• Korean journal of applied entomology
• /
• v.57 no.4
• /
• pp.393-399
• /
• 2018

A loop-mediated isothermal amplification (LAMP) primer set (WBPH-65) was designed for the species-specific detection of white-backed planthopper (WBPH) Sogatella furcifera based on the full-length sequence of the internal transcribed spacer 2 (ITS2) (KC417469.1). The WBPH-65 primer set consists of six primers (total 165 bp), F3 (18 bp), B3 (18 bp), FIP (43 bp), BIP (40 bp), LF (21 bp), and LB (25 bp). After the LAMP reaction of three rice planthoppers, S. furcifera, Nilaparvata lugens, and Laodelphax striatellus, with the WBPH-65 primer set for 60 min at $65^{\circ}C$, the LAMP products were observed in the genomic DNA of S. furcifera only. According to the DNA amount of S. furcifera and incubation duration at $65^{\circ}C$, the difference of fluorescence relative to the negative control (0 ng) was clearly observed in a 40-min incubation with 10 and 100 ng or in case of 60-min incubation with 0.01, 0.1, 1, 10, and 100 ng. There was little difference in fluorescence between the negative control and all the other DNAs tested in 20- and 30-min incubations. On the other hand, the WBPH-65 primer set without LF and LB primers showed little amplification in the genomic DNAs of the three rice planthoppers, S. furcifera, N. lugens, and L. striatellus in a 60-min incubation. These results suggest that all six primers (F3, B3, FIP, BIP, LF, and BF) are necessary for the WBPH-65 primer set to detect S. furcifera within a 60-min incubation, and is able to discriminate S. furcifera from at least N. lugens and L. striatellus.

• Research in Plant Disease
• /
• v.22 no.2
• /
• pp.72-80
• /
• 2016

Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.

• Research in Plant Disease
• /
• v.26 no.4
• /
• pp.264-271
• /
• 2020

We have developed a simultaneous diagnostic method that can identify both the species of thrips and tomato spotted wilt virus (TSWV) that are problematic in chrysanthemum plants. This is a method of amplifying DNA by performing reverse transcription-polymerase chain reaction by simultaneously adding primers specific to TSWV coat protein (N) gene and primers specific to the internal transcribed spacer 2 region of Frankliniella occidentalis and F. intonsa using total nucleic acid extracted from one thrips. The sizes of DNA fragments for TSWV, F. occidentalis, and F. intonsa were 777, 287, and 367 bp, respectively. These results showed species identification of thrips and whether thrips carrying TSWV can be simultaneously confirmed. Further usefulness of the simultaneous diagnostic method was made from greenhouse survey at chrysanthemum greenhouses in Taean (Chungcheongnam-do) and Changwon (Gyeongsangnam-do) to investigate the identification of thrips species and the rate of thrips carrying TSWV. Of thrips collected from the greenhouses, 83.7% thrips was F. occidentalis and 72.9% F. occidentalis carried TSWV in Taean. Similarly, the diagnostic method showed that 92.2% thrips was F. occidentalis and 84.0% F. occidentalis carried TSWV in Changwon. These results confirm that F. occidentalis is a dominant thrips species and the thrips species plays a crucial role in the transmission of TSWV in chrysanthemum plants in the greenhouses. Taken together, this study showed a simple diagnostic method for thrips identification and epidemiological studies of the timing and spread of TSWV through thrips in chrysanthemum greenhouses in South Korea.

• Horticultural Science & Technology
• /
• v.33 no.3
• /
• pp.409-419
• /
• 2015

This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.

• Research in Plant Disease
• /
• v.27 no.4
• /
• pp.172-179
• /
• 2021

This study aimed to assess the disease incidence and distribution of toxigenic in Korean triticale. The pathogen of triticale that cause Fusarium head blight were isolated from five different triticale cultivars that cultivated in Suwon Korea at 2021 year. The 72 candidate were classified as a Fusarium asiaticum by morphology analysis and by ITS1, TEF-1α gene sequence analysis. And the results of pathogenicity with 72 isolates on seedling triticale, 71 isolates were showed disease symptom. Also, seven out of 71 Fusarium isolates were inoculated on the wheat, to test the pathogenicity on the different host. The results showed more low pathogenicity on the wheat than triticale. The results of analysis of toxin type with 72 isolates, 64.6% isolates were produced nivalenol type toxin and other 4.6% and 30.8% isolates were produce 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol, respectively. To select fungicide for control, the 72 Fusarium isolates were cultivated on the media that containing four kinds fungicide. The captan, hexaconazole, and difenoconazole·propiconazole treated Fusarium isolates were not showed resistance response against each fungicide. However, six isolates out of 72 isolates, showed resistance response to fludioxonil. This study is first report that F. asiaticum causes Fusarium head blight disease of triticale in Korea.