Key Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, China.
Int J Mol Sci. 2022 May 3;23(9):5084. doi: 10.3390/ijms23095084.
WRINKLED1 (WRI1), an APETALA2/ethylene-responsive-element-binding protein (AP2/EREBP) subfamily transcription factor, plays a crucial role in the transcriptional regulation of plant fatty acid biosynthesis. In this study, was overexpressed in the soybean cultivar 'Dongnong 50' using -mediated transformation to generate three transgenic lines with high seed oil contents. PCR and Southern blotting analysis showed that the T-DNA was inserted into the genome at precise insertion sites and was stably inherited by the progeny. Expression analysis using qRT-PCR and Western blotting indicated that and driven by the CaMV promoter were significantly upregulated in the transgenic plants at different developmental stages. Transcriptome sequencing results showed there were obvious differences in gene expression between transgenic line and transgenic receptor during seed developmental stages. KEGG analysis found that the differentially expressed genes mainly annotated to metabolic pathways, such as carbohydrated metabolism and lipid metabolism. A 2-year single-location field trial revealed that three transgenic lines overexpressing (GmWRI1a-OE) showed a stable increase in seed oil content of 4.97-10.35%. Importantly, no significant effect on protein content and yield was observed. Overexpression of changed the fatty acid composition by increasing the linoleic acid (C18:2) content and decreasing the palmitic acid (C16:0) content in the seed. The three GmWRI1a-OE lines showed no significant changes in agronomic traits. The results demonstrated that the three overexpression lines exhibited consistent increases in seed oil content compared with that of the wild type and did not significantly affect the seed yield and agronomic traits. The genetic engineering of will be an effective strategy for the improvement of seed oil content and value in soybean.
WRINKLED1(WRI1)是 APETALA2/ethylene-responsive-element-binding protein(AP2/EREBP)亚家族转录因子,在植物脂肪酸生物合成的转录调控中发挥着关键作用。本研究通过 -介导转化,在大豆品种‘东农 50’中过表达 ,生成了三个种子油含量较高的转基因株系。PCR 和 Southern 印迹分析表明,T-DNA 精确插入到基因组中,并稳定遗传给后代。qRT-PCR 和 Western 印迹分析表明,在不同发育阶段,由 CaMV 启动子驱动的 和 显著上调。转录组测序结果表明,在种子发育阶段,转基因株系和转基因受体之间的基因表达存在明显差异。KEGG 分析发现,差异表达基因主要注释到代谢途径,如碳水化合物代谢和脂质代谢。为期 2 年的单点田间试验表明,过表达 的三个转基因株系(GmWRI1a-OE)的种子油含量稳定增加了 4.97-10.35%。重要的是,蛋白质含量和产量没有显著变化。过表达 改变了脂肪酸组成,增加了种子中的亚油酸(C18:2)含量,降低了棕榈酸(C16:0)含量。三个 GmWRI1a-OE 株系在农艺性状上没有显著变化。结果表明,与野生型相比,三个 过表达株系的种子油含量均有一致增加,且对种子产量和农艺性状没有显著影响。 的遗传工程将是提高大豆种子油含量和价值的有效策略。