Identification and molecular characterisation of hordoindolines from barley grain

• Published in: Plant molecular biology 2001-12, Vol.47 (6), p.785-794
• Main Authors: Darlington, H F, Rouster, J, Hoffmann, L, Halford, N G, Shewry, P R, Simpson, D J
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Bacteria; Barley; Base Sequence; Cultivars; DNA, Complementary - chemistry; DNA, Complementary - genetics; DNA, Plant - chemistry; DNA, Plant - genetics; Gene Expression Regulation, Plant; Grain; Hordeum - genetics; Hordeum vulgare; hordoindolines; Microbiology; Molecular Sequence Data; Plant Proteins - genetics; Protein Isoforms - genetics; RNA, Messenger - genetics; RNA, Messenger - metabolism; Seeds - genetics; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Wheat
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1023/A:1013691530675

Grain texture in barley is an important quality character as soft-textured cultivars have better malting quality. In wheat, texture is considered to be determined by the puroindolines, a group of basic hydrophobic proteins present on the surface of the starch granule. Hard wheats have been proposed to lack puroindoline a or to have mutant forms of puroindoline b which do not bind to the granule surface. Analysis of six barley cultivars (three soft-textured and three hard) showed that all contained proteins homologous to wheat puroindoline b, but PCR analysis failed to show any differences in amino acid sequences similar to those which have been proposed to determine textural differences in wheat. Southern blot analysis showed two hordoindoline b genes which were isolated and shown to encode proteins with 94% sequence identity. Expression of hordoindoline b mRNA occurred in the starchy endosperm and aleurone layer of the developing seed, but not in the embryo. Analysis of seven soft- and six hard-textured barley varieties showed that all contained hordoindoline a except two hard varieties (Sundance, Hart) which were subsequently shown to both lack hordoindoline a mRNA. It was therefore concluded that there is not a clear relationship between the presence of hordoindoline a and grain texture in barley.