• Title/Summary/Keyword: Meristem tip culture

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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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Development of a Reliable Technique to Eliminate Sweet potato leaf curl virus through Meristem Tip Culture Combined with Therapy of Infected Ipomoea Species

  • Cheong, Eun-Ju;Hurtt, Suzanne;Salih, Sarbagh;Li, Ruhui
    • Korean Journal of Plant Resources
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    • v.23 no.3
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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.

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

  • Singh, Shweta;Hazra, Sulekha
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.333-340
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    • 2009
  • Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

In Vitro Mass Propagation and Soil Adjastment of Zanthoxylum piperitum var. inerme Makino through Apical Meristem Culture (生長點 培養에 依한 민초피나무(Zanthoxylum piperitum var. inerme Makino)의 器內 大量 增殖 및 土壤 活着)

  • Jeong, Woo-Gyu;Lee, Sang-Rae
    • Korean Journal of Plant Resources
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    • v.6 no.2
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    • pp.171-179
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    • 1993
  • This study was conducted to investigate the effect of growth regulators and medium composition on the growth of each stage in apical meristem culture for mass propagation of Zanthoxylum piperitum var. inerme Makino. The source material, shoot tip segments were taken from three-years old graft trees. Apical meristems were cultured in vitro on basal MS, GD, WS, half strength MS(1/2MS) and half strength GD(1/2GD) media supplemented with various concentrations for growth regulators(BA, IBA) and inorganic nutrients. The results summarized are as follows: 1. In culture establishment stage, ratio of culture establishment was 96.7% and the best resuit was obtained using MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 2. In shoot multitication stage, both shoot multiplication and growth were achieved in average 5.6cm. These results were obtained on in MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 3. In roothing stage, phloroglucinol(PG) acted as IBA synergist in root initiation. The most faverable combinations for root development was half-strength MS medium supplemented with 162mg/l PG and 0.2mg/l IBA, and ratio of rooting was 58.0%. 4. In Vitro formed plantlets were transplanted to paper pots in greenhouse with 85% of relative humidity. 96% of survival rate was obtained from artificial soil mix having same volume of sand, vermiculite, peat, and soil.

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Effects of the Phytohormones on the Organ Differentiation and the Callus Induction from the Meristem Tip and the Segments of the Leaf and Stem of Potato by in vitro Culture (기내배양시 몇가지 생장조절물질이 감자의 생장점 및 경엽조직편으로부터의 Callus 및 기관분화에 미치는 영향)

  • 김충수;조재성;최창열
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.26 no.4
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    • pp.344-349
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    • 1981
  • This study was conducted to define the effect of 2.4-D, NAA, Benzyladenine, and basic mediums on the callus induction and the organ differentiation from the meristem tips and the stem and leaf segments of the potato. Benzyladenine promoted the induction and growth of shoot from the meristem tip of potato but inhibited initiation of roots and induction of callus. At higher concentration of NAA than 0.5 ppm and of 2.4-D than 1.0 ppm the shoots were not initiated but the callus was induced from the meristem. The callus growth was significantly promoted on the medium containing NAA than 2.4-0. The initiation and growth of the shoots from the potato meristem was significantly increased in the medium containing 2.4-D and BA, or NAA and BA, compared with those containing BA, NAA or 2.4-D alone. The callus was more easily induced from the stem segments than the leaf segments of potato. And the 2.4-D was more effective for the induction and growth of the callus than the NAA. MS medium diluted its concentration to 1/2 was more suitable for the initiation and growth of the shoots from the potato meristem than the MS standard medium. For the initiation and growth of the shoots from the potato meristem, the most desirable medium was the diluted MS medium containing 1.0 ppm BA and 0.1 ppm NAA or 0.1 ppm 2.4-D.

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Elimination of Grapevine fleck virus from infected grapevines 'Kyoho' through meristem-tip culture of dormant buds (휴면아 경정 배양법을 통한 포도 '거봉' 에서 Grapevine fleck virus의 제거)

  • Kim, Mi Young;Cho, Kang Hee;Chun, Jae An;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.401-408
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    • 2017
  • Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.

Effects of Cultural Conditions on the In Vitro Propagation of Plantlets derived from Apical Meristem in Ipomoea batatas L. (고구마 정단분열조직 유래 식물체의 기내 증식에 미치는 배양조건의 영향)

  • Eun, Jong-Seon;Park, Jong-Suk;Kim, Young-Seon
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.37-40
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    • 2002
  • Sweet potato is a crop vegetatively propagated by vine cuttings, an ineffective method for maintaining pathogene-free stock plants. As an alternative method, single-node cultures of virus-free plantlets derived from apical meristem in sweet potato (cv. Yulmi) was examined. Effective pH range, sugar concentration and nodal order were investigated to establish an in vitro mass propagation system with high quality virus-free stock plantlets to farmhouse. Although the plantlets grew at wide range of pH, the most effective pH of the medium was 4.8 in single-node cultures. High sugar concentration of 60∼80 g/L resulted in increased growth response in shoot length, root length, number of node, leaf area and fresh and dry weight of shoot and root, whereas reducing sugar contents below 6% was showed reduced growth response. The first node including meristem tip was the best for the rapid growth of plantlets and the other nodes also showed a very similar growth response. Uniform plantlet can be obtained massively at the same time by culture of single node except for the first node including meristem tip. In conclusion, the most effective pH range and sugar concentration of medium for the growth of plantlets via single-node cultures was 4.8, 60∼80 g/L respectively. The first node was the best for the rapid propagation of plantlets in nodal order.

