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Comprehensive Proteomics Analysis of Laticifer Latex Reveals New Insights into Ethylene Stimulation of Natural Rubber Production.

Wang X, Wang D, Sun Y, Yang Q, Chang L, Wang L, Meng X, Huang Q, Jin X, Tong Z - Sci Rep (2015)

Bottom Line: Moreover, we found that ethylene improves the generation of small rubber particles.Functional classification of ERLPs revealed that enzymes involved in post-translational modification, carbohydrate metabolism, hydrolase activity, and kinase activity were overrepresented.Phosphoproteomics analysis identified 59 differential phosphoproteins; notably, specific isoforms of rubber elongation factor and small rubber particle protein that were phosphorylated mainly at serine residues.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Biology and Genetic Resources for Tropical Crops, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, China.

ABSTRACT
Ethylene is a stimulant to increase natural rubber latex. After ethylene application, both fresh yield and dry matter of latex are substantially improved. Moreover, we found that ethylene improves the generation of small rubber particles. However, most genes involved in rubber biosynthesis are inhibited by exogenous ethylene. Therefore, we conducted a proteomics analysis of ethylene-stimulated rubber latex, and identified 287 abundant proteins as well as 143 ethylene responsive latex proteins (ERLPs) with mass spectrometry from the 2-DE and DIGE gels, respectively. In addition, more than 1,600 proteins, including 404 ERLPs, were identified by iTRAQ. Functional classification of ERLPs revealed that enzymes involved in post-translational modification, carbohydrate metabolism, hydrolase activity, and kinase activity were overrepresented. Some enzymes for rubber particle aggregation were inhibited to prolong latex flow, and thus finally improved latex production. Phosphoproteomics analysis identified 59 differential phosphoproteins; notably, specific isoforms of rubber elongation factor and small rubber particle protein that were phosphorylated mainly at serine residues. This post-translational modification and isoform-specific phosphorylation might be important for ethylene-stimulated latex production. These results not only deepen our understanding of the rubber latex proteome but also provide new insights into the use of ethylene to stimulate rubber latex production.

No MeSH data available.


Related in: MedlinePlus

Proteome profiles and MS identification of main latex proteins.Protein bands of total latex (TL), C-serum (CS), lutoids (LO), and rubber particles (RP) from the D-3 and E-3 plants (from left to right) were identified by MS (a). Total latex proteins labeled with Cy5 on the pH 4–7 range 2-DE gels were visualized by a Typhoon scanner. The main protein spots were excised and identified by MS, which are marked with numbers in different colors (b). The protein identities are presented in Fig. S2 and Table S1.
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f2: Proteome profiles and MS identification of main latex proteins.Protein bands of total latex (TL), C-serum (CS), lutoids (LO), and rubber particles (RP) from the D-3 and E-3 plants (from left to right) were identified by MS (a). Total latex proteins labeled with Cy5 on the pH 4–7 range 2-DE gels were visualized by a Typhoon scanner. The main protein spots were excised and identified by MS, which are marked with numbers in different colors (b). The protein identities are presented in Fig. S2 and Table S1.

Mentions: To determine the protein profile of latex, the main proteins on both 1-DE and 2-DE gels were subjected to mass spectrometry (MS) analysis. The main bands of different latex fractions (Fig. 2a) were excised for in-gel digestion, and 35 proteins were positively identified. Then, the abundant protein spots (Vol% > 0.01) on the 2-DE gel (Fig. 2b) were subjected to MS analysis. Finally, 252 proteins were identified (Fig. S1; Table S1). Among them, 121 proteins were identified from H. brasiliensis, and 36 proteins were from Ricinus communis (Fig. S2a; Table S1). Radial chart analysis revealed that more than 90% of the proteins were distributed between the cyclical line 0.6 and 1.2 (Fig. S2b). These latex proteins belonged to 16 functional groups, the largest of which included 38 proteins involved in post-translational modification, followed by the categories of carbohydrate transport and metabolism, energy production, ribosomal structure and biogenesis (Fig. S1; Fig. S2c). Gene ontology (GO) pathway and biological process analysis verified that most of the proteins identified were involved in carbohydrate metabolism (Fig. S2d). An additional 19 proteins were related to abiotic stress responses, followed by 17 proteins involved with glycolysis. By GO cell biological analysis, 42 protein species were found to be overrepresented in the cytoplasm (Fig. S2e), in agreement with the fact that rubber latex is present in the cytoplasm of laticifers in Hevea phloem15.


