• Title/Summary/Keyword: multiple shoot formation

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An Improved Method of Organogenesis from Cotyledon Callus of Acacia sinuata (Lour.) Merr. using Thidiazuron

  • Shahzad Anwar;Ahmad Naseem;Anis Mohammad
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.15-19
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    • 2006
  • An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

Callus Formation and Plant Regeneration from Immature Embryos of Eleutherococcus senticosus (가시오갈피 미숙배 배양으로부터 Callus 형성 및 식물체 재분화)

  • Yu, Chang-Yeon;Kim, Jae-Kwang;Ahn, Sang-Deuk
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.49-55
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embryos in Eleutherococcus senticosus. Immature embryos from seeds were removed under the microscope and placed on MS, $MSB_5\;and\;B_5$ medium containing several plant growth regulators. While the calli were well formed on media containing 2 mg/l of 2, 4-D, 2 mg/l of 2, 4-D and 0.7 mg/l of TDZ, shoot regeneration was better on MS medium with combinations of high concentrations of TDZ and low concentrations of 2, 4-D. Treatment of 2, 4-D alone was better than treatment of TDZ alone in callus induction, but plant regeneration was reversed. The results of callus formation and shoot regeneration on $MSB_5\;and\;B_5$ media were similar to those of MS media. The rate of callus formation was nearly 100% when 2, 4-D was added to $B_5$, medium on concentration of 2 mg/l or 0.7 mg/l. TDZ showed very significant effect on the formation of multiple shoots.

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Multiple Shoot Formation from Flower Stalk Tissue Culture of Phalaenopsis Hybrids (팔레높시스 화경절편조직 배양에 의한 다신초 형성)

  • Choi, Jin Young;Koh, Jae Chul
    • FLOWER RESEARCH JOURNAL
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    • v.17 no.3
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    • pp.165-171
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    • 2009
  • This experiment was conducted to extend utility of in-vitro culture material stalk and to mass produce uniform plants in Phalaenopsis. The optimal concentration of hyponex medium and plant hormone were examined. The most effective concentration of the hyponex in the flower stalk tissue culture of Phalaenopsis was $4mg{\cdot}L^{-1}$. The most effective concentration of TDZ and BA on the formation of multiple shoots in Phalaenopsis flower stalk culture was $0.3mg{\cdot}L^{-1}$ and $5mg{\cdot}L^{-1}$, respectively. TDZ was more effective for formation of the shoots and multiple shoots than BA at the basic medium with hyponex $4g{\cdot}L^{-1}$.

Correlation between in vitro Plant Regeneration of Apple Cotyledon and Light Condition, Polarity of Explant (사과 자엽의 기내 식물체 재분화와 배양 광조건 및 절편체 극성과의 관계)

  • 안현주;예병우;임용표;신용억
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.19-23
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    • 2000
  • The experiment was carried out to investigate the influence of light condition and polarity of explant on adventitious shoot formation from cotyledon of apple in vitro The treatment of light culture after 10days dark treatment showed effective result than other treatments on the rate of adventitious shoot formation on light intensity treatments and also the treatment of blue light treatment after 4days dark treatment showed more effective result than other treatments in light quality treatments. The polarity of explant influence on adventitious shoot formation. Adventitious multiple shoot formation occured at the proximal end of an excised cotyledon. In other words. shoot organogenesis occured at the proximal cut surfaces of both proximal and distal explants rather than at the distal cut surface of proximal explants.

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In vitro Shoot Propagation Derived from Stem and Shoot Tip in Hovenia dulcis var. koreana Nakai by Plnat Growth Regulators and Light Resources (식물생장조절제 및 광원처리에 따른 헛개나무 줄기와 경정유래 신초의 기내증식)

  • Park, Mi-Young;Wang, Fengbo;Eom, Seok-Hyun;Lee, Seung-Woo
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.1
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    • pp.47-53
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    • 2012
  • This study was conducted to examine effects of plant growth regulators and light resources on the formation of multiple shoot and plant regeneration of Hovenia dulcis var. koreana Nakai. Stem and shoot tip were cultured on MS medium or WPM supplemented with various plant growth regulators. At the single treatment, the highest shoot formation was obtained when stem explants were cultured on WPM supplemented with kinetin $1.0mg{\cdot}L^{-1}$. MS medium containing NAA 0.1 and TDZ $0.1mg{\cdot}L^{-1}$ gave the best results for shoot induction rate and shoot growth in combination treatments. Of the BAP and kinetin tested, BAP $0.5mg{\cdot}L^{-1}$ on WPM was found to be more effective for shoot growth from shoot tip. Under white fluorescent light treatment, shoot growth was much higher than blue, red LED treatments. Root induction from in vitro growth of plantlet was the best on WPM supplemented with $1.0mg{\cdot}L^{-1}$ IBA. The results suggest that selection of plant growth regulators and light resources could be important factor to achieve an efficient in vitro growth.

