Abstract

Lysine is the most limiting essential amino acid in cereals, and efforts have been made over the decades to improve the nutritional quality of these grains by limiting storage protein accumulation and increasing lysine content, while maintaining desired agronomic traits. The single lys3 mutation in barley has been shown to significantly increase lysine content but also reduces grain size. Herein, the regulatory effect of the lys3 mutation that controls storage protein accumulation as well as a plethora of critically important processes in cereal seeds was investigated in double mutant barley lines. This was enabled through the generation of three hordein double-mutants by inter-crossing three single hordein mutants, that had all been backcrossed three times to the malting barley cultivar Sloop. Proteome abundance measurements were integrated with their phenotype measurements; proteins were mapped to chromosomal locations and to their corresponding functional classes. These models enabled the prediction of previously unknown points of crosstalk that connect the impact of lys3 mutations to other signalling pathways. In combination, these results provide an improved understanding of how the mutation at the lys3 locus remodels cellular functions and impact phenotype that can be used in selective breeding to generate favourable agronomic traits.

Highlights

  • Barley is an important cereal grown mainly for feed and malting

  • The coeliac disease (CD)-mutant line showed the substantial increase in total fatty acid, with an 82.2% increase as compared to the

  • The highest increase in total Triacyl Glycerol (TAG) content was observed in the CDmutant (127.3% increase), followed by the BC-mutant (110.1% increase); no significant change was observed in the BD-mutant (Table 1)

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Summary

Introduction

Barley is an important cereal grown mainly for feed and malting. The major seed storage proteins in barley, the hordeins, are elicitors of coeliac disease (CD) – a condition that adversely affects ∼1% of the world population (∼70 million people). There is no current treatment other than strict adherence to a life-long gluten-free (GF) diet. In the diploid barley genome, there are four types of hordeins present: B-, C-, D- and γ-hordeins. The B- and C-hordeins are the primary classes representing >90% of the hordeins in barley (Kreis et al, 1983; Shewry et al, 1985). The B- and C-hordeins are encoded by the Hor-2 and Hor-1 loci, respectively, and both loci are located on the short arm of chromosome 1. The high molecular weight glutenin orthologous D-hordein genes are located at Hor-3 (1H long arm) and the S-rich γ-hordeins present at Hor-5 (1H short arm) (Shewry, 1993). The B- and C-hordeins comprise two multigene families consisting

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