title: HvPap-1 C1A Protease and HvCPI-2 Cystatin Contribute to Barley Grain Filling and Germination.
abstract: Proteolysis is an essential process throughout the mobilization of storage proteins in barley (Hordeum vulgare) grains during germination. It involves numerous types of enzymes, with C1A Cys proteases the most abundant key players. Manipulation of the proteolytic machinery is a potential way to enhance grain yield and quality, and it could influence the mobilization of storage compounds along germination. Transgenic barley plants silencing or over-expressing the cathepsin F-like HvPap-1 Cys protease show differential accumulation of storage molecules such as starch, proteins, and free amino acids in the grain. It is particularly striking that the HvPap-1 artificial microRNA lines phenotype show a drastic delay in the grain germination process. Alterations to the proteolytic activities in the over-expressing and knock-down grains associated with changes in the level of expression of several C1A peptidases were also detected. Similarly, down-regulating cystatin Icy-2, one of the proteinaceous inhibitors of the cathepsin F-like protease, also has important effects on grain filling. However, the ultimate physiological influence of manipulating a peptidase or an inhibitor cannot be always predicted, since the plant tries to compensate the modified proteolytic effects by modulating the expression of some other peptidases or their inhibitors.
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Mercedes
authors.identifier: 0000-0002-3545-987X
authors.initials: M
authors.lastname: Diaz-Mendoza
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Jose D
authors.identifier: 0000-0002-2614-557X
authors.initials: JD
authors.lastname: Dominguez-Figueroa
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Blanca
authors.initials: B
authors.lastname: Velasco-Arroyo
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Ines
authors.initials: I
authors.lastname: Cambra
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Pablo
authors.initials: P
authors.lastname: Gonzalez-Melendi
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Angeles
authors.initials: A
authors.lastname: Lopez-Gonzalvez
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Antonia
authors.initials: A
authors.lastname: Garcia
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Goetz
authors.identifier: 0000-0002-5539-3097
authors.initials: G
authors.lastname: Hensel
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Jochen
authors.identifier: 0000-0001-7080-7983
authors.initials: J
authors.lastname: Kumlehn
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.).
authors.forename: Isabel
authors.identifier: 0000-0001-9865-902X
authors.initials: I
authors.lastname: Diaz
authors.affiliation: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid, Campus Montegancedo, 28223-Pozuelo de Alarcon, Madrid, Spain (M.D.-M., J.D.D.-F., B.V.-A., I.C., P.G.-M., I.D., M.M.); Centre for Metabolomics and Bioanalysis, Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte 28668, Madrid, Spain (A.L.-G., A.G.); andPlant Reproductive Biology, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466 Stadt Seeland/OT Gatersleben, Germany (G.H., J.K.) m.martinez@upm.es.
authors.forename: Manuel
authors.identifier: 0000-0002-7826-5872
authors.initials: M
authors.lastname: Martinez
mesh_terms: D015891:Cystatins / Q000235:genetics / Q000378:metabolism; D002523:Edible Grain / Q000196:embryology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D015870:Gene Expression; D020868:Gene Silencing; D018525:Germination; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D035683:MicroRNAs / Q000235:genetics; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D059748:Proteolysis; D055314:Seed Storage Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015891:Cystatins; D035683:MicroRNAs; D010940:Plant Proteins; D055314:Seed Storage Proteins; D010447:Peptide Hydrolases