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열처리와 생장점 배양 및 항바이러스제 처리에 의한 포도 GLRaV-3의 무독화효과
김현란,정재동,박진우,최용문,임명순,Kim, Hyun-Ran,Chung, Jae-Dong,Park, Jin-Woo,Choi, Yong-Mun,Yiem, Myoung-Soon 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.2
Grapevine leafroll-associated virus 3(GLRaV-3) is one of the most severe pathogens for viral diseases found in Korea. This study was conducted to establish the virus-free stock production system for the virus disease control. The effects of thermotherapy, merestem culture and chemotheratpy to eliminate the GLRaV-3 in gratevines were tested. Thermotherapy at 37$\pm$2$^{\circ}C$ for 6∼8 weeks combined with 0.5∼1.0mm size of meristem culture method was the most effective for virus elimination. Thermotherapy alone was not effective. In chemotheratpy, DHT and Amantadine (20, 40mg/L) treatment in medium was more effective than Ribavirin to eliminate the GLRaV-3 in grapevine. However, Ribavirin spraying to potted was not available for virus elimination. Therefore, virus-free stock production system using the thermotherapy combined with shoot apical meristem culture was the most effective in grapevine.
Identification of Grapevine leafroll-associated virus 3 Ampelovirus fromGrapevines in Korea
김현란,이신호,이봉춘,김영태,박진우 한국식물병리학회 2004 Plant Pathology Journal Vol.20 No.2
Grapevine leaf roll-associated virus 3 (GLRaV-3) is one of the most important viral diseases of grapevine in the world. In this study, GLRaV-3 Ampelovirus was identified from grapevines in Korea by analyzing viral coat protein size, nucleotide, and amino acid sequences. The molecular weight of viral coat protein from virusinfected in vitro plantlets was determined by western blot using a commercial GLRaV-3 polyclonal antibody. Western blot analysis showed a coat protein of about 43 kDa. RT-PCR product of about 942 bp which encoded the coat protein (CP) gene was amplified with specific primers. When the viruses existed at low titers in the host plant, the dsRNA had very specific template in RTPCR amplification of fruit tree viruses. Especially, small-scale dsRNA extraction method was very reliable and rapid. Sequence analysis revealed that the CP of the GLRaV-3 Ko consisted of 942 bp nucleotide, which encoded 314 amino acid residues. The CP gene of GLRaV-3 Ko had 98.9% nucleotide sequence and 98.7% amino acid sequence identities with earlier reported GLRaV-3. This is the first report on molecular assay of GLRaV-3 Ampelovirus identified from Korea. The GLRaV-3 Ko CP clone would be very useful for breeding of virus resistant grapevines.
김현란,이신호,이동혁,박진우,김정수 한국식물병리학회 2006 Plant Pathology Journal Vol.22 No.1
Apple scar skin, one of the most destructive diseases affecting apple, is caused by Apple scar skin viroid (ASSVd). Fruit dappling appeared on several cultivars in Korea and has been distributed to major cultivated areas since 2001. ASSVd was identified from infected fruits by using nucleic acid sequence-based amplification with electrochemiluminescence (NASBA-ECL). NASBA-ECL method was faster and hundredfold more sensitive than reverse transcription-polymerase chain reaction (RT-PCR) for ASSVd detection in apple leaves/ stems. ASSVd was rapidly transmitted to the entire tree in the second year after artificial inoculation. The ASSVd could be transmitted efficiently by using contaminated pruning scissors to both lignified stems (60 to 70%) and green shoots (20 to 40%) of apple tree and young plants. Dipping of contaminated scissors in 2% sodium hypochlorite solution effectively prevented viroid transmission. In the ASSVd-infected fruits, the viroid was easily detected from fruit skin, seed coat, and embryo. Moreover, embryo and endosperm separately excised from the ASSVd-infected seeds were ASSVd positive in NASBA-ECL assay. Seedlings germinated from ASSVd-positive seeds showed 7.7% infection rate., which indicated that ASSVd is seed-borne.