Metabolomics

, 13:51 | Cite as

The use of metabolome analysis to identify the cause of an unexplained disease of Japanese gentians (Gentiana triflora)

  • Hideyuki Takahashi
  • Hiroshi Abe
  • Kohei Fujita
  • Ken-Taro Sekine
Original Article
  • 180 Downloads

Abstract

Introduction

Gentian spotted bleaching disease (GSBD), a novel disease of unknown etiology, affects Gentiana triflora plants that are cultivated as ornamental flowers in Japan. This disease leads to the production of necrotic leaf spots, a delay in flowering, and has thus become a serious problem for gentian production.

Objectives

The objective of this study was to identify the cause of GSBD in G. triflora by analyzing differences between healthy and GSBD-affected leaves.

Method

Selected metabolite concentrations in healthy and GSBD-affected leaves were quantified using capillary electrophoresis and liquid chromatography-mass spectrometry, and statistically significant differences in metabolite concentrations were assessed. GSBD-affected metabolic pathways were identified followed by examination of pathway-related gene expression and enzyme activities. Furthermore, the effects of root hypoxia on metabolite concentrations and gene expression were investigated.

Results

We found that concentrations of Calvin cycle intermediates and ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) activity were significantly lower in GSBD-affected leaves, whereas sucrose cleavage and Ala accumulation were enhanced. Since these metabolic changes are frequently observed in plants exposed to hypoxia, the expression of hypoxia-responsive genes was investigated. Expression levels of hypoxia-responsive genes were higher in GSBD-affected plants than in the controls. Furthermore, root hypoxia induced similar symptoms and metabolic changes as those observed in GSBD-affected plants.

Conclusion

Our results indicate that GSBD was likely induced by root hypoxia and that metabolome analysis is an effective tool for identifying the cause of plant disease with unknown etiologies.

Keywords

Gentiana triflora Hypoxia Mass spectrometry Physiological disorder Targeted metabolome analysis 

Supplementary material

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Supplementary material 5 (XLSX 59 KB)

