• Title/Summary/Keyword: callus development

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Multi-secondary Somatic Embryogenesis and Plant Regeneration from Hypocotyl Cultures of Alfalfa (Medicago sativa L.) (알팔파의 하배축으로부터 다량의 이차 체세포배 발생과 식물체 재분화)

  • Won, S.H.;Lee, B.H.;Kim, K.Y.;Lee, H.S.;Lee, H.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.273-280
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    • 1999
  • Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

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The Study on Tissue-Cultured Echinacea purpurea Adventitious Roots Extract for Application as a Cosmetic Ingredient (조직 배양한 에키네시아 추출물에 관한 효능 연구)

  • Park, Chang-Min;Joung, Min-Seok;Choi, Jong-Wan;Paek, Kee-Yoeup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.2
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    • pp.137-142
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    • 2008
  • Echinacea purpurea, an indian traditional plant medicine has been widely used as herbal remedy for the treatment of disease such as colds or other infections. However, Echinacea purpurea extracts recently have been applied as a cosmetic ingredient for skin care. We artificially cultured Echinacea purpurea by using the bioreactor culture system for this study. We induced callus from Echinacea purpurea and separated adventitious roots, harvested and extracted after cultured in bioreactors. Previously, several studies have been reported on anti-oxidant and immuno-enhancing effects of Echinacea purpurea extract but other efficacies were not well known. In this study, we investigated the whitening, anti-wrinkle and anti-oxidant effects to know applicable value of tissue-cultured Echinacea purpurea adventitious roots extract(TCEPARE) as a cosmetic ingredient. TCEPARE did not show cytotoxicity until a concentration of 2% and showed the anti-oxidative effect in DPPH and NBT tests. Also, the extract decreased tyrosinase expression in a dose-dependent manner and inhibited melanin synthesis in B16 melanoma cells. TCEPARE reduced protein levels of MMP-1, 2 secreted in culture medium or in cell lysates. From these results we suggest that TCEPARE has potential benefits applicable as to cosmetic ingredient for skin care products.

Development of Bialaphos-Resistant Transgenic Rice Using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 bialaphos 저항성 형질전환 벼의 개발)

  • 이효연;이춘환;김호일;한원동;최지은;김진호;임용표
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.283-288
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    • 1998
  • The bialaphos is a potent inhibitor of glutamine synthease in higher plants and is used as a non-selective herbicide. We have used the bialaphos resistant gene(Bar) encoding for an acetyltransferase isolated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice(Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefaciens EHA101 carring a plasmid pGPTV-HB containing genes for hygromycin resistance (HygR) and Bar. Transgenic plants showing in vitro resistance to 50 mg/L hygromycin and 10 mg/L bialaphos were obtained by using a two-step selection/regeneration procedure. Transformation efficiency of rice was about 30% which was as high as reported in other dicotyledons. Progenies ($\textrm{T}_{1}$ generation) derived from primary transformant of 17 lines were segregated with a 3 resistant : 1 sensitive ratio in medium containing hygromycin and bialaphos. Stable integration of Bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from $\textrm{T}_{2}$ progenies. Transgenic plants ($\textrm{T}_{3}$) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.

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Plant regeneration through multiple-shoot induction and ex vitro rooting in Vaccinium oldhamii Miq. (정금나무(Vaccinium oldhamii Miq.)의 다신초 유도 및 기외발근을 통한 식물체 재분화)

  • Yun, Ayoung;Kim, Tae Dong;Kim, Ji Ah;Lee, Na Nyum;Cheong, Eun Ju;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.82-89
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    • 2022
  • In vitro techniques were developed for propagating Vaccinium oldhamii using shoots with apical buds. Explants having an apical bud were cultured on Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, and 5.0 mg/L of each zeatin, thidiazuron, 6-benzylaminopurine (BA), and 6-(γ,γ-dimethylallylamino)purine (2-iP) in order to induce multiple shoots. Among the tested treatments, the 2.0 mg/L of 2-iP proved to be most suited for the multiplication and growth of shoots; the multiple shoot induction rate was 100.0%, the average number of shoots was 7.4 per explant, and the average shoot length was 51.7 mm. The in vitro elongated shoots were rooted on half-strength MS medium containing various concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA). However, overall callus overgrowth was observed in all treatments and resulted in necrosis and abnormal shoot growth in root formation. A low concentration (0.5 mg/L) of IBA was appropriate for normal root development and the in vitro rooting rate was 30%. Ex vitro treatments on root formation using various concentrations of IBA with Talc powder and two types of rooting substrates (Flexi-Plugs or Horticultural soil) were examined. The ex vitro rooting rate (80%) and length of roots (32.9 mm) were obtained when the cut ends of the shoots were treated with 1.0 mg/L IBA and cultivated in Horticultural soil for 2 months. These findings suggest that ex vitro rooting is the more effective method for improving root formation in Vaccinium oldhamii than in vitro rooting.

