Xun Hongwei, Zhang Xue, Yu Jiamiao, Pang Jinsong, Wang Shucai, Liu Bao, Dong Yingshan, Jiang Lili, Guo Dongquan
Jilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, 130033, Changchun, China.
Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, 130024, Changchun, China.
Transgenic Res. 2021 Dec;30(6):799-810. doi: 10.1007/s11248-021-00266-7. Epub 2021 Jun 11.
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.
组织特异性启动子的表征对于研究特定组织/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，我们利用已发表的基于RNA测序的大豆转录组数据并结合逆转录-聚合酶链反应（RT-PCR）分析，筛选了组织特异性表达的基因。我们克隆了三个基因GmADR1、GmBTP1和GmGER1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。我们培育了转基因拟南芥植株，并通过GUS染色和RT-PCR分析检测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染色检测启动子表达。我们发现GmADR1和GmBTP1在拟南芥和大豆中均呈现根特异性表达模式，并且GmGER1的启动子在转基因拟南芥植株中表现出叶特异性模式。为了通过转基因手段测试这些启动子在大豆改良中的潜在效用，我们通过培育转基因大豆植株，利用GmADR1启动子驱动大豆中一个抗盐基因GmCaM4的表达。我们发现与未转化的野生型相比，转基因植株的耐盐性显著增强，这表明通过转基因手段引入内源启动子可以驱动功能基因在大豆特定组织和器官中的表达。
