• The Plant Pathology Journal
• /
• 제26권2호
• /
• pp.194-197
• /
• 2010

Most orchids are propagated by tissue culture. To survey the viral infection of tissue cultured Orchids, total RNA was extracted from in vitro Cymbridium and Phalaenopsis spp. collected from companies producing tissue-cultured orchids, and RT-PCR analysis was conducted with primer pairs specific to Cymbidium mosaic virus (CymMV) and Odontoglossum ring spot virus(ORSV), which are infecting wide range of orchid genera. The bulb size of Cymbidium infected with CymMV and ORSV was compared with healthy one at 10 months after planting in vitro orchids in the glasshouse. The CymMV or ORSV infection in 97 Cymbidium and 55 Phalaenopsis plants was 84.5 and 89.1 %, respectively. Mixed infection was found in 52.6 and 47.3% of Cymbidium and Phalaenopsis tested, whereas virus-free orchids were 15.5 and 10.9%, respectively. The CymMV and ORSV reduced the bulb size by 2.7-50% depending on the cultivars of Cymbidium. The both viruses caused yellowing, mottle and mosaic with or without necrosis in 4 Cymbidium cultivars.

• The Plant Pathology Journal
• /
• 제36권5호
• /
• pp.509-514
• /
• 2020

Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

• 식물병연구
• /
• 제28권2호
• /
• pp.92-97
• /
• 2022

Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63℃ for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.

• Journal of Plant Biotechnology
• /
• 제49권3호
• /
• pp.193-206
• /
• 2022

Astringent persimmon (Diospyros kaki Thunb.) is an important fruit crop in Korea; it possesses significant medicinal potential. However, knowledge regarding the pathogens affecting this crop, particularly, viruses and viroids, is limited. In the present study, reverse transcription-polymerase chain reaction (RT-PCR) and high-throughput transcriptome sequencing (HTS) were used to investigate the viruses and viroids infecting astringent persimmons cultivated in Korea. A one-step multiplex RT-PCR (mRT-PCR) method for the simultaneous detection of the pathogens was developed by designing species-specific primers and selecting the primer pairs via combination and detection limit testing. Seven of the sixteen cultivars tested were found to be infection-free. The RT-PCR and HTS analyses identified two viruses and one viroid in the infected samples (n = 51/100 samples collected from 16 cultivars). The incidence of single infections (n = 39/51) was higher than that of mixed infections (n = 12/51); the infection rate of the Persimmon cryptic virus was the highest (n = 31/39). Comparison of the monoplex and mRT-PCR results using randomly selected samples confirmed the efficiency of mRT-PCR for the identification of pathogens. Collectively, the present study provides useful resources for developing disease-free seedlings; further, the developed mRT-PCR method can be extended to investigate pathogens in other woody plants.

• 한국작물학회지
• /
• 제59권4호
• /
• pp.498-504
• /
• 2014

Potato virus Y (PVY) and potato leafroll virus (PLRV) are among the most damaging potato viruses and prevalent in most potato growing areas. In this study, cryopreservation was used to eradicate PVY and PLRV using two cryogenic methods. Potato shoot tips proliferated in vitro were cryopreserved through droplet-vitrification and encapsulation-vitrification using plant vitrification solution 2 (PVS2; 30% glycerol + 15% dimethyl sulfoxide + 15.0% ethylene glycol + 13.7% sucrose) and modified PVS2. Both cryogenic procedures produced similar rates of survival and regrowth, which were lower than those from shoot tip culture alone. The health status of plantlets regenerated from shoot tip culture alone and cryopreservation was checked by reverse transcription-polymerase chain reaction. The frequency of virus-free plants regenerated directly from highly proliferating shoot tips reached 42.3% and 48.6% for PVY and PLRV, respectively. In comparison, the frequency of PVY and PLRV eradication after cryopreservation was 91.3~99.7% following shoot-tip culture. The highest cryopreserved shoot tip regeneration rate was observed when shoot tips were 1.0~1.5 mm in length, but virus eradication rates were very similar (96.4~99.7%), regardless of shoot tip size. This efficient cryotherapy protocol developed to eliminate viruses can also be used to prepare potato material for safe long-term preservation and the production of virus-free plants.

• 한국응용곤충학회지
• /
• 제19권2호
• /
• pp.67-72
• /
• 1980

우리나라에 발병하고 있는 생강모자이크 바이러스병의 기생범위, 물리화학적 성질, 혈청검정 및 전자현미경적 관찰을 조사하였다. 생강의 초기병징은 황록반문을 일으키고 후기에는 위축현상 및 괴경의 소구화를 이룬다. 포장이병율은 약 $43\%$이었다. 즙액전염, 접구전염(core grafting)이 이루어지며, 동부, 오이 Chenopodium amaranticolar, N. tabaccum var. Havana를 비롯하여 23종의 CMV감수성 식물에 병증을 나타낸다. 희석한계점은 $10^{-4}\~10^{-5}$ 범위이고, 온도한계활성점는 $65\~70^{\circ}C$사이이다. 혈청반응은 CMV 항혈청과 양성으로 나타난다. 전자현미경에 의한 바이러스립자의 크기는 $28\~32m\mu$의 구형이다. 이병엽육조직의 세포질 속에는 구형바이러스입자의 밀집상과 유리되어 있는 입자도 관찰되었다. 이상을 종합하여 볼 때 생강에 모자이크병을 일으키는 바이러스는 CMV군에 속하는 바이러스로 믿어진다.

