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  • Author or Editor: Y.-H. Kim x
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Effects of reduced osmotic potential on somatic embryos of celery (Apium graveolens L.) were studied in an attempt to understand and improve their tolerance to partial desiccation. Embryos responded similarly to application of high osmoticum (384 mOs/kg H2O vs. 190 mOs/kg H2O in the control), achieved either by manipulation of sucrose or polyethylene glycol concentrations (PEG). Treatments of high osmotic concentration applied during the last 2 days of the embryo production cycle increased embryo survival and conversion after partial desiccation. The most striking effect of the high osmotic concentrations was the 4-fold increase in proline, while a 2-fold increase was obtained with 1 μm ABA alone. Application of high osmotica decreased reducing sugars, increased sucrose, but did not affect starch content of embryos; of these responses, only the change in sucrose was similar to that induced by ABA. Osmotic treatments did not affect total fatty acid content in the embryos compared to the 2-fold increase induced by ABA. Chemical name used: abscisic acid (ABA).

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The development of genetic transformation systems has led to remarkable progress in the area of plant molecular biology. This has included the introduction of useful traits, such as resistance to viruses, herbicides, and insects. Transformed plant cells can be selected, using chimeric genes that confer resistance to toxic drugs, such as kanamycin, hygromycin, streptomycin, gentamycin, and bleomycin. Expression of these chimeric genes in the transformed cells confers the ability to survive and proliferate on the selective medium, while non-transformed cells die. In this study, we report a simple and efficient system to regenerate Chinese cabbage plants and study of the effects of plant growth regulators, AgNO3, initial dark treatment, various antibiotics, and herbicide on shoot induction from hypocotyl or cotyledon of Chinese cabbage. Shoots were induced at various combinations of naphtalene acetic acid (NAA) and benzyladenine (BA) levels. The best combination of plant growth regulators was 2.0 mg/L NAA and 1.0 mg/L BA for cotyledon, and 1.0 mg/L NAA and 5.0 mg/L BA for hypocotyl. The experiment investigating the effect of AgNO3 demonstrated that 16.7 mg/L AgNO3 was effective for inducing shoot regeneration from both of explants. Three to five days of initial dark treatments had significant effects for increasing the number of regenerated shoots; however, different growth regulator combinations showed various responses to duration of dark treatments. The effects of kanamycin, hygromycin, cefatoxime, carbenicillin and phosphinothricin (PPT) on shoot induction from cotyledon and hypocotyl were tested. Shoot induction was completely inhibited by kanamycin at 10 mg/L, hygromycin at 5 mg/L, PPT at 5 mg/L or higher, but not by carbenicillin and cefatoxime.

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This experiment was conducted to identify the effect of various growth retardants on the growth of Aerides japonicum in vitro. Paclobutrazol was found the most effective retardant for reducing the leaf growth of seedling. Ancymidol and uniconazole also showed retarding effects on leaf growth of one, whereas Daminozide didn't. When growth retardants were added to culture medium, leaf length of seedlings was gradually shortened and leaf width became wider than that of control. However, root length was shorter and number of roots and root diameter were greatly increased. On the contrary, at 0.05 and 0.1 ppm uniconazole, growth of leaf and root were enhanced. It was showed that the possibility of using as an additive for good growth of Aerides japonicum seedling in vitro. The activity of GA-like substances was higher in the portion in which growth of seedlings were promoted. It was identified by anatomical observations that the number of stomata and thickness of cell layer in leaf were increased by treatment of retardants.

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Three Korean cultivars, Pungkak, Kalmi, and Subi, were crossed with PI 201234, which has resistance to P. capsici. A backcross breeding program was initiated to incorporate the Phytophthora resistance into the Korean cultivars, but the level of resistance decreased as the backcross round increased. Highly resistant plants occurred frequently in the BC1F2 populations but were rare in the BC2F1 populations. Resistant plants selected in BC1F3 populations had nearly enough recovery of the growth and fruit characteristics of the Korean recurrent parents. Crosses were made between resistant selections in each BC1F2 population. The F1 hybrids showed a considerably increased level of resistance.

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Somatic embryogenesis was initiated from in vitro-grown leaf explants of rose using an induction period of 4 weeks on MS basal medium supplemented with auxin followed by several subcultures on MS basal medium with cytokinin. `4th of July' showed the highest regeneration frequency (24.4%) on 5.3 μm NAA followed by culture on medium containing 18.2 μm zeatin. `Tournament of Roses' produced somatic embryos when cultured for 4 weeks on medium containing dicamba, 2.3 μm followed by three subcultures on medium containing 18.2 μm zeatin. Embryogenic callus matured on MS media containing 0.5 μm NAA, 6.8 μm zeatin, and 2.9 μm GA3. Long-term cultures were established for both cultivars. Somatic embryos germinated on MS medium containing IBA and BA. Silver nitrate (58.8 μm) enhanced shoot formation and germination of somatic embryos. Plants derived from somatic embryos were acclimatized and successfully established in the greenhouse.

