• Title/Summary/Keyword: callus development

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Development of rice(Oryza sativa L.) transformation system to improve callus utilization (캘러스 활용도를 향상시키기 위한 벼(Oryza sativa L.) 형질전환 시스템 구축)

  • Park, Ji-Sun;Moon, Ki-Beom;Ha, Jang-Ho;Jang, Ji-Young;Kim, Mi-Jin;Jeon, Jae-Heung;Park, Sang-Un;Kim, Hyun-Soon
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.170-179
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    • 2017
  • Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

Propagation of Bell Pepper (Capsicum annuum var. grossum) through tissue culture

  • Senarath, WTPSK;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.131-132
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    • 2003
  • Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

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In vitro plant regeneration from axillary buds of Hibiscus syriacus L.

  • Jeon, Seo-Bum;Kang, Seung-Won;Kim, Wan-Soon;Lee, Gung-Pyo;Kim, Sun-Hyung;Seo, Sang-Gyu
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.174-178
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    • 2009
  • Presently, we report a simple, reproducible and high frequency plant regeneration in Hibiscus syriacus L. using axillary buds. H. syriacus was regenerated from axillary buds directly or through a callus phase. Regenerated shoots were directly induced from young and fresh axillary buds cultured on Murashige and Skoog medium (MS) supplemented with 0.01 mg/L of the growth regulator thidiazuron (TDZ) after 2 weeks of culture. Directly induced shoots were transferred to hormone-free MS medium and root development was observed after 6 weeks. On the other hand, old and stale axillary buds were regenerated to shoots via callus induction on MS medium containing 0.01–2 mg/L TDZ after 4 weeks. A TDZ concentration of 0.01 mg/L was most effective in callus formation. Green callus was transferred to MS medium containing 0.01 mg/L α-naphthalene acetic acid (NAA) and 0.5 mg/L benzylaminopurine (BA). After 4 weeks, callus had developed into multiple shoots. Plantlets were formed from 10 week cultures of single shoots on hormone-free MS medium. Regenerated plantlets were cultured on MS medium for one month and then transferred to pots containing garden soil. Potted plants were acclimatized for one month and grown to maturity under greenhouse conditions. The present study has shown that various concentrations of plant growth regulator can be effective for in vitro plant regeneration of H. syriacus. The direct and indirect regeneration protocol presented here will be useful for understanding the manipulation and propagation of H. syriacus.

Effects of Growth Regulators on Plant Regeneration in Shoot-Tip-Derived Embryogenic Callus Cultures of Sweet Potato (Ipomoea batatas) (고구마 경단 유래 배발생 캘러스로부터 식물체 재분화에 미치는 생장조절제의 영향)

  • ;Shozo FUJIOKA
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.281-286
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    • 1994
  • The hormonal regulation of organ differentiation was investigated in the tissue culture of sweet potato. Embryogenic callus was induced from shoot tips cultured on MS medium supplemented with 1 mg/L 2,4-D. When embryogenic callus was transferred to medium containing 0.1 mg/L GA$_4$, it proliferation was stimulated. The callus gave rise to plantlets when cultured on medium containing 0.1 mg/L BA. Addition of 0.1 mg/L jasmonic acid or 0.01 mg/L brassinolide to medium was effective for the development of healthy normal plantlets.

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Cytohistological Study of Abnormal Cell Division of Arabidopsis Stem Infected with Geminivirus (Geminivirus에 감염된 Arabidopsis 줄기의 이상세포분열에 관한 세포조직학적 연구)

  • 박종범;이석찬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.153-158
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    • 1998
  • The internal structures of Arabidopsis thaliana infected with beet curly top virus (BCTV) were studied by light microscopy. Hyperplasia was observed in the inflorescence stems of Arabidopsis thaliana ecotype Sei-O at 2 weeks after BCTV-Logan inoculation and callus was induced on symptomatic tissues at 4 weeks after virus inoculation. The infection processes were revealed as follows: hyperplasia of phloem tissue, necrosis of hyperplastic phloems, lacuna formation of necrotic tissues, elongation and enlargement of cortex and epidermal cells surrounding the lacuna formed phloem tissues, induction of cell division in the enlarged cortex and epidermal cells, and induction of callus tissue. Callus formation on Arabidopsis was caused by the virus infection, and virus inclusion body was observed in both phloem and callus tissue by azure-A staining.

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Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration from Seed Culture of Reed (갈대(Phragmites communis Trinius.)의 종자배양에 있어서 식물생장조절물질이 캘러스 유도와 식물체 재분화에 미치는 영향)

  • Kim, Yong-Goo;Kim, Kyung-Hee;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.229-234
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    • 2011
  • In order to develop an efficient, reliable and reproducible tissue culture system for reed (Phragmites communis Trinius), an efficient plant regeneration system via callus induction was established using mature seeds as explants. MS medium containing 1 mg/L 2,4-D and 0.5 mg/L BA was optimal for callus induction from mature seeds. The highest frequency (88.7%) of callus formation was obtained in 1.0 mg/L 2,4-D. The highest plant regeneration frequency (59.6%) was found when the embryogenic calli were subcultured on MS medium supplemented with 100 mg/L myo-inositol, whereas, adding of plant growth regulators was not so promising in this case. Our result would be useful for development of transgenic reed plants.

