• Title/Summary/Keyword: Regeneration frequency

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Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

  • Bhuiyan, Mohammed Shafi Ullah;Min, Sung-Ran;Choi, Kwan-Sam;Lim, Yong-Pyo;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.137-143
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    • 2009
  • An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

High-frequency Plant Regeneration from Mature Seed-derived Callus Cultures of Orchardgrass (오차드그래스 성숙종자로부터 캘러스 유도 및 고효율 식물체 재분화)

  • Lee, Sang-Hoon;Lee, Dong-Gi;Kim, Jin-Soo;Lee, Byung-Hyun
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.341-346
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    • 2003
  • In an effort to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), an efficient and high-frequency plant regeneration system from seed-derived calli was established. Embryogenic calli induced on MS medium containing 3mg/L 2,4-D and 0.1mg/L BA had significantly improved regeneration ability. Plant regeneration rate was 92% when embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among three kinds of medium, MS and N6 medium were optimal for embryogenic callus induction and plant regeneration, respectively. Ho difference in callus induction frequency was observed among four cultivars of orchardgrass, however, "Roughrider" cultivar showed higher regenerability with the frequency of 61%. Addition of maltose to the regeneration medium as a carbon source dramatically increased regeneration frequency up to 69%. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of orchardgrass through genetic transformation.

High Frequency Somatic Embryogenic Callus Induction and Plant Regeneration from Various Indica Rice Genotypes

  • Hoque Md. Enamul;Mansfield John W.
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.257-262
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    • 2006
  • The paper evaluated the behavior of in vitro culture responses from a diverse set of Indica rice (Oryza sativa L.) genotypes. Significant differences were found in embryogenic callus induction frequency, callus growth and plant regeneration frequency when mature embryos of 11 cultivars, breeding lines and land races were compared. Genotype as well as plant growth regulator influenced the plant regeneration frequency. Callus induction frequency was not correlated with callus growth as well as plant regeneration frequency. The regenerated plants could grow to normal, fertile plants after they were successfully established in soil.

The identification of optimum condition for direct regeneration in black raspberry

  • Ran, Choi-Heh;Park, Pill-Jae;Lee, Hee-Kwon;Joong, Yun-Song;Lee, In-Sok
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.163-167
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    • 2008
  • Adventitious buds appeared within 2 weeks on the base of the petiole explants and increased for two months. A maximum of regeneration (15.6%) was obtained on the medium containing $1.5\;{\mu}M$ TDZ in combination with $1\;{\mu}M$ IBA. To know which explants are the best for the induction of regeneration, three explants such as leaf, petiole and leaf-petiole were used. Among the explant types, the leaf-petiole explant was significantly more effective than leaf and petiole for promoting adventitious shoots, with leaf-petiole inducing at the highest regeneration frequency (33.7%). The regeneration frequency of adventitious shoots in the leaf-petiole explants was significantly affected by leaf size and the position of explants. The leaf-petiole smaller than 5 mm leaf in width was induced at the highest regeneration frequency (68.9%). The smaller leaf sizes, the greater regeneration frequency. Also when the leaves are nearer to the shoot tip, the regeneration frequency is higher. When the rooted micro-shoots were transferred to the soil after growing for 6 weeks in the media, the survival rate was 90%.

Callus Induction and Plant Regeneration from Mature Embryos in Oat

  • Lee, Byung-Moo;Kim, Kyung-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.5
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    • pp.352-355
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    • 2002
  • Mature embryos of five oat genotypes were cultured to develop an efficient method of callus induction and plant regeneration. Murashige and Skoog(MS) and N6 media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin were used for callus induction. Percentage of callus induction showed significant among the combinations of plant growth regulators. Callus induction showed high efficiency in medium containing 3 mg/$\ell$ of 2,4-D. The high frequency of callus induction was obtained in Gwiri37. For plant regeneration, calli induced from mature embryos were transferred onto MS and N6 media supplemented with combinations of 6-benzyladenine (BA) and naphthaleneacetic acid (NAA) for 5 weeks. Percentage of plant regeneration showed high in MS medium containing 0.2 mg/$\ell$ of NAA and 1 mg/$\ell$ of BA. The callus initiation medium affected the subsequent plant regeneration. Treatment with 3 mg/$\ell$ of 2,4-D, and 3 mg/$\ell$ of 2,4-D and 3 mg/$\ell$ of kinetin in callus induction media showed high frequency for plant regeneration. Plant regeneration frequency among the genotypes showed significant. Especially, Gwiri37 showed high regeneration frequency. Regenerated shoots were treated with 200, 350 and 500 mg/$\ell$ of indole-3-butyric acid (IBA) transferred onto half-strength MS medium without plant growth regulators. Treatment of shoots with IBA induced root formation rapidly.

