Global metabolite profiles of rice brown planthopper-resistant traits reveal potential secondary metabolites for both constitutive and inducible defenses
Brown planthopper (BPH) is a phloem feeding insect that causes annual disease outbreaks, called hopper burn in many countries throughout Asia, resulting in severe damage to rice production. Currently, mechanistic understanding of BPH resistance in rice plant is limited, which has caused slow progression on developing effective rice varieties as well as effective farming practices against BPH infestation.
To reveal rice metabolic responses during 8 days of BPH attack, this study examined polar metabolome extracts of BPH-susceptible (KD) and its BPH-resistant isogenic line (IL308) rice leaves.
Ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS) was combined with multi-block PCA to analyze potential metabolites in response to BPH attack.
This multivariate statistical model revealed different metabolic response patterns between the BPH-susceptible and BPH-resistant varieties during BPH infestation. The metabolite responses of the resistant IL308 variety occurred on Day 1, which was significantly earlier than those of the susceptible KD variety which showed an induced response by Days 4 and 8. BPH infestation caused metabolic perturbations in purine, phenylpropanoid, flavonoid, and terpenoid pathways. While found in both susceptible and resistant rice varieties, schaftoside (1.8 fold), iso-schaftoside (1.7 fold), rhoifolin (3.4 fold) and apigenin 6-C-α-l-arabinoside-8-C-β-l-arabinoside levels (1.6 fold) were significantly increased in the resistant variety by Day 1 post-infestation. 20-hydroxyecdysone acetate (2.5 fold) and dicaffeoylquinic acid (4.7 fold) levels were considerably higher in the resistant rice variety than those in the susceptible variety, both before and after infestation, suggesting that these secondary metabolites play important roles in inducible and constitutive defenses against the BPH infestation.
These potential secondary metabolites will be useful as metabolite markers and/or bioactive compounds for effective and durable approaches to address the BPH problem.
KeywordsLC-HRMS Multi-block principal component analysis Metabolite profiling Brown planthopper resistance Thai Jasmine rice Oryza sativa
This work was financially supported by Platform Technology Program (P-12-01893, P-16-50339 and P-18-50973), National Center for Genetic Engineering and Biotechnology (BIOTEC, Thailand). The PhD scholarship to Umaporn Uawisetwathana was awarded from Graduate and Professional Development Division, National Science and Technology Development Agency (NSTDA, Thailand).
NK, TT, AV, CE and RG conceived and designed research. WK and UU conducted experiments. UU, OPC contributed analytical tools. UU and YX analyzed data. UU, NK, IN, RG, YX, OPC, CE and TP wrote the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
This article does not contain any studies with human subjects.
- Abubakirov, N. K. (1980). New phytoecdysones. In S. N. Ananchenko (Ed.), Frontiers of bioorganic chemistry and molecular biology. Oxford: Pergamon.Google Scholar
- Allwood, J. W., Woznicki, T. L., Xu, Y., Foito, A., Aaby, K., Sungurtas, J., et al. (2019). Application of HPLC–PDA–MS metabolite profiling to investigate the effect of growth temperature and day length on blackcurrant fruit. Metabolomics, 15, 12. https://doi.org/10.1007/s11306-018-1462-5.CrossRefPubMedPubMedCentralGoogle Scholar
