• Title/Summary/Keyword: root regeneration

Search Result 382, Processing Time 0.03 seconds

Expression of the Green Fluorescent Protein (GFP) in Tobacco Containing Low Nicotine for the Development of Edible Vaccine

  • Kim Young-Sook;Kim Mi-Young;Kang Tae-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yang Moon-Sik
    • Journal of Plant Biotechnology
    • /
    • v.7 no.2
    • /
    • pp.97-103
    • /
    • 2005
  • This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.

Plant Regeneration by Anther Culture of Lilium asiatic hybrid 'Gran Paradiso' (아시아틱 백합 ( Lilium asiatic hybrid 'Gran Paradiso' )의 약배양에 의한 식물체 재생)

  • 고정애
    • Korean Journal of Plant Tissue Culture
    • /
    • v.26 no.1
    • /
    • pp.1-6
    • /
    • 1999
  • In order to obtain plantlet derived by anthers, the anthers of Lilium asiatic hybrid 'Gran Paradiso' were cultured on Murashige and Skoog's medium supplemented with various combinations of auxin and cytokinin. The most suitable pollen stage of anther culture for the callus induction was 3 days before anthesis at the early to late binucleate stage. Organogenic calli were induced on MS medium supplemented with 5.0 mg/L 2,4-D alone and the combination of 1.0 mg/L 2,4-D and 1.0 mg/L kinetin, however, the combination of NAA and BA was more effective than that of 2,4-D and kinetin on plant regeneration through organogenesis. Shoots were formed from the induced callus on the medium with 0.5 mg/L NAA and 1.0 mg/L BA after 180 days of culture. Multiple shoots with 3-4 leaves, roots, and bulblets were formed on the medium with the combination of 2.0 mg/L NAA and 2.0 mg/L BA after 250 days of culture. The chromosome from root tip of the regenerated plantlet showed the diploid (2n=2x=24). Diploid plants were transferred to the pots and all plants were flowered in two years.

  • PDF

Factors Affecting the Production of In Vitro Plants from the Nodal Pieces of Chinese Yam (Dioscorea Opposita Thunb)

  • Shin, Jong-Hee;Kim, Sang-Kuk;Kwon, Jung-Bae;Lee, Bong-Ho;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
    • /
    • v.6 no.2
    • /
    • pp.97-102
    • /
    • 2004
  • This study was carried out to establish The regeneration of healthy seedlings from the nodal segment culture of Chinese yam (Dioscorea opposita cv. Danma), cultivated in Korea. Different explants such as leaves, petioles, roots and nodal pieces, excised from the in vitro grown seedlings of Chinese yam, were cultured on MS medium supplemented with various combinations of growth regulators. All the growth regulators used induced plantlet regeneration from the nodal segments at a high frequency, while there was no induction of shoot or callus from leaf, petiole or root tissues. The medium supplemented with 0.01mg/L NAA, 0.5mg/L BA, 0.5-1.0mg/L kinetin and without plant growth regulator was effective for shoot development of buds from the nodal segment culture. The concentration of BA and NAA was an important factor in the bud induction of buds from the nodal segments of Chinese yam. Nodal segments cultured on the medium containing 1.0mg/L NAA and 0.5-1.0mg/L BA gave the best response to bud formation. The addition of GA$_3$ to the culture medium suppressed shoot induction and growth, while it increased microtuber formation. The shoot growth and microtuber formation were also affected by medium strength and solidity. The MS basal medium containing 1 g/L gelrite was suitable for microtuber formation from the nodal segment of Chinese yam.

Correlation between In vitro Plant Regeneration and Polarity with Boxthorn (Lycium chinense Mill.) Seedlings (구기자나무 유식물체를 이용한 식물체의 재분화와 극성과의 관계)

  • Kwon Hye-Kyoung;Cho Yi-Yun;Yoon Eui-Soo
    • Korean Journal of Plant Resources
    • /
    • v.19 no.2
    • /
    • pp.308-314
    • /
    • 2006
  • This experiment was conducted out to investigate the effect of plant growth regulators on callus and shoot formation. The callus formation was effective on 1/2 MS medium containing 2,4-D, while shoot formation was suppressed. Shoot formation and differentiation were the highest in combination 0.1 mg/L of IAA and 0.1 mg/L of BA. The polarity of explants was investigated from cotyledon, which excised 20% of each basal and terminal parts. Formation of shoot was induced from excised ends of the basal part. In excised ends of the basal part, callus was induced vigorously and shoots were produced lately. Root induction was easily achieved in 1/3 MS medium from the adventitious shoot and more than 90% of regenerated plantlets acclimatized successfully and flowered normally.

