Protein Targeting to Starch 1 is essential for starchy endosperm development in barley

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  • ery398

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    Yingxin Zhong, Department of Molecular Biology and Genetics, Aarhus University, Flakkebjerg, Forsøgsvej, Slagelse, Denmark., National Technique Innovation Center for Regional Wheat Production/Key Laboratory of Crop Physiology and Ecology in Southern China, Ministry of Agriculture/National Engineering and Technology Center for Information Agriculture, Nanjing Agricultural University, Andreas Blennow, Department of Plant and Environmental Sciences, University of Copenhagen, Olivia Kofoed-Enevoldsen, Department of Plant and Environmental Sciences, University of Copenhagen, Dong Jiang, National Technique Innovation Center for Regional Wheat Production/Key Laboratory of Crop Physiology and Ecology in Southern China, Ministry of Agriculture/National Engineering and Technology Center for Information Agriculture, Nanjing Agricultural University,
  • Kim Henrik Hebelstrup

Plant starch is the main energy contributor to the human diet. Its biosynthesis is catalyzed and regulated by co-ordinated actions of several enzymes. Recently, a factor termed Protein Targeting to Starch 1 (PTST1) was identified as being required for correct granule-bound starch synthase (GBSS) localization and demonstrated to be crucial for amylose synthesis in Arabidopsis. However, the function of its homologous protein in storage tissues (e.g. endosperm) is unknown. We identified a PTST1 homolog in barley and it was found to contain a crucial coiled-coil domain and carbohydrate-binding module. We demonstrated the interaction between PTST1 and GBSS1 by fluorescence resonance energy transfer (FRET) in barley endosperm. By tagging PTST1 with the fluorophore mCherry, we observed that it is localized in the stroma of barley endosperm amyloplasts. PTST1 overexpression in endosperm increased endogenous gbss1a gene expression and amylose content. Gbss1a and ptst1 mutants were generated using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-related protein 9 (Cas9)-based targeted mutagenesis. Homozygous gbss1a mutants showed a waxy phenotype. Grains of ptst1 mutants did not accumulate any starch. These grains dried out during the desiccation stage and were unable to germinate, suggesting that PTST1 is essential for development of starchy endosperm and viable grains.

OriginalsprogEngelsk
TidsskriftJournal of Experimental Botany
Vol/bind70
Tidsskriftsnummer2
Sider (fra-til)485-496
Antal sider12
ISSN0022-0957
DOI
StatusUdgivet - 7 jan. 2019

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