Key Laboratory of Plant Molecular Physiology, Institute of Botany, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing, 100093, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Theor Appl Genet. 2023 Mar 13;136(3):47. doi: 10.1007/s00122-023-04250-z.
Plastidial α-glucan phosphorylase is a key factor that cooperates with plastidial disproportionating enzyme to control short maltooligosaccharide mobilization during the initiation process of starch molecule synthesis in developing rice endosperm. Storage starch synthesis is essential for grain filling. However, little is known about how cereal endosperm controls starch synthesis initiation. One of core events for starch synthesis initiation is short maltooligosaccharide (MOS) mobilization consisting of long MOS primer production and excess MOS breakdown. By mutant analyses and biochemical investigations, we present here functional identifications of plastidial α-glucan phosphorylase (Pho1) and disproportionating enzyme (DPE1) during starch synthesis initiation in rice (Oryza sativa) endosperm. Pho1 deficiency impaired MOS mobilization, triggering short MOS accumulation and starch synthesis reduction during early seed development. The mutant seeds differed significantly in MOS level and starch content at 15 days after flowering and exhibited diverse endosperm phenotypes during mid-late seed development: ranging from pseudonormal to shrunken (Shr), severely or excessively Shr. The level of DPE1 was almost normal in the PN seeds but significantly reduced in the Shr seeds. Overexpression of DPE1 in pho1 resulted in plump seeds only. DPE1 deficiency had no obvious effects on MOS mobilization. Knockout of DPE1 in pho1 completely blocked MOS mobilization, resulting in severely and excessively Shr seeds only. These findings show that Pho1 cooperates with DPE1 to control short MOS mobilization during starch synthesis initiation in rice endosperm.
质体 α-葡聚糖磷酸化酶是与质体分相酶协同作用的关键因子,控制着水稻胚乳淀粉分子合成起始过程中短直链麦芽寡糖的动员。贮藏淀粉合成对于籽粒灌浆至关重要。然而,关于谷物胚乳如何控制淀粉合成起始的了解甚少。淀粉合成起始的核心事件之一是短直链麦芽寡糖(MOS)的动员,包括长 MOS 引物的产生和过量 MOS 的分解。通过突变体分析和生化研究,我们在此提出了在水稻(Oryza sativa)胚乳淀粉合成起始过程中质体 α-葡聚糖磷酸化酶(Pho1)和分相酶(DPE1)的功能鉴定。Pho1 缺陷会损害 MOS 的动员,导致早期种子发育过程中 MOS 积累和淀粉合成减少。突变体种子在花后 15 天的 MOS 水平和淀粉含量差异显著,在中晚期种子发育过程中表现出不同的胚乳表型:从假正常到皱缩(Shr),严重或过度 Shr。PN 种子中的 DPE1 水平几乎正常,但 Shr 种子中的 DPE1 水平显著降低。Pho1 中的 DPE1 过表达仅导致饱满的种子。DPE1 缺陷对 MOS 动员没有明显影响。Pho1 中的 DPE1 敲除完全阻断了 MOS 的动员,仅导致严重和过度 Shr 的种子。这些发现表明,Pho1 与 DPE1 合作控制水稻胚乳淀粉合成起始过程中短 MOS 的动员。