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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

DIGE analysis of rubber latex proteins upon ethylene treatment.Proteins obtained from rubber trees treated with ethylene and ddH2O for 3 and 5 days (E-3, E-5; D-3, D-5) were labeled with different fluorescence dyes (Cy3 or Cy5). The combined images of each sample with internal standard (IS, labeled with Cy2) are presented. The differential spots (D1-D143) identified by MS are numbered with different colors (red, up-regulated upon ethylene; green, down-regulated).
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f3: DIGE analysis of rubber latex proteins upon ethylene treatment.Proteins obtained from rubber trees treated with ethylene and ddH2O for 3 and 5 days (E-3, E-5; D-3, D-5) were labeled with different fluorescence dyes (Cy3 or Cy5). The combined images of each sample with internal standard (IS, labeled with Cy2) are presented. The differential spots (D1-D143) identified by MS are numbered with different colors (red, up-regulated upon ethylene; green, down-regulated).

Mentions: 2-D DIGE was performed to identify ERLPs. Typical DIGE gels for each treatment and the combined images are presented (Fig. 3a–h). The global protein patterns on DIGE gels with ∼1750 ± 38 spots remained consistent, and 143 protein spots (termed D1-D143) with a more than 1.3-fold change (confidence > 95%) following ethylene treatment were positively identified by MS (Fig. 3; Fig. S3; Table S2). In 3-day treated latex, 41 proteins had increased, while 43 had decreased. The top five induced proteins were mitogen-activated protein kinase (MAPK), aspartic proteinase precursor, enolase, regulator of ribonuclease, and caffeic acid O-methyltransferase (CAMT). Following 5 days of ethylene treatment, a greater number of differential proteins was observed. Among them, 50 proteins, including rubber elongation factor isoforms, Hev b7.02, esterase and proteasome, had increased, while 55 had decreased. Notably, although most spots containing rubber biosynthesis-related proteins, such as REF and SRPP, did not change significantly, some of their family members or isoforms were sharply increased by ethylene treatment. With regard to REF, 14 isoforms were significantly changed after ethylene treatment, including 10 that were sharply induced (D25, 27, 30, 32, 65, 86, 96, 139, 142 and 143). With regard to SRPP, at least 5 protein isoforms (D52, 66, 67, 99 and 104) increased. However, glucanase and hevein - two key activators of lutoid-mediated rubber particle aggregation (RPA) and sequential latex coagulation19 -were decreased upon ethylene treatment (Fig. 3; Fig. S3; Table S2). The decrease of accumulation of glucanase and hevein in ethylene-treated latex may inhibit RPA, thus maintaining latex flow.


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)

DIGE analysis of rubber latex proteins upon ethylene treatment.Proteins obtained from rubber trees treated with ethylene and ddH2O for 3 and 5 days (E-3, E-5; D-3, D-5) were labeled with different fluorescence dyes (Cy3 or Cy5). The combined images of each sample with internal standard (IS, labeled with Cy2) are presented. The differential spots (D1-D143) identified by MS are numbered with different colors (red, up-regulated upon ethylene; green, down-regulated).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: DIGE analysis of rubber latex proteins upon ethylene treatment.Proteins obtained from rubber trees treated with ethylene and ddH2O for 3 and 5 days (E-3, E-5; D-3, D-5) were labeled with different fluorescence dyes (Cy3 or Cy5). The combined images of each sample with internal standard (IS, labeled with Cy2) are presented. The differential spots (D1-D143) identified by MS are numbered with different colors (red, up-regulated upon ethylene; green, down-regulated).
Mentions: 2-D DIGE was performed to identify ERLPs. Typical DIGE gels for each treatment and the combined images are presented (Fig. 3a–h). The global protein patterns on DIGE gels with ∼1750 ± 38 spots remained consistent, and 143 protein spots (termed D1-D143) with a more than 1.3-fold change (confidence > 95%) following ethylene treatment were positively identified by MS (Fig. 3; Fig. S3; Table S2). In 3-day treated latex, 41 proteins had increased, while 43 had decreased. The top five induced proteins were mitogen-activated protein kinase (MAPK), aspartic proteinase precursor, enolase, regulator of ribonuclease, and caffeic acid O-methyltransferase (CAMT). Following 5 days of ethylene treatment, a greater number of differential proteins was observed. Among them, 50 proteins, including rubber elongation factor isoforms, Hev b7.02, esterase and proteasome, had increased, while 55 had decreased. Notably, although most spots containing rubber biosynthesis-related proteins, such as REF and SRPP, did not change significantly, some of their family members or isoforms were sharply increased by ethylene treatment. With regard to REF, 14 isoforms were significantly changed after ethylene treatment, including 10 that were sharply induced (D25, 27, 30, 32, 65, 86, 96, 139, 142 and 143). With regard to SRPP, at least 5 protein isoforms (D52, 66, 67, 99 and 104) increased. However, glucanase and hevein - two key activators of lutoid-mediated rubber particle aggregation (RPA) and sequential latex coagulation19 -were decreased upon ethylene treatment (Fig. 3; Fig. S3; Table S2). The decrease of accumulation of glucanase and hevein in ethylene-treated latex may inhibit RPA, thus maintaining latex flow.

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