A proteomic approach to studying plant response to crenate broomrape (Orobanche crenata) in pea (Pisum sativum)

• Published in: Phytochemistry (Oxford) 2004-06, Vol.65 (12), p.1817-1828
• Main Authors: Ángeles Castillejo, M, Amiour, Nardjis, Dumas-Gaudot, Eliane, Rubiales, Diego, Jorrı́n, Jesús V
• Format: Article
• Language: English
• Subjects: Broomrape; Databases, Protein; Electrophoresis, Gel, Two-Dimensional - methods; Electrophoresis, Polyacrylamide Gel; Gene Expression; Genotype; Life Sciences; MALDI-TOF; mass spectrometry; matrix-assisted laser desorption ionization time-of-flight mass spectrometry; Orobanche - physiology; Orobanche crenata; Parasitic plants; Pea; Pea proteomics; peas; Peptide Mapping; Peptide mass fingerprinting; Phytopathology and phytopharmacy; Pisum sativum; Pisum sativum - genetics; Pisum sativum - microbiology; plant proteins; Plant Proteins - analysis; plant response; Plant Roots - metabolism; proteome; Proteomics; qualitative analysis; quantitative analysis; Root proteomics; roots; seed germination; Silver Staining; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Two-dimensional gel electrophoresis; Vegetal Biology
• ISSN: 0031-9422; 1873-3700
• DOI: 10.1016/j.phytochem.2004.03.029

By using a proteomic approach, including two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry, a number of proteins differently expressed in broomrape-infected susceptible and partially resistant pea roots have been identified. Crenate broomrape (Orobanche crenata) is a parasitic plant that threatens legume production in Mediterranean areas. Pea (Pisum sativum) is severely affected, and only moderate levels of genetic resistance have so far been identified. In the present work we selected the most resistant accession available (Ps 624) and compared it with a susceptible (Messire) cultivar. Experiments were performed by using pot and Petri dish bioassays, showing little differences in the percentage of broomrape seed germination induced by both genotypes, but a significant hamper in the number of successfully installed tubercles and their developmental stage in the Ps 624 compared to Messire. The protein profile of healthy and infected P. sativum root tissue were analysed by two-dimensional electrophoresis. Approximately 500 individual protein spots could be detected on silver stained gels. At least 22 different protein spots differentiated control, non-infected, Messire and Ps 624 accessions. Some of them were identified by MALDI-TOF mass spectrometry and database searching as cysteine proteinase, β-1,3-glucanase, endochitinase, profucosidase, and ABA-responsive protein. Both qualitative and quantitative differences have been found among infected and non-infected root extracts. Thus, in the infected susceptible Messire genotype 34 spots were decreased, one increased and three newly detected, while in Ps 624, 15 spots were increased, three decreased and one newly detected. In response to the inoculation, proteins that correspond to enzymes of the carbohydrate metabolism (fructokinase, fructose-bisphosphate aldolase), nitrogen metabolism (ferredoxin-NADP reductase) and mitochondrial electronic chain transport (alternative oxidase 2) decreased in the susceptible check, while proteins that correspond to enzymes of the nitrogen assimilation pathway (glutamine synthetase) or typical pathogen defence, PR proteins, including β-1,3-glucanase and peroxidases, increased in Ps 624. Results are discussed in terms of changes in the carbohydrate and nitrogen metabolism an induction of defence proteins in response to broomrape parasitism.