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Mass spectrometry: the indispensable tool for plant metabolomics of colourless chlorophyll catabolites

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Abstract

Senescence and ripening of plant tissues engage the pheophorbide a oxygenase pathway, reducing the chlorophyll content to inactive chlorophyll catabolite products, termed phyllobilins. These products are open-macrocycle derivatives, but present different structural features related to species-dependent enzyme activity. This review encompasses a brief outline of the chlorophyll catabolism pathway, a detailed description of the structural motifs of known phyllobilins, giving details of how mass spectrometry provides hints for the characterization of phyllobilins. The structural approach for the identification of phyllobilins requires several spectroscopic methodologies to reach a complete structural identification, including UV–visible spectroscopy, circular dichroism, nuclear magnetic resonance and mass spectrometry. Among these techniques, mass spectrometry presents several advantages for showing the structural features of phyllobilins, through acquisition of accurate mass, elemental composition, and detection of product ions, which provide valuable structural information. The combination of mass spectra with data-managing and in silico prediction tools greatly enhances the comprehensive building of the phyllobilin structure, and the resolving of the intriguing puzzle of enzymatic and chemical reactions involved in chlorophyll catabolism. Indeed, some strategies based on structural constraints that phyllobilins present, with recent developments in software prediction tools are proposed to foster the unravelling of phyllobilin structures.

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Abbreviations

PaO:

Pheophorbide a oxygenase

NCC:

Non-fluorescent chlorophyll catabolite

RCCR:

Red chlorophyll catabolite reductase

FCC:

Fluorescent chlorophyll catabolite

DFCC:

Dioxobilin-type fluorescent chlorophyll catabolite

DNCC:

Dioxobilin-type non-fluorescent chlorophyll catabolite

DESI:

Desorption electrospray ionization

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Acknowledgements

This work was supported by the Comisión Interministerial de Ciencia y Tecnología (CICYT-EU, Spanish and European Government, AGL 2015-63890-R). All the authors contributed equally to the performance and writing of this review.

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Authors and Affiliations

  1. Food Phytochemistry Department, Instituto de la Grasa (CSIC), Campus Universitario, Building 46, 41013, Seville, Spain

    María Roca & Antonio Pérez-Gálvez

  2. Laboratory of Mass Spectrometry, Instituto de la Grasa (CSIC), Campus Universitario, Building 46, 41013, Seville, Spain

    José J. Ríos

Authors
  1. María Roca
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  2. José J. Ríos
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Corresponding author

Correspondence to Antonio Pérez-Gálvez.

Appendix

Appendix

Figures corresponding to the tandem mass spectra of some representative phyllobilins (see Figs. 5, 6 and 7).

Fig. 5
figure 5

MS2 of [M + H]+ at m/z = 807.3447 Da (Zm-NCC2 and its structural equivalents in Table 1). Some of the structural features arise from the characteristic fragmentations described in Table 2: 32 Da, methylation at O84 (at m/z = 775 Da [M + H-MeOH]+); 123 Da, ring D presents the 181,182-vinyl arrangement (at m/z = 683 Da [M + H-ring D]+); 155 Da, ring A is hydroxylated at C32; 285 Da, presence of ring D-β-glucopyranoyl

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Fig. 6
figure 6

MS2 of [M + H]+ at m/z = 731.29237 Da (Ej-NCC2 in Table 1). Some of the structural features arise from the characteristic fragmentations described in Table 2: 88 Da, The structure presents a malonyl group and it is hydroxylated at C32 (at m/z = 643 Da [M + H-malonyl]+); 209 Da, ring D presents the 181,182-vinyl arrangement and it is hydroxylated at C32 (at m/z = 522 Da [M + H-ring D-malonyl]+)

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Fig. 7
figure 7

MS2 of [M + H]+ at m/z = 667.2974 Da (UCC and its structural equivalents in Table 1). Some of the structural features arise from the characteristic fragmentations described in Table 2: 18 Da, presence of hydroxyl group (at m/z = 649 Da [M + H-H2O]+); 32 Da, structure is methylated at O84 (at m/z = 649 Da [M + H-MeOH]+); 157 Da, ring D presents the 181,182-dihydroxyethyl arrangement (at m/z = 510 Da [M + H-ring D]+)

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Roca, M., Ríos, J.J. & Pérez-Gálvez, A. Mass spectrometry: the indispensable tool for plant metabolomics of colourless chlorophyll catabolites. Phytochem Rev 17, 453–468 (2018). https://doi.org/10.1007/s11101-017-9543-z

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