- Biais, B., Allwood, J. W., Deborde, C., Xu, Y., Maucourt, M., Beauvoit, B., et al. (2009). 1H NMR, GC-EI-TOFMS, and data set correlation for fruit metabolomics: Application to spatial metabolite analysis in melon. Analytical Chemistry, 81, 2884–2894. https://doi.org/10.1021/ac9001996.CrossRefPubMedGoogle Scholar
- Broadhurst, D., Goodacre, R., Reinke, S. N., Kuligowski, J., Wilson, I. D., Lewis, M. R., et al. (2018). Guidelines and considerations for the use of system suitability and quality control samples in mass spectrometry assays applied in untargeted clinical metabolomic studies. Metabolomics, 14, 72. https://doi.org/10.1007/s11306-018-1367-3.CrossRefPubMedPubMedCentralGoogle Scholar
- Carrizo, D., Chevallier, O. P., Woodside, J. V., Brennan, S. F., Cantwell, M. M., Cuskelly, G., et al. (2017). Untargeted metabolomic analysis of human serum samples associated with exposure levels of Persistent organic pollutants indicate important perturbations in Sphingolipids and Glycerophospholipids levels. Chemosphere, 168, 731–738. https://doi.org/10.1016/j.chemosphere.2016.11.001.CrossRefPubMedGoogle Scholar
- Chaubey, M. K. (2018). Role of phytoecdysteroids in insect pest management: A review. Journal of Agronomy, 17, 1–10.Google Scholar
- Csábi, J., Martins, A., Sinka, I., Csorba, A., Molnár, J., Zupkó, I., et al. (2016). Synthesis and in vitro evaluation of the antitumor potential and chemo-sensitizing activity of fluorinated ecdysteroid derivatives. MedChemComm, 7, 2282–2289. https://doi.org/10.1039/C6MD00431H.CrossRefGoogle Scholar
- Dunn, W. B., Broadhurst, D., Begley, P., Zelena, E., Francis-Mcintyre, S., Anderson, N., et al. (2011). Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nature Protocols, 6, 1060–1083.CrossRefGoogle Scholar
- Grayer, R. J., Harborne, J. B., Kimmins, F. M., Stevenson, P. C., & Wijayagunasekera, H. N. P. (1994). Phenolics in rice phloem sap as sucking deterrents to the brown planhopper, Nilapavata lugens (381st ed.). Leuven: International Society for Horticultural Science (ISHS).Google Scholar
- Hamilton, M. L., Kuate, S. P., Brazier-Hicks, M., Caulfield, J. C., Rose, R., Edwards, R., et al. (2012). Elucidation of the biosynthesis of the di-C-glycosylflavone isoschaftoside, an allelopathic component from Desmodium spp. that inhibits Striga spp. development. Phytochemistry, 84, 169–176. https://doi.org/10.1016/j.phytochem.2012.08.005.CrossRefPubMedGoogle Scholar
- Hao, P.-Y., Feng, Y.-L., Zhou, Y.-S., Song, X.-M., Li, H.-L., Ma, Y., et al. (2018). Schaftoside interacts with NlCDK1 protein: A mechanism of rice resistance to brown planthopper, Nilaparvata lugens. Frontiers in Plant Science. https://doi.org/10.3389/fpls.2018.00710.CrossRefPubMedPubMedCentralGoogle Scholar
- Harrison, H., Mitchell, T. R., Peterson, J. K., Wechter, W. P., Majetich, G. F., & Snook, M. E. (2008). Contents of caffeoylquinic acid compounds in the storage roots of sixteen sweetpotato genotypes and their potential biological activity. Journal of the American Society for Horticultural Science, 133, 492–500.CrossRefGoogle Scholar
- Hauck, O. K., Scharnberg, J., Escobar, N. M., Wanner, G., Giavalisco, P., & Witte, C.-P. (2014). Uric acid accumulation in an arabidopsis urate oxidase mutant impairs seedling establishment by blocking peroxisome maintenance. The Plant Cell, 26, 3090. https://doi.org/10.1105/tpc.114.124008.CrossRefPubMedPubMedCentralGoogle Scholar
- Heinrichs, E. A. (1979). Brown planthopper: Threat to rice production in Asia. In E. A. Heinrichs (Ed.), Chemical control of the brown planthopper. Los Baños: International Rice Research Institute.Google Scholar
