Skip to main content
Log in

A simultaneous extraction method for metabolome and lipidome and its application in cry1Ac and sck-transgenic rice leaf treated with insecticide based on LC–MS analysis

  • Original Article
  • Published:
Metabolomics Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Abstract

Abiotic stress caused by insecticide treatment is an interesting and challenging topic in plant research. Here a simultaneous extraction method with methyl tert-butyl ether for metabolomics and lipidomics analysis by using rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry was developed and validated. The extraction efficiency of lipidome and metabolome based on the current developed method showed an obvious improvement compared with literatures. About 300 metabolites were identified in rice leaf. Method validation was performed and analytical properties including the linearity (R2, 0.991–1.000), repeatability (over 87 % peaks with CV < 20 % accounting for over 92 % of total response) and recovery (74.4–119.6 %) were satisfactory. The method was then applied to investigate time-course changes caused by insecticide treatment in transgenic rice with cry1Ac and sck genes and its wild counterpart. Antioxidants including ferulic acid and sinapic acid were down-regulated at 24 h after insecticide treatment. Signaling metabolites including salicylic acid and nicotinamide adenine dinucleotide were significantly up-regulated at 12–24 h after treatment in transgenic rice. More flavonoids were significantly changed in transgenic plants. Results indicated that gene transformation affected the metabolic response of rice to insecticide stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Agati, G., Matteini, P., Goti, A., & Tattini, M. (2007). Chloroplast-located flavonoids can scavenge singlet oxygen. New Phytologist, 174(1), 77–89.

    Article  CAS  PubMed  Google Scholar 

  • Asensi-Fabado, M. A., & Munne-Bosch, S. (2010). Vitamins in plants: occurrence, biosynthesis and antioxidant function. Trends in Plant Science, 15(10), 582–592.

    Article  CAS  PubMed  Google Scholar 

  • Bijlsma, S., Bobeldijk, L., Verheij, E. R., Ramaker, R., Kochhar, S., Macdonald, I. A., et al. (2006). Large-scale human metabolomics studies: A strategy for data (pre-) processing and validation. Analytical Chemistry, 78(2), 567–574.

    Article  CAS  PubMed  Google Scholar 

  • Bollina, V., Kumaraswamy, G. K., Kushalappa, A. C., Choo, T. M., Dion, Y., Rioux, S., et al. (2010). Mass spectrometry-based metabolomics application to identify quantitative resistance-related metabolites in barley against Fusarium head blight. Molecular Plant Pathology, 11(6), 769–782.

    CAS  PubMed  Google Scholar 

  • Brazier-Hicks, M., Evans, K. M., Gershater, M. C., Puschmann, H., Steel, P. G., & Edwards, R. (2009). The C-glycosylation of flavonoids in cereals. Journal of Biological Chemistry, 284(27), 17926–17934.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brodeur, J., & Dubois, K. P. (1963). Comparison of acute toxicity of anticholinesterase insecticides to weanling and adult male rats. Proceedings of the Society for Experimental Biology and Medicine, 114(2), 509–511.

    Article  CAS  PubMed  Google Scholar 

  • Buseman, C. M., Tamura, P., Sparks, A. A., Baughman, E. J., Maatta, S., Zhao, J., et al. (2006). Wounding stimulates the accumulation of glycerolipids containing oxophytodienoic acid and dinor-oxophytodienoic acid in Arabidopsis leaves. Plant Physiology, 142(1), 28–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chang, Y., Zhao, C., Wu, Z., Zhou, J., Zhao, S., Lu, X., et al. (2012). Chip-based nanoflow high performance liquid chromatography coupled to mass spectrometry for profiling of soybean flavonoids. Electrophoresis, 33(15), 2399–2406.

