• Title/Summary/Keyword: plant calluses

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Cytological and Morphological Characterization of Anther Derived Plants from Sweet Pepper (Capsicum annuum L.) cv. 'Special'

  • Shrestha, Surendra Lal;Luitel, Binod Prasad;Lee, Taek Jong;Kang, Won Hee
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.431-438
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    • 2010
  • Anthers of sweet pepper $F_1$ cultivar 'Special' were cultured on Dumas De Vaulx (C medium), supplemented with $0.1mgL^{-1}$ 2, 4-D and $0.1mg{\cdot}L^{-1}$ kinetin with 3% sucrose, and 0.32% phytagel. The calluses obtained were further sub-cultured on Murashige and Skoog (MS) medium without growth regulators for regeneration. Regenerated plantlets were grown in plastic pots under plastic house and characterized their cytological and morphological characters in spring, 2008. Twenty percent plantlets were identified as haploid plants after chromosome and ploidy analysis. Haploid plants contained 12 chromosomes, high stomatal density with small stomatal length as compared to diploid plants. Stomatal length in haploids was 23.3% smaller than diploids. Haploid plants were characterized as small leaf and petiole size, poor vigor, thin stem and short plant height, short internodes and small flower buds, fruit size and fruit weight as compared to diploid plants and most of the haploid fruits were seedless. SP55, SP62, SP68, SP72 and SP77 are found high yielding double haploids with high total soluble content (8.6, 8.7, 9.2, 9.1 and $9.8^{\circ}Brix$, respectively) and desirable fruit shape, and recommended them to exploit as inbred lines for heterosis breeding.

Effects of Some Plant Growth Regulators on Protein Biosynthesis of Carrot Cells (당근 세포의 단백질 생합성에 대한 몇가지 식물생장조절제의 영향)

  • Yoo, Ki-Jung;Park, Chang-Kyu;Song, Tae-Chul
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.187-195
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    • 1985
  • Electrophoretic studies of protein extracts from carrot calluses suspension-cultured on the media containing kinetin, BA, IAA, NAA or $GA_3$ at the levels of $10^{-6},\;10^{-5},\;10^{-4}M$, respectively, were performed to identify polypeptides and proteins regulated by auxin, cytokinin or GA. Fifteen bands of polypeptide(s) were observed in the callus cultured in the control medium devoid of growth regulators, and their molecular weights were $18._4,\;20._2,\;20._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1,\;and\;59._9\;KD$, respectively. The synthesis of polypeptide appeared to be promoted in two bands by kinetin, in six bands by BA, in one band by IAA, in two bands by NAA, and in four bands by $GA_3$, while inhibited in five bands by kinetin, in three bands by BA, in four bands by IAA, in three bands by NAA and in three bands by $GA_3$. The polypeptides of $40._3\;KD\;42._2\;KD$ seemed to be regulated by cytokinins, and those of $44._1\;KD,37._4\;KD,\;and\;56._6\;KD$ by auxins. The proteins of three bands with relative mobilities of 0.56, 0.84, and 0.92, respectively, increased in the calluses cultured on the media containing kinetin, IAA, $GA_3$, NAA or BA, compared to the control, but it was difficult to identify the proteins specific for each growth regulator.

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Establishment of Efficient Callus Induction System with Picloram Hormone in Lily Plants (Picloram을 이용한 효율적인 백합 캘러스 유도 체계 확립)

  • Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.3
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    • pp.605-608
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    • 2021
  • Lily is one of the most important 5 cut flowers in international flower market and lilies are distributed in Asia, Eurasia and North America. To develop a new lily cultivar, in addition to hybridization, mutation and selection methods, biotechnological techniques including tissue culture are also required. Establishment of tissue culture system is one of the requirement for the breeding program in Lily. Among many fields of plant tissue culture, establishment of regeneration system via embryogenic calluses are studied in many crops. In this study, research was carried out to decide the proper concentration of picloram which is used for the induction of embryogenic calluses. As a result, 3 different types of callused were observed after 3-4 weeks. They were CEC (compact embryogenic callus), FEC (friable embryogenic callus) and white callus type. 1.0 mg /l of picloram showed the best result for the production of embryogenic callus, however, due to its higher rate of browning in this concentration, 0.75 mg/l of picloram was selected as a proper concentration of picloram for the induction of CEC and FEC in Lily. These results can be contributed to the establishment of both regeneration system and mass propagation in lily in the future.

