Arabidopsis is wildly used as a model plant and camelina is increasingly used for oilseed research and applications. Although the Arabidopsis genome has been sequenced for two decades, the functions of many lipid‐related genes and their regulators have not been well characterized. Improvements in the efficiency and accuracy of gene investigations are key to effective discovery of gene function and downstream bioengineering of plant oil quantity and quality. In this study, a visible marker was used to quickly identify transgenic T₁ seeds and a method has been developed to phenotype fatty acid compositions and oil content of single T₁ seeds. A whole seed direct transmethylation method was first optimized with multiple seeds and incubation at 85°C for 2 hours in a transmethylation solvent (5% H₂SO₄ in methanol with 30% toluene cosolvent) is recommended. Based on this method, a single Arabidopsis seed mini‐transmethylation (SAST) method has been established in a 1.5 ml GC sample vial with 200 μl transmethylation solvent. Characteristics of the method were evaluated and it was used to phenotype transgenic T₁ seeds expressing AtFAD2 or RcWRI1. Our results indicate that fatty acid composition of T₁ individual seeds are consistent with those of pools of multiple seeds from higher generations. However, oil content per individual seed varied substantially and therefore pooling five seeds is recommended for phenotyping oil content of T₁ seeds. Additionally, a whole camelina single‐seed direct transmethylation was evaluated and results confirm its feasibility. The suitability of partial seed analysis of camelina was investigated but variation in composition of different seed tissues limits this approach.

中文翻译：

---

通过对单一转基因 T1 拟南芥和亚麻荠种子的脂肪酸组成和油含量进行快速表型分析，加速基因功能发现。


拟南芥被广泛用作模式植物，亚麻荠越来越多地用于油籽研究和应用。尽管拟南芥基因组测序已有二十年历史，但许多脂质相关基因及其调节因子的功能尚未得到很好的表征。提高基因研究的效率和准确性是有效发现基因功能和植物油数量和质量的下游生物工程的关键。在这项研究中，使用可见标记来快速识别转基因 T ₁种子，并开发了一种方法来对单个 T ₁种子的脂肪酸组成和油含量进行表型分析。首先对多个种子优化了整个种子直接转甲基方法，并建议在转甲基溶剂（5% H ₂ SO ₄的甲醇溶液和 30% 甲苯共溶剂）中于 85°C 孵育 2 小时。基于该方法，在装有 200 μl 转甲基溶剂的 1.5 ml GC 样品瓶中建立了单拟南芥种子微型转甲基化 (SAST) 方法。评估了该方法的特征，并将其用于表达AtFAD2或RcWRI1的转基因T ₁种子的表型。我们的结果表明，T ₁单个种子的脂肪酸组成与来自更高世代的多个种子库的脂肪酸组成一致。然而，每个种子的含油量差异很大，因此建议合并五颗种子来对 T ₁种子的含油量进行表型分析。此外，还对整个亚麻荠单种子直接转甲基进行了评估，结果证实了其可行性。 研究了亚麻荠部分种子分析的适用性，但不同种子组织的组成变化限制了这种方法。