• Plant Resources
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• 제7권3호
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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.

• 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.

• 한국자원식물학회지
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• 제17권3호
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• pp.331-338
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• 2004

본 연구에서는 카네이션과 고구마의 무병 종묘를 생산하고자 정단분열조직배양을 실시하였으며, 생산된 배양종묘의 바이러스 감염여부를 진단하기 위해 ELISA방법을 이용하여 주요 바이러스를 검정하였다. 카네이션의 정단분열조직 배양에 공시 한 'Roland' 등 4품종 중 투명화 현상이 없이 가장 정상적인 잎이 발생하고 절간신장이 이루어지며 뿌리의 발생도 양호한 생장반응을 보인 품종은 석죽계통인 'Giant Gipsy' 이었으며 MS기본배지에 NAA 0.05 ㎎/L와 BA 0.1㎎/L 혹은 kinetin 1.0㎎/L와 조합한 구에서 shoot의 재생이 양호하였다. 배양7주일 후에는 explant 1개체에서 평균 5-6개, 많은 것은 10개체 이상의 multiple shoot가 형성되었고 기내 마디삽목법에 의하여 대량증식할 수 있었다. 투명화현상이 심한 품종은 'Casha'와 'Desio'로 나타났는데 투명화 현상은 kinetin 첨가보다는 BA첨가구에서 현저하게 증가하였다. 카네이션 기내 배양종묘의 바이러스 감염여부는 ELISA방법을 이용하여 CarMV와 CarRSV를 신속하게 다량의 시료를 진단할 수 있었다. 그 결과 정단분열조직의 배양을 통해 65.2%에서 무병 주가 생산되었음을 확인할 수 있었다.

• 식물조직배양학회지
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• 제27권4호
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• pp.299-307
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• 2000

A large amount of information is currently available for somatic embryogenesis of plants. However, one common problem related to somatic embryos is that the conversion rate can be low for some species. Apical meristems are responsible for post-embryonic growth of the embryo. The low percentage observed is most likely a result of poor apical meristem development or defects in the meristem organization during somatic embryogenesis. In flowering plants, apical meristems are well developed by the late heart stage of zygotic embryo development. In conifers, such as white spruce, apical meristems are formed at the pre-cotyledon stage. Thus, apical meristem development occurs very early, prior to the maturation stage of embryo development. Once formed, meristems are stably determined. In the somatic embryo, as exemplified by white spruce, since embryo development is not synchronous, tissue differentiation including apical meristem formation occurs throughout the“maturation”stage. Different apical meristem organizations can be found among different individuals within a population. In contrast to their zygotic counterparts, the apical meristems appear not to be stably determined as their organization, as the shoot apical meristem especially, can be easily modified or disrupted. Precocious germination seldom results in functional plantlets. All these observations suggest that the conditions for somatic embryo maturation have not been optimized or are not suitable for meristem formation and development. The following strategies could improve meristem development and hence conversion: 1. Simulate in ouuio conditions to promote meristem development prior to the“maturation”treatment.2. Prevent deterioration of apical meristem organization during somatic embryo maturation.3. Promote further meristem development during embryo germination.

• 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.

• 한국약용작물학회지
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• 제5권3호
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• pp.186-190
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• 1997

This study was carried out to investigate the effect of polyamines, salt strength. sucrose and gelling agents on the regeneration of plantlets by meristem culture of Aloe arborescens Mill. and Aloe vera L.. Shoot multiplication was more effective when 10mg/ l spermine in Aloe arborescens and 1mg/ l spermidine in Aloe vera added into MS medium than when other polyamines were treated into media. A quarter strength of MS medium was effective for rooting of shoots regenerated. Higher concentration of sucrose (45g/ l) was more effective for shoot regeneration. Addition of 4g/ l gelrite into the medium was effective for induction of multiple shoots from Aloe than that of agar or other concentrations of gelrite. When plantlets regenerated from meristem culture were transferred to pot. survival rate of plantlets was 80% on perlite and was 95% on vermiculite. respectively.

• 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.

• 한국자원식물학회지
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• 제7권1호
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• pp.17-22
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• 1994

This study was carried out to investigate the optimum medium and concentrations of growth regulators for induction of multiple shoot by meristem culture of floe otorefcenf Mill. MS medium supple-mented with 3${\mu}{\textrm}{m}$ TDZ was effective for induction of multiple shoot. Shoot multiplication was more ef-fective when 2mg/1 BA combined with 0.Img/1 IAA than when only BA were treated on medium. Halfstrength of MS medium supplemented with 2mg/L IAA was effective for rooting of shoots regenerated.When plantlets regenerated from meristem culture were transferred to pot, survival rate of plantletswas 80% on perlite and was 95% on vermiculite, respectively.

• The Plant Pathology Journal
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• 제33권3호
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• pp.288-295
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• 2017

Fig mosaic disease (FMD) is a viral disease that spreads in all Tunisian fig (Ficus carica L.) orchards. RT-PCR technique was applied to leaf samples of 29 fig accessions of 15 fig varieties from the fig germplasm collection of High Agronomic Institute (I.S.A) of ChattMariem, to detect viruses associated to FMD. Analysis results show that 65.5% of the accessions (19/29) and 80.0% (12/15) of the fig varieties are infected by FMD-associated viruses. From all fig accessions, 41.4% of them are with single infection (one virus) and 24.1% are with multi-infections (2 virus and more). Viruses infecting fig leaf samples are Fig mosaic virus (FMV) (20.7%), Fig milde-mottle-associated virus (FMMaV) (17.25%), Fig fleck associated virus (FFkaV) (3.45%), and Fig cryptic virus (FCV) (55.17%). A reliable protocol for FCV and FMMaV elimination from 4 local fig varieties Zidi (ZDI), Soltani (SNI), Bither Abiadh (BA), and Assafri (ASF) via in vitro culture of 3 meristem sizes was established and optimized. With this protocol, global sanitation rates of 79.46%, 65.55%, 68.75%, and 70.83% respectively for ZDI, SNI, BA, and ASF are achieved. For all sanitized varieties, the effectiveness of meristem culture for the elimination of FCV and FMMaV viruses was related to meristem size. Meristem size 0.5 mm provides the highest sanitation rates ranging from 70% to 90%.

• 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.