College of Life Sciences, Jilin Normal University, Siping, China.
College of Life Sciences, Jilin Academy of Agricultural Sciences, Changchun, China.
PLoS One. 2024 Sep 12;19(9):e0307706. doi: 10.1371/journal.pone.0307706. eCollection 2024.
Soybean is one of the most important food crops, breeding salt-tolerant soybean varieties is of great significance to alleviate soybean shortage. In this study, the F-box protein family homologous gene GmFBX322 was cloned from the soybean variety Williams 82 and overexpressed in the Shennong 9 soybean variety to further study and explore the physiological mechanism of soybean salt tolerance. GmFBX322 was constructed on the vector pTF101:35S, and integrated into the genome of Shennong 9 soybean variety by Agrobacterium EHA101-mediated cotyledonary node transformation technology, and 4 overexpressed transgenic lines were obtained, molecular assays were performed on the transformed plants. The expression of GmFBX322 was detected by qRT-PCR and it was found that the leaves of the 4 transgenic lines increased by 2.49, 2.46, 2.77, 2.95 times compared with the wild type; after salt treatment for 12 hours, it was found that the expression of wild type Shennong 9 Inducible expression of GmFBX322. After 72 hours of salt treatment, the leaves of wild-type Shennong 9 soybean plants showed obvious wilting and chlorosis, while the leaves of GmFBX322 plants overexpressing GmFBX322 showed no obvious changes. The leaves were taken at 0, 6, 12, 24, and 48 hours of salt stress to determine the antioxidant activity. Ability and osmotic adjustment level, etc. The results showed that the catalase activity in the leaves of the transgenic lines 2265, 2267, 2269, and 2271 was 2.47, 2.53, 3.59, 2.96 times that of the wild-type plant after 48 hours of salt treatment; the soluble sugar content was 1.22, 1.14, and 1.22 of the wild-type plant. 1.14, 1.57 times; the proline content is 2.20, 1.83, 1.65, 1.84 times of the wild type. After comparing the physiological indicators determined by the experiment, the transgenic lines performed better than the control group, indicating that overexpression of GmFBX322 can enhance the salt tolerance of soybean plants. To verify the function of GmFBX322 gene related to stress resistance, add it to the candidate gene of stress resistance, and provide scientific basis for the selection and breeding of salt-tolerant varieties.
大豆是最重要的粮食作物之一，培育耐盐大豆品种对于缓解大豆短缺具有重要意义。本研究从大豆品种 Williams 82 中克隆了 F-box 蛋白家族同源基因 GmFBX322，并在 Shennong 9 大豆品种中过表达，以进一步研究和探索大豆耐盐的生理机制。GmFBX322 构建在载体 pTF101:35S 上，并通过农杆菌 EHA101 介导的子叶节转化技术整合到 Shennong 9 大豆品种的基因组中，获得了 4 个过表达的转基因株系，对转化植株进行了分子检测。通过 qRT-PCR 检测 GmFBX322 的表达，发现 4 个转基因株系的叶片分别比野生型增加了 2.49、2.46、2.77、2.95 倍；经过 12 小时盐处理后，发现野生型 Shennong 9 可诱导表达 GmFBX322。经过 72 小时盐处理后，野生型 Shennong 9 大豆植株叶片明显萎蔫、黄化，而过表达 GmFBX322 的 GmFBX322 植株叶片无明显变化。在盐胁迫 0、6、12、24 和 48 小时时取叶片，测定抗氧化活性。能力和渗透调节水平等。结果表明，转基因株系 2265、2267、2269 和 2271 在盐处理 48 小时后，叶片中的过氧化氢酶活性分别是野生型植物的 2.47、2.53、3.59 和 2.96 倍；可溶性糖含量分别为野生型植物的 1.22、1.14、1.22 和 1.14 倍；脯氨酸含量分别为野生型的 2.20、1.83、1.65 和 1.84 倍。通过比较实验测定的生理指标，转基因株系的表现优于对照组，表明过表达 GmFBX322 可以增强大豆植株的耐盐性。为了验证 GmFBX322 基因与抗逆性相关的功能，将其添加到抗逆候选基因中，为耐盐品种的选择和选育提供科学依据。
