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<p>Salinity is a major abiotic factor that adversely affects soybean yield and seed quality. In <i>Arabidopsis thaliana</i>, the transcription factor RAP2-4 has been shown to modulate numerous downstream genes associated with light, drought, salt, and oxidative stress responses. In soybean, RAP2-4 has been predicted to function in drought and salt tolerance, yet its regulatory role has not been experimentally confirmed. This study investigated the expression dynamics and regulatory influence of the soybean <i>GmRAP2-4</i> gene on salt stress responsive genes in transgenic tobacco. A construct carrying <i>GmRAP2-4</i> was introduced into tobacco plants, and transcript levels of <i>GmRAP2-4</i> and selected downstream targets were quantified <i>via</i> real-time RT-PCR. Under salt stress, <i>GmRAP2-4</i> expression in transgenic lines increased 2.04- to 5.01-fold compared with wild-type (WT) plants and 1.81- to 13.39-fold relative to non-stressed conditions (<i>p</i> &lt; 0.05). Lines T05, T09, and T018 showed notable induction of downstream genes, with <i>NtSODFe</i> upregulated to 2.33- to 3.63-fold, <i>NtCLC</i> to 11.72- to 26.40-fold, and <i>NtP5CS</i> to 1.27- to 2.41-fold (<i>p</i> &lt; 0.05). <i>In silico</i> docking analysis demonstrated that the AP2 domain of RAP2-4 binds specifically to cis-regulatory motifs within the promoters of these genes, with the AP2 and NtP5CS promoter complex displaying the highest predicted stability. Elevated proline accumulation in the T09 line corresponded with strong <i>NtP5CS</i> activation, suggesting that <i>GmRAP2-4</i> may directly regulate <i>NtP5CS</i> expression to enhance proline biosynthesis, thereby contributing to improved salt tolerance in transgenic tobacco.</p>

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A novel Glycine max DREB transcription factor, GmRAP2 4, regulates tobacco salt stress response genes: in vitro and in silico evidence

  • 期刊论文

  • 发表日期：

    2025年10月23日

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  • Luong Trong Vu,
  • Giang Thu Nguyen,
  • Yen Thi Hai Nguyen,
  • Hung Duc Nguyen,
  • Thu Thi Mai Lo,
  • Thuy Thi Thu Vu,
  • Lan Thi Ngoc Nguyen,
  • Quan Huu Nguyen,
  • Tan Quang Tu,
  • Mau Hoang Chu

摘要

Salinity is a major abiotic factor that adversely affects soybean yield and seed quality. In Arabidopsis thaliana, the transcription factor RAP2-4 has been shown to modulate numerous downstream genes associated with light, drought, salt, and oxidative stress responses. In soybean, RAP2-4 has been predicted to function in drought and salt tolerance, yet its regulatory role has not been experimentally confirmed. This study investigated the expression dynamics and regulatory influence of the soybean GmRAP2-4 gene on salt stress responsive genes in transgenic tobacco. A construct carrying GmRAP2-4 was introduced into tobacco plants, and transcript levels of GmRAP2-4 and selected downstream targets were quantified via real-time RT-PCR. Under salt stress, GmRAP2-4 expression in transgenic lines increased 2.04- to 5.01-fold compared with wild-type (WT) plants and 1.81- to 13.39-fold relative to non-stressed conditions (p < 0.05). Lines T05, T09, and T018 showed notable induction of downstream genes, with NtSODFe upregulated to 2.33- to 3.63-fold, NtCLC to 11.72- to 26.40-fold, and NtP5CS to 1.27- to 2.41-fold (p < 0.05). In silico docking analysis demonstrated that the AP2 domain of RAP2-4 binds specifically to cis-regulatory motifs within the promoters of these genes, with the AP2 and NtP5CS promoter complex displaying the highest predicted stability. Elevated proline accumulation in the T09 line corresponded with strong NtP5CS activation, suggesting that GmRAP2-4 may directly regulate NtP5CS expression to enhance proline biosynthesis, thereby contributing to improved salt tolerance in transgenic tobacco.