具不同蔗糖合成酶 RSuS3 表現量之轉殖水稻建立及性狀分析

Production and Characterization of Transgenic Rice Plants Expressing Various Abundance of Rice Sucrose Synthase Isoenzyme RSuS3

張德清(Te-Ching Chang)

指導教授 ： 王愛玉

共同指導教授 ： 劉麗飛(Li-Fei Liu)

國立臺灣大學/生命科學院/微生物與生化學研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.10040

摘要

蔗糖合成酶催化蔗糖與 UDP形成果糖與 UDP-glucose 的可逆反應。在水稻中，此酵素是由至少六種 RSus 基因所表現，其中 RSus3 主要表現於發育中的種子。本論文的目的在建構具有不同 RSus3表現量的轉殖水稻，以探討 RSus3 的生理功能。
將預期可造成水稻各組織中恆常性表現 RSus3、於穀實中過量表現 RSus3 及於穀實中抑制 RSus3 表現的三種質體 pActinSuS3、pGluB1SuS3 及 pSuS3RNAi，經由農桿菌法轉殖到水稻。pActinSuS3 及 pGluB1SuS3 的 T0 及 T1 轉殖株，以 Southern 分析後發現，這些轉殖株帶有一個或二個外來的 RSus3 基因，且能夠在葉子偵測到其表現。分析 T1 植株充實期種子發現，轉殖株 RSus3 的表現比未轉殖之台農 67 號水稻早，轉殖株 RSus3 mRNA 的量也比未轉殖之台農 67 號水稻多。此外，轉殖株 T2 種子的千粒重及平均內外穎長有大於未轉殖之台農 67 號水稻的趨勢。

關鍵字

水稻蔗糖合成酶農桿菌基因轉殖

並列摘要

Sucrose synthase catalyzes the reversible conversion of sucrose and UDP into fructose and UDP-glucose. The enzyme is encoded by at least six RSus genes in rice. One of the genes, RSus3, is predominatly expressed in the developing seeds. The objective of this study is to construct transgenic rice plants having different expression levels of RSuS3 in order to investigate the biological function of RSus3.
Plasmids pActinSuS3, pGluB1SuS3 and pSuS3RNAi, which are expected to respectively result in constitutive expression of RSus3 in various rice tissues, over-expression of RSus3 in rice seed, and inhibition of RSus3 expression in rice seed were transformed into rice by the Agrobacterium-mediated method. Southern analysis of the T0 and T1 transgenic plants transformed with pActinSuS3 or pGluB1SuS3 revealed that one or two copies of RSus3 genes were introduced into these transgenic plants. Expression of RSus3 was detected in leaves. Analysis of ripening seeds of the T1 transgenic plants revealed that the expression of RSus3 in the developing seeds of these transgenic plants occurred earlier than the untransformed TNG67 rice plant. The abundance of RSus3 mRNA in the seeds of transgenic plants was also higher than the untransformed plant. Moreover, the weight of 1000 grains and the average length of grains of transgenic T2 seeds are larger than the untransformed TNG67.

並列關鍵字

rice sucrose synthase ； agrobacterium mediated gene transformation

參考文獻

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