Abstract

Within the cereal grain, the endosperm and its nutrient reserves are critical for successful germination and in the context of grain utilization. The identification of molecular determinants of early endosperm development, particularly regulators of cell division and cell wall deposition, would help predict end-use properties such as yield, quality, and nutritional value. Custom microarray data have been generated using RNA isolated from developing barley grain endosperm 3 d to 8 d after pollination (DAP). Comparisons of transcript abundance over time revealed 47 gene expression modules that can be clustered into 10 broad groups. Superimposing these modules upon cytological data allowed patterns of transcript abundance to be linked with key stages of early grain development. Here, attention was focused on how the datasets could be mined to explore and define the processes of cell wall biosynthesis, remodeling, and degradation. Using a combination of spatial molecular network and gene ontology enrichment analyses, it is shown that genes involved in cell wall metabolism are found in multiple modules, but cluster into two main groups that exhibit peak expression at 3 DAP to 4 DAP and 5 DAP to 8 DAP. The presence of transcription factor genes in these modules allowed candidate genes for the control of wall metabolism during early barley grain development to be identified. The data are publicly available through a dedicated web interface (https://ics.hutton.ac.uk/barseed/), where they can be used to interrogate co- and differential expression for any other genes, groups of genes, or transcription factors expressed during early endosperm development.

Highlights

  • Within the cereal grain, the endosperm and its nutrient reserves are critical for successful germination and in the context of grain utilization

  • Developing barley grains were collected from 3 d after pollination (DAP) to 8 DAP and examined by transverse sectioning, toluidine blue staining, and light microscopy to determine the temporal sequence of developmental events

  • Under the growth conditions used in this study, the syncytial phase of endosperm development continued until approximately 3 DAP, at which point the nuclei were evenly arranged at the periphery of the embryo sac

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Summary

Introduction

The endosperm and its nutrient reserves are critical for successful germination and in the context of grain utilization. Comparisons of transcript abundance over time revealed 47 gene expression modules that can be clustered into 10 broad groups Superimposing these modules upon cytological data allowed patterns of transcript abundance to be linked with key stages of early grain development. The total amount of starch, storage proteins, and to a lesser extent lipids and minerals, determine the nutritional value and quality of the grain, both for the human diet and in grain processing. These factors, together with the speed with which the reserves are released to support seedling growth after germination, are important determinants of seedling vigor and crop establishment. The callosic material that was abundant in the early cell walls is replaced by arabinoxylans and (1,3;1,4)-b-glucans, which are the major noncellulosic wall polysaccharides in mature grains (Fincher, 1975; Bacic and Stone, 1981)

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