• Title/Summary/Keyword: anther-derived callus

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Embryogenesis and Regeneration from Anther Cultures of Bupleurum falcatum L. (시호의 약배양의 통한 배발생 및 재분화)

  • 권순태;정형진;김길웅
    • Journal of Life Science
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    • v.11 no.4
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    • pp.340-345
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    • 2001
  • Effects of growth regulators(2,4-D, IAA, NAA, BA and kinetin) and chilling treatment on callus induction. embryogenesis and regeneration through anther cultures of B. falcatum L. were examined. Frequency of callus induction and embryogenesis was effectively increased by the treatment of chilling at 5$^{\circ}C$ for 10 days before anther inoculation. Optimal level of growth regulator for callus induction and embryogenesis from anther was 2,4-D 1.0 mg/L in Murashige and Skoog(MS) basal medium supplemented with 30 g/L sucrose, 8 g/L agar. Frequency of embryogenesis from anther derived callus was increased up to 48% or 45% by addition of IAA 0.1+ kinetin 1.0 mg/L of IAA 0.1+ BA 1.0 mg/L in MS medium, respectively, Optimal medium for obtaining green callus was MS basal supplemented with BA 1.0mg/L. Addition of auxins(IAA or NAA) inhibited the formation of green callus from anther derived callus.

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Selection of Rice Primary Pollen Callus with Improved Cold Tolerence (벼 꽃가루 캘러스의 저온처리에 의한 내연성 기내선발)

  • 양세준;오병근
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.35-39
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    • 1994
  • Is obtain cell lines showing high level of rice cold tolerance, direct in vitro selection through cold stress on primary pollen callus derived from anther culture was carried out Genotypic difference in callus formation and plant regeneration was recognized Rates of albino was increased along the duration of cold stress. Reciprocal effects were not noticed in anther culturability There was no variants related to rice leaf discoloration in pollen derived lines from parental varieties, regardless of days of cold stress. The regeneration and recombination of rice leaf discoloration in 146 pollen-derived lines, 70 pollen-derived lines from cold stress at $0^{\circ}C$ for 10 days, and 830 F$_2$ plants presented normal distribution curves with skewness in tolerance and no significant difference among 3 populations. Direct in vitro selection for rice cold tolerance through cold stress on primary pollen callus derived from anther culture, therefore, was revealed ineffective as a in vitro technology.

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Selection of Herbicide Tolerant Cell lines from $\gamma$-ray-Irradiated Cell Cultures in Rice (Oryza sativa L. cv. Ilpumbyeo)

  • Bae, Chang-Hyu;Lee, Young-Ill;Lim, Yong-Pyo;Seo, Yong-Won;Lee, Do-Jin;Yang, Deuk-Chun;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.4 no.3
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    • pp.123-127
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    • 2002
  • Herbicide tolerant rice (Oryza sativa L. cv. Ilpumbyeo) cell lines were selected from $\gamma$-ray-irradiated anther-derived cell cultures. The anther-derived cell clusters were small (300 to 400 ${\mu}{\textrm}{m}$ in diameter) and uniform ones that were screened by miracloth filtering. The cell suspensions were very efficient to plate one layer onto agar medium and to screen target cell lines. Herbicide tolerant cell lines were selected by 5 mg/L cyhalofop butyl (CHB) treatment by using the small cell suspensions on agar N6 medium containing 1 mg/L 2,4-D and 0.2 mg/L kinetin. Of the cell lines, one line (CHB-1) showed stable tolerance at 10 mg/L concentration after 6-month culture without herbicide suspension. Growth stability of CHB-1 was similar to that of control cell line on 10 mg/L CHB containing medium. In this experiment we established herbicide tolerant cell line selection system by using anther-derived uniform-cell suspensions with $\gamma$-ray-irradiation.

