Department of Plant Production Engineering and Genetics, Razi University. Kermanshah, Iran.
Biotechnology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Cell Mol Biol (Noisy-le-grand). 2024 Jul 28;70(7):243-251. doi: 10.14715/cmb/2024.70.7.35.
Oil seeds now make up the world's second-largest food source after cereals. In recent years, the medicinal- oil plant Camelina sativa has attracted much attention for its high levels of unsaturated fatty acids and low levels of saturated fatty acids as well as its resistance to abiotic stresses. Improvement of oil quality is considered an important trait in this plant. Erucic acid is one of the fatty acids affecting the quality of camelina oil. Altering the fatty acid composition in camelina oil through genetic manipulation requires the identification, isolation, and cloning of genes involved in fatty acid biosynthesis. The Fatty Acid Elongase 1 (FAE1) gene encodes the enzyme β-ketoacyl CoA synthase (KCS), a crucial enzyme in the biosynthesis of erucic acid. In this study, the isolation and cloning of the FAE1 gene from Camelina sativa were conducted to construct an antisense structure. The molecular homology modeling of DFAE1 proteins using the SWISS-MODEL server on ExPASy led to the generation of the 3D structures of FAE1 and DFAE1 proteins. The GMQE values of 0.44 for FAE1 and 0.08 for DFAE1 suggest high accuracy in the structural estimation of these genes. The fragments were isolated from the DNA source of the genomic Soheil cultivar with an erucic acid content of about 3% (in matured seeds) using PCR. After cloning the FAE1 gene into the Bluescript II SK+ vector and sequencing, the resulting fragments were utilized to construct the antisense structure in the pBI121 plant expression vector. The approved antisense structure was introduced into the Camelina plant using the Agrobacterium-mediated method, with optimization of tissue culture and gene transfer conditions. This approach holds potential to advance our knowledge of fat biosynthesis, leading to potential improvements in oil quality in Camelina sativa.
油料作物现在是仅次于谷物的世界第二大粮食来源。近年来，药用油用植物荠蓝因其高含量的不饱和脂肪酸和低含量的饱和脂肪酸以及对非生物胁迫的抗性而备受关注。改善油质被认为是该植物的一个重要特性。芥酸是影响荠蓝油质量的脂肪酸之一。通过遗传操作改变荠蓝油的脂肪酸组成，需要鉴定、分离和克隆参与脂肪酸生物合成的基因。脂肪酸延长酶 1（FAE1）基因编码酶β-酮酰基辅酶 A 合酶（KCS），这是芥酸生物合成中的关键酶。在这项研究中，从荠蓝中分离和克隆了 FAE1 基因，构建了反义结构。使用 ExPASy 上的 SWISS-MODEL 服务器对 DFAE1 蛋白进行分子同源建模，生成了 FAE1 和 DFAE1 蛋白的 3D 结构。FAE1 和 DFAE1 基因的 GMQE 值分别为 0.44 和 0.08，表明这些基因结构估计的准确性很高。使用 PCR 从含有约 3%（成熟种子）芥酸的基因组 Soheil 品种的 DNA 源中分离出片段。将 FAE1 基因克隆到 Bluescript II SK+载体并测序后，将得到的片段用于在 pBI121 植物表达载体中构建反义结构。通过农杆菌介导的方法将经过批准的反义结构引入荠蓝植物中，并优化了组织培养和基因转移条件。这种方法有望提高我们对脂肪生物合成的认识，从而有可能改善荠蓝的油质。
