Glucocorticoid-inducible gene expression in rice
Glucocorticoid-inducible gene expression in rice
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DOI：
10.1007/s004250100583
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发表时间：
2001-07-01
期刊：
影响因子：
4.3
通讯作者：
Meijer, AH
中科院分区：
文献类型：
--
作者：
Ouwerkerk, PBF;de Kam, RJ;Meijer, AH
我们研究了基于糖皮质激素受体的可诱导基因表达系统在单子叶模式植物水稻(Oryza sativa L.)中的应用。这个系统最初是由T.Ayama和N.H.Chua[(1997)植物J 11：605-612]开发的，是基于嵌合转录激活因子GVG的。由酵母Gal4DNA结合区、VP16激活区和糖皮质激素受体结构域组成。为了在水稻上应用，我们设计了一个优化的双元载体系列(PINDEX)，并用fl-葡萄糖苷酸酶(GUSA)报告基因对其进行了测试。GUS的表达受到严格控制，相对较低浓度(1-10um)的糖皮质激素地塞米松(DEX)能够诱导GUS活性达到与强大的花椰菜花叶病毒(CaMV)35S启动子相当的水平。在水培条件下，地塞米松能被组培苗或成熟植株的根有效吸收，并诱导整个植株的GUS活性。地塞米松诱导的GUS在所有品系及其TI后代中的表达模式是一致的。用10-100um DEX诱导1-4天或用1um-DEX诱导2周，对水稻组织培养苗的表型没有影响。这已经足以达到接近最大的GUS活性。然而。10 MU地塞米松诱导2周后，出现生长迟缓和发育缺陷。由于这些影响的严重程度在不同的品系之间有所不同，我们可以选择表型温和的品系作为激活系，用于与目标植物杂交。
We have studied the use of a glucocorticoid receptor-based inducible gene expression system in the monocotyledonous model plant rice (Oryza sativa L.). This system, originally developed by T. Aoyama and N.-H. Chua [(1997) Plant J 11: 605-612], is based on the chimaeric transcriptional activator GVG. consisting of the yeast Gal4 DNA-binding domain, the VP16 activation domain and the glucocorticoid receptor domain. For application in rice, we designed an optimized binary vector series (pINDEX) and tested this with the fl-glucuronidase (gusA) reporter gene. GUS expression was tightly controlled and relatively low concentrations (1-10 muM) of the glucocorticoid hormone dexamethasone (DEX) were able to induce GUS activities to levels comparable to those conferred by the strong cauliflower mosaic virus (CaMV) 35S promoter. DEX was taken up efficiently by the roots of tissue-cultured plantlets or mature plants in hydroponic culture, and induced GUS activity throughout the whole plant. DEX-induced GUS expression patterns were consistent in all lines and their TI progeny. The phenotype of tissue-cultured rice plantlets was not affected when inductions with 10-100 muM DEX were limited to 1-4 days or when 2-week inductions were performed with I muM DEX. which was already sufficient to reach near-maximal GUS activity. However. 2-week inductions with 10 muM DEX caused growth retardation and developmental defects. As the severity of these effects varied between different lines, we could select lines with a mild phenotype for future use as activator lines in crosses with 'target', plants.