A low-starch barley mutant, risø 16, lacking the cytosolic small subunit of ADP-glucose pyrophosphorylase, reveals the importance of the cytosolic isoform and the identity of the plastidial small subunit.
To provide information on the roles of the different forms of ADP-glucose pyrophosphorylase (AGPase) in barley (Hordeum vulgare) endosperm and the nature of the genes encoding their subunits, a mutant of barley, Risø 16, lacking cytosolic AGPase activity in the endosperm was identified. The mutation specifically abolishes the small subunit of the cytosolic AGPase and is attributable to a large deletion within the coding region of a previously characterized small subunit gene that we have called Hv.AGP.S.1. The plastidial AGPase activity in the mutant is unaffected. This shows that the cytosolic and plastidial small subunits of AGPase are encoded by separate genes. We purified the plastidial AGPase protein and, using amino acid sequence information, we identified the novel small subunit gene that encodes this protein. Studies of the Risø 16 mutant revealed the following. First, the reduced starch content of the mutant showed that a cytosolic AGPase is required to achieve the normal rate of starch synthesis. Second, the mutant makes both A- and B-type starch granules, showing that the cytosolic AGPase is not necessary for the synthesis of these two granule types. Third, analysis of the phylogenetic relationships between the various small subunit proteins both within and between species, suggest that the cytosolic AGPase single small subunit gene probably evolved from a leaf single small subunit gene.
Johnson
Philip E
PE
John Innes Centre, Norwich Research Park, Colney, Norfolk NR4 7UH, United Kingdom.
Patron
Nicola J
NJ
Bottrill
Andrew R
AR
Dinges
Jason R
JR
Fahy
Brendan F
BF
Parker
Mary L
ML
Waite
Darren N
DN
Denyer
Kay
K
D000245:Adenosine Diphosphate Glucose / Q000378:metabolism; D000595:Amino Acid Sequence; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D051599:Glucose-1-Phosphate Adenylyltransferase; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D001467:Hordeum / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009154:Mutation; D009713:Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D018087:Plastids / Q000201:enzymology; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000648:ultrastructure; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013213:Starch / Q000096:biosynthesis*
D018076:DNA, Complementary; D007527:Isoenzymes; D000245:Adenosine Diphosphate Glucose; D013213:Starch; D005964:Glucosyltransferases; C022699:sucrose synthase; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase