Downregulation of a chloroplast-targeted beta-amylase leads to a starch-excess phenotype in leaves.
A functional screen in Escherichia coli was established to identify potato genes coding for proteins involved in transitory starch degradation. One clone isolated had a sequence very similar to a recently described chloroplast-targeted beta-amylase of Arabidopsis. Expression of the gene in E. coli showed that the protein product was a functional beta-amylase that could degrade both starch granules and solubilized amylopectin, while import experiments demonstrated that the beta-amylase was imported and processed into pea chloroplasts. To study the function of the protein in transitory starch degradation, transgenic potato plants were generated where its activity was reduced using antisense techniques. Analysis of plants reduced in the presence of this beta-amylase isoform showed that their leaves had a starch-excess phenotype, indicating a defect in starch degradation. In addition, it was shown that the antisense plants degraded only 8-30% of their total starch, in comparison with 50% in the wild type, over the dark period. This is the first time that a physiological role for a beta-amylase in plants has been demonstrated.
Scheidig
Andreas
A
Max Planck Institute of Molecular Plant Physiology, Willmitzer Department, Am Mühlenberg 1, 14476 Golm, Germany.
Fröhlich
Anja
A
Schulze
Silke
S
Lloyd
James R
JR
Kossmann
Jens
J
D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015536:Down-Regulation; D004926:Escherichia coli; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D017343:Genes, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D010641:Phenotype; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D021381:Protein Transport; D011198:Solanum tuberosum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D013213:Starch / Q000378:metabolism*; D001614:beta-Amylase / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D013213:Starch; D001614:beta-Amylase