Vol.:(0123456789) 1 3
J Plant Res
DOI 10.1007/s10265-017-0923-7
REGULAR PAPER
Synthesis and degradation of long-chain base phosphates affect
fumonisin B
1
-induced cell death in Arabidopsis thaliana
Daiki Yanagawa
1,2
· Toshiki Ishikawa
3
· Hiroyuki Imai
1,2
Received: 10 October 2016 / Accepted: 7 December 2016
© The Botanical Society of Japan and Springer Japan 2017
LCBK1-OX and -KD plants showed resistance and sensi-
tivity to FB
1
, respectively, whereas free LCB and LCBP
levels in FB
1
-treated LCBK1-OX and -KD plants were
moderately different to those in FB
1
-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 FB
1
.
Keywords Fumonisin B
1
· Long-chain base kinase ·
Long-chain base phosphates · LC-MS/MS · Sphingolipid
catabolism
Abbreviations
CaMV35S Cauliflower mosaic virus 35S
Cer Ceramide
DPL LCBP lyase
d17:1 C
17
-E-4-sphingenine
d17:1-P C
17
-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
FB
1
Fumonisin B
1
GlcCer Glucosylceramide
GIPC Glycosyl inositolphosphoceramide
HPLC High-performance liquid chromatography
HR Hypersensitive response
LC Liquid chromatography
LCB Long-chain base
Abstract Fumonisin B
1
(FB
1
), 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 FB
1
toxicity in plants is connected with preventing the
accumulation of free LCBs through their phosphorylation.
Here a workflow for the extraction, detection and quantifi-
cation 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 degrada-
tion of LCBPs, LCB kinase (LCBK1), LCBP phosphatase
(SPP1) and lyase (DPL1) on FB
1
-induced cell death. As
expected, large accumulations of saturated free LCBs,
dihydrosphingosine and phytosphingosine, were observed
in the FB
1
-treated leaves. On the other hand, a high level of
sphingenine phosphate was found in the FB
1
-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 FB
1
-induced cell death determined by trypan blue stain-
ing. The FB
1
-treated leaves in dpl1 plants showed severe
cell death and the elevation of free LCBs and LCBPs.
Electronic supplementary material The online version of this
article (doi:10.1007/s10265-017-0923-7) contains supplementary
material, which is available to authorized users.
* Hiroyuki Imai
imai@konan-u.ac.jp
1
Department of Biology, Graduate School of Natural Science,
Konan University, Kobe 658-8501, Japan
2
The Institute for Integrative Neurobiology, Konan University,
Kobe 658-8501, Japan
3
Graduate School of Science and Engineering, Saitama
University, Saitama 338-8570, Japan