• Title/Summary/Keyword: in vitro plantlet

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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.

Propagation of Endangered Species, Daphne pseudomezereum var. koreana via in vitro Bud Culture (멸종위기종 두메닥나무(Daphne pseudomezereum var. koreana)의 줄기 기내배양을 통한 식물체 생산)

  • Chu, Yerin;Park, Sanghee;Cheong, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.109 no.2
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    • pp.189-194
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    • 2020
  • Daphne pseudomezereum var. koreana is native to Korea and is distributedin Kangwon-do, Jeollabuk do, and Gyeongsang-do. This economically valuable species has experienced a dramatic decrease in natural habitat due to climate change and is difficult to cultivate. In this study, we investigate a mass propagation method for D. pseudomezereum through in vitro culture and genetic resource preservation.WPM medium was better than the MS medium for shoot growth. As a result, we compared the shoot number and length of apical (W/AP) and non-apical shoots (W0/AP) with BA and GA3 treatments in WPM medium. Their shoots and length grew well in both BA 8ìM + GA38ìM-treated apical shoot and without-apical shoot. NAA did not effectively induce rooting of the in vitro plantlet.

Effect of BA and NAA on Adventitious Bud Induction From In Vitro Germinant Eucalyptus pellita (Eucalyptus pellita 기내발아묘의 부정아 유도에 미치는 BA 및 NAA 효과)

  • Kim Ji-Ah;Moon Heung-Kyu;Kang Ho-Duck
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.201-207
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    • 2005
  • This study was conducted to examine the effect of BA and NAA on adventitious bud induction from in vitro germinants E. pellita. The capacity of adventitious bud formation greatly depends on juvenility and explants origin; the more juvenile materials are the better ability to form adventitious buds even in in vitro raised plantlets. In case of in vitro germinants, 7 day old plantlets showed a better morphological response than did 14 day old ones in the induction of adventitious buds. The capacity to show morphological response was in decreasing order : cotyledons> petioles> roots. Ho adventitious buds formed when root segments were used as culture material. And optimum medium appeared to be MS + 0.5 mg/L BA and 0.2 mg/L NAA. Adventitious buds could be developed into multiple shoots and regenerated normal plantlets on DKW medium plus 0.2 mg/L BA and 0.01 mg/L NAA.

Adventitious Shoots Regeneration from Seed Explants of Xanthoceras sorbifolium

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.58-58
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    • 2020
  • Xanthoceras sorbifolium Bunge (yellowhorn) is a woody tree in the soapberry family, Sapindaceae, native to northern China. This species has been identified as a major woody bioenergy plant for bio-diesel production because of high oil content in seed. But the flowers do not bear fruit well while the many flowers blooming. This study was performed to regenerate in vitro plantlet using adventitious shoot formation. To establish the protocol of plant regeneration, adventitious shoots formation rate in the culture of cotyledon of immature zygotic embryos was 68.6% in 1/2 MS medium with 0.5 mg l-1 BA and 3% sucrose (w/v). In the culture of cotyledons of mature zygotic embryos, induction of adventitious shoots was needed to contain high sucrose in pre-culture medium and the frequency of shoot induction was 64.4%. Multiple shoots were induced in 0.5 mg l-1 TDZ, and rooting of shoot was induced 4.0 mg l-1 IBA. Flow cytometry analysis revealed that all the regenerated plantlets were diploid.

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In vitro propagation of endangered species, Hylotelephium ussuriense (Kom.) H. Ohba (멸종위기종 둥근잎꿩의비름 (Hylotelephium ussuriense (Kom.) H. Ohba)의 기 내 증식)

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Kim, Ji-Ah;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.38-43
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    • 2014
  • To establish the system of in vitro plant regeneration, the different explants (stem with axillary bud and stem without axillary bud) of Hylotelephium ussuriense were cultured on the Murashige and Skoog's medium containing 6-benzylaminopurine (BA) and indolebutyric acid (IBA). The adventitious shoot induction was more effective in the stem with axillary bud explants than the stem without axillary bud explants, and was the best on MS medium containing 3.0 mg/L BA and 0.01 mg/L IBA. Frequency of plantlet growth was not significantly treated on MS and sucrose. Total chlorophyll contents under ventilation treatment were higher than those in control (non-ventilation). This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

Effect of Basal Medium and Plant Growth Regulator on in vitro Plant Regeneration from Axillary Buds of Walnut New Cultiver "Sinlyeong"

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.15-15
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    • 2019
  • The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.

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Plant Regeneration through Callus of Korean Native Seosanjong of Zingiber officinale Rosc.