• Journal of Plant Biotechnology
• /
• 제5권2호
• /
• pp.101-105
• /
• 2003

To eradicate several viruses such as PVX, PVY, and PLRV which often cause considerable damages to the growth and yields of potatoes, several stems including shoot tips were excised from the potato plants grown for 50 days and electric shock was treated. Shoot tips excised from electric-shocked stems were transferred into the medium supplemented with antiviral compound, ribavirin to examine the combinatorial effect. When treated only with 20 mg/L ribavirin, PVX concentration in the regenerated plant-lets was slowly decreased as repeating sub-culture and finally, it took 32 weeks to reach completely PVX-free stock. With an electric shock treatment (10 mA electric current), all the replicates became free from PVY. However, PLRV was not completely eradicated from 94P70-4 and 93P29-3 lines even by treating with 10 mA electric shock. In this case, both electric shock and antiviral compound treatments in axillary buds from the stem segment were successful in eradicating viral contamination.

• Journal of Plant Biotechnology
• /
• 제46권3호
• /
• pp.228-235
• /
• 2019

사과(Malus pumila)는 국내에서 가장 경제적으로 중요한 과수 중의 하나이다. 하지만 사과 바이러스 감염은 생산량을 감소시키고 수확량 손실과 과일 품질 저하와 같은 심각한 문제를 야기한다. 국내에 감염된 사과 바이러스 및 비로이드 종류는 Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV)와 Apple scar skin viroid (ASSVd) 등이 알려져 있다. 사과는 바이러스나 비로이드에 감염되어 있어도 대체로 이상한 징후가 발견되지 않아 바이러스로 인해 피해가 많았다. 본 연구는 사과 왜성대목 M.9 및 M.26의 무독묘 생산을 위하여 고온처리($37^{\circ}C$, 6주), 화학처리(Ribavirin) 및 생장점 배양하여 바이러스 제거 처리를 하였다. 바이러스 검출에 일반적으로 사용되는 방법은 효소면역 측정법(ELlSA)과 중합효소연쇄반응(RT-PCR)을 이용하였는데, RT-PCR은 ELlSA방법보다 10 ~ 30% 더 민감하였다. 사과 왜성대목 바이러스 검정 결과, 바이러스 제거 효율은 생장점 배양이 가장 높았다. 생장점 배양 후 바이러스 무병묘의 획득율은 30 ~ 40%로 높게 나타났다. 생장점 배양에서 사과 왜성대목 M.9은 ACLSV, ASPV 및 ASGV의 비율이 각각 45%, 60%, 50%로 높았고, 사과 왜성대목 M.26에서는 ACLSV, ASPV 및 ASGV의 감염율은 각각 40%, 55%, 55%였다. 이상의 결과, 사과 왜성대목에서 무독묘를 생산할 수 있는 가장 효과적인 방법은 생장점 배양에 의한 것으로 판단되었다.

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2003년도 추계학술대회 발표논문 초록집
• /
• pp.161-161
• /
• 2003

• 식물병연구
• /
• 제19권4호
• /
• pp.319-325
• /
• 2013

Soybean yellow mottle mosaic virus(SYMMV)와 Soybean yellow common mosaic virus(SYCMV) 병 발생은 최근에 우리나라에 보고되었지만 두 바이러스 병의 전국적인 분포에 대한 조사가 이루어지지 않아 본 연구팀은 2012년에 영세 농가를 중심으로 전국적인 조사를 실시하였다. 전국에 걸쳐 채집한 682점의 시료를 SYMMV, SYCMV, 그리고 Soybean mosaic virus(SMV) 특이 프라이머를 이용하여 진단한 결과, SMV 102점, SYMMV 116점, SYCMV는 17점이 PCR 양성으로 나타났다. SYMMV와 SYCMV의 복합감염은 검출되지 않았으나, SMV와 SYMMV 혹은 SMV와 SYCMV 복합감염은 각각 1건과 5건이 확인되었다. 처음으로 실시한 영세농가의 콩 바이러스 병 조사를 근거로 추정할 때, 바이러스의 발생은 자가 채종으로 인한 종자전염으로부터 발생되었을 것으로 추정된다. 따라서 바이러스 병의 예방을 위해서는 감염종자 관리 및 무병종자 보급이 무엇보다 필요함을 알 수 있다.