    Article  CAS  PubMed  Google Scholar 

  • Chen, S., Hoene, M., Li, J., Li, Y., Zhao, X., Haring, H. U., et al. (2013). Simultaneous extraction of metabolome and lipidome with methyl tert-butyl ether from a single small tissue sample for ultra-high performance liquid chromatography/mass spectrometry. Journal of Chromatography A, 1298, 9–16.

    Article  CAS  PubMed  Google Scholar 

  • Falcone Ferreyra, M. L., Rius, S. P., & Casati, P. (2012). Flavonoids: biosynthesis, biological functions, and biotechnological applications. Frontiers in Plant Science, 3, 1–15.

    Google Scholar 

  • Gammulla, C. G., Pascovici, D., Atwell, B. J., & Haynes, P. A. (2010). Differential metabolic response of cultured rice (Oryza sativa) cells exposed to high- and low-temperature stress. Proteomics, 10(16), 3001–3019.

    Article  CAS  PubMed  Google Scholar 

  • Garcia-Hernandez, J. L., Nolasco, H., Troyo-Dieguez, E., Jones, H. G., & Ortega-Rubio, A. (2001). The effects of selected insecticides on the superoxide dismutase activity in hot pepper plants (Capsicum annuum L. cv. Ancho San Luis). Phyton, International Journal of Experimental Botany, 2001, 67–74.

    Google Scholar 

  • Giavalisco, P., Li, Y., Matthes, A., Eckhardt, A., Hubberten, H. M., Hesse, H., et al. (2011). Elemental formula annotation of polar and lipophilic metabolites using (13) C (15) N and (34) S isotope labelling, in combination with high-resolution mass spectrometry. Plant Journal, 68(2), 364–376.

    Article  CAS  PubMed  Google Scholar 

  • Gika, H. G., Theodoridis, G. A., Wingate, J. E., & Wilson, I. D. (2007). Within-day reproducibility of an HPLC-MS-Based method for metabonomic analysis: Application to human urine. Journal of Proteome Research, 6(8), 3291–3303.

    Article  CAS  PubMed  Google Scholar 

  • Hu, C., van Dommelen, J., van der Heijden, R., Spijksma, G., Reijmers, T. H., Wang, M., et al. (2008). RPLC-Ion-Trap-FTMS method for lipid profiling of plasma: Method validation and application to p53 mutant mouse model. Journal of Proteome Research, 7(11), 4982–4991.

    Article  CAS  PubMed  Google Scholar 

  • Hunt, L., Lerner, F., & Ziegler, M. (2004). NAD: new roles in signalling and gene regulation in plants. New Phytologist, 163(1), 31–44.

    Article  CAS  Google Scholar 

  • Karanth, S., Liu, J., Olivier, K., & Pope, C. (2004). Interactive toxicity of the organophosphorus insecticides chlorpyrifos and methyl parathion in adult rats. Toxicology and Applied Pharmacology, 196(2), 183–190.

    Article  CAS  PubMed  Google Scholar 

  • Kennedy, C. W. (2002). Phytotoxicity in pearl millet varies among in-furrow insecticides. Crop Protection, 21(9), 799–802.

    Article  CAS  Google Scholar 

  • Lin, W., & Oliver, D. J. (2008). Role of triacylglycerols in leaves. Plant Science, 175(3), 233–237.

    Article  CAS  Google Scholar 

  • Lindsey, K., Pullen, M. L., & Topping, J. F. (2003). Importance of plant sterols in pattern formation and hormone signalling. Trends in Plant Science, 8(11), 521–525.

    Article  CAS  PubMed  Google Scholar 

  • Liu, C., Hao, F., Hu, J., Zhang, W., Wan, L., Zhu, L., et al. (2010). Revealing different systems responses to brown planthopper infestation for pest susceptible and resistant rice plants with the combined metabonomic and gene-expression analysis. Journal of Proteome Research, 9(12), 6774–6785.

