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A cDNA-AFLP based strategy to identify transcripts associated with avirulence in Phytophthora infestans
Jun Guo1, Rays H Y Jiang, Lars G Kamphuis
1Plant Sciences Group, Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, NL-6709 PD Wageningen, The Netherlands.
Fungal Genetics and Biology : FG & B
|February 4, 2006
Summary
This study used cDNA-AFLP and bulked segregant analysis to identify avirulence (Avr) gene transcripts in Phytophthora infestans. The efficient strategy identified potential Avr genes linked to late blight resistance.
Area of Science:
- Plant Pathology
- Genomics
- Molecular Biology
Background:
- cDNA-AFLP is a standard method for transcriptome profiling.
- Phytophthora infestans causes late blight, a significant plant disease.
Purpose of the Study:
- To identify avirulence (Avr) gene transcripts in Phytophthora infestans using a segregating F1 population.
- To develop an efficient strategy for discovering Avr-associated transcriptome markers.
Main Methods:
- Utilized cDNA-AFLP for expression profiling.
- Employed bulked segregant analysis (BSA) on germinated cyst cDNA from F1 progeny with defined avirulence phenotypes.
- Performed homology searches and database mining on transcript-derived fragment (TDF) sequences.
- Confirmed co-segregation of TDFs with AVR phenotypes using RT-PCR.
Main Results:
- Screened over 30,000 transcript-derived fragments (TDFs).
- Identified 99 Avr-associated TDFs and TDFs with opposite patterns.
- Found four TDFs that co-segregated 100% with specific AVR phenotypes.
- Identified two likely candidates for Avr4 and two associated with the Avr3b-Avr10-Avr11 locus.
Conclusions:
- The combined cDNA-AFLP/BSA strategy is effective for identifying Avr-associated transcriptome markers.
- This approach can complement positional cloning efforts for discovering Avr genes.
- The study successfully identified potential Avr gene candidates in Phytophthora infestans.
