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Tuber-specific cphA expression to enhance cyanophycin production in potatoes
Authors:Maja Hü  hns,Katrin Neumann &dagger  ,Tina Hausmann,Friederike Klemke,Wolfgang Lockau,Uwe Kahmann,Lilya Kopertekh,Dorothee Staiger,Elfriede K. Pistorius,Jens Reuther,Eva Waldvogel,Wolfgang Wohlleben,Martin Effmert,Holger Junghans,Katja Neubauer,Udo Kragl,Kerstin Schmidt,Jö  rg Schmidtke, Inge Broer
Affiliation:Agrobiotechnology, University of Rostock, Rostock, Germany;
Biochemie der Pflanzen, Humboldt-Universität zu Berlin, Berlin, Germany;
Biologie VIII: Molekulare Zellphysiologie, Universität Bielefeld, Bielefeld, Germany;
Zentrum für ultrastrukturelle Diagnostik im IIT, Bielefeld, Germany;
Julius-Kühn-Institute, Braunschweig, Germany;
Department of Microbiology/Biotechnology, Institute of Microbiology, University of Tuebingen, Tuebingen, Germany;
NORIKA GmbH, Gross Lüsewitz, Germany;
Technical Chemistry, University of Rostock, Rostock, Germany;
BioMath GmbH, Gross Lüsewitz, Germany
Abstract:The production of biodegradable polymers that can be used to substitute petrochemical compounds in commercial products in transgenic plants is an important challenge for plant biotechnology. Nevertheless, it is often accompanied by reduced plant fitness. To decrease the phenotypic abnormalities of the sprout and to increase polymer production, we restricted cyanophycin accumulation to the potato tubers by using the cyanophycin synthetase gene ( cph ATe) from Thermosynechococcus elongatus BP-1, which is under the control of the tuber-specific class 1 promoter (B33). Tuber-specific cytosolic (pB33- cph ATe) as well as tuber-specific plastidic (pB33-PsbY- cph ATe) expression resulted in significant polymer accumulation solely in the tubers. In plants transformed with pB33- cph ATe, both cyanophycin synthetase and cyanophycin were detected in the cytoplasm leading to an increase up to 2.3% cyanophycin of dry weight and resulting in small and deformed tubers. In B33-PsbY- cph ATe tubers, cyanophycin synthetase and cyanophycin were exclusively found in amyloplasts leading to a cyanophycin accumulation up to 7.5% of dry weight. These tubers were normal in size, some clones showed reduced tuber yield and sometimes exhibited brown sunken staining starting at tubers navel. During a storage period over of 32 weeks of one selected clone, the cyanophycin content was stable in B33-PsbY- cph ATe tubers but the stress symptoms increased. However, all tubers were able to germinate. Nitrogen fertilization in the greenhouse led not to an increased cyanophycin yield, slightly reduced protein content, decreased starch content, and changes in the amounts of bound and free arginine and aspartate, as compared with control tubers were observed.
Keywords:biodegradable polymer    cyanophycin    polyaspartate    Solanum tuberosum    tuber-specific    plastid
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