@article{OrzechowskiSitnickaGrabowskaetal.2021, author = {Orzechowski, Slawomir and Sitnicka, Dorota and Grabowska, Agnieszka and Compart, Julia and Fettke, J{\"o}rg and Zdunek-Zastocka, Edyta}, title = {Effect of short-term cold treatment on carbohydrate metabolism in potato leaves}, series = {International journal of molecular sciences}, volume = {22}, journal = {International journal of molecular sciences}, number = {13}, publisher = {MDPI}, address = {Basel}, issn = {1422-0067}, doi = {10.3390/ijms22137203}, pages = {17}, year = {2021}, abstract = {Plants are often challenged by an array of unfavorable environmental conditions. During cold exposure, many changes occur that include, for example, the stabilization of cell membranes, alterations in gene expression and enzyme activities, as well as the accumulation of metabolites. In the presented study, the carbohydrate metabolism was analyzed in the very early response of plants to a low temperature (2 degrees C) in the leaves of 5-week-old potato plants of the Russet Burbank cultivar during the first 12 h of cold treatment (2 h dark and 10 h light). First, some plant stress indicators were examined and it was shown that short-term cold exposure did not significantly affect the relative water content and chlorophyll content (only after 12 h), but caused an increase in malondialdehyde concentration and a decrease in the expression of NDA1, a homolog of the NADH dehydrogenase gene. In addition, it was shown that the content of transitory starch increased transiently in the very early phase of the plant response (3-6 h) to cold treatment, and then its decrease was observed after 12 h. In contrast, soluble sugars such as glucose and fructose were significantly increased only at the end of the light period, where a decrease in sucrose content was observed. The availability of the monosaccharides at constitutively high levels, regardless of the temperature, may delay the response to cold, involving amylolytic starch degradation in chloroplasts. The decrease in starch content, observed in leaves after 12 h of cold exposure, was preceded by a dramatic increase in the transcript levels of the key enzymes of starch degradation initiation, the alpha-glucan, water dikinase (GWD-EC 2.7.9.4) and the phosphoglucan, water dikinase (PWD-EC 2.7.9.5). The gene expression of both dikinases peaked at 9 h of cold exposure, as analyzed by real-time PCR. Moreover, enhanced activities of the acid invertase as well as of both glucan phosphorylases during exposure to a chilling temperature were observed. However, it was also noticed that during the light phase, there was a general increase in glucan phosphorylase activities for both control and cold-stressed plants irrespective of the temperature. In conclusion, a short-term cold treatment alters the carbohydrate metabolism in the leaves of potato, which leads to an increase in the content of soluble sugars.}, language = {en} } @article{MalinovaKoesslerOrawetzetal.2019, author = {Malinova, Irina and K{\"o}ssler, Stella and Orawetz, Tom and Matthes, Ulrike and Orzechowski, Slawomir and Koch, Anke and Fettke, J{\"o}rg}, title = {Identification of two Arabidopsis thaliana plasma membrane transporters able to transport glucose 1-phosphate}, series = {Plant \& cell physiology}, volume = {61}, journal = {Plant \& cell physiology}, number = {2}, publisher = {Oxford University Press}, address = {Oxford}, issn = {0032-0781}, doi = {10.1093/pcp/pcz206}, pages = {381 -- 392}, year = {2019}, abstract = {Primary carbohydrate metabolism in plants includes several sugar and sugar-derivative transport processes. Over recent years, evidences have shown that in starch-related transport processes, in addition to glucose 6-phosphate, maltose, glucose and triose-phosphates, glucose 1-phosphate also plays a role and thereby increases the possible fluxes of sugar metabolites in planta. In this study, we report the characterization of two highly similar transporters, At1g34020 and At4g09810, in Arabidopsis thaliana, which allow the import of glucose 1-phosphate through the plasma membrane. Both transporters were expressed in yeast and were biochemically analyzed to reveal an antiport of glucose 1-phosphate/phosphate. Furthermore, we showed that the apoplast of Arabidopsis leaves contained glucose 1-phosphate and that the corresponding mutant of these transporters had higher glucose 1-phosphate amounts in the apoplast and alterations in starch and starch-related metabolism.