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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.


Comparison of protein and RNA patterns in ethylene-treated latex.The 65 identified proteins were assayed by RT-PCR, DIGE, and iTRAQ to compare their gene and protein expression patterns in the D-0, D-3, E-3, D-5 and E-5 plants (a–c). The actin gene from the rubber tree was used as a reference. Then, their relative changed ratios (E-3/D-3 and E-5/D-5) at both protein level (gray for data from DIGE; black for data from iTRAQ) and mRNA level (white) are presented right aside the semi-quantitative RT-PCR photographs. The dot line in each rectangle graph represents a 1.0 ratio value. The relative changed ratios at protein level over (a), near (b), and below (c) the 1.0 ratio line are classified into different groups. Finally, Western blotting for the 22 latex proteins was performed to determine the general protein accumulation in D-0, D-3, E-3, D-5 and E-5 plants (d). The approximately molecular weight (kDa) of the target bands is indicated with an arrow. Abbreviations, antibody information, and primers used for RT-PCR are listed in Supplemental Fig. S2, Fig. S6, and Fig. S7, respectively.
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f7: Comparison of protein and RNA patterns in ethylene-treated latex.The 65 identified proteins were assayed by RT-PCR, DIGE, and iTRAQ to compare their gene and protein expression patterns in the D-0, D-3, E-3, D-5 and E-5 plants (a–c). The actin gene from the rubber tree was used as a reference. Then, their relative changed ratios (E-3/D-3 and E-5/D-5) at both protein level (gray for data from DIGE; black for data from iTRAQ) and mRNA level (white) are presented right aside the semi-quantitative RT-PCR photographs. The dot line in each rectangle graph represents a 1.0 ratio value. The relative changed ratios at protein level over (a), near (b), and below (c) the 1.0 ratio line are classified into different groups. Finally, Western blotting for the 22 latex proteins was performed to determine the general protein accumulation in D-0, D-3, E-3, D-5 and E-5 plants (d). The approximately molecular weight (kDa) of the target bands is indicated with an arrow. Abbreviations, antibody information, and primers used for RT-PCR are listed in Supplemental Fig. S2, Fig. S6, and Fig. S7, respectively.

Mentions: To further evaluate the correlation between the protein and transcript abundance, 65 ERLPs were selected for RT-PCR to assess gene expression patterns. The results demonstrated that the expression patterns of most detected proteins and their genes changed similarly upon ethylene treatment. Compared with H2O treated plants (D-3 and -5), only 13 genes were induced by ethylene (E-3 and -5), while 27 genes were inhibited (Fig. 7a–c). The other 25 transcripts either did not change or changed less than the level of their encoded proteins upon ethylene treatment. Among the down-regulated genes, were several enzymes known to be key factors in rubber biosynthesis, including HMGR, HMGS, MEVD, MEVK, REF, SRPP, FADS and CIPT (Fig. 7a–c).


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)

Comparison of protein and RNA patterns in ethylene-treated latex.The 65 identified proteins were assayed by RT-PCR, DIGE, and iTRAQ to compare their gene and protein expression patterns in the D-0, D-3, E-3, D-5 and E-5 plants (a–c). The actin gene from the rubber tree was used as a reference. Then, their relative changed ratios (E-3/D-3 and E-5/D-5) at both protein level (gray for data from DIGE; black for data from iTRAQ) and mRNA level (white) are presented right aside the semi-quantitative RT-PCR photographs. The dot line in each rectangle graph represents a 1.0 ratio value. The relative changed ratios at protein level over (a), near (b), and below (c) the 1.0 ratio line are classified into different groups. Finally, Western blotting for the 22 latex proteins was performed to determine the general protein accumulation in D-0, D-3, E-3, D-5 and E-5 plants (d). The approximately molecular weight (kDa) of the target bands is indicated with an arrow. Abbreviations, antibody information, and primers used for RT-PCR are listed in Supplemental Fig. S2, Fig. S6, and Fig. S7, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Comparison of protein and RNA patterns in ethylene-treated latex.The 65 identified proteins were assayed by RT-PCR, DIGE, and iTRAQ to compare their gene and protein expression patterns in the D-0, D-3, E-3, D-5 and E-5 plants (a–c). The actin gene from the rubber tree was used as a reference. Then, their relative changed ratios (E-3/D-3 and E-5/D-5) at both protein level (gray for data from DIGE; black for data from iTRAQ) and mRNA level (white) are presented right aside the semi-quantitative RT-PCR photographs. The dot line in each rectangle graph represents a 1.0 ratio value. The relative changed ratios at protein level over (a), near (b), and below (c) the 1.0 ratio line are classified into different groups. Finally, Western blotting for the 22 latex proteins was performed to determine the general protein accumulation in D-0, D-3, E-3, D-5 and E-5 plants (d). The approximately molecular weight (kDa) of the target bands is indicated with an arrow. Abbreviations, antibody information, and primers used for RT-PCR are listed in Supplemental Fig. S2, Fig. S6, and Fig. S7, respectively.
Mentions: To further evaluate the correlation between the protein and transcript abundance, 65 ERLPs were selected for RT-PCR to assess gene expression patterns. The results demonstrated that the expression patterns of most detected proteins and their genes changed similarly upon ethylene treatment. Compared with H2O treated plants (D-3 and -5), only 13 genes were induced by ethylene (E-3 and -5), while 27 genes were inhibited (Fig. 7a–c). The other 25 transcripts either did not change or changed less than the level of their encoded proteins upon ethylene treatment. Among the down-regulated genes, were several enzymes known to be key factors in rubber biosynthesis, including HMGR, HMGS, MEVD, MEVK, REF, SRPP, FADS and CIPT (Fig. 7a–c).

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.