This study was carried out to develop an efficient transformation protocol via particle bombardment with PLBs (protocorm-like bodies) in Phalaenopsis. To achieve this aim, osmoticum treatment and an increasing shooting chances in particle bombardment process were applied for this study. In addition, pCAMBIA3301: ORE7 vector which contains a herbicide-resistance bar gene as a selectable marker and ORE7 gene as a gene of interests were employed. With regard to the increasing chances of shooting in particle bombardment, double shooting was the best results with 1.5 ~ 2.5 times higher than those of a single or triple shooting treatment in the productioon of PPT (D-L-phosphinothricin)-resistant PLBs. However, regeneration rate of shoots in double shooting was not high as a single shooting. Further, double shooting showed 35~40% higher than that of a single shooting in the frequency of browning. Regarding effects of different osmotic treatments, combination of 0.2 M sorbitol with 0.2 M mannitol showed the best results in transformation efficiency, regeneration of transformants and reduction of browning. Putative transgenic Phalaenopsis plants were analyzed by PCR analysis and confirmed the presence of bar and ORE 7 gene. Also, real-time PCR was conducted by using 21 transgenic plants and showed only 4 plants had one copy of transgene; whereas, the other 17 plants had more than 2 copies of transgene. Transgenic phalaenopsis plants produced in this study were transferred to pots and flowered normally without morphological variations in flower and leaf.
Over the past decades, carbon dioxide concentration of the atmosphere of the world has increased significantly, and thereby the greenhouse effect has become a social issue. To solve this problem, new renewable energy sources including solar, hydrogen, geothermal, wind and bio-energy are suggested as alternatives. Among these new energy sources, bio-energy crops are widely introduced and under rapid progress. For example, corn and oilseed rape plants are used for the production of bio-ethanol and bio-diesel, respectively. However, grain prices has increased severely because of the use of corn for bio-ethanol production. Therefore, non-edible switchgrass draws attention as an alternative source for bio-ethanol production in USA. This review describes the shortage of fossil energy and an importance of switchgrass as a bio-energy crop. Also, some characteristics of its major cultivars are introduced including growth habit, total output of biomass yields. Furthermore, biotechnological approaches have been conducted to improve the productivity of swichgrass using tissue culture and genetic transformation.
Lily is an important cut-flower in Korea and world as well due to it's a variety of flower colors and various sizes of flowers. To develop elite lily cultivars, conventional breeding techniques have been used so far. However, an introduction of tissue culture system in mass propagation of bulbs and regeneration of shoots with a high efficiency is prerequisite at this moment. Especially, growth of bulbs and shoots as well as reduction of browning is critical factor to proliferate bulbs with shoots of superior lines or cultivar in lily. For this purpose, we tried to test whether ascorbic acid, citric acid and silver nitrate in medium to facilitate growth of bulbs and shoots as well as reduction of browning of bulb scales in lily. As a result, ascorbic acid and silver nitrate showed no significant differences compared to control plants in the growth of bulbs and shoots. When bulb scales were treated with 150 mg․l-1 of citric acid, formations of shoots and bulbs showed best result. While, bulb scale treated with 100 mg․l-1 of citric acid showed the growth of shoot and root as well as increasing of fresh weight compared to other treatments. Regarding the reduction of browning, 150 mg․l-1 of ascorbic acid showed the best result with the less than 2%. Although more experiments with several commercial varieties are needed in the future to establish mass propagation of bulbs of lily, results obtained in this study are supposed to provide the basic knowledge and contribution in tissue culture system of lily.
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