Studies on the Anther Culture of Prunus mume S. et Z. and the Other Three Species (Prunus mume S. et Z. 외(外) 삼종(三種)의 약배양(葯培養)에 관(關)한 연구(硏究))

  • Kim, Jai Saing
    • Journal of Korean Society of Forest Science
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    • v.31 no.1
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    • pp.1-7
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    • 1976
  • When haploid plant would be appeared by the anther culture, the large quantity of young plant multiplied maternal inheritance and the same pure line rapidly in the short length of time, which will be effected to cut down much expences efforts and time for the production of seeds or seedlings. Therefore, the development of the technique for this would be much profited in the country industry. In the late of a few years studies were early attempted in this field, but up this time there were a few success of plants only and none of perennial plant. In this status of the country condition required earnestly for the development of the green industry, this researcher attempted to culture the anther of late uninucleate microspore or early binucleate microspore of the Prunus mume and other three psecies, economic trees estimated specially economic, on the place of Modified Murashige and Skoog's medium supliment with Kinetine, 2.4-D, and N.A.A for inducing haploid plants. The obtained results were as follows: 1. 2,000 anthers were cultured and there were shown that 2N callus in Prunus mume had 82 as 4.1%, 2N callus in Prunus tomentosa 15 as 0.8%, 2 N callus in Prunus salisina 75 as 4% 2. N callus had shown 40 as 2% from Prunus armeniaca var. ansu only, and the other trees showed all 2N callus. 3. Callus had appeared in every tree but 2N callus appeared was all filaments and there showed from only connective tissue N callus appeared was all from anther locule inside. 4. Then Prunus armeniaca var. ansu only was not callus of somatic anther tissue origin, but as there was callus origined from microspore which was changed in to swollen microspores or polynucleate microspores, it was certained to need haploid plant.

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Histochemical and Cytochemical Studies on the Secondary Xylem Formation during Grafting Process of Pinus thunbergii (접목 과정에서 형성된 신생 2차목부의 조직학적, 세포화학적 연구)

  • Kim, Jong Sik;Lee, Kwang Ho;Kim, Yoon Soo
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.6
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    • pp.1-11
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    • 2006
  • Grafted tissues were investigated using various microscopic techniques. Pinus thunbergii was used as stock and scion and autografted by cleft graft method. Histochemically, grafting processes can be proceeded by four stages: 1) formation of necrotic layer, 2) proliferation of callus, 3) development of neo-cambium from callus, and 4) restoration of new vascular xylem. Necrotic la yer composed of pectin and lignin was gradually degraded during grafting process and disappeared when new union was formed between stock and scion. A large number of starch and lipid bodies in the cytoplasm were also gradually degraded during grafting process and disappeared at the grafting interface. Nucleus and plasmodesmata were not changed. Bubble-like callus was generated from all living parenchyma cells and from the callus. The tracheary elements differentiated from the callus had either reticulate or pit-like thickenings in the secondary walls with bordered pits. Secondary cell wall thickening occurred toward filing to the void parts between reticulated secondary wall. Tracheids formed in the secondary xylem were short with irregular wall thickness. New secondary xylem cells with swirled shapes, which developed in graft union were oriented horizontally and obliquely to axis of the stem.

Anther Culture of Cornus officinalis Sieb. et Zucc. (산채막(山菜莫)의 약배양(約培養)에 환한 연구(硏究))

  • Kwon, Tae-Oh;Kim, Tai-Soo;Lee, Seung-Yeob
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.3
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    • pp.233-239
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    • 1995
  • The effects of media, growth regulators, low-temperature treatments, culture temperature and light were investigated to improve the callus induction and growth in the anther culture of Comus officinalis Sieb. et Zucco. The frequency of callus induction was more effective on WPM medium than MS medium, and it was highest as 54% in WPM medium supplemented with Img/L NAA. Callus growth was stimulated on MS medium supplemented with 2mg/L NAA. Effect of temperature and light on the callus induction and growth was highest as 62% in the treatment for 16/8 hrs. (light/dark) at $25^{\circ}C$ Ef­fect of low - temperature treatment on callus induction was highest as 19. 5% in the treatment for 36 hrs. at $4^{\circ}C$. For organization, green cells and rootings were promoted on MS medium supplemented with O. 5mg/L 2,4-D and 1 mg/L kinetin. The prevention of callus browning was effective on the medium con­taining $3{\sim}5mg/L$ ABA or 5mg/L $AgNO_3$. The supplement of ABA or $AgNO_3$,were maintained callus ac­tivity for 4-5 weeks and they were promoted the development of green cells.

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