Effects of Media and its Components on Callus Induction and Plant Differentiation in Rice Anther Culture (배 약배양에서 배지조성이 Callus 및 식물체분화에 미치는 영향)

  • 손재근;오병근;이수관
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.3
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    • pp.271-276
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    • 1985
  • Effects of media and its components on callus induction and plant regeneration were studied to increase the cultural efficiency in rice anther culture. The N$_{6}$ basic medium gave better results in callus induction than those of MS or Miller. The medium used for callus induction affects the plant regeneration. The frequency of plant regeneration from callus grown on Miller basic medium was lower than those of N$_{6}$ or MS. Most of calli derived from anthers, above 90%, were induced from 20 days to 40 days after anther inoculation. The cultural efficiency of modified N$_{6}$ basic medium which was composed of 31.5mM KNO$_3$ and 1.75mM(NH$_4$)$_2$SO$_4$ as nitrogen sources was higher than those of N$_{6}$ basic medium. Combination of NAA and Kinetin showed better results than that of 2, 4-D only in cultural efficiency. Effect of DL-alanine on callus induction in Indica variety, IR40, showed better response in the anthers pretreated for 6 days at 12$^{\circ}C$.

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Embryogenesis and plant regeneration of Panax ginseng Meyer via anther culture and ploidy assessment using flow cytometry (인삼 약 배양을 통한 배 발생과 식물 재분화 및 유세포 분석기를 이용한 배수성 검정)

  • Jung-Woo Lee;Kyong-Hwan Bang;Dong-Hwi Kim;Jang-Uk Kim;Young-Chang Kim;Ick-Hyun Jo
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.19-26
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    • 2023
  • Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

Effect of Genotype of Donor Plants on the Success of Anther Culture in Sweet Pepper (Capsicum annuum L.)

  • Shrestha, Surendra Lal;Kang, Won-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.6
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    • pp.506-512
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    • 2009
  • To study the effect of genotype of donor plants on anther culture, anthers of nine hybrid cultivars (Derby, Special, Bossanova, Minipaprika, Fiesta, Boogie, Phenlene, Kufrah, and Clarity) of sweet pepper (Capsicum annuum L.) were cultured in a petridish containing C medium (Sibi, Dumas De Vaulx medium) supplemented with 0.1 mg/L 2,4-D and 0.1 mg/L kinetin, 3% sucrose and 0.32% Phytagel. The cultures were incubated in the dark at $35^{\circ}C$ for seven days, and then cultured at $25^{\circ}C$ with a photoperiod of 16 hr. daylength for 40 days. Frequency of callus formation and plant regeneration was varied among cultivars. Callus formation was ranged from 6% in Phenlene to 69.8% in Kufrah. The highest percentage of regenerated plantlets was obtained in cv. Phenlene (2.67%) followed by Bossanova (2.41%). Result of ploidy analysis; chromosome number observation and flowcytometry analysis, showed that haploid plants could be developed from all of these hybrid cultivars except cv. Fiesta, where highest percentage of haploid plants were obtained in Minipaprika (40%) followed by cv. Bossanova (36.1%). Haploid plants derived from these hybrid cultivars contained single set of chromosome (12 in numbers), higher stomata density (numbers), and smaller sized stomata as compare to diploid plants. The mean length of stomata was 26.9 ${\mu}m$ in haploid plants and 35.7 ${\mu}m$ in diploids.

Effects of Gelling Agent Brands and Concentration on Rice Anther Culture (배지응고제의 종류와 농도에 따른 벼 약배양 효율)