References

  1. Abdel-Latif, A. (2008). Activity of sucrose synthase and acid invertase in wheat seedlings during a cold-shock using micro plate reader assays. Australian Journal of Basic and Applied Sciences, 2, 53–56.Google Scholar
  2. Armengaud, P., Sulpice, R., Miller, A. J., Stitt, M., Amtmann, A., & Gibon, Y. (2009). Multilevel analysis of primary metabolism provides new insights into the role of potassium nutrition for glycolysis and nitrogen assimilation in Arabidopsis roots. Plant Physiology, 150, 772–785.CrossRefPubMedPubMedCentralGoogle Scholar
  3. Ashraf, M., & Harris, P. J. C. (2013). Photosynthesis under stressful environments: An overview. Photosynthetica, 51, 163–190.CrossRefGoogle Scholar
  4. Atsumi, G., Tomita, R., Kobayashi, K., & Sekine, K. T. (2013). Establishment of an agroinoculation system for broad bean wilt virus 2. Archives of Virology, 158, 1549–1554.CrossRefPubMedGoogle Scholar
  5. Atsumi, G., Tomita, R., Yamashita, T., & Sekine, K. T. (2015). A novel virus transmitted through pollination causes ring-spot disease on gentian (Gentiana triflora) ovaries. The Journal of General Virology, 96, 431–439.CrossRefPubMedGoogle Scholar
  6. Bailey-Serres, J., & Voesenek, L. A. (2008). Flooding stress: acclimations and genetic diversity. Annual Review of Plant Biology, 59, 313–339.CrossRefPubMedGoogle Scholar
  7. Bieniawska, Z., Paul Barratt, D. H., Garlick, A. P., Thole, V., Kruger, N. J., Martin, C., et al. (2007). Analysis of the sucrose synthase gene family in Arabidopsis. The Plant Journal, 49, 810–828.CrossRefPubMedGoogle Scholar
  8. Cantu, M. D., Mariano, A. G., Palma, M. S., Carrilho, E., & Wulff, N. A. (2008). Proteomic analysis reveals suppression of bark chitinases and proteinase inhibitors in citrus plants affected by the citrus sudden death disease. Phytopathology, 98, 1084–1092.CrossRefPubMedGoogle Scholar
  9. Chaves, M. M., Flexas, J., & Pinheiro, C. (2009). Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Annals of Botany, 103, 551–560.CrossRefPubMedGoogle Scholar
  10. Chen, Y., Chen, X., Wang, H., Bao, Y., & Zhang, W. (2014). Examination of the leaf proteome during flooding stress and the induction of programmed cell death in maize. Proteome Science, 12, 33.CrossRefPubMedPubMedCentralGoogle Scholar
  11. Ciereszko, I., & Kleczkowski, L. A. (2005). Expression of several genes in volved in sucrose/starch metabolism as affected by different strategies to induce phosphate deficiency in Arabidopsis. Acta Physiologiae Plantarum, 27, 147–155.CrossRefGoogle Scholar
  12. Cramer, G. R., Urano, K., Delrot, S., Pezzotti, M., & Shinozaki, K. (2011). Effects of abiotic stress on plants: A systems biology perspective. BMC Plant Biology, 11, 163.CrossRefPubMedPubMedCentralGoogle Scholar
  13. Else, M. A., Janowiak, F., Atkinson, C. J., & Jackson, M. B. (2009). Root signals and stomatal closure in relation to photosynthesis, chlorophyll a fluorescence and adventitious rooting of flooded tomato plants. Annals of Botany, 103, 313–323.CrossRefPubMedGoogle Scholar
  14. Gimenez, C., Mitchell, V. J., & Lawlor, D. W. (1992). Regulation of photosynthetic rate of two sunflower hybrids under water stress. Plant Physiology, 98, 516–524.CrossRefPubMedPubMedCentralGoogle Scholar
  15. Herbers, K., Meuwly, P., Frommer, W. B., Metraux, J. P., & Sonnewald, U. (1996). Systemic acquired resistance mediated by the ectopic expression of invertase: Possible hexose sensing in the secretory pathway. The Plant cell, 8, 793–803.CrossRefPubMedPubMedCentralGoogle Scholar
  16. Hsu, F. C., Chou, M. Y., Peng, H. P., Chou, S. J., & Shih, M. C. (2011). Insights into hypoxic systemic responses based on analyses of transcriptional regulation in Arabidopsis. PLOS ONE, 6, e28888.CrossRefPubMedPubMedCentralGoogle Scholar
  17. Imamura, T., Higuchi, A., Sekine, K. T., Yamashita, T., & Takahashi, H. (2015). High concentrations of sucrose induce overwintering bud formation in gentian plantlets cultured in vitro. Plant Biotechnology, 31, 97–104.CrossRefGoogle Scholar
  18. Kim, J. Y., Mahe, A., Brangeon, J., & Prioul, J. L. (2000). A maize vacuolar invertase, IVR2, is induced by water stress. Organ/tissue specificity and diurnal modulation of expression. Plant Physiology, 124, 71–84.CrossRefPubMedPubMedCentralGoogle Scholar
  19. Kobayashi, K., Atsumi, G., Iwadate, Y., Tomita, R., Chiba, K., Akasaka, S., et al. (2013). Gentian Kobu-sho-associated virus: A tentative, novel double-strand RNA virus that is relevant to gentian Kobu-sho syndrome. Journal of General Plant Pathology, 79, 56–63.CrossRefGoogle Scholar
  20. Kobayashi, K., Atsumi, G., Yamaoka, N., & Sekine, K. T. (2012). Sequencing-based virus hunting and virus detection. Japan Agricultural Research Quarterly, 46, 123–128.CrossRefGoogle Scholar
  21. Kocal, N., Sonnewald, U., & Sonnewald, S. (2008). Cell wall-bound invertase limits sucrose export and is involved in symptom development and inhibition of photosynthesis during compatible interaction between tomato and Xanthomonas campestris pv vesicatoria. Plant Physiology, 148, 1523–1536.CrossRefPubMedPubMedCentralGoogle Scholar
  22. Koch, K. (2004). Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology, 7, 235–246.CrossRefPubMedGoogle Scholar
