• Title/Summary/Keyword: 재분화

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In vitro Regeneration and Genetic Stability Analysis of the Regenerated Green Plants in Japanese Blood Grass (Imperata cylindrica 'Rubra') (홍띠 기내 재생과 재생 녹색식물체의 유전적 안정성)

  • Kang, In-Jin;Lee, Ye-Jin;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.34 no.2
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    • pp.156-165
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    • 2021
  • The in vitro regeneration was established, and the genetic stability among the mother plants (control) and the micropropagated green plants was evaluated using ISSR markers in Imperata cylindrica 'Rubra', Poaceae which containing important bioenergy plants. Green shoots were multiply induced from growing point culture via callus on MS medium supplemented with 0.01 mg/L NAA and 2 mg/L BA, and the shoots were proliferated on the MS medium with rooting. Rooted plantlets were transplanted to the pot with 100% survival rate. Using ISSR markers, somaclonal variation was analyzed in eight mother plants (control), ten green-regenerant cultivated at culture room (ReR) and ten green-regenerant cultivated at field condition (ReF). All ISSRs produced a total of 97 bands, and the scorable bands varied from one to seven with an average of 4.4 bands per primer. The polymorphism rate of ReRs and ReFs was 4.1% and 3.1% respectively, showing higher rate than that of control (0%). The genetic similarity matrix (GSM) among all accessions ranged from 0.919 to 1.0 with a mean of 0.972. According to the clustering analysis, ReFs and mother plants were divided into two independent groups. The results indicate that no clear genetic diversity was detected among regenerated plants, and ISSR markers were useful tool for identification of somaclonal variation of regenerants.

Efficient plant regeneration through callus induction from the hypocotyl of Perilla frutescens L var. Dayu ('다유들깨'품종의 하배축에서 캘러스를 통한 고효율 식물재분화)

  • Ruyue Xu;Ji-Hi Son;Hong-Gyu Kang;Hyeon-Jin Sun;Hyo-Yeon Lee
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.248-254
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    • 2023
  • This study was conducted to establish an efficient plant regeneration system in 'Dayu', a Korean variety of Perilla frutescens developed for seed oil production, in conjunction with the previously studied variety 'Namcheon'. The healthiest callus was formed on the hypocotyl explants cultured on a medium containing 0.1 mg/L NAA and 0.5 mg/L BA, outperforming the leaf and cotyledon samples. In both dark and long-day conditions, Dayu consistently exhibited significantly higher shoot regeneration rates compared with Namcheon. The highest shoot regeneration rates in Dayu were observed from the hypocotyl explants cultured on 0.1 mg/L NAA and 0.5 mg/L BA media, with shoot regeneration rates of 84.4% and 86.7% under dark and long-day conditions, respectively. Various combinations of plant growth regulators were tested to establish the optimal shoot regeneration conditions for Dayu hypocotyl explants. The results demonstrated that the highest shoot regeneration rate (90%) was achieved when 0.5 mg/L of BA was added to the medium without NAA. Among the regenerated shoots, 70.5% were normal plants, while 19.3% were abnormal. The addition of NAA or an increase in its concentration led to a higher occurrence of abnormal plants. After the regenerated shoots were transferred to 1/2 MS medium, roots were observed within 10-15 days. By day 30, they had developed into complete plants. The results obtained from the regeneration experiments with the perilla variety Dayu can valuably inform molecular breeding reliant on transformation techniques such as genome-editing and genetic modification technology.

유용 2차 대사산물의 생산을 위한 약용식물의 증진과 형질전환 I. 기내배양을 통한 병풀 엽병의 식물체 재분화

  • Kim, Kwang-Su;Paek, Yun-Woong;Ko, Kyung-Min;Hwang, Sung-Jin;Im, Hyung-Tag;Hwang, Baik
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.211-215
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    • 1995
  • 병풀(Centella asiatica)의 엽병 조직절편에서 유도된 배발생 캘러스로부터 체세포배 발생에 의하여 소식물체의 재분화를 이루었다. 엽병 조직절편을 1mg/L 2,4-D와 1mg/L kinetin이 조합 처리된 MS 기본배지에 치상하여 85%의 효율로 배발생 캘러스를 유도할 수 있었으며 이와 같은 배발생 캘러스를 5mg/L NAA와 1mg/L kinetin이 첨가된 배지로 옮겼을 때 체세포배의 형성은 87%까지 이루어졌다. 체세포배는 기본배지의 농도를 절반으로 줄이고 0.2 mg/L NAA와 0.2 mg/L kinetin이 첨가된 배지조건에서 기관분화를 거쳐 소식물체로 재분화되었다.

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Plant Regeneration From Mature Seed of Domestic Italian Ryegrass Cultivar (국내개발 이탈리안 라이그라스 품종 성숙종자의 식물체 재분화)

  • Kim, Kyung-Hee;Kim, Yong-Goo;Heo, Sung-Hyun;Lee, Ki-Won;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.3
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    • pp.235-242
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    • 2011
  • In order to improve forage characteristics of Italian ryegrass by genetic transformation, an efficient callus induction from mature seed and optimal plant regeneration system were established using a domestic cultivar 'Kospeed'. Addition of 5 mg/L of 2,4-D showed highest frequency of embryogenic callus induction from mature seeds. N6 medium showed higher frequency of both callus induction and plant regeneration as compared with MS and SH medium. The highest plant regeneration frequency 67% was obtained when embryogenic calli were transferred to N6 medium containing 1 mg/L 2,4-D and 5 mg/L BA. Supplementation of regeneration medium with sucrose at 30 g/L level maximized regeneration frequency as compared to the other concentrations. These data would be very helpful for molecular breeding of domestic Italian ryegrass cultivar through genetic transformation.

