The WRINKLED1 (WRI1) and LAFL [LEAFY COTYLEDON1 (LEC1), ABSCISIC ACID INSENSITIVE3 (ABI3), FUSCA3 (FUS3), and LEC2] transcription factors play essential roles in governing seed development and oil biosynthesis. To gain a comprehensive understanding of the transcriptional regulation of WRI1 and LAFL, we conducted genome-wide association studies for the expression profiles of WRI1 and LAFL in developing seeds at 20 and 40 days after flowering (DAF) using 302 rapeseed (Brassica napus) accessions. We identified a total of 237 expression quantitative trait nucleotides (eQTNs) and 51 expression QTN-by-environment interactions (eQEIs) associated with WRI1 and LAFL. Around these eQTNs and eQEIs, we pinpointed 41 and 8 candidate genes with known transcriptional regulations or protein interactions with their expression traits, respectively. Based on RNA-seq and ATAC-seq data, we employed the XGBoost and Basenji models which predicted 15 candidate genes potentially regulating the expression of WRI1 and LAFL. We further validated the predictions via tissue expression profile, haplotype analysis, and expression correlation analysis, and verified the transcriptional activation activity of BnaC03.MYB56 (R2R3-MYB transcription factor 56) on the expression of BnaA09.LEC1 by dual-luciferase reporter and yeast one-hybrid assays. BnaA10.AGL15 (AGAMOUS-LIKE 15), BnaC04.VAL1 (VIVIPAROUS1/ABSCISIC ACID INSENSITIVE3-LIKE 1), BnaC03.MYB56, and BnaA10.MYB56 were co-expressed with WRI1 and LAFL at 20 DAF in M35, a key module for seed development and oil biosynthesis. We further validated the positive regulation of MYB56 on seed oil accumulation using Arabidopsis (Arabidopsis thaliana) mutants. This study not only delivers a framework for future eQEI identification but also offers insights into the developmental regulation of seed oil accumulation.

中文翻译：

---

甘蓝型油菜种子发育过程中转录因子基因WRI1和LAFL的转录调控


WRINKLED1 (WRI1) 和 LAFL [LEAFY COTYLEDON1 (LEC1)、ABSCISIC ACID INSENSITIVE3 (ABI3)、FUSCA3 (FUS3) 和 LEC2] 转录因子在控制种子发育和油生物合成中发挥着重要作用。为了全面了解 WRI1 和 LAFL 的转录调控，我们使用 302 个油菜籽 (Brassica napus) 材料，对开花后 20 天和 40 天种子发育 (DAF) 中 WRI1 和 LAFL 的表达谱进行了全基因组关联研究。 。我们总共鉴定了 237 个与 WRI1 和 LAFL 相关的表达数量性状核苷酸 (eQTN) 和 51 个表达 QTN 与环境的相互作用 (eQEI)。围绕这些 eQTN 和 eQEI，我们分别确定了 41 个和 8 个候选基因，这些基因具有已知的转录调控或与其表达特征的蛋白质相互作用。基于 RNA-seq 和 ATAC-seq 数据，我们采用 XGBoost 和 Basenji 模型预测了 15 个可能调节 WRI1 和 LAFL 表达的候选基因。我们通过组织表达谱、单倍型分析和表达相关性分析进一步验证了预测，并通过双荧光素酶报告基因和酵母验证了BnaC03.MYB56（R2R3-MYB转录因子56）对BnaA09.LEC1表达的转录激活活性单杂交测定。 BnaA10.AGL15 (AGAMOUS-LIKE 15)、BnaC04.VAL1 (VIVIPAROUS1/ABSCISIC ACID INSENSITIVE3-LIKE 1)、BnaC03.MYB56 和 BnaA10.MYB56 与 WRI1 和 LAFL 在 M35（M35 的关键模块）中于 20 DAF 共表达种子发育和油生物合成。我们利用拟南芥（Arabidopsis thaliana）突变体进一步验证了MYB56对种子油积累的正向调节。 这项研究不仅为未来的 eQEI 识别提供了框架，而且还提供了对种子油积累的发育调控的见解。