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標題: 基因工程調控洋桔梗開花時間以解決高溫生長簇生化之問題(Ⅲ)
Manipulation of Flowering Time to Solve the Problem for Production of Rosette in Eustoma Grandiflorum
作者: 楊長賢
關鍵字: 生物技術, 園藝;應用研究
To face the effects potentially caused by attending WTO, the development and enhancement offlower industry is necessary for Taiwan’s agriculture. In plants, the transition from vegetative toreproductive stage is controlled by endogenous and environmental factors such as age, hormone,day length and temperature. These pathways regulate the expression of FT, SOC1, LFY and AP1to promote flowering. Exposed to low temperature, termed vernalization, activates VIN3 thatrepressed the expression of FLC and FVP, and induced the expression of AGL24 and AGL19 toaccelerate flowering. Eustoma grandiflorum is a popular and important plant in flower market dueto its abundant floral colors, shape and longevity. However, seedlings of E. grandiflorum grownunder high temperature produced rosette that limit its flower production and caused a significantdecrease of its market value. Therefore, to find a way by using biotechnology to solve thisproblem is urgent and definitely will benefit to the agriculture. The purpose of this project is toselect heat-tolerant, non rosette-forming E. grandiflorum through transgenic approach. Full lengthcDNA for EgFT、EgLFY and EgAP1 (EgMADS3/6) will be isolated and characterized from E.grandiflorum. Furthermore, the transgenic system for E. grandiflorum has been successfully setup in our laboratory. Ectopic expression of MADS box genes from orchid (OMADS1) in E.grandiflorum generated phenotype by flowering early. These results revealed the possibility forgenerating heat-tolerant E. grandiflorum through transgenic approach. In the project, EgFT,EgLFY, and EgAP1 (EgMADS3/6) will be transferred into Arabidopsis and E. grandiflorum forfunctional analysis. Transgenic E. grandiflorum will be selected to test the rosette forming at hightemperature. Moreover, EgSVP、EgFLC、EgVIN3 will also be cloned and characterized.Transgenic E. grandiflorum over-expression of EgVIN3 or down regulation of EgSVP、EgFLC byRNA interference will be generated and then examined the rosette forming at high temperature.All transgenic lines of E. grandiflorum shown the repression of the rosette formation in hightemperature will be further characterized, evaluated, and subjected into field test and finally intoflower market. Furthermore, EST library specific for high or low temperature will be constructedfor E. grandiflorum. Genes specifically expressed at high temperature will be sequenced andanalyzed to investigate the clues to rosette formation. Finally, the heat-tolerant, nonrosette-forming E. grandiflorum will be generated.

台灣加入世界貿易組織後,花卉產業是台灣農業的發展重點。花卉產業由於政府資源的投入,期望將臺灣轉變成亞太農業生物科技的重鎮。植物由營養期轉變為生殖期的過程受到內在與外在因子的影響,包括植物年紀、荷爾蒙、光照與溫度。這些途徑影響FT、SOC1、LFY 與AP1 的表現,因此促進植物進入生殖時期。而低溫的春化作用造成阿拉伯芥早開花是因為低溫促進VIN3 的表現,VIN3 再經由3 個不同路徑-抑制FLC、FVP 及促進AGL24 或AG19 的表現,因此使阿拉伯芥提早進入生殖期。洋桔梗在日本屬於高級花卉,而台灣在2006 年栽培面積已高達65 公頃,出口價值為2,300 萬,其花型與花色種類多、切花吸水性佳且耐儲運、周年均可供貨,因此是深具市場潛力的新興花卉。洋桔梗在產業栽培上最主要的問題是高溫會造成苗株簇生化 (rosette),因此延長栽培時間,增加生產成本。所以,培育高溫環境下不會簇生化的品種是洋桔梗育種上的首要目標。本實驗室對洋桔梗開花誘導的研究深感興趣,已選殖出EgFT、EgLFY、EgAP1 (EgMADS3/6)之基因全長。本實驗室亦成功建立洋桔梗的基因轉殖技術,將文心蘭OMADS1 或阿拉伯芥FYF 基因轉殖至洋桔梗中大量表現後,轉基因植物都會出現提早開花的性狀,這些結果對本計劃擬培育克服高溫簇生化的洋桔梗品種有很大的信心。本計劃先將選殖出全長的EgFT、EgLFY 與EgAP1(EgMADS3/6)基因轉殖入阿拉伯芥測試,再轉殖入洋桔梗大量表現後,以高溫環境測試轉基因植物抗簇生化的情形。本計劃同時要選殖洋桔梗春花作用的關鍵基因EgSVP、EgFLC、EgVIN3 等,經由以RNA 干擾技術來抑制EgSVP或EgFLC 基因的表現,或是大量表現EgVIN3 後,將這些轉基因洋桔梗以高溫環境來測試植物簇生化的情況。上述本計畫中所有洋桔梗轉基因植物若通過簇生化的測試實驗,亦即高溫生長下仍正常開花不會簇生化,則可立刻進行田間測試,並經評估後推廣到市場。本計劃同時要將高溫 (33/28℃) 與涼溫 (23/18℃) 培育之洋桔梗小苗進行EST library 之建立,經由序列分析在高溫育苗時特異表現之cDNA clones 後,選殖春花作用相關的基因或其他造成簇生化的基因,來深入研究洋桔梗簇生化的分子機制。最終亦將這些基因轉殖到洋桔梗中進行簇生化的測試實驗,培育出高溫不簇生的洋桔梗。
其他識別: NSC99-2622-B005-001-CC2
Appears in Collections:生物科技學研究所

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