蝴蝶蘭因為具有花形優美且花期長的特性，因而成為廣受歡迎的觀賞植物之一。大部分的蘭花花形皆展現出一特化的花瓣，即唇瓣，在生物功能上乃扮演昆蟲降落的平台、或製造營養物質以提供獎賞給授粉者，亦或進行擬態以欺騙授粉者，藉此吸引授粉者前來拜訪進而達到傳宗接代的目的。然而，目前對於參與在蝴蝶蘭唇瓣發育的基因功能鑑定以及花部形態與功能基因表達之關聯仍尚未釐清。由蘭花微矩陣(microarray) 實驗結果中發現一AP2/EREBP轉錄因子在姬蝴蝶蘭唇瓣大量表現，命名為PeEREBP_PHL1。經即時定量RT-PCR的檢測分析證實此基因會在唇瓣、合蕊柱及花梗軸大量表現。此外，經由序列比對以及分析其在AP2/EREBP家族的親緣演化，PeEREBP_PHL1與角質層形成相關的SHINE clade 之同源基因 (At5g25190) 相似，具有保守的AP2 domain 及不完整的SHINE domains (middle motif與C-terminal motif)。利用阿拉伯芥過量表現PeEREBP_PHL1，結果發現在轉殖植株表現出與含有完整SHINE domains的SHINE clade基因高表達之後一樣的表現型，且有角質層摺疊 (cuticular folds) 與臘質沉積在輪狀葉的表面。此外，在蝴蝶蘭對PeEREBP_PHL1進行病毒誘導基因靜默後，不僅造成花部表皮細胞大小的改變，唇瓣表面的cuticular folds與角質層也有減少的現象，顯示PeEREBP_PHL1確實參與調控蝴蝶蘭唇瓣之角質形成。進一步分析目前市面上之大唇蘭花 (具花瓣化唇瓣) ，結果發現PeEREBP_PHL1於花瓣化唇瓣的基因表現不僅明顯下降96%，其表皮細胞形態亦轉形如同花瓣表皮細胞，且在細胞表面不具cuticular folds的形成。綜合以上，本篇論文乃首次證實蝴蝶蘭之SHINE同源基因雖不具完整SHINE domains，但仍具類似於SHINE的功能參與在角質層的形成。本研究結果也提供我們對於蝴蝶蘭唇瓣發育與角質層形成的基因轉錄調節機制有更進一步的了解。

Phalaenopsis orchid becomes one of the most popular ornamental plants owing to its elegant floral morphology and long florescence duration. Most orchids represent a highly evolved petal, labellum or lip, that offers a landing platform for pollinators or function in mimicry in rewarding or deceptive orchids. However, the key factors for lip development and the relationship between the function of genes and floral morphological structures remain uncertain. Previously, a member of AP2/EREBP family, PeEREBP_PHL1, with enhanced expression in lip in microarray analysis was identified in our laboratory. In this study, the full-length cDNA sequence of PeEREBP_PHL1 was cloned and its phylogenetic relationship was analyzed. PeEREBP_PHL1 gene showed a high expression in lip, column and pedicle, and to a less-level in sepal and petal by using real-time reverse transcription- polymerase chain reaction (RT-PCR). The phylogenetic tree shows that PeEREBP_PHL1 is close to At5g25190, a SHINE clade homolog. The SHINE clade is reported to control cuticle formation, but the At5g25190 lacks intact SHINE domains and lacks SHINE functions. Heterologous overexpression of PeEREBP_PHL1 in Arabidopsis showed phenocopied overexpression of the AtSHNs, showing shiny leaves on the adaxial surface, and not only the cuticular folds but also the wax deposition were observed in the epidermis of rosette leaves. Knockdown expression of PeEREBP_PHL1with virus-induced gene silencing (VIGS) by using Cymbidium mosaic virus (CymMV) vector resulted in morphological alterations in epidermal cells and reductions in cuticular folds in the lip of Phalaenopsis. Furthermore, the plants with petal-like lip, the Big Foot ‘TH.365’, were also examined in this study. The expression of PeEREBP_PHL1was significantly reduced with a 96% reduction in the P. Big Foot ‘TH.365’. In addition, the cell morphology of the P. Big Foot ‘TH.365’ also shows that the lip epidermis is transformed into petal epidermis, which lacks cuticular folds on the surface. Consequently, this study is the first report demonstrating that the PeEREBP_PHL1 lacks entire of the SHINE domains can still exert SHINE functions in Arabidopsis. These results provide new insights into transcriptional mechanisms mediating floral cuticle formation in Phalaenopsis.

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