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Identification and Characterization of Phosphoproteins in Somatic Embryogenesis Acquisition during Oil Palm Tissue Culture - PubMed

  • ️Tue Jan 01 2019

Identification and Characterization of Phosphoproteins in Somatic Embryogenesis Acquisition during Oil Palm Tissue Culture

Suvichark Aroonluk et al. Plants (Basel). 2019.

Abstract

Somatic embryogenesis during oil palm tissue culture is a long process. The identification of the proteins that control this process may help to shorten the time of oil palm tissue culture. We collected embryogenic callus and somatic embryos at the globular, torpedo, and cotyledon maturation stages, as well as from plantlets, for total protein extraction. An enrichment column was used to enrich the phosphoproteins, which were subjected to tryptic enzyme digestion. Each sample was analyzed with nano-liquid chromatography-tandem mass spectrometry (nano LC-MS/MS). A total of 460 phosphoproteins were identified and analyzed. The functional characterization of phosphoproteins were observed as highest in the metabolic process, protein/nucleotide/ion binding, and membrane component. The different phosphoproteins are involved in the control of vegetative growth, cellular differentiation, cell morphogenesis, and signaling roles in plants. The Quantitative Real-Time Reverse Transcription-PCR technique (qPCR) was successfully used to verify the expression of genes, and the results were consistent with the level of protein expression from nano-LC-MS/MS. The E3 ubiquitin-protein ligase and sister chromatid cohesion PDS5 were specifically expressed only in the somatic embryo and plantlet, and these could be used as protein biomarkers to determine the oil palm somatic embryo maturation stage. This study sheds light on the protein phosphorylation mechanism that regulates somatic embryogenesis transition during oil palm tissue culture.

Keywords: Elaeis guineensis; micropropagation; nano-liquid chromatography–tandem mass spectrometry; proteomics.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1

Elaeis guineensis Jacq cv “Tenera” developmental stages of somatic embryogenesis. (a) embryogenic callus (bar = 0.25 cm), (b) globular (bar = 0.25 cm), (c) torpedo (bar = 0.25 cm) (d) cotyledonary (bar = 0.25 cm), and (e) plantlet (bar = 0.25 cm) stages.

Figure 2
Figure 2

Overlap of identified phosphoproteins in callus (blue), globular (pink), torpedo (brown), cotyledon (yellow) and plantlet (green) stages.

Figure 3
Figure 3

Biological function and subcellular localization of 460 identified phosphoproteins during somatic acquisition and plant regeneration. (a) Assignment of the 460 identified phosphoproteins to biological process categories with the use of gene ontology (GO) classification. (b) Assignment of the 460 identified phosphoproteins to molecular function categories with the use of GO classification. (c) Assignment of the 460 identified phosphoproteins to subcellular localization categories with the use of GO classification.

Figure 4
Figure 4

mRNA expression patterns derived from differentially expressed phosphoproteins.

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