  23. Krasensky, J., & Jonak, C. (2012). Drought, salt, and temperature stress-induced metabolic rearrangements and regulatory networks. Journal of Experimental Botany, 63, 1593–1608.CrossRefPubMedPubMedCentralGoogle Scholar
  24. Kreuzwieser, J., Hauberg, J., Howell, K. A., Carroll, A., Rennenberg, H., Millar, A. H., et al. (2009). Differential response of gray poplar leaves and roots underpins stress adaptation during hypoxia. Plant Physiology, 149, 461–473.CrossRefPubMedPubMedCentralGoogle Scholar
  25. Mehta, P., Jajoo, A., Mathur, S., & Bharti, S. (2010). Chlorophyll a fluorescence study revealing effects of high salt stress on Photosystem II in wheat leaves. Plant Physiology and Biochemistry, 48, 16–20.CrossRefPubMedGoogle Scholar
  26. Motohashi, R., & Myouga, F. (2015). Chlorophyll fluorescence measurements in Arabidopsis plants using a pulse-amplitude-modulated (PAM) fluorometer. Bio-protocol, 5, e1464. http://www.bio-protocol.org/e1464.Google Scholar
  27. Nakatsuka, T., Yamada, E., Saito, M., Fujita, K., & Nishihara, M. (2013). Heterologous expression of gentian MYB1R transcription factors suppresses anthocyanin pigmentation in tobacco flowers. Plant Cell Reports, 32, 1925–1937.CrossRefPubMedGoogle Scholar
  28. Narsai, R., Rocha, M., Geigenberger, P., Whelan, J., & van Dongen, J. T. (2011). Comparative analysis between plant species of transcriptional and metabolic responses to hypoxia. The New phytologist, 190, 472–487.CrossRefPubMedGoogle Scholar
  29. Nouri, M. Z., Moumeni, A., & Komatsu, S. (2015). Abiotic stresses: insight into gene regulation and protein expression in photosynthetic pathways of plants. International Journal of Molecular Sciences, 16, 20392–20416.CrossRefPubMedPubMedCentralGoogle Scholar
  30. Rocha, M., Licausi, F., Araujo, W. L., Nunes-Nesi, A., Sodek, L., Fernie, A. R., et al. (2010). Glycolysis and the tricarboxylic acid cycle are linked by alanine aminotransferase during hypoxia induced by waterlogging of Lotus japonicus. Plant Physiology, 152, 1501–1513.CrossRefPubMedPubMedCentralGoogle Scholar
  31. Sturm, A., & Tang, G. Q. (1999). The sucrose-cleaving enzymes of plants are crucial for development, growth and carbon partitioning. Trends in Plant Science, 4, 401–407.CrossRefPubMedGoogle Scholar
  32. Takahashi, H., Fujita, K., Yoshida, C., & Nishihara, M. (2016). Metabolite profiling reveals the involvement of aberrant metabolic changes in Gentiana triflora seed showing poor germination. Journal of Horticultural Science & Biotechnology, 91, 148–155.CrossRefGoogle Scholar
  33. Takahashi, H., Imamura, T., Konno, N., Takeda, T., Fujita, K., Konishi, T., et al. (2014). The gentio-oligosaccharide gentiobiose functions in the modulation of bud dormancy in the herbaceous perennial Gentiana. The Plant cell, 26, 3949–3963.CrossRefPubMedPubMedCentralGoogle Scholar
  34. Takahashi, H., Imamura, T., Miyagi, A., & Uchimiya, H. (2012). Comparative metabolomics of developmental alterations caused by mineral deficiency during in vitro culture of Gentiana triflora. Metabolomics, 8, 154–163.CrossRefGoogle Scholar
  35. Takahashi, H., Takahara, K., Hashida, S. N., Hirabayashi, T., Fujimori, T., Kawai-Yamada, M., et al. (2009). Pleiotropic modulation of carbon and nitrogen metabolism in Arabidopsis plants overexpressing the NAD kinase2 gene. Plant Physiology, 151, 100–113.CrossRefPubMedPubMedCentralGoogle Scholar
  36. Takahashi, H., Watanabe, A., Tanaka, A., Hashida, S. N., Kawai-Yamada, M., Sonoike, K., et al. (2006). Chloroplast NAD kinase is essential for energy transduction through the xanthophyll cycle in photosynthesis. Plant and Cell Physiology, 47, 1678–1682.CrossRefPubMedGoogle Scholar
  37. Tauzin, A. S., & Giardina, T. (2014). Sucrose and invertases, a part of the plant defense response to the biotic stresses. Frontiers in Plant Science, 5, 293.CrossRefPubMedPubMedCentralGoogle Scholar
  38. Wang, G., Kong, F., Zhang, S., Meng, X., Wang, Y., & Meng, Q. (2015). A tomato chloroplast-targeted DnaJ protein protects Rubisco activity under heat stress. Journal of experimental botany, 66, 3027–3040.CrossRefPubMedGoogle Scholar
  39. Waraich, E. A., Ahmad, R., Halim, A., & Aziz, T. (2012). Alleviation of temperature stress by nutrient management in crop plants: a review. Journal of Soil Science and Plant Nutrition, 12, 221–244.CrossRefGoogle Scholar
  40. Yanagisawa, H., Tomita, R., Katsu, K., Uehara, T., Atsumi, G., Tateda, C., et al. (2016). Combined DECS analysis and next-generation sequencing enable efficient detection of novel plant RNA viruses. Viruses, 8(3), 70CrossRefPubMedPubMedCentralGoogle Scholar
  41. Yang, J. Y., Zheng, W., Tian, Y., Wu, Y., & Zhou, D. W. (2011). Effects of various mixed salt-alkaline stresses on growth, photosynthesis, and photosynthetic pigment concentrations of Medicago ruthenica seedlings. Photosynthetica, 49, 275–284.CrossRefGoogle Scholar
  42. Yordanova, R. Y., & Popova, L. P. (2007). Flooding-induced changes in photosynthesis and oxidative status in maize plants. Acta Physiologiae Plantarum, 29, 535–541.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Hideyuki Takahashi
    • 1
  • Hiroshi Abe
    • 2
  • Kohei Fujita
    • 1
  • Ken-Taro Sekine
    • 1
  1. 1.Iwate Biotechnology Research CenterKitakamiJapan
  2. 2.Iwate Agricultural Research CenterKitakamiJapan

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