Overexpression of <i>NtIPMS</i> reduces tobacco seed germination under cold stress by influencing amino acids and reactive oxygen species

Yongzhi Niu; Chengjing Wang; Zepeng Wu; Dandan Wang; Wenlong Suo; Zhoufei Wang; Yunye Zheng; Yongzhi Niu; Chengjing Wang; Zepeng Wu; Dandan Wang; Wenlong Suo; Zhoufei Wang; Yunye Zheng

doi:10.48130/seedbio-0024-0004

Figure 1.

Overexpression of NtIPMS reduces seed germination under cold stress in tobacco. (a) Representative images of seed germination under normal (25 °C) and cold stress (15 °C) conditions. Scale bars = 1 or 5 mm. (b) Comparison of germination percentage and percentage of seedling establishment in Yunyan 87 (WT) and NtIPMS-OE lines. Each point represents the mean ± standard deviation. n = 3. * and ** denote significant differences determined using Student's t-test at p < 0.05 and p < 0.01, respectively.

Figure 2.

Expression patterns of NtIPMS during seed germination under normal (25 °C) and cold stress (15 °C) conditions in tobacco. (a) Relative expression level of NtIPMS in Yunyan 87 tobacco during (a) the early germination stage and (b) the late germination stage determined via quantitative RT-PCR. NtEF1a gene was used as the internal control. Each column represents the mean ± standard deviation n = 3.

Figure 3.

Comparison of amino acids between Yunyan 87 (WT) and NtIPMS-OE-1 lines after 10 d germination under normal (25 °C) and cold stress (15 °C) conditions in tobacco. Each column represents the mean ± standard deviation. n = 3. Different letters indicate significant differences determined using Duncan's multiple range test: p < 0.05.

Figure 4.

NtIPMS regulates seed germination involving amino acid pathway under cold stress (15 °C) in tobacco. (a) Representative images of seed germination by exogenous amino acid (Asp, Cys, and Pro) treatment for 10 d in Yunyan 87 (WT) and NtIPMS-OE-1 lines under cold stress. Scale bars = 1 mm. (b) Comparison of radicle length between WT and NtIPMS-OE-1 lines under cold stress after amino acids treatment. Each column represents the mean ± standard deviation. n = 3. Different letters indicate significant differences determined using Duncan's multiple range test: p < 0.05.

Figure 5.

NtIPMS regulates seed germination involving ROS pathway under cold stress (15 °C) in tobacco. (a) Representative images of NBT staining for O₂^•− content and DAB staining for H₂O₂ content. Scale bars = 1 mm. (b) Comparison of O₂^•− and H₂O₂ content between Yunyan 87 (WT) and NtIPMS-OE-1 line under normal (25 °C) and cold stress (15 °C) conditions. (c) Representative images of seed germination by exogenous H₂O₂ treatment for 10 d in Yunyan 87 (WT) and NtIPMS-OE-1 line under cold stress. Scale bars = 1 mm. (d) Comparison of radicle length between WT and NtIPMS-OE-1 lines under cold stress after H₂O₂ treatment. Each column represents the mean ± standard deviation, n = 3. Different letters indicate significant differences determined using Duncan's multiple range test: p < 0.05.

Figure 6.

Comparison of the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) between Yunyan 87 (WT) and NtIPMS-OE-1 lines under normal (25 °C) and cold stress (15 °C) conditions. Each column represents the mean ± standard deviation, n = 3. Different letters indicate significant differences determined using Duncan's multiple range test: p < 0.05.