• Title/Summary/Keyword: leaf discs

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Comparative Analysis of Resistance to Antibiotics in Populus alba$\times$P. glandulosa Transformed by nptII or hpt Gene (NPTII 및 HPT 유전자가 삽입된 현사시의 항생제에 대한 저항성 분석)

  • 이은정;노은운;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.243-248
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    • 2001
  • This study was peformed to find out the optimal conditions for the selection of transformed cells using already established transgenic plants. Several transgenic poplar (Populus alba$\times$P giandulosa) lines carrying npt II or hpt gene as a selectable marker were tested against kanamycin or hygromycin. Two culture explants, leaf discs and nodes, were compared regarding their sensitivity to the antibiotics. When leaf discs of untransformed control plants were cultured on callus inducing media in the presence of varying levels of kanamycin or hygromycin, only those cultured on the media containing lower than 50 mg/L kanamycin or 2 mg/L hygromycin formed callus. However, much higher concentration of kanamycin was needed to suppress the growth of axillary buds of untransformed plants. On the other hand, hygromycin at the concentration of 5 mg/L effectively suppressed shoot growth of untransformed plants. Root induction from untransformed plants could also be suppressed at the concentration of 50 mg/L kanamycin or 5 mg/L hygromycin. The transgenic plants showed resistance to 100 mg/L kanamycin or 50 mg/L hygromycin in the growth of callus, shoots, and roots. Hygromycin appeared to be more efficient in selecting untransformed cells than kanamycin.

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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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Changes in Isozyme Patterns of Peroxidase and Esterase during Regeneration of Pepper (Capsicum annuum L.) (고추 (Capsicum annuum L.)의 재분화에 따른 Peroxidase와 Esterase 동위효소의 양상 변화)

  • 정현숙
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.9-15
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    • 1992
  • The plantlet was regenerated on MS medium containing BAP (2 mg/I) and 1M (1 mg/I) from leaf discs of pepper after 3 weeks of culture. And then, we investigated the activity of peroxidase and esterase and the pattern of their isozymes from leaf, stem and root in order to observe physiological and biochemical changes on the developemental stage, respectively. The peroxidase was expressed with tissue specificity because peroxidase activity according to the developemental stage of the tissue was not only highest in the leaf of the pepper at 10 days after it germinated but also 2 new bands of its isozyme were found in pI 7.2 and pI 5.2. However, a new pI 3.4 band was found in the leaf and root of the pepper after 14 days of germination, and in the stem was found out pI 5.2 band. As regeneration of leaf dises was progressed, its peroxiase activity was increased about 80% more than that of control after 14 days of culture and new pI 3.2 and 6.5 bands of it isozyme were found. The results suggested that peroxidase would be connected with regeneration of pepper. Also, esterase activity was increased about 50% more than that of control after 14 days of culture, the pattern of esterase isozyme was shown to be 3 cathodic bands and 1 anodic band after 7 days of culture.ulture.

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Quality Control of Photosystem II during Photoinhibition

  • Yamamoto, Yasusi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.55-58
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    • 2002
  • The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

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A High-Efficiency Direct Somatic Embryogenesis System for Strawberry(Fragaria x ananassa Duch.) Cultivar Chandler

  • Husaini, Amjad M.;Aquil, Samina;Bhat, Mukhtar;Qadri, Tabassum;Kamaluddin, Kamaluddin;Abdin, Malik Zainul
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.107-110
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    • 2008
  • A high-efficiency, reproducible somatic embryogenesis system for strawberry cultivar Chandler was developed. Thirty-one somatic embryos per explant(max no.) were recorded in leaf discs which were cultured on medium containing MS salts+$B_5$ vitamins+2% glucose+4.0 mg $1^{-1}$TDZ(Thidiazuron) and incubated at $10{\pm}1^{\circ}C$ under darkness for one week followed by three weeks under 16-h photoperiod. The scanning electron microscopic(SEM) ontogeny revealed the normal development of somatic embryos from globular to heart-shaped and dissection microscopy from torpedo-shaped to cotyledonary-stage embryos. The maximum germination percentage of 48% could be obtained on MS medium containing kinetin(1.0 mg $1^{-1}$) and the maximum survival percentage(79%) of plantlets after four weeks was found to be in the mixture of vermiculite, peatmoss, and soilrite(1:1:1).

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Influences of Temperature and Light on the Herbicidal Activity of Bleaching Herbicides (Bleaching Herbicides의 제초활성에 영향을 미치는 온도 및 광의 영향)

  • Kim, J.S.;Na, J.Y.;Cho, K.Y.
    • Korean Journal of Weed Science
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    • v.9 no.3
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    • pp.230-237
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    • 1989
  • This research was carried out to investigate the influences of temperature and light on the herbicidal activity of oxyfluorfen, oxadiazon and paraquat. Increased temperature from 10 to $35^{\circ}C$ resulted in increase of herbicidal activity in whole plants or leaf discs treated with herbicides. It seemed that temperature affected herbicide penetration into and reaction to the action site rather than appearance process of herbicidal activity (maybe membrane peroxidation after being absorbed. The activity of compounds tested increased with increased light intensity. Paraquat showed similar activities regardless of light qualities but oxyfluorfen and oxadiazon showed the highest activities in blue light spectrum, indicating that they seemed to be closely related to chlorophyll biosynthesis rather than carotenoid biosynthesis or electron transport systems of photosynthesis and respiration.

