Sequence distribution for molecular function was visualized at the ontology level 2 with a cutoff of 5. Interpro scan was also run and the results were merged with the annotations as described in the Blast2Go tutorial [40]. In order to identify proteins involved in foliar SDS, the protein profiles of xylem sap samples collected from the F.
There were no qualitative or obvious quantitative differences between the crude xylem sap samples of the F. Sizes of the molecular weight marker in kDa are shown at the right side. Infected, xylem sap from soybean plants infected with F. Differences between xylem sap samples of the F.
Five F. All five proteins possessed N-terminal secretory signal peptide sequences Table 1. One of these proteins showed high sequence similarity to the cerato-platanin toxin, which was found to be the most abundant F.
Identified peptides of these five proteins are presented in Table S1. To determine if any of the five proteins Table 1 identified from the xylem sap of the F. Ninety four proteins were identified in the cell-free F.
Forty-four proteins with more than one peptide are presented in Table 2. A lipoprotein was found to be the most abundant protein in the CF proteome. Of the 44 reported proteins, 37 shown to have predicted secretary signal peptides Table 2. These proteins included cerato-platanin and FvTox1, with cerato-platanin toxin being the second most abundant protein Table 2.
All five proteins identified from the xylem sap of the F. The CF proteome contained proteins that showed high identity to cutinase, catalase, sporozite P67 surface antigen, eukaryotic type carbonic anhydrase, alpha amylase inhibitor, and laminin. Glycosyl hydrolases belonging to four different families were also identified Table 2. Over soybean proteins were identified from the xylem saps collected from F.
The high number of protein detection could be due to the slightly high percentage of false discovery rate FDR. The FDR percentage ranged from 2—3. Details of the Glycine max accession numbers, GO annotations, as well as the biological replications from which these proteins were identified are provided Table S3.
However, at least two peptides were detected only for soybean proteins Table 3. Fifty of these proteins possessed predicted N-terminal secretory signal peptides. The most abundant proteins commonly found in the xylem saps of both F. Gamma-glutamyl hydrolase, 50S ribosomal proteins, trypsin and protease inhibitor protein, and peroxidases were some of the soybean proteins commonly found in sap samples of both F.
This study identified six soybean proteins that were only detected in xylem sap of F. These proteins were detected in at least two out of the five biological replications of xylem sap samples collected from either F. Only one infection induced proteins contained a predicted N-terminal secretory signal. Several stress related proteins were detected in the xylem sap of the infected plants but not in the sap of the uninfected plants.
Pathogenicity related family 5 PR5 protein was the most abundant of the six proteins identified from sap of the infected plants. Serine carboxypeptidase, disulfide oxidoreductase, and citrate synthase were among the proteins only detected in the sap of the healthy, F. To determine the type metabolic activities in the soybean xylem saps, the sequences of the putative soybean xylem sap proteins detected in xylem sap samples of both F. Most of these proteins were involved in more than one biological process and had more than one molecular function Tables S5.
Based on the sequence distribution with a cutoff of 5, majority of the common xylem sap proteins showed binding and catalytic activities Figure 3A.
Among the proteins that were assigned with enzyme commissions EC , 44 metabolic pathways including phenylalanine metabolism, glycolysis, methane metabolism, carbon fixation, and phenylpropanoid biosynthesis were observed Figure 3B , Table S6. The soybean xylem sap in general shown to be active in both carbon and amino acid metabolisms. A Percentage of proteins in different functional categories at ontology level 2, with a cutoff of 5.
Only the prominent pathways with a sequence cutoff of 3 are reported here. Sudden death syndrome is considered to be caused by one or more toxins released by F. We have shown that FvTox1 is a major virulence factor that is involved in foliar SDS in soybean [13] , [16] , [17].
The foliar SDS was not completely absent in fvtox1 infected soybean plants [17] suggesting that additional toxins may be involved in foliar SDS development; and it is conceivable that such toxins may be present in xylem sap of infected soybean plants. Therefore, the main objective of this investigation was to identify those possible F.
In addition, we investigated if there are changes in proteomes of xylem sap of soybean plants following F. In this investigation we identified five F. This observation strengthens the previous speculations that the pathogen uses the vascular system to transport host-selective toxins to the above ground plant parts to cause foliar SDS.
