Proteomic analysis of Masson pine with high resistance to pine wood nematodes

Abstract

Pine wilt disease is a dangerous pine disease globally. We used Masson pine (Pinus massoniana) clones, selected through traditional breeding and testing for 20 years, to study the molecular mechanism of their high resistance to pine wood nematodes (PWN,Bursaphelenchus xylophilus). Nine strains of seedlings of genetically stable Masson pine screened from different families with high resistance to PWN were used. The same number of sensitive clones were used as susceptible controls. Total proteins were extracted for tandem mass tag (TMT) quantitative proteomic analysis. The key proteins were verified by parallel reaction monitoring (PRM). A threshold of upregulation greater than 1.3-fold or downregulation greater than 0.3-fold was considered significant in highly resistant strains versus sensitive strains. A total of 3491 proteins were identified from the seedling tissues, among which 2783 proteins contained quantitative information. A total of 42 proteins were upregulated and 96 proteins were downregulated in the resistant strains. Functional enrichment analysis found significant differences in the proteins with pectin esterase activity or peroxidase activity. The proteins participating in salicylic acid metabolism, antioxidant stress reaction, polysaccharide degradation, glucose acid ester sheath lipid biosynthesis, and the sugar glycosaminoglycan degradation pathway were also changed significantly. The PRM results showed that pectin acetyl esterase, carbonic anhydrase, peroxidase, and chitinase were significantly downregulated, while aspartic protease was significantly upregulated, which was consistent with the proteomic data. These results suggest that Masson pine can degrade nematode-related proteins by increasing protease to inhibit their infestation, and can enhance the resistance of Masson pine to PWN by downregulating carbon metabolism to limit the carbon available to PWN or for involvement in cell wall components or tissue softening. Most of the downregulated proteins are supposed to act as an alternative mechanism for latter enhancement after pathogen attacks. The highly resistant Masson pine, very likely, harbors multiple pathways, both passive and active, to defend against PWN infestation.

Comparative Proteome Profiling of Extracellular Vesicles from Three Growth Phases of Haematococcus pluvialis under High Light and Sodium Acetate Stresses

“…To acquire insight into the proteomic profile of HpEVs, the identified proteins were screened in several databases and analyzed using bioinformatics methods, as described in previous reports [57][58][59][60]. Briefly, hierarchical clustering analysis was carried out using the R Package pheatmap (v.2.0.3 https://cran.r-project.org/web/packages/cluster/, accessed on 3 March 2020), and the heat map was produced by a dendrogram depicting the extent of similarity in the protein expression among samples.…”

Proteomics research in forest trees: A 2012-2022 update

“…These works have revealed the importance of peptidases, hydrolases, and antioxidant proteins in overcoming the defense mechanisms deployed by different hosts ( Silva et al., 2021 ; Cardoso et al., 2022 ). A recent study used Masson pine ( P. massoniana ) clones selected through traditional breeding over 20 years and screened for different resistance to pine wilt ( Gao et al., 2022 ). Comparative Tandem Mass Tagged (TMT) based quantitative proteomic analysis combined with parallel reaction monitoring (PRM) identified proteins related to SA metabolism, the antioxidant system, polysaccharide degradation, and lipid biosynthesis to change significantly during the infestation process.…”

Molecular Defense Response of Pine Trees (Pinus spp.) to the Parasitic Nematode Bursaphelenchus xylophilus

“…This is supported by the differential expression of distinct RLK/RLP and NLR receptor genes after PWN inoculation when comparing resistant and susceptible P. pinaster plants, as well as the downregulation of NLRs and resistance genes in susceptible P. massoniana [ 38 , 41 ]. Furthermore, two NLR receptor proteins were constitutively more expressed in resistant P. massoniana plants [ 43 ]. The activation of the ETI usually takes place in plants adapted to the pathogen or pest, implying a coevolution of the two organisms and often an arms race [ 28 ].…”