Promoter Activity of a Soybean Gene Encoding a Seed Maturation Protein, GmPM9

大豆種子成熟蛋白GmPM9基因之調節機制

李佩芳(Pei-Fang Lee) ； 邢禹依(Yue-Ie C.Hsing) ； 週德源(Teh-Yuan Chow)

《Botanical Bulletin of Academia Sinica》 41卷3期 (2000/07) Pp. 175-182

https://doi.org/10.7016/BBAS.200007.0175

摘要

在種子發育過程中，有一類蛋白質會大量累積於成熟後期，故被稱為LEA(late embryogenesis abundant)蛋白質，此類蛋白質也稱為種子成熟蛋白，在單子葉及雙子葉植物中皆已廣泛地被發表。本文利用β-glucuronidase(GUS)為報導基因，探討大豆LEA基因GmPM9之啟動子在阿拉伯芥中之調控表現。將不等長度之啟動子區域片段接上GUS基因，轉殖入阿拉伯芥中，得知各啟動子片段皆表現GUS活性於轉殖阿拉伯芥之成熟種子及子葉與上胚軸部份。對轉殖植株施以PEG及NaCL處理，可誘導GUS活性表現於葉片，但離層酸(ABA)處理則無法誘導表現。於種子發育期間，各啟動子片段皆於開花十天後開始累積GUS活性於種子中，并在開花十四天後種子成熟時達到最高值。文中數據顯示，全長之啟動子區域(-966至+91bp)可獲得最高之GUS表現值，而最短之啟動子片段(-114bp至+91bp)已足以使報導基因表現於種中。基於上述結果，我們認為GmPM9之啟動子具有種子專一性，適合將其利用於研究種子專一性之課題上。

關鍵字

β-glucuronidase(GUS) ； 種子成熟蛋白 ； 啟動子 ； 轉殖阿拉伯芥

並列摘要

Late embryogenesis abundant (LEA) proteins are synthesized during the late stages of seed development and have been widely reported in monocot and dicot plants. In order to understand the developmental regulation of the promoter of a soybean gene encoding LEA protein GmPM9, we fused a series of fragments of the promoter region to the β-glucuronidase (GUS) reporter gene (uirA) and transformed the resulting constructs (pZP966, pZP572, pZP51O, pZP294 and pZPll4) into Arabidopsis thaliana. GUS enzyme activities were detected in mature seeds and cotyledons and hypocotyls of seedlings in transgenic plants containing any one of the five constructs; they were not detected in other tissues at different developmental stages. During seed development, GUS began to appear at 10 days after flowering (DAF) and increased rapidly to a maximum in the mature seeds at 14 DAF. The longest promoter construct (pZP966) enabled the transgenic plants to exhibit the highest GUS activity, whereas the shortest construct (pZP1 14) was sufficient to direct the expression of the GUS gene at a detectable level. The above findings indicate that the promoter of gGmPM9 can be used to express desirable genes in seeds during late seed maturation. The expression of the GUS gene could be induced in the leaves of the transgenic plants containing pZP966 by salt and desiccation, but not by ABA treatment.

並列關鍵字

β-glucuronidase ； Late embryogenesis abundant ； Promoter ； Transgenic Arabidopsis.