Comparative phosphoproteomic analysis reveals differentially phosphorylated proteins regulate anther and pollen development in kenaf cytoplasmic male sterility line
Cytoplasmic male sterility (CMS) is widely used in plant breeding and represents a perfect model to understand cyto-nuclear interactions and pollen development research. Protein phosphorylation is ubiquitous and is involved in the regulation of diverse cellular processes. To reveal the possible mechanism of CMS and pollen development in kenaf, we performed an iTRAQ-based comparative phosphoproteome analysis in the anthers of a CMS line and wild-type plant (Wt). Whole transcriptome unigenes of kenaf as the reference genome, we identified a total of 3045 phosphorylated sites on 1640 peptides corresponding to 974 unique proteins. 292 of the peptides which corresponding to 247 unique proteins were differentially phosphorylated (fold change ≥ 1.20 with P value< 0.05) between these two materials. 113 and 134 proteins were characterized as up-regulated or down-regulated phosphorylated, respectively. An evaluation of the phosphoproteome and proteomic results indicated that the most significantly phosphorylated proteins were not associated with abundant changes at the protein level. Bioinformatics analysis demonstrated that many of these proteins were involved in various biological processes which may play key roles in pollen development, including carbohydrate metabolism, energy metabolism, transport, gene expression regulation, signal transduction, and cell cycle control. Our results provide insight into the CMS mechanism and pollen development in kenaf from a protein phosphorylation perspective.
KeywordsKenaf Pollen Cytoplasmic male sterility (CMS) Phosphoproteome iTRAQ
This work was supported by the National Natural Science Foundation of China (Grant no. 31560341) and the earmarked fund for Modern Agro-industry Technology Research System (CARS-16-E14). We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.
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Conflict of interest
The authors declare that they do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.
- Bourbon HM, Aguilera A, Ansari AZ, Asturias FJ, Berk AJ, Bjorklund S, Blackwell TK, Borggrefe T, Carey M, Carlson M, Conaway JW, Conaway RC, Emmons SW, Fondell JD, Freedman LP, Fukasawa T, Gustafsson CM, Han M, He X, Herman PK, Hinnebusch AG, Holmberg S, Holstege FC, Jaehning JA, Kim Y-J, Kuras L, Leutz A, Lis JT, Meisterernest M, Naar AM, Nasmyth K, Parvin JD, Ptashne M, Reinberg D, Ronne H, Sadowski I, Sakurai H, Sipiczki M, Sternberg PW, Stillman DJ, Strich R, Struhl K, Svejstrup JQ, Tuck S, Winston F, Roeder RG, Kornberg RD (2004) A unified nomenclature for protein subunits of mediator complexes linking transcriptional regulators to RNA polymerase II. Mol Cell 14(5):553–557. https://doi.org/10.1016/j.molcel.2004.05.011 CrossRefPubMedGoogle Scholar
- Ischebeck T, Valledor L, Lyon D, Gingl S, Nagler M, Meijón M, Egelhofer V, Weckwerth W (2014) Comprehensive cell-specific protein analysis in early and late pollen development from diploid microsporocytes to pollen tube growth. Mol Cell Proteomics 13(1):295–310. https://doi.org/10.1074/mcp.m113.028100 CrossRefPubMedGoogle Scholar
- Jorrin-Novo JV (2014) Plant proteomics methods and protocols. In: Jorrin-Novo JV, Komatsu S, Weckwerth W, Wienkoop S (eds) Plant proteomics: methods and protocols. Humana Press, Totowa, pp 3–13. https://doi.org/10.1007/978-1-62703-631-3_1
- Laporte MM, Galagan JA, Prasch AL, Vanderveer PJ, Hanson DT, Shewmaker CK, Sharkey TD (2001) Promoter strength and tissue specificity effects on growth of tomato plants transformed with maize sucrose-phosphate synthase. Planta 212(5):817–822. https://doi.org/10.1007/s004250000433 CrossRefPubMedGoogle Scholar
- Ma H (2005) Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants. Annu Rev Plant Biol 56(1):393–434. https://doi.org/10.1146/annurev.arplant.55.031903.141717 CrossRefPubMedGoogle Scholar
- Quévillon HM, Zeitler DM, Hauptmann J, Bruckmann A, Fressigné L, Danner J, Piquet S, Strieder N, Engelmann JC, Jannot G, Deutzmann R, Simard MJ, Meister G (2017) Phosphorylation of argonaute proteins affects mRNA binding and is essential for microRNA-guided gene silencing. EMBO J. https://doi.org/10.15252/embj.201696386 Google Scholar
- Song L, Liu Z, Tong J, Xiao L, Ma H, Zhang H (2015) Comparative proteomics analysis reveals the mechanism of fertility alternation of thermosensitive genic male sterile rice lines under low temperature inducement. Proteomics 15(11):1884–1905. https://doi.org/10.1002/pmic.201400103 CrossRefPubMedGoogle Scholar
- Wisniewski JR, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Meth 6(5):359–362. http://www.nature.com/nmeth/journal/v6/n5/suppinfo/nmeth.1322_S1.html
- Xie HT, Wan ZY, Li S, Zhang Y (2014) Spatiotemporal production of reactive oxygen species by NADPH oxidase is critical for tapetal programmed cell death and pollen development in Arabidopsis. Plant Cell 26(5):2007–2023. https://doi.org/10.1105/tpc.114.125427 CrossRefPubMedPubMedCentralGoogle Scholar
- Yang CW, González-Lamothe R, Ewan RA, Rowland O, Yoshioka H, Shenton M, Ye H, Donnell E, Jones JDG, Sadanandom A (2006) The E3 ubiquitin ligase activity of Arabidopsis; plant U-BOX17 and its functional tobacco homolog ACRE276 are required for cell death and defense. Plant Cell 18(4):1084CrossRefPubMedPubMedCentralGoogle Scholar
- Zhai Q, Yan L, Tan D, Chen R, Sun J, Gao L, Dong M-Q, Wang Y, Li C (2013) Phosphorylation-coupled proteolysis of the transcription factor MYC2 is important for jasmonate-signaled plant immunity. PLoS Genet 9(4):e1003422. https://doi.org/10.1371/journal.pgen.1003422 CrossRefPubMedPubMedCentralGoogle Scholar
- Zhou RY, Zhang X, Zhang JQ, Wei HX (2008) A breakthrough in kenaf cytoplasmic male sterile lines breeding and heterosis utilization. Sci Agric Sin 41(1):314 (in Chinese) Google Scholar
- Zhu LM, Ai SY, Zhou RY (2007) A cytological study on microsporogenesis of cytoplasmic male sterile lines in kenaf (Hibiscus cannabinus L.). Acta Agronom Sinica 33(6):999–1003 (in Chinse)Google Scholar