Anatomical Studies on Root Formation in Hypocotyl and Epicotyl Cuttings of Woody Plants (임목(林木)의 배축(胚軸) 및 유경삽수발근(幼茎揷穗発根)의 해부학적(解剖学的) 연구(研究))

  • Choi, Man Bong
    • Journal of Korean Society of Forest Science
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    • v.52 no.1
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    • pp.1-30
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    • 1981
  • The origin and development of adventitious roots was studied using hypocotyl and epicotyl cuttings of 34 species, 24 genus of woody plants. These cuttings obtained from young seedlings cultured in vials containing distilled water only. The several characteristics of cuttings materials studied are shown in Table 1. The results are summerized as follows: 1. The circumference shapes of cross-sections of hypocotyl and epicotyl cuttings can be divided into six categories, namely, round, irregular round, ellipse, irregular ellipse, square, and triangle. Species differences within a genus did not show any difference of hypocotyl and epicotyl cross-sections shape, however, a noticeable variation among genus or higher taxa. 2. The arrangements of vascular bundles in the cross-sections of hypocotyls or epicotyls were almost all collateral types and generally showed generic characteristics differing one to the other. However, there were some variations between species within the genus. Six models of vascular bundle arrangement were proposed for all the above speices. 3. The rooting portions of hypocotyl and epicotyl cuttings in this experimental materials can be grouped as follows: (1) Interfascicular parenchyma; (Thuja orientalis. T. orientalis for. sieboldii, Acer microsieboldianum, A. palmatum, A. saccharinum, Cercis chinensis, Lespedeza bicolor, Magnolia obovata, M. sieboldii, Mallotus japonicus, Staphylea bumalda) (2) Cambial and phloem parenchyma: (Chamaecyparis obtusa, C. pisifera, Albizzia julibrissin, Buxus microphylla var. Koreana, Cereis chinensis, Euonymus japonica, Firmiana platanifolia, Lagerstroemia indica, Ligustrum salicinum, L. obtusifolium, Magnolia kobus, M. obovata, Mallotus japonicus, Morus alba, Poncirus trifoliata, Quercus myrsinaefolia, Rosa polyantha, Styrax japonica, Styrax obassia) (3) Primary ray tissues; (Euonymus japonica, Styrax japonica) (4) Leaf traces; (Quercus acutissima, Q. aliena) (5) Cortex parenchyma; (Ailanthus altissima) (6) Callus tissues; (Castanea crenata, Quercus aliena, Q. myrsinaefolia, Q. serrata) 4. As a general tendency throughout the species studied, in hypocotyl cuttings, the adventitious root primordia were originated from the interfascicular parenchyma tissue, however, leaf traces and callus tissues were contributed to the root primordia formation in epicotyl cuttings. The hypocotyl cuttings of Ailanthus altissima exhibited a special performance in the root primordia formation, this means that cortex parenchyma was participated to the origin tissue. And in Firmiana platanifolia, differening from the other most species, the root primordia were formed at the phloem parenchyma adjacent outwardly to xylem tissue of vascular bundle system as shown photo. 48. 5. All the easy-to, or difficult-to root species developed adventitious roots in vials filled with distilled water. In the difficult-to-root species, however, root formations seemed to be delayed because they almost all had selerenchyma or phloem fiber which gave some mechanical hindrance to protrusion of root primordia. On the other hand, in the easy-to-root species they seemed to form them more easily because they did not have the said tissues. The rooting portions between easy-to-root and difficult-to-root species have not clearly been distinguished, and they have multitudinous variations. 6. The species structured with the more vascular bundles in number compared with the less vascular bundles exhibited delayed rooting. In the cuttings preparation, the proximal end of cuttings was closer to root-to-stem transition region, the adventitious root formation showed easier. 7. A different case occured however with the mature stem cuttings, in both the needle-leaved and the broad-leaved species. In the hypocotyl cuttings, parenchymatous tissues sited near the vascular bundles become the most frequent root forming portions in general and relevant distinctions between both species were hardly recognizable. 8. In the epicotyl cuttings, root primordia originated mainly in leaf traces in connection with cambial and phloems or callus tissues itself. In the hypocotyl cuttings, interfascicular parenchyma was the most frequent portion of the root primordia formation. The portions of root primordia had more connection with vascular cambium system, as the tissues were continuing to be developed.

