• Title/Summary/Keyword: in vitro rooting

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Effect of Plant Growth Regulators on Bulblet Formation and Plant Regeneration in Fritillaria thunbergii Miq. (패모조직배양에서 생장조절 물질이 자구형성 및 식물체 재생에 미치는 영향)

  • Park, Chul-Hyoung;Ryu, Jeom-Ho;Han, Kwang-Soo;Doo, Hong-Soo;Choi, Sun-Young
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.119-125
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    • 1996
  • To improve the efficiency of mass propagation in vitro of Fritillaria thunbergii, bulb scale and nodes were cultured in LS medium supplemented with the combination of 2, 4-D and kinetin or NAA and BA. The number and size of bulb, the number of adventitious shoot, the ratio of callus formation, rooting, and the effects of light and dark on the culture, plant regeneration from calli, and the gelling substances were investigated. The combination of 2, 4-D and kinetin in media was more effective than the media of NAA and BA for the bulblet formation. The media supplemented with 2 mg/L 2, 4-D and 1 mg/L kinetin, $1{\sim}2\;mg/L$ 2, 4-D without kinetin and $1{\sim}3\;mg/L$ BAA only were effective in the adventitious shoot development. Callus formation and root formation, respectively were effective in the medium supplemented with 2mg/L 2, 4-D and 1mg/L kinetin. In bulb formation, the medium with 5 mg/L kinetin was effective, and the most of bulbs were formed from the axillary bud of node part. In bulb formation, shoot growth, callus and root formation, the light culture for 16 hours per day was better than that in the dark culture. Bulb was nicely formed in the medium with 0. 2 mg/L 2, 4-D, 1 mg/L kinetin. The medium without hormone was most effective for plant regeneration. The phytagel was more effective than agar in the medium as the gelling agent.

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Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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Effects of the Agar and Auxins Concentrations on Vitrification in Tissue Culture of Gypsophila paniculata L. cv. 'Bristol Fairy' (숙근(宿根)안개초의 조직배양(組織培養)에 있어서 Agar 농도(濃度) 및 Auxins의 농도(濃度)가 Vitrification에 미치는 영향(影響))

  • Chun, Chae Ki;Choi, Sang Tai;Park, In Hwan;Shin, Hak Ki
    • Current Research on Agriculture and Life Sciences
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    • v.6
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    • pp.37-41
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    • 1988
  • The present work deals with the effect of agar and auxins concentrations on vitrification in tissue culture of Gypsophila paniculata L. cv. 'Bristol Fairy' in vitro. The results were summarized as follows. 1. Plant growth, that is, plant height, fresh weight and branching were decreased as increasing agar concentration. On the other hand, addition effect of IAA 1.0mg/l+NAA 0.5mg/l and IAA 2.0mg/l+NAA 1.0mg/l on the plant height were increased strikingly. 2. Addition effect of auxins on the days to rooting were little. And the root development showed same tendency as plant growth. 3. The rate of non-vitrified plants were gradually increased as rising agar concentration. But the addition of agar 1.5g/l in the medium resulted in poor growth. 4. From these results, it was found that following media were the most effective for increasing of non-vitrified and good plant growth in Gypsophila paniculata L. tissue culture.

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Adventitious Shoot Formation and Plant Regeneration from Explants of Solanum nigrum L. (까마중(Solanum nigrum L.)의 유식물 절편체에서 부정아 형성 및 식물체 재분화)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.277-284
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    • 2012
  • In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

Selection of a Triploid Poplar by Flow Cytometric Analysis and Growth Characteristics of its in vitro Grown Plants (유세포 분석을 통한 현사시나무 3배체 선발 및 계통별 기내생장 특성)

  • Bae, Eun-Kyung;Lee, Hyoshin;Lee, Jae-Soon;Noh, Eun-Woon
    • Journal of Korean Society of Forest Science
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    • v.101 no.2
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    • pp.291-296
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    • 2012
  • Triploids are a useful tool for biomass production and molecular breeding of trees with a long life span. Triploids of the poplar 'Hyunsasi' (Populus alba ${\times}$ P. glandulosa) have been developed by crossing between female diploids and a male tetraploid. The tetraploid was developed around the 1970s at Korea Forest Research Institute by colchicine-induced chromosome doubling. Seedlings of the $F_1$ generation were analyzed using flow cytometry to verify their ploidy status. The mean relative fluorescence index of 3 F1 poplars, labeled as Line- 1, Line-17, Line-18, were approximately 1.5 times higher than those of diploid poplars, and the results clearly indicated that they were triploids. The phenotype of the F1 poplars included larger leaves and thicker stem than diploids, and abnormal leaf morphology, especially in the triploid 'Line-18'. Three triploid lines developed roots more slowly and had less roots than diploid. However, 3 poplar cytotypes (2x, Line-1, Line-17) rooted within 10 days on MS medium. In contrast, compared with the 3 cytotypes, the Line-18 showed about 80% and 70% in the rooting rate and the number of roots. The triploid poplars could be directly utilized for biomass production and with their sterility, they could serve as basic material for genetic transformation. In addition, flow cytometric analysis proved to be an effective and reliable method for screening forest trees for their ploidy level.

