Isolation and Characterization of OsGSTL1 Promoter from Rice

HU Ting-Zhang , WANG Wei-Ping , CAO Kai-Ming , XIA Mian , WANG Xi-Ping
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Abstract

OsGSTL1 gene was isolated from the rice genomic library. Semi-quantitative RT-PCR analysis demonstrated that the expression of the OsGSTL1 in rice was not induced by chlorsulfuron, ethylene, abscisic acid, salicylic acid, and methyl jasmonate. In order to investigate the cis-elements of OsGSTL1 promoter, the promoter regions with different lengths were fused to the β-glucuronidase (GUS) reporter gene. All constructs were transformed into onion epidermal cells or A. thaliana plants to detect the expression patterns. In onion epidermal cells, the 160 bp fragment and longer ones were functional for directing GUS expression. In transgenic A. thaliana, the 2 155 bp upstream region of OsGSTL1 gene directed the GUS expression only in cotyledon after germination, but not in the root of young seedlings. In the later seedling, the 2 155 bp upstream region of OsGSTL1 gene directed GUS expression in roots, stems, and leaves. However, the GUS gene directed by a 1 224 bp upstream fragment is expressed in all the checked tissues. These results suggest that the spatiotemporal expression response elements of OsGSTL1 existed in the 5′-upstream region between −2 155 and −1 224 bp.

水稻OsGSTL1启动子的分离与鉴定
从水稻基因组文库中分离到OsGSTL1基因。半定量RT-PCR分析表明,氯磺隆、乙烯、脱落酸、水杨酸和茉莉酸甲酯对水稻中OsGSTL1的表达没有诱导作用。为了研究OsGSTL1启动子的顺式元件,我们将不同长度的启动子区域融合到β-葡萄糖醛酸酶(GUS)报告基因上。将所有构建体转化到洋葱表皮细胞或拟蓝植物中,检测其表达模式。在洋葱表皮细胞中,160 bp及更长的片段具有指导GUS表达的功能。在转基因拟南芥中,OsGSTL1基因上游2 155 bp的区域在萌发后仅在子叶中指导GUS的表达,而在幼苗的根中没有GUS的表达。在幼苗后期,OsGSTL1基因上游2 155 bp区域指导GUS在根、茎和叶中的表达。而GUS基因上游1 224 bp片段在所有组织中均有表达。这些结果表明,OsGSTL1的时空表达响应元件存在于−2 155 ~−1 224 bp的上游5′区。
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