• Title/Summary/Keyword: Virus-free plant

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Virus Free Stock Production by In vitro Stem Cutting of Shoot Tip Cultures of Grapes (포도 경정배양에서 얻은 유묘의 기내삽목에 의한 무병묘 생산)

  • 서정해;정재동;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.81-85
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    • 2001
  • The experiment was conducted to know the effect of plant growth regulators on axillary bud elongation from in vitro stem cutting and the possibility of virus-free stock production. Axillary buds were well elongated in 3/4 strength MS medium supplemented with 0.1 or 0.5 mg/1 BA and 0.05 mg/1 NAA. Transferred plantlets could be established well in vermiculite and peat moss mixture (3:1, v/v) compare to other mixtures. In virus indexing, all the varieties of mother plants were infected by GLRV Ⅲ. Infected percentages of the three varieties were ranged from 30% to 75%. But negative response was revealed against the other species of virus, GLRV Ⅰ, GFLV and ArMV. Plantlet of 'Schuyler' and 'Muscat of Alexandria', which were cultured in vitro, showed positive response against GLRV Ⅲ and infected percentage of the former was 37.5% but the latter, 12.5%. On the other hand, that of 'Campbell Early' negativiely responded against all the species of virus indexed.

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Detection and Quantification of Apple Stem Grooving Virus in Micropropagated Apple Plantlets Using Reverse-Transcription Droplet Digital PCR

  • Kim, Sung-Woong;Lee, Hyo-Jeong;Cho, Kang Hee;Jeong, Rae-Dong
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.417-422
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    • 2022
  • Apple stem grooving virus (ASGV) is a destructive viral pathogen of pome fruit trees that causes significant losses to fruit production worldwide. Obtaining ASGV-free propagation materials is essential to reduce economic losses, and accurate and sensitive detection methods to screen ASGV-free plantlets during in vitro propagation are urgently necessary. In this study, ASGV was sensitively and accurately quantified from in vitro propagated apple plantlets using a reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assay. The optimized RT-ddPCR assay was specific to other apple viruses, and was at least 10-times more sensitive than RT-real-time quantitative PCR assay. Furthermore, the optimized RT-ddPCR assay was validated for the detection and quantification of ASGV using micropropagated apple plantlet samples. This RT-ddPCR assay can be utilized for the accurate quantitative detection of ASGV infection in ASGV-free certification programs, and can thus contribute to the production of ASGV-free apple trees.

Elimination of SPFMV from Virus-infected Sweet Potato Plants through Apical Meristem Culture

  • Kim, Young-Seon;Jeong, Jae-Hun;Park, Jong-Suk;Eun, Jong-Seon
    • Plant Resources
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    • v.7 no.3
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    • pp.200-205
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    • 2004
  • Sweet potato infected with a viral disease (SPFMV) showed irregular chlorotic patterns, so called feathering associated with faint or distinct ring spots that have purple-pigmented borders. SPFMV was eliminated from sweet potato plants using meristem tip culture. MS medium supplemented with BAP (2mg/L) and NAA (0.05 mg/L) was used for shoot proliferation and 1/2 MS medium for rooting of the plants. Highest percentage of regenerated plants (60%) was obtained from the optimum size (0.3-0.5mm) meristem tips. Of these, 60% plants were found negative for SPFMV by RT-PCR. Virus detection by RT-PCR was found to be a reliable method. Meristem-tip culture to produce SPFMV-free quality sweet potato and virus detection by RT-PCR is an efficient, time saving and reliable method for production of SPFMV-free tissue culture raised plants.

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In vitro micrografting for production of Indian citrus ringspot virus (ICRSV)-free plants of kinnow mandarin (Citrus nobilis Lour × C. deliciosa Tenora)

  • Singh, B.;Sharma, S.;Rani, G.;Hallan, V.;Zaidi, A.A.;Virk, G.S.;Nagpal, A.
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.137-143
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    • 2008
  • Production of Indian citrus ringspot virus (ICRSV)-free plants from an infected plant of kinnow mandarin (Citrus nobilis Lour ${\times}$ C. deliciosa Tenora) is reported. The shoot apices of different sizes (0.2-1.0 mm) excised from the ICRSV-infected plant were micrografted onto decapitated rootstock seedlings of rough lemon (C. jambhiri). Micrograft survival depended on the size of shoot apex and the sucrose concentration of the culture medium. Increase in scion size from 0.2 to 0.7 mm resulted in an increase in micrografting success rate from 30.55 to 51.88%. Further, micrograft survival obtained with 0.2 mm was improved from 30.55 to 38.88% by increasing sucrose concentration in the culture media from 5 to 7.5%. The micrografted plants were tested for ICRSV using ELISA and RT-PCR. All plants raised from 0.2-mm scion were found negative with both ELISA and RT-PCR whereas only 20% of the ELISA negative plants raised from 0.3-mm scion were found negative for ICRSV with RT-PCR. The outcome of this research is the successful establishment, acclimatization and virus testing of micrografted plants.