Development of Plant Regeneration and Genetic Transformation System from Shoot Apices of Sorghum bicolor (L.) Moench

  • Syamala, D.;Devi, Prathibha
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.77-85
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    • 2004
  • Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

Plant Regeneration and Mutagenesis from Organogenic Callus of Dianthus Distributed in Gangwon Province (강원지역 패랭이꽃속의 캘러스로부터 식물체 재분화와 돌연변이체 유발)

  • Chang, Mi-Young;Hong, Sung-Won;Kim, Joon-Chul
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.73-80
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    • 2003
  • Useful Dianthus species were collected and selected from two native and seven foreign species distributed in Gangwon province. For in vitro breeding,. callus was induced from the explants of apical meristem, leaf, stem and the in vitro adventitious shoots on MS basal medium with 2.0 mg/L 2,4-D and 0.5 mg/L BA at 27$^{\circ}C$ under continuous light. After 3 weeks of culture, calli initiated the most highly from the leaf explants of D. chinensis Organogenic calli were able to be selected from the adventitious shoot-derived calli. For shoot regeneration, these organogenic calli were cultured on MS medium with the combination of 0.1 mg/L NAA+1.0 mg/L BA under continuous light. Multiple shoots were proliferated with low frequency (about 30%) from those adventitious shootderived calli. Also, shoots initiated directly from the adventitious shoot explants without callus formation at high frequency of 52% when cultured on N6 medium containing 0.1 mg/L NAA and 1.0 mg/L BA in D. gratianopol. Multiple shoots and plantlets grew well and rooted on MS medium supplemented with 0.1 mg/L NAA. Regenerants with well-developed roots were transferred to 8-cm pots containing vermiculite at 85% relative humidity and 27$^{\circ}C$ These plantlets were acclimatized in artificial soil mixture and transferred to the greenhouse for flowering with normal phenotypes. M28 Mutant line was selected with white flowers from 0.03M EMS-treated organogenic calli derived from in vitro adventitious shoot explants of D. chinensis and set seeds.

Adventitious Shoots Regeneration from Seed Explants of Xanthoceras sorbifolium

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.58-58
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    • 2020
  • Xanthoceras sorbifolium Bunge (yellowhorn) is a woody tree in the soapberry family, Sapindaceae, native to northern China. This species has been identified as a major woody bioenergy plant for bio-diesel production because of high oil content in seed. But the flowers do not bear fruit well while the many flowers blooming. This study was performed to regenerate in vitro plantlet using adventitious shoot formation. To establish the protocol of plant regeneration, adventitious shoots formation rate in the culture of cotyledon of immature zygotic embryos was 68.6% in 1/2 MS medium with 0.5 mg l-1 BA and 3% sucrose (w/v). In the culture of cotyledons of mature zygotic embryos, induction of adventitious shoots was needed to contain high sucrose in pre-culture medium and the frequency of shoot induction was 64.4%. Multiple shoots were induced in 0.5 mg l-1 TDZ, and rooting of shoot was induced 4.0 mg l-1 IBA. Flow cytometry analysis revealed that all the regenerated plantlets were diploid.

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Morphological Development and Histology of Multiple Shoots and Microbulbs of Garlic Cultured in Bioreactors (생물반응기에서 배양된 마늘 신초와 Microbulb의 형태발달 및 조직관찰)

  • Kim, Eun-Kyung;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.301-306
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    • 2004
  • Multiple shoots of garlic (Allium sativum L.) were propagated in bioreactors containing MS medium supplemented with 2% sucrose for 3 weeks. Microbulbs were induced on MS medium supplemented with 0.1mg/L NAA and 11% sucrose for 9 weeks. For multiple shoot proliferation, leaves in the shoot must be removed before cultures. When the multiple shoots were cultured without removal of leaves, more than 90% of hyperhydricity and no microbulb formation were observed. Histological observation also indicated irregular size and shape of the cells in hyperhydricity of the shoot. Microbulbs were strarted to form from the shoot after 7 weeks of culture by protuberance of adventitious shoot buds followed by inner periclinal divisions and simultaneous anticlinical division in the epidermis of meristematic bulge. Analysis of ploidy level indicated no phenotypic variations in both multiple shoots and microbulbs induced from the mother plant, suggesting genetic homogeneity among the regenerants.

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.