Table_1_Transcription Factor DOF4.1 Regulates Seed Longevity in Arabidopsis via Seed Permeability and Modulation of Seed Storage Protein Accumulation.xlsx

Seed longevity is modulated by multiple genetic factors in Arabidopsis thaliana. A previous genome-wide association study using the Elevated Partial Pressure of Oxygen (EPPO) aging assay pinpointed a genetic locus associated with this trait. Reverse genetics identified the transcription factor DOF4.1 as a novel seed longevity factor. dof4.1 loss-of-function plants generate seeds exhibiting higher germination after accelerated aging assays. DOF4.1 is expressed during seed development and RNAseq data show several putative factors that could contribute to the dof4.1 seed longevity phenotype. dof4.1 has reduced seed permeability and a higher levels of seed storage proteins mRNAs (cruciferins and napins) in developing seeds, as compared to wild-type seeds. It has been reported that mutant lines defective in cruciferins or napins present reduced seed longevity. The improved longevity of dof4.1 is totally lost in the quadruple mutant dof4.1 cra crb crc, but not in a dof4.1 line depleted of napins, suggesting a prominent role for cruciferins in this process. Moreover, a negative regulation of DOF4.1 expression by the transcription factor DOF1.8 is suggested by co-inoculation assays in Nicotiana benthamiana. Indeed, DOF1.8 expression anticorrelates with that of DOF4.1 during seed development. In summary, modulation of DOF4.1 levels during seed development contributes to regulate seed longevity.

拟南芥（Arabidopsis thaliana）的种子寿命受多种遗传因素调控。此前一项利用升高氧分压（Elevated Partial Pressure of Oxygen, EPPO）衰老试验开展的全基因组关联研究，已定位到与该性状相关的遗传位点。反向遗传学研究将转录因子DOF4.1鉴定为新型种子寿命调控因子。DOF4.1功能缺失型植株所结的种子，在加速衰老试验后展现出更高的萌发率。DOF4.1在种子发育阶段表达，转录组测序（RNAseq）数据显示，存在多个潜在因子可能参与介导DOF4.1的种子寿命表型。与野生型种子相比，DOF4.1突变体的种子通透性更低，且发育种子中储存蛋白mRNA（cruciferins与napins）的表达水平更高。已有研究表明，cruciferins或napins功能缺陷的突变体株系，其种子寿命会降低。DOF4.1突变体所提升的种子寿命，在四突变体dof4.1 cra crb crc中完全丧失，但在napins敲低的DOF4.1株系中未受影响，这表明cruciferins在该过程中发挥核心作用。此外，本氏烟草（Nicotiana benthamiana）中的共接种试验提示，转录因子DOF1.8可负调控DOF4.1的表达。事实上，在种子发育过程中，DOF1.8的表达模式与DOF4.1呈显著负相关。综上，种子发育阶段对DOF4.1表达水平的调控，可参与调节种子寿命。