High-resolution spatial and temporal analysis of phytoalexin production in oats

被引:18
作者
Izumi, Yoshihiro [2 ]
Kajiyama, Shin'ichiro [1 ,2 ]
Nakamura, Ryosuke [4 ]
Ishihara, Atsushi [3 ]
Okazawa, Atsushi [2 ]
Fukusaki, Eiichiro [2 ]
Kanematsu, Yasuo [4 ]
Kobayashi, Akio [2 ]
机构
[1] Kinki Univ, Sch Biol Oriented Sci & Technol, Wakayama 6496493, Japan
[2] Osaka Univ, Grad Sch Engn, Dept Biotechnol, Suita, Osaka 5650871, Japan
[3] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Kyoto 6068502, Japan
[4] Osaka Univ, Ctr Adv Sci & Innovat, Venture Business Lab, JST CREST, Suita, Osaka 5650871, Japan
关键词
Phytoalexin production; Oats; High-resolution spatial and temporal analysis; Laser-assisted single-cell sampling; Low-diffuse nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry (LC-ESI-MS/MS); Line-scanning fluorescence microscopy; MAGNETIC-RESONANCE-SPECTROSCOPY; LASER CAPTURE MICRODISSECTION; PLANT-DISEASE RESISTANCE; MASS-SPECTROMETRY; CELL-DEATH; MESSENGER-RNAS; ARABIDOPSIS; AVENANTHRAMIDES; METABOLISM; EXPRESSION;
D O I
10.1007/s00425-008-0887-x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
The production of oat (Avena sativa L.) phytoalexins, avenanthramides, occurs in response to elicitor treatment with oligo-N-acetylchitooligosaccharides. In this study, avenanthramides production was investigated by techniques that provide high spatial and temporal resolution in order to clarify the process of phytoalexin production at the cellular level. The amount of avenanthramides accumulation in a single mesophyll cell was quantified by a combination of laser micro-sampling and low-diffuse nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) techniques. Avenanthramides, NAD(P)H and chlorophyll were also visualized in elicitor-treated mesophyll cells using line-scanning fluorescence microscopy. We found that elicitor-treated mesophyll cells could be categorized into three characteristic cell phases, which occurred serially over time. Phase 0 indicated the normal cell state before metabolic or morphological change in response to elicitor, in which the cells contained abundant NAD(P)H. In phase 1, rapid NAD(P)H oxidation and marked movement of chloroplasts occurred, and this phase was the early stage of avenanthramides biosynthesis. In phase 2, avenanthramides accumulation was maximized, and chloroplasts were degraded. Avenanthramides appear to be synthesized in the chloroplast, because a fluorescence signal originating from avenanthramides was localized to the chloroplasts. Moreover, our results indicated that avenanthramides biosynthesis and the hypersensitive response (HR) occurred in identical cells. Thus, the avenanthramides production may be one of sequential events programmed in HR leading to cell death. Furthermore, the phase of the defense response was different among mesophyll cells simultaneously treated with elicitor. These results suggest that individual cells may have different susceptibility to the elicitor.
引用
收藏
页码:931 / 943
页数:13
相关论文
共 46 条
[1]   Reactive oxygen species: Metabolism, oxidative stress, and signal transduction [J].
Apel, K ;
Hirt, H .
ANNUAL REVIEW OF PLANT BIOLOGY, 2004, 55 :373-399
[2]   Evaluation of matrix effects in metabolite profiling based on capillary liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry [J].
Boettcher, Christoph ;
v. Roepenack-Lahaye, Edda ;
Willscher, Edith ;
Scheel, Dierk ;
Clemens, Stephan .
ANALYTICAL CHEMISTRY, 2007, 79 (04) :1507-1513
[3]   Laser capture microdissection for the analysis of gene expression during embryogenesis of Arabidopsis [J].
Casson, S ;
Spencer, M ;
Walker, K ;
Lindsey, K .
PLANT JOURNAL, 2005, 42 (01) :111-123
[4]  
Celis A, 1999, ELECTROPHORESIS, V20, P355, DOI 10.1002/(SICI)1522-2683(19990201)20:2<355::AID-ELPS355>3.0.CO
[5]  
2-N
[6]  
Clarke D, 1982, ACTIVE DEFENSE MECH, V37, P321
[8]  
DeBlock M, 1995, METHOD CELL BIOL, V49, P153
[9]   Natural products and plant disease resistance [J].
Dixon, RA .
NATURE, 2001, 411 (6839) :843-847
[10]   Laser capture microdissection [J].
EmmertBuck, MR ;
Bonner, RF ;
Smith, PD ;
Chuaqui, RF ;
Zhuang, ZP ;
Goldstein, SR ;
Weiss, RA ;
Liotta, LA .
SCIENCE, 1996, 274 (5289) :998-1001