    Article  CAS  PubMed  Google Scholar 

  • Maqbool, S. B., Riazuddin, S., Loc, N. T., Gatehouse, A. M. R., Gatehouse, J. A., & Christou, P. (2001). Expression of multiple insecticidal genes confers broad resistance against a range of different rice pests. Molecular Breeding, 7(1), 85–93.

    Article  CAS  Google Scholar 

  • Markham, J. E., & Jaworski, J. G. (2007). Rapid measurement of sphingolipids from Arabidopsis thaliana by reversed-phase high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. Rapid Communications in Mass Spectrometry, 21(7), 1304–1314.

    Article  CAS  PubMed  Google Scholar 

  • Matyash, V., Liebisch, G., Kurzchalia, T. V., Shevchenko, A., & Schwudke, D. (2008). Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. Journal of Lipid Research, 49(5), 1137–1146.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Michaelson, L. V., & Napier, J. A. (2010). Sphingolipid signaling in plants. In T. Munnik (Ed.), lipid signaling in plants (pp. 307–321). Berlin Heidelberg: Springer.

    Chapter  Google Scholar 

  • Nayak, P., Basu, D., Das, S., Basu, A., Ghosh, D., Ramakrishnan, N. A., et al. (1997). Transgenic elite indica rice plants expressing CryIAc delta-endotoxin of Bacillus thuringiensis are resistant against yellow stem borer (Scirpophaga incertulas). Proceedings of the National Academy of Sciences of USA, 94(6), 2111–2116.

    Article  CAS  Google Scholar 

  • Pilar Lopez-Gresa, M., Maltese, F., Maria Belles, J., Conejero, V., Kim, H. K., Choi, Y. H., et al. (2010). Metabolic response of tomato leaves upon different plant-pathogen interactions. Phytochemical Analysis, 21(1), 89–94.

    Article  PubMed  Google Scholar 

  • Rocha, J. M., Kalo, P. J., Ollilainen, V., & Malcata, F. X. (2010). Separation and identification of neutral cereal lipids by normal phase high-performance liquid chromatography, using evaporative light-scattering and electrospray mass spectrometry for detection. Journal of Chromatography A, 1217(18), 3013–3025.

    Article  CAS  PubMed  Google Scholar 

  • Saeed, A. I., Bhagabati, N. K., Braisted, J. C., Liang, W., Sharov, V., Howe, E. A., et al. (2006). TM4 microarray software suite. Methods in Enzymology, 411, 134–193.

    Article  CAS  PubMed  Google Scholar 

  • Sanchez, D. H., Schwabe, F., Erban, A., Udvardi, M. K., & Kopka, J. (2012). Comparative metabolomics of drought acclimation in model and forage legumes. Plant, Cell and Environment, 35(1), 136–149.

    Article  CAS  PubMed  Google Scholar 

  • Sangster, T., Major, H., Plumb, R., Wilson, A. J., & Wilson, I. D. (2006). A pragmatic and readily implemented quality control strategy for HPLC-MS and GC-MS-based metabonomic analysis. Analyst, 131(10), 1075–1078.

    Article  CAS  PubMed  Google Scholar 

  • Seiwert, B., Giavalisco, P., & Willmitzer, L. (2009). Advanced mass spectrometry methods for analysis of lipids from photosynthetic organisms. In H. Wada & N. Murata (Eds.), Lipids in photosynthesis: Essential and regulatory functions (pp. 445–461). Netherlands: Springer.

    Chapter  Google Scholar 

  • Seppanen-Laakso, T., & Oresic, M. (2009). How to study lipidomes. Journal of Molecular Endocrinology, 42(3–4), 185–190.

    CAS  PubMed  Google Scholar 

  • Shimojima, M. (2011). Biosynthesis and functions of the plant sulfolipid. Progress in Lipid Research, 50(3), 234–239.

    Article  CAS  PubMed  Google Scholar 

  • Smilde, A. K., van der Werf, M. J., Bijlsma, S., van der Werff-van-der Vat, B. J. C., & Jellema, R. H. (2005). Fusion of mass spectrometry-based metabolomics data. Analytical Chemistry, 77(20), 6729–6736.

