Abstract
BACKGROUND: Inoculation of wheat plants with Puccinia triticina (Pt) spores activates a wide range of host responses. Compatible Pt interaction with susceptible Thatcher plants supports all stages of the pathogen life cycle. Incompatible interaction with TcLr9 activates defense responses including oxidative burst and micronecrotic reactions associated with the pathogen's infection structures and leads to complete termination of pathogen development. These two contrasting host-pathogen interactions were a foundation for transcriptome analysis of incompatible wheat-Pt interaction. METHODS: A suppression subtractive hybridization (SSH) library was constructed using cDNA from pathogen-inoculated susceptible Thatcher and resistant TcLr9 isogenic lines. cDNA represented steps of wheat-brown rust interactions: spore germination, haustorium mother cell (HMC) formation and micronecrotic reactions. All ESTs were clustered and validated by similarity search to wheat genome using BLASTn and sim4db tools. qRT-PCR was used to determine transcript levels of selected ESTs after inoculation in both lines. RESULTS AND DISCUSSION: Out of 793 isolated cDNA clones, 183 were classified into 152 contigs. 89 cDNA clones and encoded proteins were functionally annotated and assigned to 5 Gene Ontology categories: catalytic activity 48 clones (54 %), binding 32 clones (36 %), transporter activity 6 clones (7 %), structural molecule activity 2 clones (2 %) and molecular transducer activity 1 clone (1 %). Detailed expression profiles of 8 selected clones were analyzed using the same plant-pathogen system. The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003), receptor with leucine-rich repeat domain (JG968955), putative serine/threonine protein kinase (JG968944), calcium-mediated signaling protein (JG968925) and 14-3-3 protein (JG968969). CONCLUSIONS: The SSH library represents transcripts regulated by pathogen infection during compatible and incompatible interactions of wheat with P. triticina. Annotation of selected clones confirms their putative roles in successive steps of plant-pathogen interactions. The transcripts can be categorized as defense-related due to their involvement in either basal defense or resistance through an R-gene mediated reaction. The possible involvement of selected clones in pathogen recognition and pathogen-induced signaling as well as resistance mechanisms such as cell wall enforcement, oxidative burst and micronecrotic reactions is discussed.
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Claims 14
Claim · model-written summary
The compatible interaction supports all stages of pathogen development in susceptible Thatcher wheat.
- Taxon
- wheat
Triticum aestivumt0 - Material
- Thatchere0
- Interactor
- Pte0pathogen
- Outcome
- all stages of the pathogen life cyclee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“Compatible Pt interaction with susceptible Thatcher plants supports all stages of the pathogen life cycle”
t0“wheat”
Claim · model-written summary
The incompatible TcLr9 interaction activates oxidative burst and micronecrotic reactions and terminates pathogen development.
- Taxon
- wheat
Triticum aestivumt0 - Material
- TcLr9e0
- Interactor
- Puccinia triticina
Puccinia triticinat0pathogen - Outcome
- complete termination of pathogen developmente0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.


Verbatim
e0“Incompatible interaction with TcLr9 activates defense responses including oxidative burst and micronecrotic reactions associated with the pathogen's infection structures and leads to complete termination of pathogen development”
t0“wheat isogenic lines differing in resistance to brown rust Puccinia triticina”
Claim · model-written summary
The SSH library yielded 183 clones classified into 152 contigs from 793 cDNA clones.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- cDNA clonese0
- Value
- 183e0
- Qualifier
- 152 contigse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“Out of 793 isolated cDNA clones, 183 were classified into 152 contigs”
t0“wheat”
Claim · model-written summary
Eighty-nine cDNA clones and encoded proteins were functionally annotated.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- cDNA clones and encoded proteinse0
- Value
- 89e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“89 cDNA clones and encoded proteins were functionally annotated”
t0“wheat”
Claim · model-written summary
Catalytic activity accounted for 48 annotated clones.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- catalytic activitye0
- Value
- 48 clones (54 %)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“catalytic activity 48 clones (54 %)”
t0“wheat”
Claim · model-written summary
Binding accounted for 32 annotated clones.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- bindinge0
- Value
- 32 clones (36 %)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“binding 32 clones (36 %)”
t0“wheat”
Claim · model-written summary
Transporter activity accounted for six annotated clones.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- transporter activitye0
- Value
- 6 clones (7 %)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“transporter activity 6 clones (7 %)”
t0“wheat”
Claim · model-written summary
Structural molecule activity accounted for two annotated clones.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- structural molecule activitye0
- Value
- 2 clones (2 %)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“structural molecule activity 2 clones (2 %)”
t0“wheat”
Claim · model-written summary
Molecular transducer activity accounted for one annotated clone.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- molecular transducer activitye0
- Value
- 1 clone (1 %)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“molecular transducer activity 1 clone (1 %)”
t0“wheat”
Claim · model-written summary
Wall-associated kinase showed strong induction after infection and large differences between resistant and susceptible interactions.
- Taxon
- wheat
Triticum aestivumt0 - Interactor
- Puccinia triticina
Puccinia triticinat0pathogen - Gene
- wall-associated kinasee0
- Outcome
- strongest inductione0
- Direction
- strongest inductione0
- Comparator
- resistant and susceptible interactionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.


Verbatim
e0“The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003)”
t0“wheat isogenic lines differing in resistance to brown rust Puccinia triticina”
Claim · model-written summary
Leucine-rich repeat receptor clones showed strong induction and large interaction differences.
- Taxon
- wheat
Triticum aestivumt0 - Gene
- receptor with leucine-rich repeat domaine0
- Outcome
- strongest inductione0
- Direction
- strongest inductione0
- Comparator
- resistant and susceptible interactionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003), receptor with leucine-rich repeat domain (JG968955)”
t0“wheat”
Claim · model-written summary
Putative serine/threonine protein kinase clones showed strong induction and large interaction differences.
- Taxon
- wheat
Triticum aestivumt0 - Gene
- putative serine/threonine protein kinasee0
- Outcome
- strongest inductione0
- Direction
- strongest inductione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003), receptor with leucine-rich repeat domain (JG968955), putative serine/threonine protein kinase (JG968944)”
t0“wheat”
Claim · model-written summary
Calcium-mediated signaling protein clones showed strong induction and large interaction differences.
- Taxon
- wheat
Triticum aestivumt0 - Gene
- calcium-mediated signaling proteine0
- Outcome
- strongest inductione0
- Direction
- strongest inductione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003), receptor with leucine-rich repeat domain (JG968955), putative serine/threonine protein kinase (JG968944), calcium-mediated signaling protein (JG968925)”
t0“wheat”
Claim · model-written summary
14-3-3 protein clones showed strong induction and large interaction differences.
- Taxon
- wheat
Triticum aestivumt0 - Gene
- 14-3-3 proteine0
- Outcome
- strongest inductione0
- Direction
- strongest inductione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“The strongest induction after pathogen infection and the biggest differences between resistant and susceptible interactions were detected for clones encoding wall-associated kinase (GenBank accession number JG969003), receptor with leucine-rich repeat domain (JG968955), putative serine/threonine protein kinase (JG968944), calcium-mediated signaling protein (JG968925) and 14-3-3 protein (JG968969)”
t0“wheat”