We, however, have to establish the functions of these putative proteinacious toxins by generating knockout mutants to establish their roles in generating foliar SDS in soybean. We failed to detect FvTox1 from the xylem sap suggesting that our system is not sensitive enough to detect all proteins or peptides of the xylem sap.
FvTox1 was localized to chloroplasts H. Brar and M. Bhattacharyya, unpublished and shown to be involved in SDS development in foliar tissues [16].
This could also be the reason for not detecting the five F. All five xylem sap F. Therefore, if there were any small nonribosomal phytotoxic peptides in the xylem sap, then those were not identified in our study. Cerato-platanin is a phytotoxin produced by the ascomycete fungus Ceratocystis fimbriata f. This Cerato-platanin toxin is considered to be a pathogen associated molecular pattern PAMP because it can induce defense related responses in the host plant [43].
Proteins belonging to the cerato-platanin protein family can enhance plant defenses by inducing defense related genes, phytoalexins synthesis, and initiating cell death [43] , [44] , [45] , [46]. We identified a cerato-platanin-like protein from both xylem sap and F. Investigation of mutants for the gene encoding this cerato-platanin-like protein will assist us in determining the function of this protein in F.
This xylem sap study, in addition to identifying candidate pathogenicity F. Beta-1,3-glucanase is a pathogenic related family 2 PR-2 type protein known to be secreted upon pathogen attack and shown to be present in the xylem sap of pathogen infected plants [26] , [47] , [48]. Cytosolic isoforms of NADP malic enzyme are known to be involved in plant defense responses [51] , and an NADP-dependent malic enzyme was detected in the xylem sap of F. The expression of pathogenicity-related Bet v I family proteins PR10 are known to be induced during wounding, abiotic stress, or pathogen infection.
The possible roles of these proteins in the soybean- F. Most of the soybean proteins identified in this study were previously reported in the xylem sap of other plant species including soybean [31] , [32] , [33].
Xylem saps have been shown to contain peroxidases, which may play a role in plugging damaged vascular tissue [52]. We observed peroxidases in the xylem saps of both infected and healthy plants. Usually peroxidases are produced in response to biotic stresses; but some peroxidases are not specific to infection. Even though found in the xylem sap, the origin of these proteins is still questionable. The xylem sap collection method could impose stress on the plants.
Detection of stress-induced proteins such as ribonucleases from both diseased and healthy plants could therefore have resulted from the stress associated with the xylem sap collection method. Ligat et al. Other studies have also suggested these proteins to be synthesized in the roots [29] , [52] , [54]. Hence it is possible the xylem sap proteins identified in this study are in fact secreted by the root tissues and loaded into the xylem sap.
Some of these sap soybean proteins could have been synthesized to defend F. Pathogenicity related PR proteins are implicated as plant defense molecules and were identified in the xylem sap of F. Further study is needed to determine if PR genes are transported long distance as a defense arsenal and involved in defending soybean against F.
This study identified five secreted F. These proteins were also found to be secreted by F. This study provides evidence that F. The presence of a protein with high similarity to the phytotoxin, cerato-platanin in xylem sap of the F. We have also detected soybean proteins in xylem saps of at least eight of the 10 replications.
Most importantly we have identified four types of pathogenicity-related defense proteins only from the xylem sap of F. Thus, this study laid the foundation for studying the molecular basis of foliar SDS development in soybean and possible defense mechanisms that may be involved in conferring immunity against F.
Peptides of five F. Peptides of F. First five proteins were identified in xylem sap of the F. Gene Ontology GO annotation of over soybean proteins identified from the xylem sap. Detection of over soybean proteins across 10 biological replications of xylem sap: i five replications of F. Waste management and agricultural products Ecogen, Inc. Solder paste, adhesive, and lubrication products E. Aromatic and organic chemicals for the flavor and fragrance industries Request form only Elco Electra Holdings Ltd.
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Campbells Diamond Special C is suitable for application via hydroponic systems, for irrigation systems fertigation in soil grown crops and for foliar application. It dissolves rapidly and completely and remains in solution ready for root adsorption or is taken in via leaf surfaces. Nutrivant requires a lower volume of spray for application, reducing the nutrient cost per hectare. It can be easily tank mixed with a large number of plant protection products.
Foliar feeds from Nutrivant fit into all fertilization programs and are chloride-free and fully soluble. Plus Maize Read more. Grapes
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