    Article  CAS  PubMed  Google Scholar 

  • Snedecor, George W., & Cochran, W. G. (1980). Statistical methods (7th ed.). Ames, IA: Iowa State University.

    Google Scholar 

  • Story, P., Hooper, M. J., Astheimer, L. B., & Buttemer, W. A. (2011). Acute oral toxicity of the organophosphorus pesticide fenitrothion to fat-tailed and stripe-faced dunnarts and its relevance for pesticide risk assessments in Australia. Environmental Toxicology and Chemistry, 30(5), 1163–1169.

    Article  CAS  PubMed  Google Scholar 

  • Szabados, L., & Savoure, A. (2010). Proline: A multifunctional amino acid. Trends in Plant Science, 15(2), 89–97.

    Article  CAS  PubMed  Google Scholar 

  • Tattini, M., Galardi, C., Pinelli, P., Massai, R., Remorini, D., & Agati, G. (2004). Differential accumulation of flavonoids and hydroxycinnamates in leaves of Ligustrum vulgare under excess light and drought stress. New Phytologist, 163(3), 547–561.

    Article  CAS  Google Scholar 

  • Vadhana, D., Carloni, M., Fedeli, D., Nasuti, C., & Gabbianelli, R. (2011). Perturbation of rat heart plasma membrane fluidity due to metabolites of permethrin insecticide. Cardiovascular Toxicology, 11(3), 226–234.

    Article  CAS  PubMed  Google Scholar 

  • Vlot, A. C., Dempsey, D. M. A., & Klessig, D. F. (2009). Salicylic acid, a multifaceted hormone to combat disease. Annual Review of Phytopathology, 47, 177–206.

    Article  CAS  PubMed  Google Scholar 

  • Vukics, V., & Guttman, A. (2010). Structural characterization of flavonoid glycosides by multi-stage mass spectrometry. Mass Spectrometry Reviews, 29(1), 1–16.

    CAS  PubMed  Google Scholar 

  • Wewer, V., Dombrink, I., Vom Dorp, K., & Doermann, P. (2011). Quantification of sterol lipids in plants by quadrupole time-of-flight mass spectrometry. Journal of Lipid Research, 52(5), 1039–1054.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang, J., Zhang, Y., Du, Y., Chen, S., & Tang, H. (2011). Dynamic metabonomic responses of tobacco (Nicotiana tabacum) plants to salt stress. Journal of Proteome Research, 10(4), 1904–1914.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, J., Zhang, L., Chang, Y., Lu, X., Zhu, Z., & Xu, G. (2012a). Alteration of leaf metabolism in Bt-transgenic rice (Oryza sativa L.) and its wild type under insecticide stress. Journal of Proteome Research, 11(8), 4351–4360.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, J., Zhang, L., Li, X., Chang, Y. W., Gu, Q., Lu, X., et al. (2012b). Metabolic profiling of transgenic rice progeny using gas chromatography-mass spectrometry: the effects of gene insertion, tissue culture and breeding. Metabolomics, 8(4), 529–539.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study has been supported by the National Grand Project of Science and Technology (2013ZX08012-002, 2014ZX08012-002, 2014ZX08001-001), the National Natural Science Foundation (21321064, 21075122).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Chunxia Zhao, Zhen Zhu or Guowang Xu.

Additional information

Yuwei Chang and Lei Zhang contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 256 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chang, Y., Zhang, L., Lu, X. et al. A simultaneous extraction method for metabolome and lipidome and its application in cry1Ac and sck-transgenic rice leaf treated with insecticide based on LC–MS analysis. Metabolomics 10, 1197–1209 (2014). https://doi.org/10.1007/s11306-014-0658-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11306-014-0658-6

Keywords

Navigation