Review on the Occurrence and Studies of Potato Viral Diseases in Korea (한국에서의 감자 바이러스병 발생과 그 연구에 대한 고찰)

  • Hahm, Young-Il
    • Research in Plant Disease
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    • v.9 no.1
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    • pp.1-9
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    • 2003
  • The occurrence of potato(Sotanum tuberosum) viral diseases caused by Potato virus X(PVX), Potato virus Y (PVY), Potato leafroll virus(PLRV), Potato vims S(PVS), Potato virus M(PVM), Potato virus A(PVA), Potato virus T(PVT), Alfalfa mosic virus(AIMV), Tobacco mosic virus(TMV), Potato mop top virus(PMTV) Tobacco rattle virus(TRV) and Potato spindle tuber viroid(PSTVd), potato witches' broom phytoplasma, have been identified so far in Korea. Major viral diseases such as PVX, PVY and PLRV had been studied more deeply, however, the others are just identified and only partially characterized since the first study on the relation between PVX nucleic acid and virus protein by Kim in 1961. The most studies on potato viral diseases are mainly focused on the problems of seed potato production. The National Alpine Agricultural Experiment Station(NAAES), since it began its activities in 1961, has given special attention to this problem by doing studies to identify, characterize and control potato virus diseases. This effort resulted in the development of new potato virus detection methods as a basis for elaborating new method of control, such as the production of seed potato free of virus and the selection of new virus-resistant transgenic potatoes. The further studies of potato viral diseases required would be fallowings: the continuous monitoring for the occurrence of identified or not identified potato viruses in Korea, the isolation of resistant viral genes, the development of control method for the non-persistently transmitted viruses like PVY, special vectors such as nematode and fungus transmitted viruses, TRV and PMTV and the development of control methods against potato viral diseases by viral cross protection, therapy, transgenic plant, and the use of the agents or molecules, such as virus inhibitors and antiviral proteins, etc., blocking viral replication.

Incidence of Viral Infection of Main Garlic Growing Areas of Gangwon Province in Korea (강원도 마늘 주산지 재배마늘의 바이러스 감염 실태)

  • Shin, Ji-Eun;Lee, Yeong-Gyu;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.233-236
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    • 2013
  • This research was conducted to investigate the incidence of viral infection in Samcheok-shi and Jeongseon-gun, main garlic growing areas of Gangwon province in Korea. Four primers sets (GLV, LYSV, GCLV, OYDV) were used for RT-PCR to test of 100 samples collected from each location. Infection rates of GLV, LYSV, GCLV and OYDV of garlic samples from Samcheok-shi and Jeongseon-gun were 95, 95, 92 and 33%, respectively. All garlic samples tested in this research were infected at least one virus. Coinfection of 3 kinds of virus (GLV, LYSV and GCLV) and 4 kinds of virus (GLV, LYSV, GCLV and OYDV) were 60 and 25%, respectively. While infections of GLV, LYSV and GCLV were evenly found in both two regions, OYDV infection of garlic samples collected from Jeongseon-gun was higher than Samcheok-shi. Viral infections at garlic fields of whole country and Gangwon province are very serious now. Therefore, it is very necessary to develop technic for virus-free bulb and renew seed bulb of garlic.

Combining ex vitro thermotherapy with shoot-tip grafting for elimination of virus from potted apple plants (기외 열처리와 경정접목을 이용한 사과 폿트묘에서의 바이러스 제거)

  • Chun, Jae An;Gwon, Jiyeong;Lee, Seon Gi
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.222-229
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    • 2022
  • Apples are the most grown fruit crops in the fruit industry of Korea. However, virus or viroid infection such as apple mosaic virus (ApMV), apple stem grooving capillovirus (ASGV), apple stem pitting virus (ASPV), apple chlorotic leaf spot virus (ACLSV), apple scar skin viroid (ASSVd) causes fruit yield reduction and poor fruit quality. Therefore, in this study, we examined to established an efficient virus-free system to eliminate the most infected ASGV virus in domestic apple orchard. We investigated that the shoot growth rate and the virus removal rate in ASGV infected potted apples that were treated with heat treatment in a growth chamber (constant temperature/humidity device) maintained at 36℃, 38℃ and 40℃ for 4 weeks. Here we found that the shoot growth rate was the highest in the heat treatment group (36℃) and the virus was removed in the middle and top of the shoot but not in the bottom. The virus was did not removed in the 38℃ and 40℃ heat treatment group in all section of shoots, and the heat treatment group (40℃) died after 4 weeks of heat treatment without growth of shoots. We performed in vivo shoot-tip grafting using the shoot-tip of potted apple heat-treated at 36 ℃, and we also investigated the viability and virus removal rate, which showed 94% viability and 20% virus removal rate. Collectively, our results suggest that it would be possible to produce the virus-free apple plants through heat treatment and shoot-tip grafting.