A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)

  • Beyaz, Ramazan;Darcin, E. Selcen;Aycan, Murat;Kayan, Mustafa;Yildiz, Mustafa
    • Journal of Plant Biotechnology
    • /
    • v.43 no.2
    • /
    • pp.240-247
    • /
    • 2016
  • In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

The Effect Of Platelet - Derived Growth Factor And Insulin - Like Growth Factor On The Guided Tissue Regeneration In The Treatment Of Human Furcation Involvement (Platelet - derived growth factor-BB와 Insulin Iike gowth factor-1이 e- PTFE를 이용한 치근 이개부의 조직유도재생에 미치는 영향)

  • Ju, Ae-Ra;Kim, Sung-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
    • /
    • v.26 no.1
    • /
    • pp.80-88
    • /
    • 1996
  • The aim of the present investigation was to see the effect of combined use of PDGF BB and IGF -1 on the guided tissue regeneration(GTR) using barrier membrane in the treatment of human furcation involvement. Twelve patients with initially diagnosed as having moderate to advanced adult periodontitis with mandibular class II buccal furcation defects have been wer selected. Initial scaling and root planing has been performed and baseline data consisting of probing depths and attachment levels have been recorded prior to surgical procedures. The GTR procedures using either barrier membrane(control : ePTFE) alone or together with the application of PDGF - BB and IGF -l(experimental : ePTFE+PDGF/IGF) have been done under the routine guidelines. During the surgery, the distance from CEJ either to the bottom of the bone defects(CEJ - BD) or to the bone crest(CEJ-BC) were measured. Horizontal distance to the deepest area in the furcal defects were measured from the reference line connection the most prominent bony walls of the two buccal roots. 6 months following the GTR therapy, all the measurements were made repeatedly. The probing attachment gain of the experimental and the control grous were 2.14mm and l.07mm, respectively with no statically significnant difference. Amont of vertical bone fill in the experimental and the control groups were 2.43mm and 2.29mm, rexpectively. Amonut of horizontal bone fill were 2.86mm in the experimental group and 2.17mm in the control group, respectively. However, there were no significant differences in the amount of bone fill(both vertical and horizontal)between the two groups.

  • PDF

Effect of plant growth regulators on plant regeneration from the Sedum rotundifolium D. Lee (둥근잎꿩의비름(Sedum rotundifolium D. Lee)의 식물체 재분화에 미치는 식물생장조절제의 영향)

  • Kwon, Hye-Kyoung;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
    • /
    • v.37 no.1
    • /
    • pp.84-88
    • /
    • 2010
  • To establish the system of In vitro plant regeneration, the floral bud and leaf explants of Sedum rotundifolium were cultured on the MS media supplemented with different concentration of 2,4-D, NAA, and BA. The callus induction was more effective in the floral explants than the leaf explants, and was the best on MS medium containing 1.0 or 2.0 mg/L 2,4-D and 1.0 mg/L BA. The highest numbers of shoots were regenerated when callus were cultured on MS medium containing 2.0 mg/L 2,4-D and 1.0 mg/L BA for 8 weeks. The normal root formation from shoot was effective on the MS medium containing IAA alone. The regenerated plantlets were transferred to the pot and acclimatized successfully.

Direct somatic embryogenesis, plant regeneration and genetic transformation of Panax ginseng

