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標題: | 基因轉殖水稻之農藝性狀及花粉流佈評估 Agronomic Traits and Pollen Flow Assessment of Transgenic Rice |
作者: | Ching-Shan Tseng 曾清山 |
指導教授: | 林彥蓉 |
關鍵字: | 農藝性狀,生物安全性評估,緩衝區,綠籬,花粉性狀,花粉媒介基因流佈,基因轉殖水稻, Agronomic traits,Biosafety assessment,Buffer zone,Green fence,Pollen characters,Pollen-mediated gene flow,Transgenic rice, |
出版年 : | 2012 |
學位: | 博士 |
摘要: | 近年來應用基因轉殖技術所培育出之基因轉殖作物,其種類及栽培面積逐年上升,然而生物安全議題和對環境風險的影響也受到越來越多的關注。本研究利用台農67號 (Oryza sativa L. ssp japonica, TNG 67) 的3個基因轉殖水稻為材料,分別為轉殖澱粉普魯楠糖水稻 (APU) 、轉殖豬乳鐵蛋白水稻 (LAC) 和轉殖植酸酵素水稻 (AAN) ,進行農藝性狀之調查、基因轉殖水稻外源基因藉由花粉媒介的基因流佈頻率之田間評估和防範基因轉殖水稻花粉污染的策略,以瞭解基因轉殖水稻在商業化生產種植前,對目前已有之水稻耕作及環境無不良影響。
在田間評估試驗中,進行為期一年二期作的農藝及產量性狀評估,結果顯示轉殖澱粉普魯楠糖水稻、轉殖豬乳鐵蛋白水稻和轉殖植酸酵素水稻的農藝性狀與其受體品種台農67號水稻相比,普遍表現較差,三個轉殖品系每公頃產量皆低於台農67號水稻。在雜草化評估試驗中,包括種子生產量、不同溫度下種子最終發芽率與平均發芽時間及不同溫控下之幼苗生長勢,目前所得數據未顯示三個轉殖品系比台農67號水稻具有較大的優勢,由此推測,本研究三個轉殖品系轉變成雜草的風險性極小。 基因轉殖水稻花粉性狀評估試驗中,結果顯示轉殖澱粉普魯楠糖水稻的花粉數量、花粉離體發芽率及花粉活力均低於台農67號水稻。基因轉殖水稻花粉流佈田間試驗中,結果顯示在交叉種植設計中,以轉殖豬乳鐵蛋白水稻的花粉流佈頻率最高為7.82%,其次台農67號水稻及轉殖植酸酵素水稻分別為7.13%和5.91%,最低為轉殖澱粉普魯楠糖水稻只有3.18%。在間隔種植設計中,則以TNG 67的花粉流佈頻率最高為2.8%,其次LAC及AAN轉殖品系分別為2.32%和1.53%,最低為APU轉殖品系只有0.54%。整體而言三個轉殖品系的花粉流佈頻率平均為3.55%,低於其非基因轉殖的TNG 67的4.97%。 水稻花粉流佈之田間試驗中,貢獻親台農糯73號水稻的雜交種子頻率受到接受親台農67號水稻種植距離和方向所影響,花粉流佈頻率隨著種植距離增加而下降。在3 m內,8個方向皆有檢測到雜交種子,在相距1 m時其花粉流佈平均頻率為1.68%,在2 m及3 m處分別為0.74%及0.61%。在距離5 m處仍有6個方向測得雜交種子,在距離30 m和35 m處僅有1個地方檢測到雜交種子,花粉流佈頻率平均從0.33%到0.01%,超過40 m並未發現有任何雜交種子。在南方及東南方35 m內有顯著地測得較高的雜交種子,花粉流佈頻率平均分別為0.78%和0.73%,最遠在南方35 m處仍可以檢測到雜交種子。然而相對地在東北、西北及北方僅測到較少的雜交種子,在東北方及北方5 m以後未檢測到任何雜交種子。 有無緩衝區設置對水稻花粉流佈之評估試驗中,結果顯示有無緩衝區設置的效果受到距離及風向所影響。在順風狀況下,位於下風南方與東南方7 m之內,有無緩衝區設置對花粉流佈頻率差異不大,然而在9 m處其花粉流佈頻率有顯著的差別。緩衝區設置南方與東南方9 m處的花粉流佈頻率分別為0.83%和0.84%,而無緩衝設置則分別為1.53%和1.65%。在逆風狀況下,位於上風北方與東北方1、3和5 m處有緩衝區設置的花粉流佈頻率高於無緩衝區設置,但在7 m以後有緩衝區設置的花粉流佈頻率則明顯低於無緩衝區設置,顯示在上風處之風速較小的情況下,5 m的緩衝區設置仍可減少花粉流佈頻率。 利用2 m高、0.6 m寬之朱槿當綠籬設置對水稻花粉流佈之評估試驗中,在台農糯73號水稻並未發現有來自貢獻親轉殖植酸酵素水稻的雜交種子,其花粉流佈頻率為0%。顯示在本試驗條件下,利用朱槿當綠籬可以有效地防止台農糯73號水稻受到相鄰田區轉殖植酸酵素水稻花粉的污染。 In recent years, more and more transgenic plants are being developed and cultivated for the commercial around the world. However, safety issues and environmental risks arisen by transgenic crops still evoke serious considerations. Three transgenic rice lines, line AAN, LAC, and APU, which were the phytase-, lactoferrine-, and amylopullulanase-gene transformed cv. Tainung 67 (Oryza sativa L. ssp japonica, TNG 67 ), respectively, were employed for investigation of agronomic traits of the transgenic lines, pollen-mediate gene flow in fields, and the strategy of preventing pollution of transgenic pollens The aim of study is to comprehend whether there is any negative influence on current rice cultivation and environments prior to commercialize transgenic rice or not. Field trials of evaluating agronomic traits of three transgenic strains and TNG 67 were carried out in two crops seasons, the first crop season and the second crop seasons of 2004. Based on the evaluation of seed production, final germination rate and average germination time, and seedling growth under various temperatures, the three transgenic lines performed worse in agronomic traits and weediness assessment than TNG 67 did. All three important factors accounting for capabilities of pollen flow, quantity, germination rate, and viability of pollen were lower in the transgenic rice line, APU, than in the non-transgenic rice, TNG67. The mean frequency of outcrossed