- Heong, K. L., & Hardy, B. (2009). Planthopper: New treats to the sustainability of intensive rice production systems in Asia. Los Banos (Philippines): International Rice Research Institute.Google Scholar
- Jairin, J., Teangdeerith, S., Leelagud, P., Kothcharerk, J., Sansen, K., Yi, M., et al. (2009). Development of rice introgression lines with brown planthopper resistance and KDML105 grain quality characteristics through marker-assisted selection. Field Crops Research, 110, 263–271. https://doi.org/10.1016/j.fcr.2008.09.009.CrossRefGoogle Scholar
- Kamolsukyeunyong, W., Ruengphayak, S., Chumwong, P., Kusumawati, L., Chaichoompu, E., Jamboonsri, W., et al. (2019). Identification of spontaneous mutation for broad-spectrum brown planthopper resistance in a large, long-term fast neutron mutagenized rice population. Rice, 12, 1–26. https://doi.org/10.1186/s12284-019-0274-1.CrossRefGoogle Scholar
- Kang, K., Yue, L., Xia, X., Liu, K., & Zhang, W. (2019). Comparative metabolomics analysis of different resistant rice varieties in response to the brown planthopper Nilaparvata lugens Hemiptera: Delphacidae. Metabolomics, 15, 62. https://doi.org/10.1007/s11306-019-1523-4.CrossRefPubMedPubMedCentralGoogle Scholar
- Kessler, A., & Baldwin, I. T. (2002). Plant responses to insect herbivory: The emerging molecular analysis. Annual Review of Plant Biology. https://doi.org/10.1146/annurev.arplant.53.100301.135207.CrossRefPubMedGoogle Scholar
- Kuijpers, T. F. M., Narváez-Cuenca, C.-E., Vincken, J.-P., Verloop, A. J. W., Van Berkel, W. J. H., & Gruppen, H. (2012). Inhibition of enzymatic browning of chlorogenic acid by sulfur-containing compounds. Journal of Agricultural and Food Chemistry, 60, 3507–3514. https://doi.org/10.1021/jf205290w.CrossRefPubMedGoogle Scholar
- Kusumawati, L., Chumwong, P., Jamboonsri, W., Wanchana, S., Siangliw, J. L., Siangliw, M., et al. (2018). Candidate genes and molecular markers associated with brown planthopper (Nilaparvata lugens Stål) resistance in rice cultivar Rathu Heenati. Molecular Breeding, 38, 88. https://doi.org/10.1007/s11032-018-0847-5.CrossRefGoogle Scholar
- Li, X., Grand, L., Pouleriguen, T., Queneau, Y., Da Silva, P., Rahbé, Y., et al. (2016). Synthesis of new dicinnamoyl 4-deoxy quinic acid and methyl ester derivatives and evaluation of the toxicity against the pea aphid Acyrthosiphon pisum. Organic & Biomolecular Chemistry, 14, 2487–2497. https://doi.org/10.1039/C5OB02483H.CrossRefGoogle Scholar
- Li, Z., Xue, Y., Zhou, H., Li, Y., Usman, B., Jiao, X., et al. (2019). High-resolution mapping and breeding application of a novel brown planthopper resistance gene derived from wild rice (Oryza. rufipogon Griff). Rice, 12, 41. https://doi.org/10.1186/s12284-019-0289-7.CrossRefPubMedPubMedCentralGoogle 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, 6774–6785. https://doi.org/10.1021/pr100970q.CrossRefPubMedGoogle Scholar
- Martinez, V., Mestre, T. C., Rubio, F., Girones-Vilaplana, A., Moreno, D. A., Mittler, R., et al. (2016). Accumulation of flavonols over hydroxycinnamic acids favors oxidative damage protection under abiotic stress. Frontiers in Plant Science, 7, 838. https://doi.org/10.3389/fpls.2016.00838.CrossRefPubMedPubMedCentralGoogle Scholar
- Matsui, T., Ebuchi, S., Fujise, T., Abesundara, K. J. M., Doi, S., Yamada, H., et al. (2004). Strong antihyperglycemic effects of water-soluble fraction of brazilian propolis and its bioactive constituent, 3,4,5-Tri-O-caffeoylquinic acid. Biological and Pharmaceutical Bulletin, 27, 1797–1803. https://doi.org/10.1248/bpb.27.1797.CrossRefPubMedGoogle Scholar
