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Functional studies of plant hexokinases and development of genetic methods in the moss Physcomitrella patens

Ulfstedt, Mikael (2014). Functional studies of plant hexokinases and development of genetic methods in the moss Physcomitrella patens. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2014:29
ISBN 978-91-576-8006-8
eISBN 978-91-576-8007-5
[Doctoral thesis]

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Abstract

In the field of biology, model systems are frequently used for practical reasons. Model organisms possess several features that make them easy to work with in a laboratory setting. In addition they usually have a host of established genetic tools that have been developed by the research community. This thesis explores the potential of adapting methods and tools used in the yeast Saccharomyces cerevisiae to the plant model system Physcomitrella patens. We have studied the fate and integrity of plasmids transformed into Physcomitrella with the purpose of developing a working shuttle plasmids system in moss. Other methods from yeast genetics such as the use of auxotrophic strains would also be useful if adapted to the Physcomitrella model system. Experiments using Physcomitrella patens showed that a histidine auxotrophic moss strain can be complemented with the wild type gene on a plasmid and that the plasmid can be rescued back into E. coli. This indicates that shuttle plasmids, auxotrophic marker genes such as PpHIS3, cloning by complementation and perhaps even dosage suppressor screens could be used in Physcomitrella patens. Furthermore this thesis investigates the hexose phosphorylating enzyme hexokinase in Physcomitrella patens as well as in the charophyte alga Klebsormidium nitens. The characterization and localization studies performed on the eleven Physcomitrella hexokinases led to the discovery of two new types of plant hexokinases, the type C and type D hexokinases. In addition to these two new types, Physcomitrella contained several of the previously described type A and type B hexokinases. In addition to its enzymatic function, hexokinases in both plants and fungi have been implicated in glucose sensing and signaling. This thesis examines if hexokinase 1 of the microalga Klebsormidium nitens may play a similar role in glucose sensing as observed in higher plants. With this purpose in mind transgenic lines of the Arabidopsis thaliana glucose insensitive mutant gin2-1 expressing the Klebsormidium nitens hexokinase 1 were created. It was found that the K. nitens hexokinase can complement the glucose signaling defect in the gin2-1 mutant.

Authors/Creators:Ulfstedt, Mikael
Title:Functional studies of plant hexokinases and development of genetic methods in the moss Physcomitrella patens
Series/Journal:Acta Universitatis agriculturae Sueciae (1652-6880)
Year of publishing :29 April 2014
Volume:2014:29
Number of Pages:69
Papers/manuscripts:
NumberReferences
I.Murén, E., Nilsson, A., Ulfstedt, M., Johansson, M. & Ronne, H. (2009). Rescue and characterization of episomally replicating DNA from Physcomitrella. Proceedings of the National Academy of Sciences of the United States of America 106(46):19444-9
II.Nilsson, A., Olsson, T., Ulfstedt, M., Thelander, M. & Ronne, H. (2011). Two novel types of plant hexokinases in the moss Physcomitrella patens. BMC Plant Biology 11:32
III.Ulfstedt, M., Hu, G-Z., Eklund, M. & Ronne, H. A charophyte hexokinase can complement both enzymatic and glucose signalling functions in yeast and Arabidopsis thaliana hexokinase mutants. (manuscript)
IV.Ulfstedt, M., Hu, G-Z. & Ronne, H. Development and testing of an auxotrophic selection marker for use in the moss Physcomitrella. (manuscript)
Place of Publication:Uppsala
Publisher:Department of Microbiology, Swedish University of Agricultural Sciences
ISBN for printed version:978-91-576-8006-8
ISBN for electronic version:978-91-576-8007-5
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:F Plant production > F60 Plant physiology and biochemistry
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Genetics (medical genetics to be 30107 and agricultural genetics to be 40402)
(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Biological Systematics
(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Evolutionary Biology
(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 404 Agricultural Biotechnology > Plant Biotechnology
Obsolete subject words > NATURAL SCIENCES > Biology > Cell and molecular biology > Molecular biology
Agrovoc terms:bryophyta, hexokinase, algae, carbohydrate metabolism, auxotrophy, plant models
Keywords:Sugar signaling, hexokinase, Physcomitrella patens, shuttle vector, plant model system, Klebsormidium nitens, Auxotroph, HIS knockout
URN:NBN:urn:nbn:se:slu:epsilon-e-1871
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-1871
ID Code:11143
Faculty:NJ - Fakulteten för naturresurser och jordbruksvetenskap
Department:(NL, NJ) > Dept. of Microbiology (until 161231)
Deposited By: Mikael Ulfstedt
Deposited On:24 Apr 2014 12:56
Metadata Last Modified:02 Dec 2014 11:06

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