  • Jo, Man-Hyun;Ham, In-Ki;Song, Nam-Hyun;Woo, In-Shik
    • Plant Resources
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    • v.3 no.2
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    • pp.131-134
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    • 2000
  • Embryogenic callus cultures of Korean native Seosanjong of ginger(Zingiber of officinale Rosc.) were induced through stem explants taken from in vitro shoot-tip cultures. Among the four concentrations of 2,4-D tested in Murashige and Skoog medium, 0.5 and 1 mg/L of 2,4-D was most effective in inducing embryogenic callus. Leaf explants did not express any new morphogenetic response in all 2,4-D concentrations tested. Plantlets transferred to hormone-free MS medium were developed and successfully acclimatized under greenhouse.

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Root and Shoot Formation in Explant and Callus Derived from Root and Cotyledon of GinBeng(Panun ginseng C. A. Meyer) (인삼근 및 자엽 Callus의 기관분화에 관한 연구)

  • Choe, Gwang-Tae;Kim, Myeong-Won;Sin, Hui-Seok
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.35-40
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    • 1981
  • Explants of mature root tissues and calli derived from root and cotyledon of Panax ginseng were cultured in vitro on Murashige and Skoog medium supplemented with 2, 4-dichlorophen-oxyacetic acid(3,4-D), naphthaleneacetic acid(NAA), benzyladenine, and gibberellic acid to assess their capacity to regenerate organs. Root formation at high percentage (46.2-61.1%) was obtained 20-30 days after culturing on media supplemented with combinations of NAA(5 mg/l) and kinetin (1 mg/l), And calli derived from cotyledon produced numerous embryoids in media($\frac{1}{2}$MS) containing 2,4-D(0.5 mg/l) and kinetin (0.5 mg/l). Reculture of these embryoids in media($\frac{1}{2}$MS) enriched with 1 mg/l of benzyladenine and 1 mg/l of gibberellic acid resulted in more plantlet regeneration.

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Ultrastructural Characteristics of Developmental Stages During in vitro Regeneration in Citrus junos SIEB. (유자 (Citrus junos SIEB.) 의 발생단계에 따른 미세구조적 특성)

  • 박민희
    • Korean Journal of Plant Resources
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    • v.8 no.3
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    • pp.237-246
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    • 1995
  • In this study, the callus was induced and regenerated from the immature embryo and ultrastructural characteristics of developmental stages in Citrus junos SIEB, were investigated. The yellowish callus was induced by 5 to 6 week of culture of citrus. In proliferation callus after 6 weeks of culture, large vacuole was formed by fusion between adjacent small ones. In the non-embryogenic callus cultured for 12weeks, re-differentiated cells of callus showed the large nucleus with globular nucleus and amyloplast with large size of starches. In the embryogenic callus cltured for 14-16 weeks, the active exocytosis occurred in cells, secretory vesicles appeared on cell membrane and small particles from cytoplasm were released to intercelluar space. In the embryogenic callus cultured for 24 weeks, a sperical type of chloroplast bounded on cytoplasm by double membrane and typical grana was dispersed equally among matrix. In the normal plantlet after 26 weeks of culture, a lot of vessels and companion cells apperaed in the leaf cell of plantlet. In the normal plantlet after 30 weeks of culture, the immature leaf showed many small companion cells, sieve tubes and central vacuole. Also, the secondary vacuole protruded into the central vacuole and elongated chloroplasts near plasma membrane. In the matured plant habituated on the soil, palisada tissue composed of orderly arranged cells contained the nucleus in the center of the cell and large vacuoles on either side of the nucleus.

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The Factors on Somatic Embryogenesis of Soybean [Glycine max. (L.) Merrill]

  • Kim, Kyong-Ho;Kim, Hag-Sin;Oh, Young-Jin;Suh, Sug-Kee;Kim, Tae-Soo;Park, Ho-Kee;Park, Moon-Soo;Kim, Seok-Dong;Yeo, Up-Dong
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.123-128
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    • 2000
  • To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as material part and size of immature seed, 2,4-D, pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up to 22.9%) of somatic embryogenesis was obtained from the immature cotyledon, following embryonic axis and the immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25.4%) of embryogenesis was obtained from 3-4 mm length of the seed (after 25 days of flowering). The optimum concentration of 2,4-D for embryogenesis was 10 mg/L. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tips from the germinating somatic embryos, they were cultured on MS medium containing 2 mg/L indole-3-butyyic acid (IBA) and 1 mg/L 6-benzyladenine (BA). After then somatic embryos with one and three cotyledons were transferred to the growth regulator free medium. The medium exhibited the higher rate (ca. 50%) of development than the multiplication medium.

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