Identification of Differentially Expressed Genes during Initial Floral Development in Oncidium Pseudobulb

花序發育初期於文心蘭假球莖中具差異性表現基因之選殖

王芝鈺(Chih-Yu Wang) ； 徐明聖(Ming-Sheng Hsu) ； 葉開溫(Kai-Wun Yeh)

《作物、環境與生物資訊》 4卷2期 (2007/06) Pp. 135-144

https://doi.org/10.30061/CEB.200706.0005

摘要

利用抑制扣除雜交與微陣列技術，選殖花序發育初期於文心蘭假球莖中特異表現的基因。利用花序發育初期（花序長2-3公分）的假球莖，扣減具較長花序（7-8公分）的假球莖，獲得425個差異表現基因片段。又經微陣列分析，其中45個基因片段在花序發育初期的表現高於另一個時期兩倍以上，而基因片段平均大小為328bp。再經序列分析，其中31個基因片段與已知蛋白質序列相似，分別參與逆境反應、細胞生理調節、細胞壁新陳代謝、構造蛋白質及光合作用，其中也出現與開花時間相關的過氧化氫酶基因片段。但其餘14個基因片段並未與任何已知蛋白質序列相似。

關鍵字

文心蘭南西品種 ； 抑制扣除雜交 ； 微陣列 ； 開花誘導相關基因 ； 假球莖

並列摘要

We used suppression subtractive hybridization (SSH) and microarray techniques to isolate genes that increased abundantly in the pseudobulb of Oncidium Gower Ramsey during initial inflorescence development. A subtraction cDNA library was generated by using the cDNA from pseudobulbs in two different stages: the ones with initial inflorescence (2-3cm) and the other with longer inflorescence (7-8cm), and the former was subtracted against the latter. In this work, a total of 425 clones with differential expression were obtained. With microarray analysis, forty-five clones which have expression levels up regulated two-fold in the initial inflorescence stage were obtained, and they shared the average insert size of 328bp. Thirty-one ESTs showed homology to the known genes. Eight out of 31 genes were related to stress responses, including peroxidase, mannosebinding lectin precursor, metallothionein, and RD22. It is interesting that the appearance of peroxidase gene related to flowering time. Ten genes were involved in regulation, including latex-like protein, dicarboxylate transporter, profilin, allantoinase, ADP-ribosylation factor, and heat shock protein. One gene was related to cell wall metabolism, which is GTP-binding protein Rac. One gene encoded a structural protein, alpha-tubulin. Four genes were involved in protein synthesis and degradation, including ribosomal protein and ubiquitin. Seven genes were related to photosynthesis. Additionally, fourteen genes showed no homology with previously-reported sequences.

並列關鍵字

Oncidium Gower Ramsey ； Suppression subtractive hybridization SSH ； Microarray ； Flower initiation-associated genes ； Pseudobulb