial increase of SRBSDV titer in the leaf sheath together with typical symptoms (stunted growth and twisting of leaf tips) was observed at 20 days after the end of a 7-day exposure of viruliferous S. furcifera. Approximately 40% of S. furcifera acquired SRBSDV through feeding for 5 days on rice plants that were infected following exposure to viruliferous vectors for 10 to 15 days. These results suggest that rice infected by S. furcifera can be a source of SRBSDV before the next generation of S. furcifera emerges.

Three distinct groups of isolates of Tomato yellow leaf curl virus in Japan and construction of an infectious clone

Complete nucleotide sequences of eight Japanese isolates of Tomato yellow leaf curl virus (TYLCV) were determined and compared with four TYLCV isolates already reported. These isolates separated into three groups – Shizuoka (Sz), Aichi (Ai), Nagasaki (Ng) – and had 99% identities within the groups. Full-length molecules of DNA-A of group Sz consist of 2791 nt and those of group Ai contain 2787 nt. Both were closely related to TYLCV-Is.M, although those of group Ng had 2793 nt and were more closely related to TYLCV-Is. Comparison of common sequences of isolates belonging to groups Sz and Ai had substitutions of 4 nt in the intergenic region and nonsynonymous substitutions at open reading frames between the groups. None of the isolates tested had DNAβ molecules. Agroinfection of four plant species with a DNA-A dimeric infectious clone of TYLCV-SzY, a member of group Sz, resulted in systemic infection. Tomato plants then developed typical yellow leaf curl symptoms.

Quantitative Analysis of Southern rice black-streaked dwarf virus in Sogatella furcifera and Virus Threshold for Transmission.

We investigated Southern rice black-streaked dwarf virus (SRBSDV) accumulation in a vector insect, the whitebacked planthopper (Sogatella furcifera), to elucidate the association of virus accumulation in the vector with virus transmission efficiency. Real-time quantitative reverse transcription polymerase chain reaction analysis confirmed that this virus is transmitted in a persistent-propagative manner. SRBSDV was successfully transmitted by S. furcifera males in which RNA accumulation of the capsid protein gene of SRBSDV was >10(3) in the whole body of S. furcifera, indicating that the threshold accumulation of the virus RNA for virus transmission is 10(3) in an S. furcifera male. The SRBSDV detection rate in the immigrant population of S. furcifera was high in 2011 (39.5%); however, most of the insects contained fewer than 10(3) RNAs of the capsid protein gene. This result indicates that the risk of SRBSDV epidemics could be estimated from the proportion of virus-transmissible S. furcifera (i.e., S. furcifera that contained more than 10(3) RNAs of the virus capsid protein gene) rather than the SRBSDV detection rate in S. furcifera.

Ageratumyellow vein virus isolated from tomato plants with leaf curl on Ishigaki Island, Okinawa, Japan

In 2005, severe leaf curling and yellowing were observed on tomato plants on Ishigaki Island. Because the symptoms were consistent with infection by a begomovirus, we used a polymerase chain reaction (PCR) with specific primers for begomovirus DNA-A and DNA satellite component (DNA-β) and detected products of the expected sizes from symptomatic tomato plant samples. DNA sequence analyses of the PCR products revealed that the symptomatic tomato plants were associated with Ageratum yellow vein virus (AYVV) infection. We confirmed AYVV transmission from the naturally infected weed host, Ageratum conyzoides, to healthy tomato plants by the insect vector Bemisia tabaci B biotype. This report is the first of AYVV occurrence in Japan.

Quantification of DNA of citrus huanglongbing pathogen in diseased leaves using competitive PCR

A quantification system for huanglongbing pathogen using a competitive polymerase chain reaction method and image-analyzing software were developed to obtain precise results. Significant differences in the quantity of pathogen were thus determined in leaves of two citrus cultivars commonly cultivated in southern Vietnam. Less pathogen-related DNA was detected from the tissue of citrus cultivars that are believed to be more tolerant than susceptible cultivars. The quantification system will be used in studies on pathogen proliferation and movement inside citrus tissue.

Geminate particle morphology of sweet potato leaf curl virus in partially purified preparation and its serological relationship to two Begomoviruses by western blotting

Sweet potato leaf curl virus (SPLCV) is a possible member of the genus Begomovirus; however, the presence of typical geminate particles in sap has not been confirmed. We attempted to observe SPLCV virions by partially purifying the virus using an enzyme-assisted procedure. The observed virions in the partially purified preparations were typical geminate particles with a size of ca. 18×30nm. This virus preparation was subjected to western blot analysis using antisera against Bean golden mosaic virus (BGMV) and Mungbean yellow mosaic virus (MYMV). SPLCV reacted with both antisera. Specific bands appeared to be slightly larger than the 30-kDa marker protein and were considered to be SPLCV coat protein. This western blot analysis revealed for the first time a serelogical relationship between SPLCV and the two well-characterized begomoviruses.

Seasonal occurrence of Laodelphax striatellus (Hemiptera: Delphacidae) in a rice-forage crops mixed cropping area in central Kyushu, Japan

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.

Genomic Structure of a Geminivirus in the Genus Begomovirus from Yellow Vein-affected Eupatorium makinor

A virus from yellow vein-affected Eupatorium makinoi was tentatively designated as Eupatorium-infecting geminivirus (EuGV) on the basis of whitefly transmissibility, electron microscopic observation of geminate particles in sap and symptomatology. EuGV-speciflc DNA fragments were obtained by polymerase chain reaction (PCR). Based on the restriction analysis of the PCR products, EuGV was suggested to have a monopartite genome. We determined the putative, complete nucleotide sequence of the EuGV genome which is comprised of 2766 nucleotides. The EuGV sequence had two virus sense open reading frames (ORF)(V1, V2), four complementary sense ORFs (C1–C4) and a non-coding region termed the intergenic region. This genome structure is quite similar to other monopartite begomoviruses already reported. The nucleotide and amino acid sequences were compared with other begomoviruses. The results supported the conclusion that EuGV is distinct and divergent from other begomoviruses, whereas potential sequence motifs reported in other geminiviruses are well conserved.

Characterization of an isolate of sweet potato feathery mottle virus isolated from sweet potato in Shizuoka Prefecture.

Characterization of an isolate of sweet potato feathery mottle virus isolated from sweet potato in Shizuoka Prefecture. Masatoshi ONUKI, Jun-ichi SAKAI, Kaoru HANADA, Kimihiko KATOZ and Tomio USUGI Kyushu National Agricultural Experiment Station, Nishigoshi, Kumamoto 861-11. Shizuoka Agricultural Experiment Station, Iwata, Shizuoka 438. Okinawa Branch of Japan International Research Center for Agricultural Sciences, Ishigaki, Okinawa 907)