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Analysis of expression characteristics of soybean leaf and root tissue-specific promoters in Arabidopsis and soybean
Transgenic Research ( IF 2.7 ) Pub Date : 2021-06-11 , DOI: 10.1007/s11248-021-00266-7
Hongwei Xun 1, 2 , Xue Zhang 2 , Jiamiao Yu 2 , Jinsong Pang 2 , Shucai Wang 3 , Bao Liu 2 , Yingshan Dong 1 , Lili Jiang 2 , Dongquan Guo 1
Affiliation  

The characterization of tissue-specific promoters is critical for studying the functions of genes in a given tissue/organ. To study tissue-specific promoters in soybean, we screened tissue-specific expressed genes using published soybean RNA-Seq-based transcriptome data coupled with RT-PCR analysis. We cloned the promoters of three genes, GmADR1, GmBTP1, and GmGER1, and constructed their corresponding β-Glucuronidase (GUS) promoter-GUS reporter vectors. We generated transgenic Arabidopsis plants and examined the expression patterns of these promoters by GUS staining and RT-PCR analysis. We also transformed the promoter-GUS reporter vectors into soybean to obtain hairy roots, and examined promoter expression by GUS staining. We found a root-specific expression pattern of GmADR1 and GmBTP1 in both Arabidopsis and soybean, and the promoter of GmGER1 showed a leaf-specific pattern in transgenic Arabidopsis plants. To test the potential utility of these promoters in soybean improvement by transgenic means, we used the GmADR1 promoter to drive expression of a salt resistance gene in soybean, GmCaM4, by generating transgenic soybean plants. We found that the transgenic plants had significantly enhanced salt tolerance compared to non-transformed wild-type, suggesting that introducing endogenous promoters by transgenic means can drive the expression of functional genes in specific tissues and organs in soybean.



中文翻译:


拟南芥和大豆中大豆叶根组织特异性启动子表达特征分析



组织特异性启动子的表征对于研究给定组织/器官中基因的功能至关重要。为了研究大豆中的组织特异性启动子,我们使用已发表的基于大豆 RNA-Seq 的转录组数据并结合 RT-PCR 分析来筛选组织特异性表达基因。我们克隆了GmADR1GmBTP1GmGER1这三个基因的启动子,并构建了它们相应的β-葡萄糖醛酸酶( GUS )启动子-GUS报告载体。我们生成了转基因拟南芥植物,并通过 GUS 染色和 RT-PCR 分析检查了这些启动子的表达模式。我们还将启动子-GUS报告载体转化到大豆中以获得毛状根,并通过GUS染色检查启动子表达。我们在拟南芥和大豆中发现了GmADR1GmBTP1的根特异性表达模式,并且GmGER1的启动子在转基因拟南芥植物中显示出叶特异性模式。为了测试这些启动子在转基因大豆改良中的潜在效用,我们使用GmADR1启动子通过产生转基因大豆植物来驱动大豆中耐盐基因GmCaM4的表达。我们发现,与未转化的野生型相比,转基因植物的耐盐性显着增强,这表明通过转基因手段引入内源启动子可以驱动大豆特定组织和器官中功能基因的表达。

更新日期:2021-06-11
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