}, language = {en} } @article{KrasuskaCiackaOrzechowskietal.2016, author = {Krasuska, Urszula and Ciacka, Katarzyna and Orzechowski, Slawomir and Fettke, J{\"o}rg and Bogatek, Renata and Gniazdowska, Agnieszka}, title = {Modification of the endogenous NO level influences apple embryos dormancy by alterations of nitrated and biotinylated protein patterns}, series = {Planta}, volume = {244}, journal = {Planta}, publisher = {Springer}, address = {New York}, issn = {0032-0935}, doi = {10.1007/s00425-016-2553-z}, pages = {877 -- 891}, year = {2016}, abstract = {NO donors and Arg remove dormancy of apple embryos and stimulate germination. Compounds lowering NO level (cPTIO, L -NAME, CAN) strengthen dormancy. Embryo transition from dormancy state to germination is linked to increased nitric oxide synthase (NOS)-like activity. Germination of embryos is associated with declined level of biotin containing proteins and nitrated proteins in soluble protein fraction of root axis. Pattern of nitrated proteins suggest that storage proteins are putative targets of nitration. Nitric oxide (NO) acts as a key regulatory factor in removal of seed dormancy and is a signal necessary for seed transition from dormant state into germination. Modulation of NO concentration in apple (Malus domestica Borkh.) embryos by NO fumigation, treatment with NO donor (S-nitroso-N-acetyl-d,l-penicillamine, SNAP), application of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), N (omega)-nitro-l-arginine methyl ester (l-NAME), canavanine (CAN) or arginine (Arg) allowed us to investigate the NO impact on seed dormancy status. Arg analogs and NO scavenger strengthened embryo dormancy by lowering reactive nitrogen species level in embryonic axes. This effect was accompanied by strong inhibition of NOS-like activity, without significant influence on tissue NO2 (-) concentration. Germination sensu stricto of apple embryos initiated by dormancy breakage via short term NO treatment or Arg supplementation were linked to a reduced level of biotinylated proteins in root axis. Decrease of total soluble nitrated proteins was observed at the termination of germination sensu stricto. Also modulation of NO tissue status leads to modification in nitrated protein pattern. Among protein bands that correspond to molecular mass of approximately 95 kDa, storage proteins (legumin A-like and seed biotin-containing protein) were identified, and can be considered as good markers for seed dormancy status. Moreover, pattern of nitrated proteins suggest that biotin containing proteins are also targets of nitration.}, language = {en} } @article{GietlerNykielOrzechowskietal.2017, author = {Gietler, Marta and Nykiel, Malgorzata and Orzechowski, Slawomir and Fettke, J{\"o}rg and Zagdanska, Barbara}, title = {Protein carbonylation linked to wheat seedling tolerance to water deficiency}, series = {Environmental and experimental botany}, volume = {137}, journal = {Environmental and experimental botany}, publisher = {Elsevier}, address = {Oxford}, issn = {0098-8472}, doi = {10.1016/j.envexpbot.2017.02.004}, pages = {84 -- 95}, year = {2017}, abstract = {The appearance of the first leaf from the coleoptile in wheat seedlings (Triticum aestivum L.) coincides with the development of seedling susceptibility to water deficiency on the fifth day following imbibition. In dehydrated wheat seedlings, an increase in the protein carbonyl group has been observed. The coincidence of higher protein carbonylation levels with development of dehydration intolerance drew our attention. To gain more insight into the molecular basis of wheat drought tolerance, the seedling profiles of carbonylated proteins were analysed and compared. Two-dimensional gel electrophoresis (2D-PAGE) and mass spectrometry (MALDI-TOF and LC-MS/MS) were used to indicate and identify differential carbonylated proteins. Among the protein spots with at least a two-fold change in protein abundance in dehydrated seedlings in relation to control (well-watered) plants during the tolerant phase of growth, 19 carbonylated proteins increased and 18 carbonylated proteins decreased in abundance. Among 26 differentially expressed carbonylated proteins in sensitive seedlings, the abundance of 10 protein spots increased while that of 16 proteins decreased upon dehydration. We have demonstrated a link between protein carbonylation and seedling sensitivity to dehydration. The analysis of carbonylated protein profiles clearly showed that proteins with a potential role in the maintenance of dehydration tolerance in wheat seedlings are mainly linked to energy production, anti-fungal and/or insecticidal activity, or to the regulation of both protein synthesis and degradation.}, language = {en} } @article{GietlerNykielOrzechowskietal.2016, author = {Gietler, Marta and Nykiel, Malgorzata and Orzechowski, Slawomir and Zagdanska, Barbara and Fettke, J{\"o}rg}, title = {Proteomic analysis of S-nitrosylated and S-glutathionylated proteins in wheat seedlings with different dehydration tolerances}, series = {Plant physiology and biochemistry : an official journal of the Federation of European Societies of Plant Physiology}, volume = {108}, journal = {Plant physiology and biochemistry : an official journal of the Federation of European Societies of Plant Physiology}, publisher = {Elsevier}, address = {Paris}, issn = {0981-9428}, doi = {10.1016/j.plaphy.2016.08.017}, pages = {507 -- 518}, year = {2016}, abstract = {A loss of dehydration tolerance in wheat seedlings on the fifth day following imbibition is associated with a disturbance in cellular redox homeostasis, as documented