Inhibition of Starch Biosynthesis by Antisense Expression of cDNAs Encoding ADP-Glucose Pyrophosphorylase Small Subunit in Sweetpotato (고구마에서 ADP-Glucose Pyrophosphorylase Small Subunit cDNA의 Antisense 발현에 의한 전분생합성 저해)

  • Min, Sung-Ran;Bae, Jung-Myung;Harn, Chee-Hark;Jeong, Won-Joong;Lee, Young-Bok;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.277-283
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    • 2007
  • Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.

Studies on the Interspecific and Intergeneric Hybridization in Herbage Grasses I. Effects of hybrid embryo age on callus formation and plant regeneration (화본과목초의 종.속간 잡종에 관한 연구 I. 교잡* 일령에 따른 Callus형성과 식물체재분화)

  • 박병훈;김명환
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.9 no.2
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    • pp.62-67
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    • 1989
  • A breeding program in progress at Suweon Livestock Experiment Station, RDA, involves the hybridization of italian ryegrass (Lm) and several leading forage grasses in an effort to combine the nutritive, productivity and palatability qualities of Lm with the adaptive and cold resistance qual; .les of several leading grasses. In order to study the fate of the hybridization between remotely related species, immatured hybrid embryos were cultured on media. The emasculated plants of Lm were Sikem and Tetrone. Reweille, 2n=14, of perennial ryegrass (Lp.), Forager, 2n=42, of tall fescue(Fa), First, 2n=14,of meadow fescue(Fp), Potomac, 2n=28, of orchardgrass(Dg), and Richmond, 2n=42, of timothy(Ph.p) were used as pollinators. Embryos were isolated on 4, 8, 12, 16, and 20 days after pollination and cultured them on modified Murashige and Skoog media. Calluses and plantlets have been obtained after 8 days old embryos crossing between $Lm{\times}Fa$, after 12 days embryos crossing between $Lm{\times}Lp$, Dg, and Ph.p, and after 16 days embryos crossing between $Lm{\times}Fp$. Both callus and shoot formation occurred on 6 % or less of the plated embryos of $Lm{\times}Fp$, Dg, and Ph.p. Embryoderived callus forming shoots have been obtained from 4 days old embryo crossing between $Lm{\times}Dg$ which has not successfully been done anywhere as far as we know. It means that hybrid plants of species without crossability can be obtained through the use of immatured embryo culture. Some of plated embryos developed directly shoots from embryos and the others shoot-forming callus. Cross between related species showed a high frequency of directly shoot formation from plated embryos and cross between remotely related species a high frequency of callus formation.

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Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by ${\gamma}$-irradiation ($^{60}Co$) of adventitious roots

  • Zhang, Jun-Ying;Sun, Hyeon-Jin;Song, In-Ja;Bae, Tae-Woong;Kang, Hong-Gyu;Ko, Suk-Min;Kwon, Yong-Ik;Kim, Il-Woung;Lee, Jaechun;Park, Shin-Young;Lim, Pyung-Ok;Kim, Yong Hwan;Lee, Hyo-Yeon
    • Journal of Ginseng Research
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    • v.38 no.3
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    • pp.220-225
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    • 2014
  • An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

Electroporation Conditions for DNA Transfer into Somatic Embryogenic Cells of Zoysia japonica (들잔디 체세포 배발생 세포로의 DNA 전입을 위한 Electroporation 조건 구명)

