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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.


Phosphoproteomics analysis of rubber latex proteins upon ethylene treatment.The 2-DE gel for E-3 latex was stained with Pro-Q Diamond dye to detect phosphoproteins (a, red). It was then restained with SYPRO Ruby (b, green). The same gel was restained by G250 (c, blue). The combination image of Pro-Q Diamond and SYPRO Ruby are presented to demonstrate the specific phosphorylated proteins (d, merged). The latex proteins obtained from the D-0 (e-g) and D-3 (h-j) plants were also stained by Pro-Q Diamond and SYPRO Ruby. Finally, 59 phosphoproteins (P1-P59) were identified by MS (d).
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f5: Phosphoproteomics analysis of rubber latex proteins upon ethylene treatment.The 2-DE gel for E-3 latex was stained with Pro-Q Diamond dye to detect phosphoproteins (a, red). It was then restained with SYPRO Ruby (b, green). The same gel was restained by G250 (c, blue). The combination image of Pro-Q Diamond and SYPRO Ruby are presented to demonstrate the specific phosphorylated proteins (d, merged). The latex proteins obtained from the D-0 (e-g) and D-3 (h-j) plants were also stained by Pro-Q Diamond and SYPRO Ruby. Finally, 59 phosphoproteins (P1-P59) were identified by MS (d).

Mentions: Therefore, we confirmed the presence of phosphorylated proteins in rubber latex. Using Pro-Q Diamond dye, phosphospecific proteins in latex exposed to different ethylene treatment conditions were visualized and more than 200 protein spots were detected in 2-DE gels (Fig. 5). In the merged image of the Pro-Q Diamond- and SYPRO Ruby-stained gels, specific phosphorylated protein spots red in color were clearly observed. Many protein spots were visualized by Pro-Q Diamond for latex under the D-0 and D-3 treatments, indicating that these proteins were phosphorylated under control conditions. However, the phosphorylation was substantially more apparent after ethylene treatment (Fig. 5). Finally, the main spots of differentially expressed phosphoproteins were subjected to MS, and 59 differential proteins (Spots P1-P59) were positively identified (Fig. S5; Table S4). Among these phosphoproteins, REF (P19-21 and P36) was the most abundant, followed by the 7 isoforms of SRPP (P5, 6, 9, 17 and 38–40) and one glucanase isoform (P45). Other abundant phosphorylated proteins included malate dehydrogenase, cytoplasmic aldolase, linamarase, latex patatin homolog, Hev b7, and several hypothetical proteins (Fig. S5; Table S4). A large portion, including phosphoesterase, phosphoglycerate kinase, phosphoglucomutase, phospholipase, and 14-3-3 protein, had high phosphotransferase and kinase activity. These enzymes were involved in Ca2+ binding and signal transduction.


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)

Phosphoproteomics analysis of rubber latex proteins upon ethylene treatment.The 2-DE gel for E-3 latex was stained with Pro-Q Diamond dye to detect phosphoproteins (a, red). It was then restained with SYPRO Ruby (b, green). The same gel was restained by G250 (c, blue). The combination image of Pro-Q Diamond and SYPRO Ruby are presented to demonstrate the specific phosphorylated proteins (d, merged). The latex proteins obtained from the D-0 (e-g) and D-3 (h-j) plants were also stained by Pro-Q Diamond and SYPRO Ruby. Finally, 59 phosphoproteins (P1-P59) were identified by MS (d).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Phosphoproteomics analysis of rubber latex proteins upon ethylene treatment.The 2-DE gel for E-3 latex was stained with Pro-Q Diamond dye to detect phosphoproteins (a, red). It was then restained with SYPRO Ruby (b, green). The same gel was restained by G250 (c, blue). The combination image of Pro-Q Diamond and SYPRO Ruby are presented to demonstrate the specific phosphorylated proteins (d, merged). The latex proteins obtained from the D-0 (e-g) and D-3 (h-j) plants were also stained by Pro-Q Diamond and SYPRO Ruby. Finally, 59 phosphoproteins (P1-P59) were identified by MS (d).
Mentions: Therefore, we confirmed the presence of phosphorylated proteins in rubber latex. Using Pro-Q Diamond dye, phosphospecific proteins in latex exposed to different ethylene treatment conditions were visualized and more than 200 protein spots were detected in 2-DE gels (Fig. 5). In the merged image of the Pro-Q Diamond- and SYPRO Ruby-stained gels, specific phosphorylated protein spots red in color were clearly observed. Many protein spots were visualized by Pro-Q Diamond for latex under the D-0 and D-3 treatments, indicating that these proteins were phosphorylated under control conditions. However, the phosphorylation was substantially more apparent after ethylene treatment (Fig. 5). Finally, the main spots of differentially expressed phosphoproteins were subjected to MS, and 59 differential proteins (Spots P1-P59) were positively identified (Fig. S5; Table S4). Among these phosphoproteins, REF (P19-21 and P36) was the most abundant, followed by the 7 isoforms of SRPP (P5, 6, 9, 17 and 38–40) and one glucanase isoform (P45). Other abundant phosphorylated proteins included malate dehydrogenase, cytoplasmic aldolase, linamarase, latex patatin homolog, Hev b7, and several hypothetical proteins (Fig. S5; Table S4). A large portion, including phosphoesterase, phosphoglycerate kinase, phosphoglucomutase, phospholipase, and 14-3-3 protein, had high phosphotransferase and kinase activity. These enzymes were involved in Ca2+ binding and signal transduction.

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.