T. Pham, Huu Quan Nguyen, T. Nguyen, X. Dao, Danh Thuong Sy, Van-Son Le, H. Chu
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Recombinant GmDREB2 protein with a molecular weight of approximately 20 kDa was expressed in four transgenic soybean lines in the T1 generation. The GmP5CS gene was shown to have significantly (P<0.05) increased expression in the T2 transgenic soybean lines and higher than compared to non-transgenic plants with considering both in well watered condition and stressed conditions were from 1.06 to 1.31-fold and 1.37 to 1.95-fold, respectively. The proline content of four transgenic soybean lines increased 155.81% to 187.42 % after five days in drought-stress conditions and increased from 180.52 % to 233.74 % after nine days under drought-stress conditions (P<0.05). Therefore, the overexpression of GmDREB2 resulted in increasing transcription level of P5CS gene, proline accumulation and drought-stress tolerance of the transgenic soybean plants. 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引用次数: 8
摘要
脱水响应元件结合(DREB)是大豆植物在面临非生物胁迫时表达的一个植物蛋白亚家族。这些DREB蛋白也被认为可以激活抗旱基因的转录。本研究提出了Glycine max DREB2 (GmDREB2)过表达与Glycine max pyroline -5-羧酸合成酶(Glycine max pyroline -5-羧酸合成酶,GmP5CS)基因转录水平、脯氨酸积累和转基因大豆耐旱性之间关系的确定结果,作为选择高耐旱性转基因品系的依据。将GmDREB2插入植物转基因载体中,通过农杆菌介导的转化将35S-GmDREB2-cmyc构建体转移到大豆植株中。在T1代的4个转基因大豆品系中表达了分子量约为20 kDa的重组GmDREB2蛋白。GmP5CS基因在T2转基因大豆品系中的表达量显著(P<0.05)增加,且在水分充足和逆境条件下均高于非转基因植株,分别为1.06 ~ 1.31倍和1.37 ~ 1.95倍。4个转基因大豆品系的脯氨酸含量在干旱胁迫5 d后从155.81%增加到187.42%,在干旱胁迫9 d后从180.52%增加到233.74% (P<0.05)。因此,GmDREB2过表达导致转基因大豆植株P5CS基因转录水平升高,脯氨酸积累增加,抗旱能力增强。转基因株系T2代(T2-1、T2-6、T2-7和T2-8)的耐旱能力高于非转基因株系,证实了GmDREB2在大豆植株中的转化。
Overexpression of the GmDREB2 gene increases proline accumulation and tolerance to drought stress in soybean plants
The dehydration responsive element binding (DREB) is a plant protein subfamily expressed when soybean plants face abiotic stresses. These DREB proteins are also considered to activate the transcription of drought-resistant genes. In this study, we present the determined results of relationships between overexpression of Glycine max DREB2 (GmDREB2) with the transcription level of Glycine max pyrroline-5-carboxylate synthetase (GmP5CS) gene, proline accumulation and drought tolerant ability transgenic soybean plants as the basis for selection of transgenic lines with high drought tolerance. GmDREB2 was inserted into a plant transgenic vector and the 35S-GmDREB2-cmyc construct was transferred into the soybean plants by Agrobacterium-mediated transformation. Recombinant GmDREB2 protein with a molecular weight of approximately 20 kDa was expressed in four transgenic soybean lines in the T1 generation. The GmP5CS gene was shown to have significantly (P<0.05) increased expression in the T2 transgenic soybean lines and higher than compared to non-transgenic plants with considering both in well watered condition and stressed conditions were from 1.06 to 1.31-fold and 1.37 to 1.95-fold, respectively. The proline content of four transgenic soybean lines increased 155.81% to 187.42 % after five days in drought-stress conditions and increased from 180.52 % to 233.74 % after nine days under drought-stress conditions (P<0.05). Therefore, the overexpression of GmDREB2 resulted in increasing transcription level of P5CS gene, proline accumulation and drought-stress tolerance of the transgenic soybean plants. The GmDREB2 transformation into soybean plants was confirmed by the results of genetically modified lines in the T2 generation (T2-1, T2-6, T2-7, and T2-8) with higher drought tolerant ability than those of non-transgenic plants.