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Initial Root Development of Larix leptolepis Gordon Cuttings as Related to Organic Substances and Cutting Date (낙엽송(落葉松)의 삽목발근(揷木發根)에 관련된 물질(物質) 및 발근과정(發根過程))

  • Koo, Yeong Bon;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.85 no.2
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    • pp.300-308
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    • 1996
  • This study was conducted to identify factors involved in rooting of Japanese larch cuttings. Cuttings were taken every week from June 24th to July 25th before and after the 10th of July which supposed to be best time for cutting. The content of IAA, carbohydrate, nitrogen and inorganic elements such as $Mg^{{+}{+}}$, $K^+$ and $Ca^{{+}{+}}$ within cutting materials were analyzed. After the cuttings were planted, the root development was observed every week. Cuttings of easy-to-root clones were higher in IAA contents than the cuttings of hard-to-root clones at the time of cuttings were taken. There were significant differences in moisture content of cuttings taken on June 24th and July 25th between easy-to-root and hard-to-root clones, however no significant differences in moisture contents were observed between them taken on the 8th and 15th of July which supposed to be the best time for cutting. Inorganic matters and carbohydrate contents of cuttings taken from hedged trees were higher than those of non-hedged trees. The ratios K/C and C/N of cuttings taken from hedged trees were higher than those of non-hedged trees. Cuttings taken from easy-to-root clones showed higher content of carbohydrate and nitrogen than those of hard-to-root clones. The contents of inorganic elements, such as $Mg^{{+}{+}}$, $K^+$, and $Ca^{{+}{+}}$ in cutting taken from hard-to-root clones were slightly higher than that of easy-to-root clones. The ratio C/N and K/C of cuttings which were taken from easy-to-root clones have shown a similar trend of carbohydrate and nitrogen. These data indicate that there was no one dominant factor that affect rooting of cutting, but many factors such as contents of IAA, carbohydrate, nitrogen, phosphate, and ratios of C/N and K/C appeared to affect on rooting of cuttings variously. Most of root initials were formed from phloem tissue within one week after cutting, and few were formed from callus tissue. Root primordia were developed from root initials within two weeks after cutting, and most of new roots were developed from root primordia within about five weeks after cutting.

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Characteristics and Possible Early Harvesting Time of Early Maturing Soybean Cultivars in Southern Korea (남부지방에서 조생종 콩 품종의 특성과 조기수확 한계기)

  • Kim, Dong-Kwan;Son, Dong-Mo;Chon, Sang-Uk;Lee, Kyung-Dong;Kim, Kyong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.125-130
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    • 2008
  • This study analyzed the growth, seed quality, and yield of major early-maturing soybean cultivars by comparing them in order to utilize the research results in the selection of early-maturing soybean cultivars in multi-cropping farms in the Southern area. This field trial was conducted at Naju region (latitude $35^{\circ}04'N$, longitude $126^{\circ}54'E$), Jeonnam, with planting on June 15. The maturing date for Keunol-kong and Hwaseong-put-kong was found to be around September 12, which was earlier than other cultivars. Thus, there were advantages to introducing a cropping system as well as having good seed quality and high yield. On the other hand, the maturing date for Saeol-kong and Sinrok-kong was found to be around September 20, which was a little bit late; however, the seed quality of the cultivars was good and they had a high yield. Therefore, if we want to sow the following crops of soybeans around mid-September, Keunol-kong and Hwaseong-put-kong are advantageous, while for the seeding around late September, Saeol-kong and Sinrok-kong would be good. This study was also performed to identify the limitation time for early harvesting by reviewing seed quality and yield of major early-maturing soybean cultivars according to early harvesting. When harvesting Keunol-kong on September 6, which was six days earlier than the optimal harvesting time (September 12), there was no difference in seed weight, yield, or seed quality than those of the harvested at the optimum maturing time. As for Saeol-kong, when harvesting on September 18, which was six days earlier than the optimal harvesting time (September 24), there was no difference in seed weight, yield, or seed quality than those of the harvested at the optimum maturing time. Therefore, the stable limitation time for early harvesting of Keunol-kong and Saeol-kong was concluded to be six days earlier than the optimal harvesting time.

The Studies of Activity of Retrotransposon(Tos17) according to Tissue Culture Periods in Rice(Oryza sativa L.) (벼 조직배양 기간에 따른 retrotransposon(Tos17)의 활성에 관한 연구)

  • Yang, Hee-Eun;Fang, Yilan;Shin, Young-Boum;Lee, Boung-Jin;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.20 no.5
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    • pp.389-397
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    • 2007
  • Using the active-increment of Tos17 copies in the genome of Oryza sativa L., there were many studies about induction and selection of new mutants. This study mainly focuses on the induction for retrotransposon(Tos17) activity in the callus of Ilpumbyeo(Oryza sativa L.) according to varied culture period and condition. The objectives of this study are obtaining various mutants($M_1$) through plant regeneration, identification of the mutation relation with Tos17, and subsequent phenotyping of the mutants($M_2,\;M_3$). A total of 371 $M_1$ mutants was obtained. The degree of Tos17 activity obtained regeneration plants with each different culture period was evaluated by Southern blot analysis. The result showed that control Ilpumbyeo rice has 5 numbers of copies and the band numbers obtained 7, 8, 9.5, 12, 6, 13.5, 17.5 from culture period of 1, 2, 3, 5, 6, 7, 8 month, respectively. In this study, the result showed that most effectual culture period for activity of Tos17 in Ilpumbyeo rice is 5 month. Hereafter, collections and analysis of various recombination plants will act on an important factor in multiplication and preservation of $M_2$ and $M_3$ generation. And an urgent and important subject is a development of screening method for selection of diverse mutants.