Effect of BA Concentrations and Culture Methods on in Vitro Plant Multiplication from Shoot-Tip Culture of Wasabia japonica (고추냉이 정단배양에 있어서 BA 농도 및 배양방법에 따른 기내증식 효과)

  • Park, Yun-Young;Cho, Moon-Soo;Lee, Young-Deuk;Chung, Jong-Bae;Park, Shin;Jeong, Byeong-Ryong;Park, Sang-Gyu
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.1-6
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    • 2007
  • Effect of BA concentrations and culture methods on in vitro plant multiplication from shoot-tip cultures of Wasabia japonica was studied. Shoot-tips with leaf primordia and apical meristem were cultured on MS basal medium for all the experiments. Liquid medium for 2 weeks followed by semi-solid medium for 4 weeks containing 1.0 mg/L BA was the best to number of shoots (22.8) and shoot length (3.5 cm). Shoots proliferated could be divided into ca. 5 to 11 of cultures for the multiplication of plantlets. Divided plantlets showed root formation (90%) well onto MS basal medium without growth regulators like IBA and NAA. After rooting, all the plantlets transferred into the pots containing composed soil (bio-media Co., peatmoss $8{\sim}10%$, coir dust $66{\sim}70%$, zeolite $13{\sim}17%$, vermiculite $3{\sim}7%$, perlite $2{\sim}4%$) and grown well into whole plants with multiple shoots.

Studies on the Mass-Propagation of Potato (Solanum tuberosum L.) by In-vitro Culture. I. Effects of Phytohormons on the Callus Induction and the Organ Differentiation from Potato Meristem tip (기내배양에 의한 감자(Solanum tuberosum L.)의 대량번식(大量繁殖)에 관한 연구(硏究) I. 몇가지 생장조절물질(生長調節物質)이 생장점(生長點)으로부터의 Callus 및 기관분화(器官分化)에 미치는 영향(影響))

  • Kim, Choong Soo;Jo, Jae Seong;Choi, Chang Yeol
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.59-64
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    • 1980
  • These experiments were carried out to define the effects of 2.4-D, NAA and Benzyladenine on the differentiation and growth of the organs and the induction of callus from the potato meristem. The results were summarized as follows ; 1. The differentiation and growth of the shoots from the potato meristem was promoted in increased concentration of Benzyladenine but the callus was not induced on the M.S. medium containing Benzyladenine. 2. On the M.S. medium containing NAA 0.5 mg/l or higher concentration of NAA, the shoots were not initiated but the callus were induced from potato meristem. The growth of callus was promoted in increased concentration of NAA. 3. The roots were initiated from 50% of potato meristems planted on the M.S. medium containing more than 0.1 mg/l of NAA but the roots were pot initiated on the medium containing 2.4-D. 4. The shoot growth was significantly increased by increasing of 2.4-D concentration up to 0.1 mg/l, but the shoots were not initiated on the medium containing 2.4-D more than 1.0 mg/l. 5. For the induction and growth of the callus from potato meristem, NAA was more effective than 2.4-D and the most effective medium was M.S. medium supplemented with 2.0 mg/l of NAA. 6. The M.S. mediums supplemented with BA 2.0 mg/l and NAA 0.1 mg/l or BA 1.0 mg/l and 2.4-D 0.1 mg/l showed good results for entire plant regeneration from potato meristem.

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Systematic Propagation of High Quality Garlic (Allium sativum L.) Through Shoot Apical Meristem Culture 1. Organogenesis from in Vitro Cultured Shoot-tips (생장점배양에 의한 우량마늘 체계적 증식 1.생장점배 양으로부터 기관형성)

  • Lee, Eun-Mo;Lee, Young-Bok
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.161-166
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    • 1994
  • Since garlics (Allium sativum L.) are propagated through cloves, infection by virus or other pathogens may become severe problem if not using high quality seed bulbs every year resulting in the reduction of yield and bulb quality, In order to solve this problem, the establishment of virus-free bulb production and its supply system have been required because no chemicals were found to eliminate viruses from seed bulbs. This experiment was conducted to develop an effective production technique of high quality seed bulbs using shoot-tip culture. Over 90% of shoot-tips explanted on January L 1990 were survived at constant temperature of either 20, 24 or 28$^{\circ}C$, wheres 88% at alternate temperature (28/20$^{\circ}C$). The growth of shoot and root was most vigorous at constant 24$^{\circ}C$, and least at alternate temperature (28/20$^{\circ}C$) condition. When shoot-tips were explanted June 21 to August 1,1991, survival and growth of shoot-tips was most vigorous on MS medium supplymented with 0.1 mg/L NAA and 2 mg/L kinetin and least 1 mg/L Gh$_3$. The shoot-tips taken from the seed bulbs stored at 4$^{\circ}C$ for 15 to 60 days were placed on MS medium, shoot growth and in vitro bulblet formation increased slightly as affected by the increase of told treatment period at 4$^{\circ}C$.

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