seeds was significantly lower from APU (3.18%) than TNG 67 (7.13%) In the arrangement of the checker-board pattern, the mean frequency of outcrossed seeds was 7.82%, 7.13%, 5.91%, and 3.18% in LAC (the highest), TNG 67, AAN, and APU (the lowest), respectively. In the alternating row arrangement, the mean frequency of outcrossed seeds was 2.8%, 2.32%, 1.53%, and 0.54% in TNG 67 (the highest), LAC, AAN, and APU (the lowest), respectively. Overall, the mean frequency of outcrossed seeds of three transgenic lines was 3.55% which was lower than their non-transgenic variety TNG 67 was 4.97%. In the open field, the frequency of outcrossed seeds varied with the distances and directions of the pollen donor TNG 67 to the pollen recipient TNG 73. Pollen dispersal leading to outcrossing was negatively associated with distance. Outcrossed seeds were found from all 8 directions within a 3-m distance. The mean frequency of outcrossed seeds was 1.68%, 0.74%, and 0.61% at the distance of 1, 2, and 3 m, respectively. Outcrossed TNG 73 seeds were still detected in 6 directions in 5-m plots and only 1 direction in 30- and 35-m plots, and the mean outcrossed TNG 73 seeds of each direction ranged from 0.33% to 0.01%. No outcrossed TNG 73 seeds were detected in any direction farther than 40 m. The frequency of outcrossed seeds was significantly higher in plots to the south and southeast, which showed the highest mean frequencies of outcrossed seeds (0.73% and 0.78%) and in the plot located at 35 m. Nevertheless, relatively fewer outcrossed seeds were observed from the plots to the northeast, northwest, and north, and no outcrossed seeds were detected from plots to the northeast and north beyond 5 m. With and without buffer zones were used for assessing pollen-mediated gene flow in the open fields, which results also indicated that the frequency of outcrossed seeds varied by the distances and directions. In the directions with wind, the frequency of outcrossed seeds of with and without buffer zone was not significantly difference in the south and southeast directions within 7 m. However, there was significantly different frequency of outcrosses seeds between plots with and without buffer zones at 9-m distance, which frequency of outcrossed seeds with buffer zone was 0.83% (south) and 0.84% (southeast) and without buffer zone was 1.53% (south) and 1.65% (southeast), respectively. In the directions against wind, the frequency of outcrossed seeds of plots with buffer zones was higher than those without buffer zones in the north and northeast at distance of 1, 3, and 5 m. Nevertheless, the frequency of outcrossed seeds of plots with buffer zones was significantly lower than those without buffer zone beyond 7 m. This study indicated 5-m buffer zones could decrease the frequency of outcrossed seeds by pollen-mediated gene flow in the directions against wind. Chinese hibiscus (Hibiscus rosa-sinensis L.) with 2-m tall and 0.6-m thick were used for assessing pollen-mediated gene flow. We found none outcrossed with pollen from AAN, regardless of the distance between recipient plants and donor plants. Thus, Chinese hibiscus as a green fence effectively prevented TNG 73 from outcrossing with pollen from phytase-transgenic rice in neighboring fields. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66351 |
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