本研究主要探討蝴蝶蘭分生組織擬原球體(protocorm-like-bodies, PLBs)，在經藥劑Ribavirin處理後對CyMV及ORSV感染之影響。結果顯示在相同處理天數下，培殖體存活率隨著處理藥劑濃度之增加而降低；在相同的處理濃度下培殖體之存活率隨著處理天數增加而降低，不同品系間無明顯差異。在增殖率方面，在相同濃度下增殖率隨著處理天數增加而降低，Phal. Sierra Gold `Suzanne’較Phal. True Lady `F86-12’稍高。Phal. Sierra Gold `Suzanne’以100ppm處理十天可獲得13.9%之無病毒植株；Phal. True Lady `F86-12’則以250ppm處理十天可獲得88.2%之無病毒植株。另外， Phal. True Lady`F86-12’ 即使不經藥劑處理也可獲得相當高比率(61.5%-88.2%)的無病毒植株。將經ELISA檢測為無病毒植株之瓶苗出瓶栽培至中苗及大苗，持續追蹤病毒去除情形，結果顯示至中苗仍分別維持66-99%的無病毒株。 在提高去病毒率方法之可行性探討方面，將擬原球體橫切後以250ppm Ribavirin處理五天後，將存活球體誘導分化，分別移至含不同濃度Ribavirin之固體培養基，結果顯示植株增殖率及存活率隨藥劑濃度增加而降低，25ppm處理與對照組相同。
Phalaenopsis is one of Taiwan’s important orchids for export. Virus-infected Phalaenopsis have been increasing. This project developed a virucide-based method to eliminate CyMV and ORSV from infected protocormlike bodies of Phalaenopsis during micropropagation. The survival rate and propagation ability of treated PLBs decreased when ribavirin concentration were increased. Some 13.9% virus-negative plants were obtained when the PLBs of Phal.Sierra Gold`Suzanne’ were treated with 100 ppm ribavirin for 10 days. For Phal.True Lady `F86-12’, 88.2% virus-negative plants were obtained when PLBs were treated with 250 ppm ribavirin for 10 days. Otherwise, Phal. True Lady `F86-12’ also gained 61.5-88.2% virus-negative plants among PLBs cultured without ribavirin. The virus-free plants were greenhouse cultured. The data showed that these plants maintained 66% and 99% virus-free when grown to medium or large plants. Different concentrations of ribavirin, which added in the solid medium, were examed to identify its effect on the multiplication and survival ratio of the induced Phalaenopsis buds. Results showed that the multiplication and the survival ratio decreased when the medium ribavirin concentration was increased. The effect of 25 ppm ribavirin showed no difference with the check.
Keywords: Phalaenopsis, elimination of virus, protocorm-like bodies (PLB)
台糖蝴蝶蘭品種改良技術之發展 The Varieties Improvement of Phalaenopsis at Taiwan Sugar Research Institute
Phalaenopsis is one of the most valued floriculture crops in the world due to its graceful appearance and long-lasting flowering. Taiwan Sugar Corporation (TSC) has developed a Phalaenopsis business management system. The Department of Horticulture at Taiwan Sugar Research Institute (TSRI) is in charge of the R&D, this includes germplasm collection, crossing seedling raising, clonal selection, and registration of new hybrids. Flowering Phalaenopsis grown by TSC have been displayed in numerous international and domestic orchid shows, and the breeding achievements have been confirmed by numerous awards. A suitable medium has been developed for germination and growth of seeds in most crosses. Micropropagation techniques were developed for germplasm conservation and mass production of superior selected varieties. Studies on isolation, fusion, and protoplast culture were also undertaken to transfer other useful genomes into Phalaenopsis. Chromosome doubling was introduced to create autotetraploid which was useful to breed superior varieties with high homogeneity. Electrophoretic pattern of isozymes and DNA amplification fingerprinting (DAF) have been used to protect the patent right of our new varieties. DAF is also being employed to screen genetic markers. Random amplified polymorphic DNA (RAPD) analysis was used to understand the phytogenetic relationship of taxonomy and evolution among wild species of Phalaenopsis. In order to understand the floral color heredity of Phalaenopsis, crosses were made between white and red floral clones. In order to develop an early detection system for somaclonal variation during micropropagation, the institute studied somaclonal variation to examine plant characteristics, including RAPD and isozyme patterns in normal and variant somaclones of Phalaenopsis. To improve TSRI’s conventional Phalaenopsis breeding program and to create novel commercial varieties, a transgenic system has been established.
Keywords: Phalaenopsis, breeding, tissue culture, isozyme, DNA amplification fingerprinting (DAF)、random amplified polymorphic DNA (RAPD)、somaclonal variation、gene transfer
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