Abstract
Clubroot disease is one of the major diseases affecting Brassica crops, especially Chinese cabbage (Brassica rapa L. ssp. pekinensis), which is known to be highly susceptible to the disease. In this study, the obligate biotrophic protist Plasmodiophora brassicae Woronin was used to infect the roots of Chinese cabbage seedlings. The disease symptoms were noticeable at 28 and 35 days after inoculation (DAI) in the susceptible (CM) line. Using isobaric tags for relative and absolute quantitation (iTRAQ) analysis, a total of 5,003 proteins of differential abundance were identified in the resistant/susceptible lines, which could be quantitated by dipeptide or polypeptide segments. Gene ontology (GO) analysis indicated that the differentially expressed proteins (DEPs) between the susceptible (CM) and resistant (CCR) lines were associated with the glutathione transferase activity pathway, which could catalyze the combination of glutathione and other electrophilic compounds to protect plants from disease. In addition, the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the DEPs may be significantly enriched cytokinin signaling or arginine biosynthesis pathways, both of which are responses to stimuli and are plant defense reactions. The cytokinins may facilitate cell division in the shoot, resulting in the hypertrophy and formation of galls and the presentation of typical clubroot symptoms. In this study, the proteomic results provide a new perspective for creating germplasm resistance to P. brassicae, as well as a genetic basis for breeding to improve Chinese cabbage.
Highlights
Www.nature.com/scientificreports of galls, a typical clubroot symptom[16,17]
In recent years, using transcriptome analysis of broccoli, wild cabbage[38], and A. thaliana[39], the defense mechanisms of B. rapa to P. brassicae have been described, but a proteomic analysis of the response of B. rapa to P. brassicae has only been reported a limited number of times
Proteomic analysis provides a large amount of information regarding the individual proteins that are involved in specific biological responses
Summary
Www.nature.com/scientificreports of galls, a typical clubroot symptom[16,17]. The infected roots can produce resting spores that can survive for so long that standard strategies, such as antimicrobial compounds and crop rotations, cannot eliminate the pathogen[2,18]. To obtain durable and broad-spectrum resistance to P. brassicae in Chinese cabbage, resistant proteins and genes must be identified and used in the development of resistant varieties. The few existing reports include a shotgun label-free proteomic analysis of clubroot (Plasmodiophora brassicae) resistance conferred by the gene Rcr[1] in Brassica rapa[40], iTRAQ analysis of proteins profiles during the secondary stage of infection of Plasmodiophora brassicae in Chinese cabbage[41], and proteomic analysis of the interaction between Plasmodiophora brassicae and Chinese cabbage at the initial infection stage[42]. Using the A. thaliana-P. brassicae interaction pathosystem, the proteins in roots and stems from infected and non-infected plants have been analyzed[49]. Chinese cabbage lines with both resistance and ssusceptibility to P. brassicae were inoculated, and plants that developed galls underwent iTRAQ-based proteomic analysis. The results of this study may lay the foundation for the development of germplasm innovation, as well as for the development of antibody resistance to P. brassicae in Chinese cabbage
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