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題名 | Cosuppression of Tobacco Chalcone Synthase Using Petunia Chalcone Synthase Construct Results in White Flowers=利用矮牽牛苯基苯乙烯酮合成酶對轉殖菸草苯基苯乙烯酮合成酶的共同抑制作用以產生白色花朶 |
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作者姓名(中文) | 王貞觀; 陳柏言; 王幸美; 陶建英; | 書刊名 | Botanical Studies |
卷期 | 47:1 民95.01 |
頁次 | 頁71-82 |
分類號 | 434.28 |
關鍵詞 | 花色素; 苯基苯乙烯酮合成酶; 共同抑制作用; 花色之修飾; 轉殖植物; 轉殖基因導致的基因靜默; Anthocyanin; Chalcone synthase; Cosuppression; Flower color modification; Nicotiana tabacum; Transegenic plants; Transgene silencing; |
語文 | 英文(English) |
中文摘要 | 苯基苯乙烯酮合成酶(chalcone synthase,簡稱Chs;EC 2.3.1.74)是花色素(anthocyanin)生合成路徑的關鍵酵素。欲了解花朶顏色的分子調控機制,我們利用農桿菌(Agrobacterium)轉殖基因的方法,將鑲嵌質體含CaMV 35S啟動子及矮牽牛chsA cDNA轉殖至菸草。所獲得轉殖菸草中,4株轉殖株產生白色花朶,3株轉殖株產生與野生型植株相同的粉紅色花朶。薄層色層分析的結果顯示,所有白色花朶的轉殖株皆缺乏花青素 (cyanidin)。北方氏墨點分析法顯示,白色花朶轉殖株內chs mRNA的總量大幅下降;相反的,花色屬粉紅色的轉殖株,chs mRNA的總量卻顯著增加。RT-PCT分析結果顯示,花色屬白色的轉殖株,其內生型菸草chs mRNA的相對量少於轉殖矮牽牛chs mRNA。此外,植物/T-DNA接合序列分析亦排除產生白色花色的原因是由於T-DNA插入植物色素合成相關基因導致該基因喪失活性的可能性。綜合而言,本研究提供了廣泛的實驗證據,說明菸草chs基因的共同抑制作用是可以透過別種植物如矮牽牛的相似基因而發揮作用,導致轉殖菸草產生白色花朶;並清楚顯示chs mRNA表現量、花青素含量及轉殖菸草花朶顏色的關聯性。 |
英文摘要 | Chalcone synthase (CHS; EC 2.3.1.74) is a key enzyme in anthocyanin biosynthesis. In order to understand the molecular mechanism controlling flower color, tobacco plants were transformed with a chimeric construct containing expression cassettes for neomycin phosphotransferase Ⅱ (nptⅡ) selection marker and CaMV 35S promoter-driven Petunia chsA cDNA, via Agrobacterium-mediated method. Four transformants produced white flowers, while three transformants produced pink flowers similar to the untransformed parent. Thin layer chromatography analysis revealed the absence of cyanidin in all whiteflowered transformants. Northern blot analysis showed that total chs mRNA levels were greatly decreased in white-flowered transformants. By contrast, chs mRNA expression was induced in pink-flowered transformants. RT-PCR analysis showed that the relative level of endogenous tobacco chs mRNA was less than that of the transgenic Petunia chsA mRNA in white-flowered lines. In addition, plant/T-DNA junction sequence analysis excluded the possibility that insertion of T-DNA into anthocyanin genes had inactivated the anthocyanin biosynthetic pathway in white-flowered tobacco plants. Taken together, these results indicate that cosuppression of the tobacco chs gene can occur using the equivalent Petunia gene, and demonstrate a linkage between the expression level of chs mRNA, cyanidin content, and flower color in transgenic tobacco plants. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。