Comprehensive Proteomics Analysis of Laticifer Latex Reveals New Insights into Ethylene Stimulation of Natural Rubber Production.

Wang X, Wang D, Sun Y, Yang Q, Chang L, Wang L, Meng X, Huang Q, Jin X, Tong Z - Sci Rep (2015)

Proteome profiles and MS identification of main latex proteins.Protein bands of total latex (TL), C-serum (CS), lutoids (LO), and rubber particles (RP) from the D-3 and E-3 plants (from left to right) were identified by MS (a). Total latex proteins labeled with Cy5 on the pH 4–7 range 2-DE gels were visualized by a Typhoon scanner. The main protein spots were excised and identified by MS, which are marked with numbers in different colors (b). The protein identities are presented in Fig. S2 and Table S1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4562231&req=5

f2: Proteome profiles and MS identification of main latex proteins.Protein bands of total latex (TL), C-serum (CS), lutoids (LO), and rubber particles (RP) from the D-3 and E-3 plants (from left to right) were identified by MS (a). Total latex proteins labeled with Cy5 on the pH 4–7 range 2-DE gels were visualized by a Typhoon scanner. The main protein spots were excised and identified by MS, which are marked with numbers in different colors (b). The protein identities are presented in Fig. S2 and Table S1.
Mentions: To determine the protein profile of latex, the main proteins on both 1-DE and 2-DE gels were subjected to mass spectrometry (MS) analysis. The main bands of different latex fractions (Fig. 2a) were excised for in-gel digestion, and 35 proteins were positively identified. Then, the abundant protein spots (Vol% > 0.01) on the 2-DE gel (Fig. 2b) were subjected to MS analysis. Finally, 252 proteins were identified (Fig. S1; Table S1). Among them, 121 proteins were identified from H. brasiliensis, and 36 proteins were from Ricinus communis (Fig. S2a; Table S1). Radial chart analysis revealed that more than 90% of the proteins were distributed between the cyclical line 0.6 and 1.2 (Fig. S2b). These latex proteins belonged to 16 functional groups, the largest of which included 38 proteins involved in post-translational modification, followed by the categories of carbohydrate transport and metabolism, energy production, ribosomal structure and biogenesis (Fig. S1; Fig. S2c). Gene ontology (GO) pathway and biological process analysis verified that most of the proteins identified were involved in carbohydrate metabolism (Fig. S2d). An additional 19 proteins were related to abiotic stress responses, followed by 17 proteins involved with glycolysis. By GO cell biological analysis, 42 protein species were found to be overrepresented in the cytoplasm (Fig. S2e), in agreement with the fact that rubber latex is present in the cytoplasm of laticifers in Hevea phloem15.

Bottom Line: Moreover, we found that ethylene improves the generation of small rubber particles.Functional classification of ERLPs revealed that enzymes involved in post-translational modification, carbohydrate metabolism, hydrolase activity, and kinase activity were overrepresented.Phosphoproteomics analysis identified 59 differential phosphoproteins; notably, specific isoforms of rubber elongation factor and small rubber particle protein that were phosphorylated mainly at serine residues.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Biology and Genetic Resources for Tropical Crops, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou Hainan 571101, China.

ABSTRACT
Ethylene is a stimulant to increase natural rubber latex. After ethylene application, both fresh yield and dry matter of latex are substantially improved. Moreover, we found that ethylene improves the generation of small rubber particles. However, most genes involved in rubber biosynthesis are inhibited by exogenous ethylene. Therefore, we conducted a proteomics analysis of ethylene-stimulated rubber latex, and identified 287 abundant proteins as well as 143 ethylene responsive latex proteins (ERLPs) with mass spectrometry from the 2-DE and DIGE gels, respectively. In addition, more than 1,600 proteins, including 404 ERLPs, were identified by iTRAQ. Functional classification of ERLPs revealed that enzymes involved in post-translational modification, carbohydrate metabolism, hydrolase activity, and kinase activity were overrepresented. Some enzymes for rubber particle aggregation were inhibited to prolong latex flow, and thus finally improved latex production. Phosphoproteomics analysis identified 59 differential phosphoproteins; notably, specific isoforms of rubber elongation factor and small rubber particle protein that were phosphorylated mainly at serine residues. This post-translational modification and isoform-specific phosphorylation might be important for ethylene-stimulated latex production. These results not only deepen our understanding of the rubber latex proteome but also provide new insights into the use of ethylene to stimulate rubber latex production.

No MeSH data available.


Related in: MedlinePlus