• 한국작물학회지
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• 제57권3호
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• pp.254-261
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• 2012

고구마의 연작장해를 경감시키기 위하여, 연작지 토양과 심토반전 토양에서 농가묘와 바이러스 무병묘를 $70{\times}25cm$ 간격으로 재배하였다. 연작지 토양의 시비량은 $N-P_2O_5-K_2O$ = 55-63-156$kg\;ha^{-1}$과 우분퇴비 10$ton\;ha^{-1}$으로 표준시비를 하였고, 심토반전 토양은 질소비료와 퇴비만을 50% 증시하였다. 삽식 30, 120일째의 생육과 수량 및 품질특성을 조사한 결과는 다음과 같다. 1. 삽식 30일경부터 농가묘에서는 바이러스 병징이 뚜렷하였으나, 무병묘에서는 나타나지 않았다. 2. 무병묘의 수량성은 농가묘보다 심토반전 토양 15.0%, 연작지 10.5%의 증가를 보였다. 3. 심토반전 토양에서의 수량성은 연작지보다 농가묘 8.8%, 무병묘 3.2%의 증가를 보였으며, 심토반전 토양에서 바이러스 무병묘 재배는 상저비율이 농가묘(60.1%) 대비 80%로 높아져 경제성이 있었다. 4. 무병묘에서 수확한 고구마의 품질은 농가묘보다 피색이 선명하고, 고구마 모양이 좋아져 외관 품질향상에도 유리하였다. 5. 심토반전 토양 및 무병묘에서 수량증가는 30일째 엽수와 120일째 분지수와 정의 상관관계(p=0.05)가 인정되었으며, 이는 고구마 괴근형성에 초기생육이 중요하다는 것을 보여주었다.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.155-160
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• 2003

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.

• Journal of Plant Biotechnology
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• 제32권1호
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• pp.45-50
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• 2005

바이러스에 감염된 감귤나무를 생육상에서 주간 $40^{\circ}C$, 야간 $30^{\circ}C$로 열처리한 후 발생한 신초를 $0.3\~0.7\;\cal{mm}$ 로 잘라내어 기내에서 치상한 탱자 (P. trifoliata) 대목에 접목하는 열처리와 경정접목 (shoot-tip grafting)을 병행한 방법으로 무독묘 생산에 성공하였다. 감글 바이러스 즉, citrus tristeza virus (CTV), satsuma dwarf virus (SDV) and citrus tatter leaf virus (CTLV) 에 대한 무독화율은 각각 $75.7\%,\;100.0,\%\;82.6\%$ 였다. 경정접목 시 접수크기는 $0.3\;\cal{mm}$ 이하 일때 바이러스 무독화율이 높았으나 절목 후 활착율은 저하되었다. 열처리 후 경정배양 (shoot-tip culture)을 하여 1 개월 후에 활착율을 조사한 결과 Yuzu (C. junos)는 $74.6\%$, 그리고 early satsuma mandarin (Iwasagi)은 $13.3\%$를 나타내었다. 그러나 항바이러스제인 rivabirin을 첨가한 후 무독효과를 보기 위하여 후대배양 중에 온주밀감의 특성상 갈변되어 고사함으로서 무독묘를 생산하지 못했다.

• 식물조직배양학회지
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• 제25권4호
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• pp.273-276
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• 1998

생장점 배양에 의한 바이러스 무병주 생산 여부를 판단하고자 바이러스 감염실태를 조사한 결과 지황1호와 국내재래종 모두 여러 종류의 바이러스에 감염되어 있었고 주종바이러스는 potexvirus와 TMV였다. 생장점 배양시 신초형성은 kinetin 첨가 배지에서 가장 양호하였으며 2.4-D 첨가 배지에서는 캘러스로 발달하였는데 캘러스로부터 다수의 신초를 획득할 수 있었다. 기내 재분화 식물체의 투과전자현미경(TEM)과 지표식물을 이용한 바이러스 검정결과 생장점 배양 유래의 식물체들은 83.3%가 바이러스 무병주임을 확인할 수 있었다.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

• 한국자원식물학회지
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• 제23권3호
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• pp.233-241
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• 2010

In vitro elimination of Sweet potato leaf curl virus (SPLCV) from infected sweet potato is difficult due to low number of virus-free plants obtained from meristem tip culture and long growth period required for the virus detection. In this study, efficient production of the SPLCV-free sweet potato by in vitro therapy coupled with a PCR assay for virus detection was investigated. Infected shoots cultured on Murashige and Skoog medium were treated at three different temperatures for 7 weeks followed by meristem tip culture on the medium with or without ribavirin at 50 mg/L. The regenerated plantlets were tested for virus infection by a PCR assay. The results showed that the both heat- and cold-treatments, and addition of the ribavirin did not have significant effect on efficiency of the virus elimination. The meristem size, however, greatly affected the survival rate. Meristems sized over 0.4 mm survived better than smaller ones (0.2-0.3 mm). The PCR assay was approved to be a rapid, sensitive and reliable for the SPLCV detection in regenerated plantlets. Therefore, combination of cultivating meristem tips sized 0.4-0.5 mm on the medium at $22^{\circ}C$ without ribavirin and detection of SPLCV in the regenerated plantlets by the PCR assay was an efficient system for the SPLCV elimination from infected sweet potato.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.221-227
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• 2004