Embryogenic Callus Induction and Plant Regeneration in Kentucky bluegrass (Poa pratensis L.) Native to Korea (자생 왕포아풀(Poa pratensis L.)의 배발생 캘러스 유도 및 식물체 재분화)

  • 이재신;심상렬;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.277-281
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    • 2001
  • Embryogenic callus induction and plant regeneration methods were developed for native Kentucky bluegrass (Poa pratenes L.) ecotypes. Mature caryopses and immature inflorescences (20 mm in length) of 4 native ecotypes and 5 foreign cultivars were plated on MS medium (30 g/L sucrose, 3 g/L Phytagel) supplemented with 1 mg/L 2,4-D, and cultured in the dark at 24$^{\circ}C$. Most explants formed calli, but more embryogenic calli were induced from the explants of immature inflorescences than caryopses which produced mostly non-embryogenic rooty calli. In P77 ecotypes, immature inflorescence explants formed embryogenic calli with the rate of 62~95%, and those of field-grown plants were more efficient than greenhouse-grown ones in embryogenic callus induction. Plantlets were regenerated from the embryogenic calli when they were transferred to hormone-free MS medium, and grew to maturity without morphological variations in greenhouse.

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Kanamycin Concentration for Selection of 'Mcintosh Wijcik' Transgenic Apple (사과 'McIntosh Wijcik' 형질전환체 선발을 위한 Kanamycin 농도)

  • Song, Kwan Jeong;Seong, Eyn Soo
    • Horticultural Science & Technology
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    • v.18 no.6
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    • pp.811-814
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    • 2000
  • Effects of kanamycin concentration on regeneration and rooting of transgenic 'McIntosh Wijcik' were investigated to establish the efficient Agrobacterium-mediated transformation system. Relatively high regeneration frequency of explants appeared even at the high concentration of $150mg{\cdot}L^{-1}$ kanamycin, but the regeneration frequency and the number of normal shoots decreased significantly at a concentration of higher than $100mg{\cdot}L^{-1}$ kanamycin in the gelrite-gellifying medium. Rooting response varied with the transgenic lines in the agar-solidifying medium supplemented with the different concentrations of kanamycin and they were grouped with the inhibition level at $30mg{\cdot}L^{-1}$ concentration. No correlation between copy number and root response was observed. The optimum concentrations of kanamycin for the regeneration of 'McIntosh Wijcik' apple in the medium gellified with gelrite and for indirect-selection of putative transformants in the rooting medium solidified with agar were found to be $100mg{\cdot}L^{-1}$ and $30mg{\cdot}L^{-1}$ respectively.

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Introduction of rolC gene into Petunia hybrida (Petunia hybrida 세포내로의 rolC 유전자의 도입)

  • 정재동;김경민;남윤연;김창길;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.21-26
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    • 1999
  • These experiments were attempted to introduce rolC gene in the Petunia hybrida cv. Titan white by Agrobacterium mediated. The maximum frequency of shoot regeneration was obtained by 60% on MS medium containing 1.0 mg/L BA, 0.1 mg/L NAA, 200 mg/L kanamycin, 500 mg/L carbenicillin, 30 g/L sucrose, and 8 g/L agar. Kanamycin-resistant calli were selected from petunia leaf discs by cocultivation with Agrobacterium suspension cultures on MS medium. The addition of AgNO$_3$ and KMnO$_4$ in the medium increased the shoot regeneration by 31.3% from leaf disc as compared with non-treated leaf disc. Among clones exhibiting kanamycin resistance, only 3 clones were confirmed by southern hybridization analysis.

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Plantlet Regeneration Cotyledon and Petiole Cultures of Cyclamen Persicum Mill (시클라멘(Cyclamen Persicum Mill.)의 자엽과 엽병배양에 의한 식물체 재분화)

  • 은종선;고정애;김영선
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.212-216
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    • 1995
  • These experiments were carried out to examine the effect of explant sources and plant growth regulators on callus induction and plantlet differentiation. Cotyledon and petiole explants of Cyclamen persium were cultured on MS medium supplemented with different concentrations of auxins and cytokinins. Cotyledon cultured on medium containing 2, 4-D and kinetin did not form callus or shoots. But when calli induced from petiole explants on medium with 1.0 mg/L 2, 4D and 1.0 mg/L kinetin were subcultured on the same medium the formation of shoots from calli occurred after 150 days of culture. The combination of NAA and BA were more effective than that of 2, 4 D and kinetin in the formation of shoots from calli, cotyledon culture was most effective on medium with 0.2 mg/L NAA and 0.5 mg/L BA. Shoots excised from calli were rooted on medium with 1.0 mg/L NAA. The plantlets were subsequently transplanted to potting soil.

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