Abstract
Fumonisin B1 (FB1), an inducer of cell death, disrupts sphingolipid metabolism; large accumulations of de novo synthesized free long-chain bases (LCBs) are observed. However, it remains unclear whether tolerance to FB1 toxicity in plants is connected with preventing the accumulation of free LCBs through their phosphorylation. Here a workflow for the extraction, detection and quantification of LCB phosphates (LCBPs) in Arabidopsis thaliana was developed. We studied the effect of expression of genes for three enzymes involved in the synthesis and degradation of LCBPs, LCB kinase (LCBK1), LCBP phosphatase (SPP1) and lyase (DPL1) on FB1-induced cell death. As expected, large accumulations of saturated free LCBs, dihydrosphingosine and phytosphingosine, were observed in the FB1-treated leaves. On the other hand, a high level of sphingenine phosphate was found in the FB1-treated leaves even though free sphingenine was found in low amounts in these leaves. In comparison of WT and spp1 plants, the LCBP/LCB ratio is likely to be correlated with the degree of FB1-induced cell death determined by trypan blue staining. The FB1-treated leaves in dpl1 plants showed severe cell death and the elevation of free LCBs and LCBPs. LCBK1-OX and -KD plants showed resistance and sensitivity to FB1, respectively, whereas free LCB and LCBP levels in FB1-treated LCBK1-OX and -KD plants were moderately different to those in FB1-treated WT plants. Overall, the findings described here suggest that LCBP/LCB homeostasis is an important topic that participates in the tolerance of plant cells to FB1.
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Abbreviations
- CaMV35S:
-
Cauliflower mosaic virus 35S
- Cer:
-
Ceramide
- DPL:
-
LCBP lyase
- d17:1:
-
C17-E-4-sphingenine
- d17:1-P:
-
C17-E-4-sphingenine-1-phosphate
- d18:0:
-
Sphinganine (dihydrosphingosine)
- d18:0-P:
-
Sphinganine (dihydrosphingosine)-1-phosphate
- d18:1(4E) :
-
E-4-sphingenine (sphingosine)
- d18:1(4E)-P:
-
E-4-sphingenine (sphingosine)-1-phosphate
- d18:1(8Z):
-
Z-8-sphingenine
- d18:1(8Z)-P:
-
Z-8-sphingenine-1-phosphate
- d18:1(8E):
-
E-8-sphingenine
- d18:1(8E)-P:
-
E-8-sphingenine-1-phosphate
- ESI:
-
Electrospray ionization
- FB1 :
-
Fumonisin B1
- GlcCer:
-
Glucosylceramide
- GIPC:
-
Glycosyl inositolphosphoceramide
- HPLC:
-
High-performance liquid chromatography
- HR:
-
Hypersensitive response
- LC:
-
Liquid chromatography
- LCB:
-
Long-chain base
- LCBP:
-
Long-chain base 1-phosphate
- LCBK:
-
LCB kinase
- MRM:
-
Multiple reaction monitoring
- MS/MS:
-
Tandem mass spectrometry
- NBD-F:
-
4-fluoro-7-nitrobenzofurazan
- ROI:
-
Reactive oxygen intermediate
- SPP:
-
LCBP phosphatase
- t18:0:
-
4-hydroxy-sphinganine (phytosphingosine)
- t18:0-P:
-
4-hydroxy-sphinganine (phytosphingosine)-1-phosphate
- t18:1(8Z):
-
4-hydroxy-Z-8-sphingenine
- t18:1(8Z)-P:
-
4-hydroxy-Z-8-sphingenine-1-phosphate
- t18:1(8E):
-
4-hydroxy-E-8-sphingenine
- t18:1(8E)-P:
-
4-hydroxy-E-8-sphingenine-1-phosphate
- WT:
-
Wild type
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Acknowledgements
We are grateful to Prof. Ikuko Nishimura (Konan University) for helpful advice on cell death data.
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Parts of this work were supported by Special Ordinary Expense Subsidies for Private Universities from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan. This research was also supported in part by the Strategic Research Foundation Grant-aided Project for Private Universities from MEXT of Japan.
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Yanagawa, D., Ishikawa, T. & Imai, H. Synthesis and degradation of long-chain base phosphates affect fumonisin B1-induced cell death in Arabidopsis thaliana . J Plant Res 130, 571–585 (2017). https://doi.org/10.1007/s10265-017-0923-7
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DOI: https://doi.org/10.1007/s10265-017-0923-7