Protocol was established for production of virus free healthy seeds using meristem ($0.3-0.5\;\cal{mm}$ in size) culture and field management under net house condition in tomato. The isolated meristem was found well established in MS liquid medium containing $0.1\;\cal{mg}\;1^{-1}\;of\;GA_3$. For shoot and root development either from primary meristem or from nodal segment of meristem derived plants, semisolid MS medium having $0.5\;\cal{mg}\;1^{-1}$ of IBA was found most effective. The elimination of the studied viruses (ToMV, CMV, ToLCV) in meristem-derived plants was confirmed by DAS-ELISA test. For field management of the virus eradicated meristem-derived plants, use of net house was found very effective measures to check viral vector visit and eventually infection. The meristem-derived plants were vigor and high yielder than the native seed derived plants and produced healthy seeds. Due to stop vector visit, no viral symptoms were observed in both $R_1\;and\;R_2$ plants cultivated in net house condition. Starting of viral infestation was observed in $R_2$ generation when they were planted in open house condition without control of vector visit. Therefore, for management of viral diseases, use of virus free meristem derived plantlets and their subsequent cultivation in soil under net house condition without using any vector killing insecticide can be recommended for producing healthy seeds in tomato. The developed protocol for environmentally healthy tomato seed production in Bangladesh may be used in the countries having similar tropical like environment conducive for viral vector visit.

• The Plant Pathology Journal
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• 제33권4호
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• pp.382-392
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• 2017

This study reports the findings of a distinct Potato virus Y (PVY) isolate found in Northeast China. One hundred and ten samples (leaves and tubers) were collected from potato plants showing mosaic symptoms around the city of Harbin in Heilongjiang province of China. The collected tubers were planted and let to grow in a greenhouse. New potato plants generated from these tubers showed similar symptoms, except for one plant. Subsequent serological analyses revealed PVY as the causing agent of the disease. A novel PVY isolate (referred to as HLJ-C-44 in this study) was isolated from this sample showing unique mild mosaic and crisped leaf margin symptoms. The complete genome of this isolate was analyzed and determined. The results showed that HLJ-C-44 is a typical PVY isolate. Phylogenetic analysis indicated that this isolate belongs to the N-Wi strain group of PVY recombinants ($PVY^{N-Wi}$) and also shared the highest overall sequence identity (nucleotide and amino acid) with other members of this strain group. However, recombination analysis of isolate HLJ-C-44 revealed a recombination pattern that differed from that of other $PVY^{N-Wi}$ isolates. Moreover, biological assays in four different potato cultivars and in Nicotiana tabacum also revealed a different phenotypic response than that of a typical $PVY^{N-Wi}$ isolate. This data, combined, suggest that HLJ-C-44 is a novel PVY recombinant with distinct biological properties.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 추계학술대회
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• pp.33-40
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• 2002

We, Tong Yang Moolsan Co. Ltd. (TYM) set up the mass-production system for virus-free seed garlic via tissue culture technique. TYM's tissue culture technique is called as 'Multiple shoot propagation technique'. This technique can lead mass propagation of genetically homogeneous seed garlic in a short period because of its highly proliferation rate of in vitro shoots ($15^{10}$ /year). TYM researchers applied the technique to some selected garlic cultivars with superior characteristics and carried out field test of productivity in the inside and outside of the country for several years. According to the yearly results of field test with virus-free seed garlic, we ascertained that virus-free seed garlic can produce the highly yield increase (max. above $50\%$) and also can enhance the product quality. Consequently, we estimated that TYM's seed garlic will contribute to farmers with increase of income and can elevate the national position of garlic market in the world for its competitive power of technical and production cost.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.54-54
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• 2019

Apple (Malus domestica) is one of the most economically important fruits in Korea. But virus infection has decreased sustainable production of apple and caused the serious problems such as yield loss and poor fruit quality. Virus or viroid infection including Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV) and Apple scar skin viroid (ASSVd) has been also reported in Korea. In many cases, apple is infected with virus and viroid with no specific symptoms, the damage caused by the virus are unaware significantly. In our research, we tried to eliminate viruses in the rootstock for the disease-free seedlings of the apple dwarfing rootstock M.9 and M.26. The method of virus elimination was meristem culture, heat($37^{\circ}C$, 6weeks) treatment and chemistry($Ribavirin^{(R)}$) treatment. The analytical methods commonly used for the detection of virus is Enzyme-linked Immuno-Sorbent Assay(ELlSA) and Reverse Transcription-polymerase Chain Reaction(RT-PCR). RT-PCR method was more 30% sensitive than ELISA method. Efficiency of method eliminate virus appeared meristem method > heat treatment > chemistry treatment. The higher acquisition rate of disease-free seedlings is 30~40% on meristem treatment. In meristem treatment, the apple dwarfing rootstock M.9 gained infection ratio of ACLSV, ASPV and ASGV were 45%, 60% and 50% respectively. In the apple dwarfing rootstock M.26, infection ratio of ACLSV, ASPV and ASGV were 40%, 55%, 55%, respectively. Based on our results, it was found that most effective method of disease-free seedlings apple dwarfing rootstocks was by meristem treatment than heat method and chemistry treatment.