Studies on Developing Novel Erycina pusilla by Biotechnological Approach
| Autor: | Chanon Lapjit, 黃獻慶 |
|---|---|
| Rok vydání: | 2015 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 103 The purposes of this research were to manipulating the in vitro cultivation of Erycina pussila by LEDs illumination, to develop the novel, new flower color/type, and early blooming of Erycina pusilla via transformation of MADS14 (AP1-like) and CHRC genes, and to create new cultivars of Erycina pusilla by EMS and sodium azide mutagenesis and hybridization. The results indicated that compared with the fluorescent light and other LED treatments, RB+W+RGB LEDs significantly increased root and leaf number and plant high of in vitro plantlets of Erycina pusilla. RB+W+RB LEDs significantly increased the values of Fo, Fv, and Fv/Fm of in vitro plantlets after 25 weeks of cultivation, whereas significantly increased values of chlorophyll fluorescent parameters was found in the fluorescent light treatment after 50 weeks of cultivation. These results suggested RB+W+RGB LEDs and RB+W+RB LEDs could promote the quality of in vitro cultivation of Erycina pusilla. Three transformation vectors, p1305.3-CHRC, p1304-CHRC-IN, and p1301-AP1-MADS14 were transferred into the PLBs of the Erycina pusilla via Agrobacterium-mediated gene transformation. The regenerated plantlets were confirmed by PCR, RT-PCR and. Results indicated that the MADS14 gene was present in the genome of transformed Erycina pusilla plants and expressed its mRNA. Changes in the leaf and plant morphology, flower shape and color, blooming time, and multiple flower stalks were founded in the MADS14 transformed Erycina pusilla plants. This is the first report on the ectopic expression of MADS box gene in Erycina pusilla using an efficient and sophisticated gene transformation technology. In general, EMS treatments were more effective in reducing survival percentages and numbers of plant regeneration as compared to those of NaN3 treatments. Changes in profiles of flow cytometric histograms were found in several regenerated plants of EMS and NaN3 treated Erycina pulsilla PLB. In this study, 10 cultivar of Oncidiinae (♀) were crossed with Erycina pusilla (♂), in order to obtain novel Erycina pusilla. Several females tribute Oncidiinae crossed relatively easily with male Erycina pusilla but the reciprocal cross is rare. Hybrid seeds could be produced in Oncsa. Sweet Sugar x Erycina pusilla cross, and germinated under asymbiotic conditions. Changes in plant mophology and profiles of flow cytometric histograms were detected in vegetative leave of F1 plants. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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