Molecular analyses of the interaction between Arabidopsis thaliana and the endophytic fungus piriformospora indica [Elektronische Ressource] / by Bationa Shahollari

Plantphys

Sujets

• Biologie

Informations

• Publié par: friedrich-schiller-universitat_jena
• Publié le: 01 janvier 2006
• Nombre de lectures: 165
• Langue: English
• Poids de l'ouvrage: 18 Mo

Extrait

Molecular analyses of the interaction between
Arabidopsis thaliana and the endophytic fungus
Piriformospora indica





Thesis


in order to receive the academic degree doctor rerum naturalium (Dr.rer.nat.)



submitted to the

Rat der Biologisch-Pharmazeutischen Fakultät

Friedrich Schiller University Jena



by
Bationa Shahollari

Jena, Mai 2006








































Referees:

1. Prof. Dr. Jutta Ludwig-Müller
2. PD. Dr. Thomas Pfannschmidt
3. Prof. Dr. Ralf Oelmüller


Date of the Rigorosum: 14.06.06
Date of the public defence: 26.06.06 Table of Contents



Table of Contents………………………………………………………1

Manuscript Overview…………………………………………………..2

1. Introduction…………………………………………………………..9

2. Manuscripts

2.1 Manuscript I…………………………………………………….22

2.2 Manuscript II……………………………………………………36

2.3 Manuscript III…………………………………………………...44

2.4 Manuscript IV…………………………………………………...59

2.5 Manuscript V……………………………………………………96

2.6 Manuscript VI………………………………………………….112

2.7 Manuscript VII…………………………………………………127

3. Discussion ………………………………………………………….135

4. Summary……………………………………………………………147

5. Zusammenfassung………………………………………………….149

6. Literature Cited……………………………………………………..151

7. Appendix

7.1 List of publications……………………………………………..162

7.2 List of presentations……………………………………………163

7.3 Acknowledgment………………………………………………165

7.4 Curriculum Vitae……………………………………………....166

7.5 Ehrenwörtliche Erklärung zur Anfertigung der Dissertation….168

1Manuscript Overview


Manuscript I



Association of Piriformospora indica with Arabidopsis thaliana roots
represents a novel system to study beneficial plant–microbe interactions
and involves early plant protein modifications in the endoplasmic reticulum
and at the plasma membrane

Tatjana Peškan-Berghöfer, Bationa Shahollari, Pham Huong Giong, Solveig Hehl, Christine
Markert, Verena Blanke, Gerhard Kost, Ajit Varma and Ralf Oelmüller

Physiologia Plantarum (2004) 122:4, 465-477





This manuscript describes, for the first time, the newly established system to study the
beneficial interaction between P. indica, an endophytic fungus of the Sebacinaceae
family, and A. thaliana.

Dr. Tatjana Peškan-Berghöfer and Prof. Ralf Oelüller planned the experiments. Most of
the experiments were performed by Dr. Tatjana Peškan-Berghöfer. Fluorescence
measurements were performed by Verena Blanke in the Department of Professor
Gerhard Kost. Christine Markert helped us with the mass spectrometry. Pham Huong
Giong introduced us into the techniques required for the experiments on soil.
I repeated all experiments performed by Dr. Tatjana Peškan-Berghöfer. In addition, I
extracted the spots from the two-dimensional gels and identified the proteins by mass
spectrometry. The quantification of the data was performed by myself. I also performed
the cultivation of plants and inoculation with fungus, and all experiments on soil.





2Manuscript Overview


Manuscript II



Receptor kinases with leucine-rich repeats are enriched in Triton X-100
insoluble plasma membrane microdomains from plants

Bationa Shahollari, Tatjana Peskan-Berghöfer and Ralf Oelmüller

Physiologia Plantarum (2004) 122: 397-403





This manuscript describes the protein pattern of Triton X-100 insoluble plasma
membrane microdomains. The protein composition is enriched in signalling
components: receptor kinases with leucine-rich repeats, 10 other kinases, the ß subunit
of heterotrimeric G-proteins and five small GTP-binding proteins. Thus, specific
signalling components are highly enriched in plant plasma membrane microdomains
while others are excluded.