by a shift of the reduced/oxidized glutathione ratio to a more oxidized state and a significant increase in the ratio of protein thiols to the total thiol group content. Therefore, the identification and characterization of redox-sensitive proteins are important steps toward understanding the molecular mechanisms of the loss of dehydration tolerance. In the present study, proteins that were differentially expressed between fully turgid (control), dehydrated tolerant (four-day-old) and dehydrated sensitive (six-day-old) wheat seedlings were analysed. Protein spots having at least a significant (p < 0.05) two-fold change in protein abundance were selected by Delta2D as differentially expressed, identified by MALDI-TOF and LC-MS/MS, and classified according to their function. The observed changes in the proteomic patterns of the differentially S-nitrosylated and S-glutathionylated proteins were highly specific in dehydration-tolerant and-sensitive wheat seedlings. The metabolic function of these proteins indicates that dehydration tolerance is mainly related to nucleic acids, protein metabolism, and energy metabolism. It has been proven that leaf-specific thionins BTH6 and DB4, chloroplastic 50S ribosomal protein L16, phospholipase A1-II delta, and chloroplastic thioredoxin M2 are both S-nitrosylated and S-glutathionylated upon water deficiency. Our results revealed the existence of interplay between S-nitrosylation and S-glutathionylation, two redox-regulated protein posttranslational modifications that could enhance plant defence mechanisms and/or facilitate the acclimation of plants to unfavourable environmental conditions. (C) 2016 Elsevier Masson SAS. All rights reserved.}, language = {en} } @article{OrawetzMalinovaOrzechowskietal.2016, author = {Orawetz, Tom and Malinova, Irina and Orzechowski, Slawomir and Fettke, J{\"o}rg}, title = {Reduction of the plastidial phosphorylase in potato (Solanum tuberosum L.) reveals impact on storage starch structure during growth at low temperature}, series = {Plant physiology and biochemistry : an official journal of the Federation of European Societies of Plant Physiology}, volume = {100}, journal = {Plant physiology and biochemistry : an official journal of the Federation of European Societies of Plant Physiology}, publisher = {Elsevier}, address = {Paris}, issn = {0981-9428}, doi = {10.1016/j.plaphy.2016.01.013}, pages = {141 -- 149}, year = {2016}, abstract = {Tubers of potato (Solanum tuberosum L.), one of the most important crops, are a prominent example for an efficient production of storage starch. Nevertheless, the synthesis of this storage starch is not completely understood. The plastidial phosphorylase (Phol; EC 2.4.11) catalyzes the reversible transfer of glucosyl residues from glucose-1-phosphate to the non-reducing end of alpha-glucans with the release of orthophosphate. Thus, the enzyme is in principle able to act during starch synthesis. However, so far under normal growth conditions no alterations in tuber starch metabolism were observed. Based on analyses of other species and also from in vitro experiments with potato tuber slices it was supposed, that Phol has a stronger impact on starch metabolism, when plants grow under low temperature conditions. Therefore, we analyzed the starch content, granule size, as well as the internal structure of starch granules isolated from potato plants grown under low temperatures. Besides wild type, transgenic potato plants with a strong reduction in the Phol activity were analyzed. No significant alterations in starch content and granule size were detected. In contrast, when plants were cultivated at low temperatures the chain length distributions of the starch granules were altered. Thus, the granules contained more short glucan chains. That was not observed in the transgenic plants, revealing that Pho1 in wild type is involved in the formation of the short glucan chains, at least at low temperatures. (C) 2016 Elsevier Masson SAS. All rights reserved.}, language = {en} } @article{BrustOrzechowskiFettke2020, author = {Brust, Henrike and Orzechowski, Slawomir and Fettke, J{\"o}rg}, title = {Starch and Glycogen Analyses}, series = {Biomolecules}, volume = {10}, journal = {Biomolecules}, number = {7}, publisher = {MDPI}, address = {Basel}, issn = {2218-273X}, doi = {10.3390/biom10071020}, pages = {24}, year = {2020}, abstract = {For complex carbohydrates, such as glycogen and starch, various analytical methods and techniques exist allowing the detailed characterization of these storage carbohydrates. In this article, we give a brief overview of the most frequently used methods, techniques, and results. Furthermore, we give insights in the isolation, purification, and fragmentation of both starch and glycogen. An overview of the different structural levels of the glucans is given and the corresponding analytical techniques are discussed. Moreover, future perspectives of the analytical needs and the challenges of the currently developing scientific questions are included}, language = {en} }