  • Park, Yong-Eui;Yang, Deok-Chun;Park, Kwang-Tae;Soh, Woong-Young;Hiroshi Sano
    • Proceedings of the Botanical Society of Korea Conference
    • /
    • 1999.07a
    • /
    • pp.85-89
    • /
    • 1999
  • Somatic embryogendesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology. This paper describes the direct somatic embryogenesis from zygotic embryos of Panax ginseng is reversely related to normal axis growth of zygotic embryos by the experiment of various chemical treatments. Under the normal growth condition, the apical tips of embryo axis produced an agar-diffusible substance, which suppressed somatic embryo development from cotyledons. Although the cells of zygotic embryos were released from the restraint of embryo axis, various factors were still involved for somatic embryo development. Electron microscopic observation revealed that the ultrastructure of cells of cotyledon epidermis markedly changed before initiation of embryonic cell division, probably indicating reprogramming events into the cells embryogenically determined state. Polar accumulation of endogenous auxin or cell-cell isolation by plasmolysis pre-treatment is the strong inducer for the somatic embryo development. The cells for the process of somatic embryogenesis might be determined by the physiological conditions fo explants and medium compositions. Direct somatic embryos from cotyledons fo ginseng were originated eithrer from single or multiple cells. The different cellular origin of somatic embryos was originated either from single or multiple cell. The different cellular origin of somatic embryos was depended on various developmental stages of cotyledons. Immature meristematic cotyledons produced multiple cell-derived somatic embryos, which developed into multiple embryos. While fully mature cotyledons produced single cell-derived single embryos with independent state. Plasmolysis pretreatment of cotyledons strongly enhanced single cell-derived somatic embryogenesis. Single embryos were converted into normal plantlets with shoot and roots, while multiple embryos were converted into only multiple shoots. GA3 or a chilling treatment was prerequisite for germination and plant conversion. Low concentration of ammonium ion in medium was necessary for balanced growth of root and shoot of plantlets. Therefore, using above procedures, successful plant regeneration of ginseng was accomplished through direct single embryogenesis, which makes it possible to produce genetically transformed ginseng efficently.

  • PDF

Long-term radiographic evaluation of infrabony defect treated by flap operation (치주판막술에 의해 치료된 골연하 결손부의 장기적 방사선학적 변화 양사의 관찰)

  • Bae, Sang-Ryul;Park, Jin-Woo;Suh, Jo-Young;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
    • /
    • v.38 no.3
    • /
    • pp.429-436
    • /
    • 2008
  • Purpose: The goal of periodontal regenerative therapy is to replace bone, cementum, and periodontal ligament on a previously diseased root surface, which has suffered the loss of these supporting structures. To accomplish the regeneration, a number of surgical procedures have been advocated throughout the years. There seems to be a potential for some spontaneous periodontal tissue regeneration in the bottom of periodontal defect following open flap debridement alone. The aim of this study was to analyse the radiographic bone changes over 2-year after flap operation. Material and Methods: Patients attending the department of periodontics of Kyungpook National University Hospital were studied. Patients had clinical and radiographic evidence of infrabony defect(s). forty two sites of 33 patients aged 26 to 65 (mean age 45.5) were treated by flap operation with or without osseous surgery. Baseline and over 2-year follow-up radiographs were collected and evaluated for this study. Radiographic assessment includes a bone fill, bone crest change, defect resolution, and % of defect resolution. Pre- and post-treatment differences between variables (maxilla and mandible, gender, defect depth, defect angle) using the paired t-test were examined. Result: We observed 0.74 mm of bone fill, 0.66 mm of crestal resorption, 1.40 mm of defect resolution, and 27% of percentage of defect resolution. Mandible, women, deeper initial defect depth, narrower initial defect angle showed greater bone fill, defect resolution, and % of defect resolution. Conclusion: The results of this study suggest that the use of flap operation did enhance the outcome in terms of radiographically detectable bone fill. Both treatment resulted in some loss of crestal bone height.

Micropropagation through Callus Culture in Chinese Foxglove (Rehmannia glutinosa) (지황의 캘러스 배양에 의한 기내 대량증식)

  • 박충헌;성낙술;백기엽;이철희
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.3
    • /
    • pp.171-175
    • /
    • 1998
  • Chinese foxglove (Rehmannia glutinosa) is receiving much attention as one of the principal medicinal crops and the crude drug damand expands rapidly.This study was conducted to obtain the basic breeding information of Chinese foxglove. Effects of supplemental plant growth regulators were investigated on leaf tissue for proliferation. 100% callus formation, 31% plantlet regeneration and 6% root differentiation were obtained by adding 0.5 mg/L NAA and 2.0 mg/L BA. 2,4-D and Zeatin treatment also resulted in 95% increase in callus formation, but shoot was not formed. During the subculture, callus propagation rate recorded 15.4% with 0.2 mg/L NAA and 1.0 mg/L BA and plant regeneration improved on MS medium supplemented with 0.2 mg/L NAA and 0.5 mg/L kinetin. The number of shoot formed ranged from 1.7 on WPM medium to 3.4 on MS medium with 0.1 mg/L NAA and 0.5 mg/L BA. Supplementation of 1.0 g/L activated charcoal improved the In vitro plant growth.

  • PDF