| rt-pcr for detecting five distinct tospovirus species using degenerate primers and dsrna template. | rt-pcr procedures for detection of multiple species of tospovirus from plant tissues were investigated. downstream primers were designated from the 3' untranslated sequences of the s rna. an upstream primer was designated from the degenerated sequences of the nucleocapsid protein. approximately 450 bp dna fragments were detected when tomato spotted wilt virus (tswv)- or impatiens necrotic spot virus (insv)- infected tissues were examined. approximately 350 bp dna fragments were detected when wat ... | 2001 | 11445145 |
| nucleotide sequence of melon yellow spot virus m rna segment and characterization of non-viral sequences in subgenomic rna. | the nucleotide sequence of melon yellow spot virus (mysv) m rna segment was determined. the m rna segment contains one open reading frame (orf) encoding 308 amino acids (aa) in the sense orientation and another orf encoding 1,127 aa in the complementary orientation, which were homologous to the nsm protein and g1/g2 glycoprotein precursor (gp) protein, respectively. amino acid sequences identities with the other tospovirus suggested that mysv is closely related to groundnut bud necrosis virus an ... | 2006 | 16132174 |
| biological and molecular characterization of tospoviruses in thailand. | twenty-eight isolates of tospoviruses associated with tomato, pepper, cucurbits, peanut, and physalis plants collected from fields in different regions of thailand were characterized. on the basis of n gene and protein sequence relationships, three tospoviruses were identified, namely watermelon silver mottle virus (wsmov), capsicum chlorosis virus (cacv), and melon yellow spot virus (mysv). clustal analysis of selected n protein sequences showed different isolates of cacv in three distinct clad ... | 2008 | 18188501 |
| a one-step reverse transcription-polymerase chain reaction system for the simultaneous detection and identification of multiple tospovirus infections. | abstract a one-step reverse transcription-polymerase chain reaction (rt-pcr) method has been developed for the simultaneous detection and identification of multiple tospoviruses that infect plants. the rt-pcr system is composed of six primers in a single tube: a universal degenerate primer and five virus species-specific primers. amplifications resulted in an 848-bp pcr product for watermelon silver mottle virus, 709-bp for tomato spotted wilt virus, 589-bp for impatiens necrotic spot virus, 511 ... | 2005 | 18943986 |
| melon yellow spot virus: a distinct species of the genus tospovirus isolated from melon. | abstract a tospovirus-like virus recovered from netted melon was transmitted by thrips palmi in a persistent manner but had different cytopathological features from tospoviruses previously reported. viral nucleocapsid (n) was purified with two protective reagents, 2-mercaptoethanol and l-ascorbic acid, and rna extracted from the viral nucleocapsid was used for genomic analysis. the virus had a genome consisting of three single-stranded rna molecules. the open reading frame on the viral complemen ... | 2000 | 18944594 |
| serological relationship between melon yellow spot virus and watermelon silver mottle virus and differential detection of the two viruses in cucurbits. | melon yellow spot virus (mysv), a tentative member of the genus tospovirus, is considered a distinct serotype due to the lack of a serological relationship with other tospoviruses in its nucleocapsid protein (np). recently, a virus isolate collected from diseased watermelon in central taiwan (mysv-tw) was found to react with a rabbit antiserum (ras) prepared against the np of watermelon silver mottle virus (wsmov), and a monoclonal antibody (mab) prepared against the common epitope of the nss pr ... | 2010 | 20480192 |
| tomato necrotic ringspot virus, a new tospovirus isolated in thailand. | a new tospovirus isolated from naturally infected tomato plants grown in nakhon pathom province (thailand) was characterized. infected plants showed symptoms consisting of necrotic spots, necrotic ringspots and stem necrosis. this virus was detected using general antibodies that could recognize watermelon silver mottle virus (wsmov), capsicum chlorosis virus (cacv) and melon yellow spot virus (mysv). however, it did not react with specific monoclonal antibodies (mabs) to wsmov and cacv or a spec ... | 2010 | 21104282 |
| antibody array in a multiwell plate format for the sensitive and multiplexed detection of important plant pathogens. | the global seed market is considered to be an important industry with a total value of $10,543 million us dollars in 2012. because plant pathogens such as bacteria and viruses cause a significant economic loss to both producers and exporters, the seed export industry urgently requires rapid, sensitive, and inexpensive testing for the pathogens to prevent disease spreading worldwide. this study developed an antibody array in a multiwell plate format to simultaneously detect four crucial plant pat ... | 2014 | 24945525 |
| implementation of microfluidic sandwich elisa for superior detection of plant pathogens. | rapid and economical screening of plant pathogens is a high-priority need in the seed industry. crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification programs. compared to conventional bench-top detection techniques routinely employed, a microfluidic-based approach offers unique benefits to address these needs simultaneously. to our knowledge, this w ... | 2013 | 24376668 |
| multiplex detection of plant pathogens using a microsphere immunoassay technology. | plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. a novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium acidovorax avenae subsp. citrulli (aac), chilli vein-banding mottle virus (cvbmv, potyvirus), watermelon silver mottle v ... | 2013 | 23638044 |
| generation of marker-free transgenic plants concurrently resistant to a dna geminivirus and a rna tospovirus. | global threats of ssdna geminivirus and ss(-)rna tospovirus on crops necessitate the development of transgenic resistance. here, we constructed a two-t dna vector carrying a hairpin of the intergenic region (igr) of ageratum yellow vein virus (ayvv), residing in an intron inserted in an untranslatable nucleocapsid protein (np) fragment of melon yellow spot virus (mysv). transgenic tobacco lines highly resistant to ayvv and mysv were generated. accumulation of 24-nt sirna, higher methylation leve ... | 2014 | 25030413 |
| development of a protocol for the identification of tospoviruses and thrips species in individual thrips. | a protocol for identifying tospovirus and thrips species in an individual thrips sample was successfully developed. first, an individual thrips was soaked in an rna stabilization solution to preserve protein and nucleic acids and ground in a carbonate buffer containing 0.2% sodium diethyldithiocarbamate. initially, the thrips extracts were screened for tospovirus infection by dot blot analysis using antibodies to nucleocapsid (n) proteins of tospoviruses. thrips extracts with positive results by ... | 2015 | 26141731 |
| development of a multiplex rt-pcr-elisa to identify four distinct species of tospovirus. | in this study, a multiplex rt-pcr-elisa was developed to detect and differentiate four tospovirus species found in thailand, namely capsicum chlorosis virus (cacv), melon yellow spot virus (mysv), tomato necrotic ringspot virus (tnrv), and watermelon silver mottle virus (wsmov). in this system, nucleocapsid (n) gene fragments of four tospoviruses were simultaneously amplified and labeled with digoxigenin (dig) in a single rt-pcr reaction using a pair of degenerate primers binding to the same con ... | 2014 | 24642237 |
| an accurate, specific, sensitive, high-throughput method based on a microsphere immunoassay for multiplex detection of three viruses and bacterial fruit blotch bacterium in cucurbits. | to employ a microsphere immunoassay (mia) to simultaneously detect multiple plant pathogens (potyviruses, watermelon silver mottle virus, melon yellow spot virus, and acidovorax avenae subsp. citrulli) in actual plant samples, several factors need to be optimized and rigorously validated. here, a simple extraction method using a single extraction buffer was successfully selected to detect the four pathogens in various cucurbit samples (cucumber, cantaloupe, melon, and watermelon). the extraction ... | 2017 | 28502647 |