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Daminozide is a growth retardant used in potted plant production as a foliar spray to inhibit shoot elongation. It has its greatest inhibitory effect immediately after application, becoming less pronounced thereafter; continued retardation is accomplished by reapplication at 7to 14-day intervals. A model for this retardation effect is useful in developing decision support tools, as well as in optimizing (perhaps minimizing) the use of this growth retardant. Such a model, as developed and described earlier, simulates the effect of a foliar spray application of daminozide at various concentrations on various days during the production cycle. The objective of this work was to validate this model for various varieties of chrysanthemum. Using the model to simulate the effect of one application of daminozide resulted in predicted plant heights very close to the observed heights for most of the varieties tested. Of four methods used to implement the multiple-application effect, two resulted in very good simulation of the observed plant heights. In summary, the model was shown to be valid for all the varieties of chrysanthemum tested.

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Abstract

The node position from which axillary buds were isolated from shoots of rose (Rosa hybrida L.) markedly affected their growth and development in culture. Those buds nearest to and furthest from the apex either failed to develop or took the longest time to develop in culture compared to those buds in the middle portion of the stem. Benzylamino purine (BA) at low concentrations (0.03 to 0.3 mg/liter) stimulated the development of the axillary buds of ‘Gold Glow’ but not of ‘Improved Blaze’. A photon flux density (400-700 nm) of 17μE m−2 s−1 for 12 to 24 hours daily was optimum for the stimulation of shoot multiplication, while 66 μE mm−2s−1 for 12 to 24 hr was optimum for root initiation and for subsequent successful transplantation to soil of tissue culture-derived plants. A constant temperature of 21°C resulted in the highest rate of shoot multiplication and root initiation. Plants which initiated roots at 16, 21, or 26° had the highest level of transplant survival. An alteration in the temperature of the 8-hr dark period from 21° did not increase shoot multiplication, although root initiation was enhanced by lowering the night temperature to 11 or 16°. Histological analysis indicated that shoot multiplication of rose shoots occurs through the growth and development of axillary buds. The development of axillary buds is apparently under the repressive influence of the shoot apex, because physical excision of the apex or application to the shoot apex of 2,3,5-triiodobenzoic acid (TIBA) facilitated axillary bud development. Root initiation was affected markedly by the length of time that cultures had been maintained on shoot multiplication medium prior to transfer to rooting medium. This effect may be attributable to the BA in the shoot multiplication medium which may have accumulated in the tissue. If the endogenous cytokinin level is too high, root initiation may be inhibited and if it is too low the shoot undergoes senescence before it becomes cytokinin-autonomous, which occurs after root initiation.

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Fluorescent proteins (FT) have become essential, biological research tools. Many novel genes have been cloned from a variety of species and modified for effective, stable, and strong expression in transgenic organisms. Although there are many applications, FT expression has been employed most commonly at the cellular level in plants. To investigate FT expression at the whole-plant level, particularly in flowers, petunia ‘Mitchell Diploid’ [MD (Petunia ×hybrida)] was genetically transformed with seven genes encoding FTs: DsRed2, E2Crimson, TurboRFP, ZsGreen1, ZsYellow1, rpulFKz1, or aeCP597. Each gene was cloned into a pHK-DEST-OE vector harboring constitutive figwort mosaic virus 35S promoter and NOS-terminator. These plasmids were individually introduced into the genome of MD by Agrobacterium tumefaciens–mediated transformation. Shoot regeneration efficiency from the cocultured explants ranged from 8.3% to 20.3%. Various intensities of red, green, and yellow fluorescence were detected from TurboRFP, ZsGreen1, and ZsYellow1-transgenic flowers, respectively, under ultraviolet light for specific excitation and emission filters. More than 70% of plants established from the regenerated shoots were confirmed as transgenic plants. Transgenic ZsGreen1 petunia generated strong, green fluorescence in all flower organs of T0 plants including petals, stigmas, styles, anthers, and filaments. Most of the chromophores were localized to the cytoplasm but also went into the nuclei of petal cells. There was a positive linear relationship (R 2 = 0.88) between the transgene expression levels and the relative fluorescent intensities of the ZsGreen1-transgenic flowers. No fluorescence was detected from the flowers of DsRed2-, E2Crimson-, rpulFKz1-, or aeCP597-transgenic petunias even though their gene transcripts were confirmed through semiquantitative reverse transcriptase-polymerase chain reaction. T1 generation ZsGreen1 plants showed green fluorescence emission from the cotyledons, hypocotyls, and radicles, which indicated stable FT expression was heritable. Four homozygous T2 inbred lines were finally selected. Throughout this study, we demonstrated that ZsGreen1 was most suitable for generating visible fluorescence in MD flowers among the seven genes tested. Thus, ZsGreen1 may have excellent potential for better utility as a sensitive selectable marker.

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