  • 박건환;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.13-19
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    • 1998
  • We have reported previously that intact embryogenic cells can be used instead of protoplasts for electroporation-mediated transformation of zoysiagrass and rice. In this study, conditions of the tissue electroporation were examined to optimize the procedures. Embryogenic cell suspensions were established in liquid MS medium containing 2 mg/L of 2,4-D with embryogenic calluses induced from mature embryos of Z. japonica. The suspension-cultured cell clumps were electroporated with 35S-gusA expression vector DNA, and degrees of DNA introduction into the cells were determined by histological expression rates of the gusA marker gene. DNA transfer into the cell clumps occurred in wide range of voltage (100-400 V) and capacitance (10-1980 $\mu\textrm{F}$), but more in the ranges of 200-300 V and 330-800 $\mu\textrm{F}$ DNA concentrations higher than 6 $\mu\textrm{g}$/mL were adequate for GUS expression of the electroporated cells. DNA transfers were confirmed in all three embryogenic cell lines but only in one out of eleven non-embryogenic lines. Positive GUS expressions occurred with DNAs added even 20-40 h after pulse treatments. As a promoter of gusA, Act1 and Ubi1 were effective 7 and 5 times than 35S respectively in number of GUS expression units on electroporated cell clumps. Embryogenic cell clumps survived and regenerated into plantlets after pulse treatments of wide range of conditions.

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Superoxide Dismutase and Peroxidase Activity of Transformed Callus in Tomato (형질전환된 토마토 캘러스의 Superoxide Dismutase와 Peroxidase 활성)

  • 유정민;정형진;김경민;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.177-181
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    • 1998
  • This study was carried out to investigate activity difference in the superoxide dismutase (SOD) and peroxidase (POD) of tomato callus transformed with Agrobacterium containing the GUS gene. Than those of other two tomato cultivars, the hypocotyl explant of JA101 was shown to have higher POD and SOD specific activity of 23 unit/mg protein and 2,156 unit/mg protein, respectively. Relatively high frequency of callus formation was obtained from the hypocotyl explant on MS medium containing 1 mg/L 2,4-D for 30 days and its POD(47 unit/mg protein) and SOD (95,786 unit/mg protein) specific activities were higher than other 2,4-D concentration. The hypocotyl explant and callus cocultivated with Agrobacterium for 72 hours were transferred to MS medium supplemented with 1 mg/L 2,4-D, 30 mg/L kanamycin, 30 g/L sucrose and 4 g/L Gelrite. The hypocotyl explants transferred to the medium formed callus with 45.5% effeciency after 8 weeks. The transformation efficiency confirmed by GUS assay was 21.6%. POD specific activity of the transformed callus (54 unit/mg protein) were somewhat lower than the non-transformed callus (64 unit/mgg protein) and SOD specific activity of the transformed callus (30,300 unit/mg protein) were also lower than the non-transformed callus (37,077 unit/mg protein). However there was no significant difference in POD and SOD isozyme patterns between the transformed and the non-transformed calluses. From these results, it revealed that there was no difference of antioxidant enzyme activities between the transformed callus and the non-transformed callus in tomato.

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The Induction of Somatic Embryogenic Callus from Petals-Derived Callus in Rosa hybrida (국내 육성 장미 품종 꽃잎 유래 체세포배 발생 캘러스 유도)

  • Lee, Su Young;Shin, Ju Young;Lee, Young Ah;Ahn, Chang Ho;Kim, Yae Jin;Park, Pil Man;An, Hye Ryun;Lee, Ka Youn;Jung, Hyun Hwan
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.652-658
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    • 2022
  • This study was conducted to induce somatic embryogenic callus (SEC) derived from petals in rose. The petal explants of 3 cultivars ('Ice Wing', 'Orange Eye' and 'Pink Beauty') with different flower colors were placed on three types media (MS, SH and WPM) supplemented with 11 mg/L 2,4-D, respectively, and then cultured in the dark for 47 days. Calluses were formed at explants of all three cultivars. Also, 'Ice Wing', which were cultured in the SH as the basal medium, showed the highest callus formation rate. However, somatic embryos were generated from only petal-derived callus of 'Ice Wing', which were induced on the WPM as the basal medium, transferred it to SH basal medium supplemented with 3 mg/L 2,4-D, and 300 mg/L L-proline, and cultured for 5 weeks. The SEC has been proliferated every four weeks at the subculture interval. In addition, as a results of making a comparison of expression of RhSERK3 and RhSERK4, which is used as signal for generation of somatic embryo from callus in rose, between the SEC and petal-derived callus from 'Ice Wing' by RT-qPCR, the former showed 10 times higher RhSERK3 expression and 700 times higher RhSERK4 expression than the latter.