Roy J. Carver Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA 50011.
Horticultural Sciences Department, University of Florida, Gainesville, FL 32611.
Proc Natl Acad Sci U S A. 2018 Jan 2;115(1):E24-E33. doi: 10.1073/pnas.1715668115. Epub 2017 Dec 18.
Maize () mutations are beneficial for endosperm nutritional quality but cause negative pleiotropic effects for reasons that are not fully understood. Direct targets of the bZIP transcriptional regulator encoded by include and that specify pyruvate phosphate dikinase (PPDK). This enzyme reversibly converts AMP, pyrophosphate, and phosphoenolpyruvate to ATP, orthophosphate, and pyruvate and provides diverse functions in plants. This study addressed PPDK function in maize starchy endosperm where it is highly abundant during grain fill. and were inactivated individually by transposon insertions, and both genes were simultaneously targeted by endosperm-specific RNAi. accounts for the large majority of endosperm PPDK, whereas specifies the abundant mesophyll form. The mutation is seedling-lethal, indicating that C4 photosynthesis is essential in maize. RNAi expression in transgenic endosperm eliminated detectable PPDK protein and enzyme activity. Transgenic kernels weighed the same on average as nontransgenic siblings, with normal endosperm starch and total N contents, indicating that PPDK is not required for net storage compound synthesis. An opaque phenotype resulted from complete PPDK knockout, including loss of vitreous endosperm character similar to the phenotype conditioned by -. Concentrations of multiple glycolytic intermediates were elevated in transgenic endosperm, energy charge was altered, and starch granules were more numerous but smaller on average than normal. The data indicate that PPDK modulates endosperm metabolism, potentially through reversible adjustments to energy charge, and reveal that - mutations can affect the opaque phenotype through regulation of PPDK in addition to their previously demonstrated effects on storage protein gene expression.
玉米突变是有益的胚乳营养品质,但由于某些原因导致其具有负的多效性效应,这些原因还不完全清楚。由 bZIP 转录因子编码的直接靶标包括和,指定丙酮酸磷酸二激酶(PPDK)。该酶可逆地将 AMP、焦磷酸和磷酸烯醇丙酮酸转化为 ATP、正磷酸盐和丙酮酸,并在植物中提供多种功能。本研究解决了 PPDK 在玉米淀粉胚乳中的功能,在灌浆期间,它在胚乳中高度丰富。和分别通过转座子插入失活,并且这两个基因都被胚乳特异性 RNAi 靶向。负责胚乳 PPDK 的绝大部分,而指定丰富的叶肉形式。突变是幼苗致死的,表明 C4 光合作用在玉米中是必不可少的。转基因胚乳中的 RNAi 表达消除了可检测的 PPDK 蛋白和酶活性。转基因核仁的平均重量与非转基因同系物相同,胚乳淀粉和总氮含量正常,表明 PPDK 不是净储存化合物合成所必需的。完全敲除 PPDK 导致不透明表型,包括类似由 - 条件的玻璃质胚乳特征的丧失。在转基因胚乳中,多个糖酵解中间产物的浓度升高,能量电荷发生改变,淀粉颗粒的数量更多,但平均而言比正常情况下更小。这些数据表明,PPDK 调节胚乳代谢,可能通过对能量电荷的可逆调节,并且表明 - 突变可以通过调节 PPDK 来影响不透明表型,除了它们对储存蛋白基因表达的先前证明的影响之外。