Cloning and Expression Analysis of LEAFY in Phalaenopsis
碩士 === 國立屏東科技大學 === 農園生產系 === 94 === The propose of this study was to clone and analyze the expression level of a Phalaenopsis LEAFY (PLFY) gene in vegetative tissues, developmental stages of floral and floral organ in Phalaenopsis Taisuco Windian × P. Taisuco Atienyuki ‘F797’. RNA extracted from yo...
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2006
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| Online Access: | http://ndltd.ncl.edu.tw/handle/06896745018064055021 |
| Summary: | 碩士 === 國立屏東科技大學 === 農園生產系 === 94 === The propose of this study was to clone and analyze the expression level of a Phalaenopsis LEAFY (PLFY) gene in vegetative tissues, developmental stages of floral and floral organ in Phalaenopsis Taisuco Windian × P. Taisuco Atienyuki ‘F797’. RNA extracted from young inflorescence was reverse-transcribed into cDNA which was subsequently used in PCR analysis. The degenerate primers were designed based on conserved amino acid sequences and used for PCR amplification of PLFY cDNA coding region. End-to-end PCR was performed to amplify full-length PLFY cDNA. Genomic Southern blot analysis indicated that there are at least two copies of PLFY in the F797 genome. Northern blot analysis shows that PLFY expression was lower in young leaves among vegetative tissues, but increased gradually in reproductive stages until flowering. Real time RT-PCR shows that PLFY expression was controlled by circadian rhythms in all tissues. Dark/light transition induced strong PLFY expression in mature leaves. The expression of PLFY during summer was 4 hours ahead of time than in winter. After transfer from continuous light to darkness, PLFY showed high level expression at 20th hours. While, continuous light more than 8 hours inhibited PLFY expression in both mature leaves and young inflorescence.
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