- Moglia, A., Lanteri, S., Comino, C., Hill, L., Knevitt, D., Cagliero, C., et al. (2014). Dual catalytic activity of hydroxycinnamoyl-coenzyme A quinate transferase from tomato allows it to moonlight in the synthesis of both mono- and dicaffeoylquinic acids. Plant Physiology, 166, 1777–1787. https://doi.org/10.1104/pp.114.251371.CrossRefPubMedPubMedCentralGoogle Scholar
- O’kane, A. A., Chevallier, O. P., Graham, S. F., Elliott, C. T., & Mooney, M. H. (2013). Metabolomic profiling of in vivo plasma responses to dioxin-associated dietary contaminant exposure in rats: Implications for identification of sources of animal and human exposure. Environmental Science and Technology, 47, 5409–5418. https://doi.org/10.1021/es305345u.CrossRefPubMedGoogle Scholar
- Pitija, K., Kamolsukyumyong, W., Vanavichit, A., Sookwong, P., & Mahatheeranont, S. (2014). Monoterpenoid allelochemicals in resistance rice varieties against brown planthoppers, Nilaparvata Lugens (Stål). Journal of Advanced Agricultural Technologies, 1, 7. https://doi.org/10.12720/joaat.1.2.82-88.CrossRefGoogle Scholar
- Refaat, J., Desoukey, S. Y., Ramadan, M. A., & Kamel, M. S. (2015). Rhoifolin: A review of sources and biological activities. International Journal of Pharmacognosy, 2, 102–109.Google Scholar
- Ren, J., Gao, F., Wu, X., Lu, X., Zeng, L., Lv, J., et al. (2016). Bph32, a novel gene encoding an unknown SCR domain-containing protein, confers resistance against the brown planthopper in rice. Scientific Reports, 6, 37645. https://doi.org/10.1038/srep37645.CrossRefPubMedPubMedCentralGoogle Scholar
- Santos, I. C. D., Almeida, A. A. F. D., Pirovani, C. P., Costa, M. G. C., Silva, M. F. D. G. F. D., Bellete, B. S., et al. (2017). Differential accumulation of flavonoids and phytohormones resulting from the canopy/rootstock interaction of citrus plants subjected to dehydration/rehydration. Plant Physiology and Biochemistry, 119, 147–158. https://doi.org/10.1016/j.plaphy.2017.08.019.CrossRefPubMedGoogle Scholar
- Stevenson, P. C., Kimmins, F. M., Grayer, R. J., & Raveendranath, S. (1996). Schaftosides from rice phloem as feeding inhibitors and resistance factors to brown planthoppers, Nilaparvata lugens. In E. Städler, M. Rowell-Rahier, & R. Bauer (Eds.), Proceedings of the 9th International Symposium on Insect-Plant Relationships. Dordrecht: Springer.Google Scholar
- Sumner, L. W., Amberg, A., Barrett, D., Beale, M. H., Beger, R., Daykin, C. A., et al. (2007). Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics: Official Journal of the Metabolomic Society, 3, 211–221. https://doi.org/10.1007/s11306-007-0082-2.CrossRefGoogle Scholar
- Tenango, M. P., Hernández, M. S., & Hernández, E. A. (2017). Flavonoids in agriculture, flavonoids—from biosynthesis to human health. IntechOpen.Google Scholar
- Uawisetwathana, U., Graham, S., Kamolsukyunyong, W., Sukhaket, W., Klanchui, A., Toojinda, T., et al. (2015). Quantitative 1H NMR metabolome profiling of Thai Jasmine rice (Oryza sativa) reveals primary metabolic response during brown planthopper infestation. Metabolomics, 11, 1640–1655. https://doi.org/10.1007/s11306-015-0817-4.CrossRefGoogle Scholar
- Xu, Y., Correa, E., & Goodacre, R. (2013). Integrating multiple analytical platforms and chemometrics for comprehensive metabolic profiling: Application to meat spoilage detection. Analytical and Bioanalytical Chemistry, 405, 5063–5074. https://doi.org/10.1007/s00216-013-6884-3.CrossRefPubMedGoogle Scholar