I isolated plasma membrane microdomains from A. thaliana and mustard cotyledons,
separated the protein in one and two dimensional gels and determined the protein
composition of the plasma membrane microdomains by mass spectrometry. The
experiments in this manuscript were designed by me, Dr. Tatjana Peskan-Berghöfer and
Prof. Ralf Oelmüller and performed by myself.












3Manuscript Overview


Manuscript III



Expression of a receptor kinase in Arabidopsis roots is stimulated by the
basidiomycete Piriformospora indica and the protein accumulates in Triton
X-100 insoluble plasma membrane microdomains

Bationa Shahollari, Ajit Varma, Ralf Oelmuller.

J Plant Physiol. (2005) 162(8):945-58.





Manuscript III describes the accumulation of the mRNA for a receptor kinase in A.
thaliana roots, which were co-cultivated with P. indica. This represents one of the
earliest events of a plant root in response to a fungus. During the recognition period of
both organisms, the mRNA for a receptor kinase with leucine-rich repeats (LRR1) is
transiently upregulated. The kinase is located in Triton X-100-insoluble plasma
membrane microdomains. P. indica promotes growth of A. thaliana, and this promotion
was accompanied by a massive transfer of phosphate from the media to the aerial parts
of the seedlings.

All experiments, with the exception of the phosphate uptake experiment, were designed
by me and Prof. Oelmüller and performed by myself. The phosphate uptake studies
were performed in the laboratory of Professor Ajit Varma.









4Manuscript Overview


Manuscript IV



A leucine rich repeat protein is required for growth promotion and
enhanced seed production mediated by the endophytic fungus
Piriformospora indica in Arabidopsis thaliana

Bationa Shahollari, Ajit Varma, Ralf Oelmüller

(revised version, submitted to The Plant Journal).





Manuscript IV describes A. thaliana mutants which fail to respond to the P. indica. An
ethylmethane sulfonate (EMS) and the corresponding insertion line are impaired in a
leucine-rich repeat protein (LRR2) and are blind to P. indica.
Inactivation of the A. thaliana single-copy gene DMI1, which codes for an ion carrier
required for mycorrihiza formation in Legumes, does not affect the beneficial
interaction between the two symbiotic partners.

All experiments were designed by me and Prof. Oelmüller and performed by me . The
original EMS mutant was isolated by Prof. Ajit Varma. Also the Fluorescence
measurements are done by Prof. Ajit Varma.








5Manuscript Overview


Manuscript V



MATH domain proteins represent a novel protein family in Arabidopsis
thaliana, and at least one member is modified in roots during the course of
a plant–microbe interaction

Ralf Oelmüller, Tatjana Peškan-Berghöfer, Bationa Shahollari, Artan Trebicka, Irena Sherameti,
Ajit Varma

Physiologia Plantarum (2005) 124:152-166



A MATH [meprin and TRAF (tumour necrosis factor receptor-associated factor)
homology] domain-containing protein in the plasma membrane of A. thaliana roots
becomes transiently modified in response to P. indica. Since nothing is known about
MATH proteins in plants, we analysed the fifty nine genes present in the A. thaliana
genome.

The experiment described here and the design of the manuscript was planned by me and
Prof. Oelmüller. I performed the isolation of microsomes, the separation of proteins by
two-dimensional gel electrophoresis and analysed the proteins by mass spectrometry.
Dr. Artan Trebicka and Dr. Irena Sherameti isolated the chloroplasts and plastid RNA. I
generated the phylogenetic tree for the MATH proteins, with some help from Dr. Artan
Trebicka.

The modification of the MATH protein in A. thaliana roots in response to P. indica was
originally discovered by Dr. Peskan-Berghöfer. I continued with the analysis of the
MATH proteins in P. indica insensitive mutants, by running two-dimensional gels from
plasma membrane microsomes from roots. I analysed the proteins by mass
spectrometry. I also participated in the analysis of the gene family in A. thaliana.

6Manuscript Overview


Manuscript VI



Molecular analyses of the interaction between Arabidopsis roots and the
growth-promoting fun