  • 양세준;오병근
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.295-299
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    • 1998
  • To detect the effects of gelling agent brands and concentration on rice anther culture, anthers of rice(O. sativa L. japonica, cv, Nagdongbyeo) were inoculated on N6-Y1 basic media supplemented with 0.4~1.6% Bacto agar(Difco, 04140-01), Agarose(Sigma, Type 1) and 0.2~0.8% Gelrite(Kelco, 143364) as gelling agents. On 0.4% Bacto agar and Agarose media, the frequency of callus formation which was significantly decreased in proportion to gelling agent's concentration was 39% and 55%, respectively. On 0.6% Gelrite media, the frequency of callus formation which was not statistically significant among the 0.2~0.8% concentration was 44%. Calli derived from the higher concentration of gelling agents showed embryogenic with slow growth, small, whitish and hard shape compare to that of the lower concentration. The frequency of green plant regeneration was high not only in calli derived from the higher concentration but also in plant regeneration medium with the higher concentration after callus transfer. Calli derived from the higher concentration was effective to maintain the frequency of green plant regeneration up to 60 days after anther inoculation. Introduction of 0.6~0.8% Geltite for callus formation, then transferred 1.6% Bacto agar and Agarose or 0.8% Gelrite for green plant regeneration was effective to increase anther culture efficiency.

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High Frequency Production of Doubled-haploid Plants by Colchicine Application in Anther Cultures of Rice (벼 약배양에서 콜히친이 식물체 배수성에 미치는 영양)

  • 권용삼;이형규;박규환;손재근
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.37-40
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    • 2001
  • To optimize the in uitro chromosome doubling procedure in anther cultures of rice, anthers were cultured on callus induction medium with 0.001 to 0.1 mg/L colchicine for 30 days. The addition of colchicine slightly reduced callus formation and plant regeneration in comparison to the colchicine-free medium (control medium). This reduction was greater with higher concentration colchicine. Microspore-derived rice plants in control medium were found to be mainly haploid (50 to 58.8%) and doubled-haploid (31 to 40%) in anther culture of 3 Japonica and 1 Tonsil type cultivars. The application of 0.001 mg/L colchicine was increased to 54.3∼60.0% in the frequency of fertile doubled-haploid plants. These results indicate that the addition of colchicine to the callus induction medium is an efficient means to obtain doubled-haploid plants in anther cultures of rice.

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Isolation of Gamma-Induced Rice Mutants with Increased Tolerance to Salt by Anther Culture

  • Lee, In-Sok;Kim, Dong-Sub;Hyun, Do-Yoon;Lee, Sang-Jae;Song, Hi-Sup;Lim, Yong-Pyo;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.51-57
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    • 2003
  • Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

Improvement of Cultural Efficiency Using DNA Markers in Anther and Seed Culture of Rice (DNA marker를 이용한 벼의 조직배양 효율 재선)

  • Kim Kyung-Min
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.527-533
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    • 2006
  • The purpose of this study is to improve the culturability of a indica type rice cultivar, IR 36, using DNA marker associated with the ability of plant regeneration in anther and seed culture. The varietal difference of ability of callus formation and plant regeneration was investigated in anther and seed culture of 8 rice cultivars. Three japonica rice cultivars showed to have better culturability than those of tongil and indica type genotypes. But two indica/japonica lines, 'MGRI 079' and 'MGRI 036', which were selected to have good culturability in previous study showed the highest regenerability (20%) in anther culture of 8 rice cultivars. Thirty four $BC_2F_4$ lines were selected by marker screening using RZ400 for 100 $BC_2F_4$ lines derived from a cross $'MGRI\;079/IR\;36^{^*3}'$. The frequency of callus formation of 30 $BC_2F_4$ lines was higher than those of 'IR 36' in anther culture of the selected $BC_2F_4$ lines. The ability of plant regeneration of 15 lines was higher than that of 'IR 36' in the seed culture of 34 $BC_2F_4$ lines. A promising line, $BC_2F_4-28$, was selected to have better culturability in the anther and seed culture of the $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4-28$ was similar to 'IR 36'. In seed culture of 50 $BC_4F_3$ lines derived from a rice cross $'MGRI\;079/IR\;36^{^*5}'$, 11 lines including $BC_4F_3-3$ showed to have higher regenerability compared with 'IR 36'. The highest frequency of plant regeneration (11%) was obtained from $BC_4F_3-46$ in seed culture of the $BC_4F_3$ lines.