Plant Regeneration from Seed-Derived Callus in Perennial Ryegrass (Perennial ryegrass의 종자유래 캘러스로부터 식물체 재분화)

  • 원성혜;이병현;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.20 no.1
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    • pp.19-24
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    • 2000
  • This study was camed out in order to establish plant regeneration via seed-derived callus of perennialryegrass.Varietal difference in callus growth and plant regeneration was obvious between two cultivars of perennialryegrass. "Reveille" showed a relatively high capacity for plant regeneration. The MS medium was superiorto SH or Bj in callus formation and plant regeneration. The highest regeneration frequency (60%) from calluswas obtained in presence of 5 mg/ l 2,4-D and 1 mg/ t kinetin. Regeneration response varied among calluscultures initiated from the same cultivar. Regeneration frequency was the most effective in 6 weeks-old calliafter initiation and lost their regeneration capacity gradually over a period of 12 weeks.(Key words : Perennial ryegrass (Lolium perenne L.), Plant regeneration)e L.), Plant regeneration)

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Frequency improvement of protoplast fusion in coryneform bacteria (Coryne형 제균의 원형질체 융합빈도 향상)

  • 김종헌;임번삼;이세영;전문진
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.190-196
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    • 1985
  • For frequency improvement of protoplast fusion in Brevibacterium flavum, Brevibacterium lactofermentum lactofermentum and Corynebacterium glutamicum, the effect of plasma expanders on fusion and cell wall regeneration, compatison between direct and two-step selection method, tendency of fusion frequency according to pH of fusion fluid and polyethylene glycol concentration were examined. By addition of 3% polyvinyl pyrrolidone to cell wall regeneration medium, regeneration frequencies were expressed 23 (Brevibacterium lactofermentum), 10.4 (Brevibacterium flavum) and 2.7 (Corynebacterium glutamicum) times higher than those of none polyvinyl pyrrolidone medium respectively.

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Improvement of Regeneration Efficiency from Mature Embryo and Leaf Base Segment in Korean Oat Genotypes

  • Kim Kyung-Hee;Moon Jung-Hun;Lee Sang-Kyu;Lee Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.349-353
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    • 2004
  • Mature embryo and leaf base segment of Korean oat were used as materials in an experiment to check plant regeneration efficiency. MS media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin, and picloram were used for callus induction from mature embryos and leaf base segments. Three mg/l of 2,4­D and 3 mg/l of picloram in callus induction medium showed high frequency for plant regeneration from mature embryos. Leaf base segments were transferred to callus induction medium and incubated at $25^{\circ}C$ in 16/8 hr light/dark cycle for 3 weeks. Callus induction from leaf base segments of Malgwiri showed high efficiency in medium containing 3 mg/l of 2,4-D and 1 mg/l of kinetin $(91.8\%)$. In case of Samhangwiri, the combinations of phytohormones did not show significant difference. Regeneration from leaf base segments showed high frequency in shoot medium containing 1 mg/l of antiauxin, tri-iodobenzoic acid (TIBA) and 1 mg/l of 6-benzyladenine (BA). Calli induced from leaf base segments of Samhangwiri and Malgwiri in media containing 3 mg/l of 2,4-D and 3 mg/l of picloram showed high regeneration frequency. It appears that the callus initiation medium may be an important factor for subsequent plant regeneration.

High-frequency plant regeneration from transgenic rice expressing Arabidopsis thaliana Bax Inhibitor (AtBI-1) tissue cultures

  • Cho, A-Ra;Lee, Dong-Kil;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.83-87
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    • 2015
  • Genetic transformation was affected by material of explant, age of callus, and medium of regeneration. Two rice seed cultivars (Ilpum and Baekjinju) and mediums were investigated in this study for enhancing regeneration of transgenic rice expressed AtBI-1 gene encoding the Arabidopsis thaliana Bax inhibitor. Regeneration rate of Ilpum rice transformant in gelrite of 5 and 8 g were 27.4% and 18.0%, respectively. In Baekjinju, regeneration rate of transformant was 5.4% and 4.3% in 5 and 8 g gelrite, respectively. The highest number of transformant plant in this study was regenerated from Ilpum cultivar on MS medium (30.4%) and was applied for the subsequent experiment. The callus regeneration rate of transformant were 40.7% in callus infection of up-side, it was higher regeneration then in the down-side (3.9%). The regeneration rate of callus of 25 days and 35 days were 14.7% and 38.6%, respectively. The most important application of this work is in genetic transformation of rice, particularly for improvement transgenic plant tissue culture protocol with high frequency of plant regeneration.

An Improved Plant Regeneration Protocol using Cotyledonary Explants from Inbred Lines of Chinese Cabbage (Brassica rapa ssp. pekinensis)

  • Yang Zhi Hong;Jin Hua;Plaha Prikshit;Woong Bae Tae;Jiang Guo Bin;Woo Jong Gyu;Yun Han Dae;Lim Yong Pyo;Lee Hyo Yeon
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.235-239
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    • 2004
  • We studied the effect of genotype, explant, age of explant, medium (plant growth regulators and gelling agents), and standardized an efficient regeneration protocol for inbred lines of Chinese cabbage (Brassica rapa ssp. pekinensis). Of the different concentrations of NAA and BA tested, the combination of $5\;\cal{mg/L}\;BA\;and\;0.5\;\cal{mg/L}$ NAA gave the highest frequency of shoot regeneration. The cotyledonary explants had more shoot regeneration frequency ($\ge40\%$) than the hypocotyl ex-plants. Besides, the cotyledonary explants, excised from the 4-day old seedlings, showed higher shoot regene-ration ($56.7\%$). Of the three gelling agents and their concentrations used, 16 g/L agar was found to be the best for shoot regeneration. Shoot regeneration frequency in-creased significantly by supplementing the medium with $4\;\cal{mg/L}\;of\;AgNO_3$ MS medium devoid of NAA showed higher frequency of rooting in the regenerated shoots than the ones supplemented with NAA. Our improved regeneration protocol will be especially useful for the genetic transformation of Chinese cabbage inbred lines to develop transgenic hybrids.