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Control Effect of New Developed Insecticide on Two Spotted Spider Mite (Tetranychus urticae Koch) Collected from Commercial Roses (장미에서 채집된 점박이 응애(Tetranychus urticae Koch)에 대한 신규 살충제의 방제 효과)

  • Park, Hyean-Cheal;Choi, In-Soo;Kang, Jeom-Soo
    • Journal of Life Science
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    • v.13 no.2
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    • pp.150-153
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    • 2003
  • To determine the effect of newly developed insecticide on two spotted spider mite, Tetranychus Urticae, populations, ten leaf discs were sprayed with three different concentrations (0.5%, 1.0%, and 1.5%) of the tested insecticide solution, and with a water-sprayed control. The tested insecticide solution was toxic to adult female and immature of two spotted spider mite, but was relatively non-toxic to eggs. For equivalent concentrations, more adult females were killed than immature stages. There was no marked difference between the hatch of treated and untreated eggs, although a weak trend between the numbers of unhatched eggs and the insecticide concentration after 7 days is shown.

Effect of Fertilization of UV-B Sensitivity of Cucumber Plant (질소, 인산, 칼륨시비에 따른 오이의 자외선 감수성 변화)

  • Bae, Gong-Young;Lee, Yong-Beom;Park, So-Hong
    • Korean Journal of Environmental Agriculture
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    • v.16 no.3
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    • pp.227-232
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    • 1997
  • Visible injury appeared 7 days after ultraviolet-B(UV-B) irradiation, but did not show any significant decline of growth in cucumber plant. However the growth of the first leaves of fertilized plants was suppressed by UV-B irradiation. Especially the most effective growth retardiation appeared when supplied with nitrogen rather than phosphate and potassium. These results suggest that UV-B may play an important role in inhibiting nitrogen metabolism. Therefore we examined the effect of activity of nitrate reductase, and found that the nitrate reductase activity of the first leaves was increased by UV-B irradiation for 7 days and fertilization. We examined the effect of plant hormone on the inhibition of growth in the first leaves. Benzyladenine promoted the growth of discs excised from the first leaves by fertilization and without UV-B, but did not promote the growth of leaf discs from UV-B irradiated plants. We conclude that the UV-B-induced decrease in the growth of the first leaves could be related to reduction in sensitivity to plant hormones.

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Gene Transformation of Ailanthus altissima Swingle by Agrobacterium tumefaciens (외래유전자(外來遺傳子)에 의(依)한 가중나무의 형질전환(形質轉換))

  • Park, Young Goo;Huh, Kyung;Choi, Myung Suk
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.137-145
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    • 1992
  • An efficient transformation system was established for Ailanthus altissima utilizing the binary system of A. tumefaciens strain LBA4404. Callus was initiated from small portions of cambium tissue of A. altissima in vitro. Optimum regeneration was achieved with Murashige and Skoog(MS) medium containing 0.01mg/${\ell}$ 2, 4-D, 0.5mg/${\ell}$ BAP, 3%(w/v) sucrose and 0.75% agar. The multiplication of explants remarkably showed up on medium containing 1.0mg/${\ell}$ BAP. Leaf discs or internodal stem segments were inoculated with A. tumefaciens strain LBA 4404 containing the binary vector pPMB 101, which has both ${\beta}$-glucuronidase (GUS) marker gene and neomycin phosphotransferase II (NPT II) gene. Shoots had been regenerated from 24 lines out of inoculative 50 lines. Transformants were selected by their ability to grow on medium containing kanamycin sulphate (100mg/${\ell}$). Putative transformation was confirmed by GUS assays. Five GUS-positive plantlets were obtained which confirmed that this marker gene has been transferred into A. altissima.

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Selection of Transgenic Potato Plants Expressing NDP Kinase 2 Gene with Enhanced Tolerance to Oxidative Stress (NDP Kinase 2 유전자를 도입한 산화스트레스 내성 형질전환 감자의 선발)

  • Li, Tang;Kwon, Suk-Yoon;Yun, Dae-Jin;Kwak, Sang-Soo;Lee, Haeug-Soon
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.191-195
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    • 2004
  • Arabidopsis NDPK2 (AtNDPK2) is a key singaling component that regulate cellular redox state and known to enhance multiple stress tolerance when over-expressed in Arabidopsis plant (Moon et al. 2003). In order to develop transgenic potato plants with enhanced tolerance to multiple stresses, we placed an AtNDPK2 cDNA under the control of a stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter. Transgenic potato plants (cv. Superior and Atlantic) were generated using an Agrobacterium-mediated transformation system and selected on MS medium containing 100 mg/L kanamycin. Genomic Southern blot analysis confirmed the incorporation of AtNDPK2 cDNA into the potato genome. When potato leaf discs were treated with methyl viologen (MV) at 10 $\mu$M, transgenic plants showed higher tolerance to MV than non-transgenic or vector-transformed plants. The NDPK2 transgenic potato plants will be further used for analysis of stress-tolerance to multiple environmental stresses.