Abstract

Accumulation of reserve materials in filling grains involves the coordination of different metabolic and cellular processes, and understanding the molecular mechanisms underlying the interconnections remains a major challenge for proteomics. Rice (Oryza sativa) is an excellent model for studying grain filling because of its importance as a staple food and the available genome sequence database. Our observations showed that embryo differentiation and endosperm cellularization in developing rice seeds were completed approximately 6 d after flowering (DAF); thereafter, the immature seeds mainly underwent cell enlargement and reached the size of mature seeds at 12 DAF. Grain filling began at 6 DAF and lasted until 20 DAF. Dynamic proteomic analyses revealed 396 protein spots differentially expressed throughout eight sequential developmental stages from 6 to 20 DAF and determined 345 identities. These proteins were involved in different cellular and metabolic processes with a prominently functional skew toward metabolism (45%) and protein synthesis/destination (20%). Expression analyses of protein groups associated with different functional categories/subcategories showed that substantially up-regulated proteins were involved in starch synthesis and alcoholic fermentation, whereas the down-regulated proteins in the process were involved in central carbon metabolism and most of the other functional categories/subcategories such as cell growth/division, protein synthesis, proteolysis, and signal transduction. The coordinated changes were consistent with the transition from cell growth and differentiation to starch synthesis and clearly indicated that a switch from central carbon metabolism to alcoholic fermentation may be important for starch synthesis and accumulation in the developmental process.

Highlights

  • Accumulation of reserve materials in filling grains involves the coordination of different metabolic and cellular processes, and understanding the molecular mechanisms underlying the interconnections remains a major challenge for proteomics

  • After 18 d after flowering (DAF), the increase in fresh weight slowed, but the dry weight kept increasing until 20 DAF, which indicates that developing seeds enter into the desiccation phase from 18 DAF

  • The results showed that the ATP level increased from 6 DAF until 12 DAF and thereafter decreased dramatically (Fig. 1F), which suggested that the active starch synthesis consumed less ATP than early cell enlargement

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Summary

Introduction

Accumulation of reserve materials in filling grains involves the coordination of different metabolic and cellular processes, and understanding the molecular mechanisms underlying the interconnections remains a major challenge for proteomics. Our observations showed that embryo differentiation and endosperm cellularization in developing rice seeds were completed approximately 6 d after flowering (DAF); thereafter, the immature seeds mainly underwent cell enlargement and reached the size of mature seeds at 12 DAF. Proteomic Analysis of Filling Rice Grains genome sequence, which is important for acquiring knowledge about the mechanisms of seed development and starch accumulation. At approximately 20 DAF, the seed enters the desiccation phase (Ishimaru et al, 2003) These findings, taken together with the observations about cellular and physiological changes in developing seeds of other species (Olsen, 2001), suggest that coordinated cellular and metabolic changes that occur in a timely manner between different developmental events are related to the accumulation of reserves, but the underlying mechanisms are still unknown

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