Rapid and Specific Detection of Apple stem grooving virus by Reverse Transcription-recombinase Polymerase Amplification

  • Kim, Nam-Yeon;Oh, Jonghee;Lee, Su-Heon;Kim, Hongsup;Moon, Jae Sun;Jeong, Rae-Dong
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.575-579
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    • 2018
  • Apple stem grooving virus (ASGV) is considered to cause the most economically important viral disease in pears in Korea. The current PCR-based methods used to diagnose ASGV are time-consuming in terms of target detection. In this study, a novel assay for specific ASGV detection that is based on reverse transcription-recombinase polymerase amplification is described. This assay has been shown to be reproducible and able to detect as little as $4.7ng/{\mu}l$ of purified RNA obtained from an ASGV-infected plant. The major advantage of this assay is that the reaction for the target virus is completed in 1 min, and amplification only requires an incubation temperature of $42^{\circ}C$. This assay is a promising alternative method for pear breeding programs or virus-free certification laboratories.

A COMPARATIVE STUDY OF FREE AMINO ACIDS IN HEALTHY AND VIRUS DISEASED CHINESE DATE TREE (Virus에 감염된 대추나무의 병엽과 건전엽에 있어서의 유이 amino산의 정성적 비교)

  • Hong, Soon-Woo;Hah, Yung-Chil
    • Journal of Plant Biology
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    • v.4 no.1
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    • pp.9-11
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    • 1961
  • A comparative investigation of free amino acids content in healthy check and virus diseased leaves of Chinese date tree, Zyzyphus jujuqa Mill var. inermis Rhed, was carried out by authors throughout the growing season of 1959 and 1960 from June to October. The methods of qualitative analysis of free amino acids aplied in this experiment is followed by Moore and Stein. Free amino acids determined in this experiment are shown in Fig. 1 and Table 1. As the figure and the table are shown, three more amino acids such as glutamine, asparagine and histidine are detected in the diseased material. The additional amino acids which are known as diamines in diseased leaves are conspicuous. It is presumed that the diamine might be incresed by the self-reproduction of the virus in cooporation with certain enzymes which are carrying out the protein metabolism in the host protoplast in contrast with the healthy checks which is carrying out normal protein metabolism. From the histological poing of view, the facts of phloem degeneration or necrosis in diseased leaves, it seems to interrupt to move free amino acids from roots to leaves and it possibly takes place an excessive production of NH3 which is diaminated by the metabolism of nitrogen compounds in such conditioned leaves. Therefore, it is also presumed that additional diamino acids are accumulated in diseased leaves. There are no change of amino acids are accumulated in diseased leaves. There are no change of amino acids in both materials of this plant throughout the growing season qualitatively, and this result agress with the paper of Knight.

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Production System of Virus-free Apple Plants Using Heat Treatment and Shoot Tip Culture (열처리와 경정배양을 이용한 바이러스 무병 사과 생산 시스템)

  • Lee, Gunsup;Kim, Jeong Hee;Kim, Hyun Ran;Shin, Il Sheob;Cho, Kang Hee;Kim, Se Hee;Shin, Juhee;Kim, Dae Hyun
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.288-293
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    • 2013
  • In worldwide, viral diseases of apple plants has caused the serious problems like reduced production and malformation of fruits. Also, the damages of apple plants by virus and/or viroid infection (Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple mosaic virus, and Apple scar skin viroid) were reported in Korea. However there is few report about the protection approach against the infection by apple viruses. Therefore, this paper introduced the experimental protocol for the development of virus-free apple cultivars (Danhong, Hongan, Saenara, Summerdream). Apple plants were treated at $37^{\circ}C$ for 4 weeks and shoot tips were cultured in vitro. After heat treatment, the detection of apple viruses was performed by RT-PCR using virusspecific detection primers in new apple cultivars. With the heat treatments followed by in vitro shoot tip culture, the proportion of virus-free stocks of 'Danhong', 'Hongan', 'Saenara', and 'Summerdream' was 28%, 16%, 12%, and 12%, respectively. Taken together, this approach can be a good tool for production of virus-free apple stocks.