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題 名 | Agrobacterium Tumefaciens--Mediated Transformation of Phalaenopsis Orchid偄 |
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作 者 | 謝瑞旻; 陳文輝; 許宏銘; 林益賢; 蔡媦婷; 傅仰明; 詹明才; 余淑美; | 書刊名 | Taiwan Sugar |
卷 期 | 44:6 民86.11-12 |
頁 次 | 頁12-18 |
分類號 | 435.432 |
關鍵詞 | 蝴蝶蘭; 農桿腄瘤菌; Phalaenopsis; Agrobacterium tumefaciens; Transformant; Attachment; |
語 文 | 英文(English) |
英文摘要 | A cocultivation method has been developed for transforming Phalaenopsis varieties in vitro with Agrobacterium tumefaciens strain EHA105 (pMT1) . Protocormoi-like-bodies (PLBs) were used as explants. and bacterial neomycin phophotransferase II (NPT II) gene and β -glucuronidase (GUS) gene as a selection marker and reporter, respectively. Scanning electron microscopy (SEM) revealed that agrobacteria attached very well to the surface cells of PLBs, and the explants at the 10-day proliferating stage were the optimal materials for infection. Histochemical GUS activity was detected in the surface of cross sections. The results showed that a difference in GUS activities occurred among genotypes and proliferating stages of explants. Phal. Brother Mirage 'A79-69' had the lowest frequency of GUS gene expression among the four cultivars tested. Phal. True Lady 'A76-13', at the 10-12 days proliferating stage, had 50-80% frequency of GUS expression. However, the highest frequency (100%) occurred at the 10 days proliferating stage of Phal. Asian Elegance 'B79-11' and Phal. Taisuco Kaaladian 'F80-13'. The transformant was obtained from the proliferating proembryoids of the explant which were selected in regeneration medium containing 100 μ g/ml kanamycin after 30-day culture |
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