Timing for Determination in Adventitious Root Formation from In Vitro Cultured Internodal Explants of Cassava (Manihot esculenta) (카사바의 절간절편 배양에서 부정근 발생이 결정되는 시기의 판별)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.1-6
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    • 2001
  • The timing for the determination in root formation from nodal and internodal explants of cassava (Manihot esculenta Crantz, cv. MCol 22) was investigated. Nodal explants about 10 mm with an axillary bud formed adventitious roots directly on MS basal medium for 8 days of cultures. But internodal segments without an axillary bud did not develop the adventitious roots on the same medium, and most internodal segments excised from nodal explants after cultures of 5 days on MS basal medium developed adventitious roots. On the other hand, the internodal segments rooted at 90% after cultures on medium with 0.5 mg/L IBA for 5 days, with 1 mg/L IBA for 2.5 days, and with 2 mg/L IBA for 1.5 days respectively. Thus the period of culture on medium with IBA and its IBA concentration affected the rooting rate. Therefore, it is suggested that the determination for root formation occurred before the differentiation of root primordia on medium with IBA, and root inducing factors in medium were absorbed and accumulated during the period of determination for root primordium differentiation in internodal segment of cassava.

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Effects of Medium Compositions and Plant Growth Regulators on in vitro Organogenesis in Cultured Explants of Platycodon grandiflorum Species

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Kim, Hye-Rim;Moon, Young-Ja;Yoon, Ki-Hong;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.259-274
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    • 2017
  • Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

Korean Tricholoma matsutake Strains that Promote Mycorrhization and Growth of Pinus densiflora Seedlings (균근 형성과 소나무 유묘 생장이 우수한 송이 균주의 선발)

  • Jeon, Sung-Min;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.155-165
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    • 2016
  • Domestic and international production of Tricholoma matsutake has decreased owing to matsutake forests being left alone, host plant disease, forest fires, climate change, and so on. In order to identify strains that are suitable for the production of T. matsutake-inoculated seedlings, Pinus densiflora seedlings were inoculated with T. matsutake after in vitro rooting and mycorrhization was examined in the roots of T. matsutake-inoculated seedlings after 6 months. The mycorrhization rate was greater than 80% for 5 strains (NIFoS 421, 434, 1681, 1984, and 2001) out of 19 total strains. Seven strains (NIFoS 434, 441, 561, 562, 1016, 1807, and 1812) showed shoot/root ratios of less than 3.0 and had a seedling shoot biomass of 2.0 to 4.8 times higher than that of the root. Eight strains (NIFoS 441, 561, 562, 1016, 1807, 1812, 1984, and 2001) stimulated increases in shoot volume and three stains (NIFoS 441, 562, and 1812) promoted the growth of root biomass by mycorrhizal formation. In conclusion, 4 strains (NIFoS 434, 561, 1984, and 2001) out of 19 total strains tested showed higher mycorrhization rates and seedling growth than those of the other strains. We expect that the use of these four strains may contribute to T. matsutake-inoculated seedling production.

Effects of glutamine and AgNO3 on plant regeneration of Sedum sarmentosum (돌나물의 식물체 분화에 미치는 Glutamine과 AgNO3의 영향)

  • Ahn, Jeong-Ho;Kim, Hyun-Soon;Lee, Seung-Yeob
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.18-22
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    • 2009
  • This work was conducted to establish an efficient plant regeneration system for genetic transformation and the in vitro conservation of Sedum sarmentosum genetic resources. Effects of glutamine and $AgNO_3$ on plant regeneration between two genotypes were investigated using MS media supplemented with 0.2 mg/L NM and 3.0 mg/L BA. Calluses were formed on leaf explants placed on MS solid media supplemented with 3 mg/L 2,4-D and 1 mg/L BA. Calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with glutamine. The highest number of shoots per callus was 17.6 at 350 mg/L glutamine. However, calluses of Wanju local strain gave rise to no shoots under the same culture conditions. Likewise, calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with $AgNO_3$ whereas Wanju local strain sporadically produced shoots. The highest number of shoots per callus of Keumsan local stain was 16.1 at $15{\mu}M$ $AgNO_3$. Regenerated shoots were subcultured on hormone-free MS medium for rooting and shoot growth, and then 3-5 cm high plantlets were transplanted to the artificial soils comprising vermiculite and perlite, where they survived at a frequency of 88-100%. After being transplanted into upland soil:sand (1:1, v/v) in a greenhouse, regenerated plants showed a morphologically normal growth.

Improvement of Black Locust(Robinia pseudoacacia L.) Through Tissue Culture. I. Micropropagation and Somatic Embryogenesis (조직배양에 의한 아까시나무(Robinia pseudoacacia L.)의 개량 I. 대량증식과 체세포배 발생)

  • Woo, Jong Ho;Choi, Myung Suk;Joung, Eun Yi;Chung, Won Il;Jo, Jin Ki;Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.84 no.1
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    • pp.41-47
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    • 1995
  • A micropropagation system for black Locust(Robinia pseudoacacia) was established by using shoots and pin-punctured leaves of in vitro germinated seedlings. The greatest number of shoots (an average of 10.5 shoots) was obtained when shoot tips were cultured on MS medium supplemented with 1.0 mg/l BAP and 0.01 mg/l NAA. When pin-punctured leaf explants were cultured on the same medium, mean number of 13.5 shoots were produced. Shoot growth was accelerated by adding 50 mg/l of silver nitrate ($AgNO_3$), an anti-ethylene compound to the culture medium. Each shoot was excised from the mass and transferred onto half strength MS medium for rooting. Zygotic embryos at different developmental stages were cultured on LS medium supplemented with various growth regulators to induce somatic embryos. When cultured on LS medium with 1.0 mg/l 2,4-D. 14.3% of the zygotic embryos induced somatic embryos. Upon transfer onto the basal medium, somatic embryos sporadically converted into plantlets.

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