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MAPKs Signaling in Vicia Guard Cells

Author: JiangJing
Tutor: HanJinFeng；SongChunPeng
School: Henan Agricultural University
Course: Crop Cultivation and Farming System
Keywords: MAP kinase PD98059 SB203580 SB202190 ABA SA H2O2 Guard cell
CLC: Q945
Type: PhD thesis
Year: 2004
Downloads: 176
Quote: 1
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Abstract


H 2 O 2 extensively as a signaling molecule involved in plant responses to stress response, specific it produce and inactivated signaling pathways, caused widespread concern. Has proved to many stress factors that can induce plant cell abscisic acid (that Miyang23 and N22 were acid, ABA) and salicylic acid (salicylic acid, SA) concentration the two coercion messenger by induced H 2 O 2 and promote stomatal closure. In addition, plant cell memory in many mitogenic original activated protein kinase (mitogen-activated protein kinase, MAPK) kinase plays an important role in the signal transduction process plant a variety of stress reaction. In this study, based on epidermal bioanalytical microinjection technique, patch-clamp technique and confocal laser microscopy, the use of a specific protein kinase inhibitor treatment, to explore the protein kinase faba bean (Vicia faba L.) ABA and SA-induced stomatal guard cells H 2 O 2 affect the mechanism of generation and its signal transduction, and the results are as follows: 1. PD98059 through specific inhibition of ABA-induced H 2 O 2 generation, or Clear generated the H 2 O 2 accumulated reversibly blocked the ABA-induced stomatal closure with ABA and H 2 of O 2 processing epidermal strip stomatal aperture significant decline, while PD98059 treatment does not affect stomatal closure; using PD98059 respectively and ABA, H O 2 co-induced stomatal aperture did not decline; were first ABA and H 2 O 2 -induced stomatal closure, and PD98059 treatment, stomatal closure was reversed. In control experiments, ABA induced fluorescence increased rapidly; separate PD98059, PD98059 and ABA of stomatal guard cells H 2 O 2 probe fluorescence intensity, there was no increase ; PD98059 injection into a guard cells, and then treated with ABA make two same guard cell fluorescence intensity of the fluorescence-based strong contrast; PD98059 microinjection into ABA-induced fluorescence intensity of DCF (dichlorofluorescin) elevated stomatal guard cells, fluorescence intensity decreased, which has not been the same DCF fluorescence intensity in the injection side of the guard cells. DCF fluorescence intensity increased rapidly in the guard cells is induced by ABA, like SA. However, PD98059 and sA common processing, increased fluorescence intensity HZoZ probe; the SA induction DCF fluorescence intensity increased after PD98059 treatment had no effect on the fluorescence intensity of DCF; PD98059 injection into a guard cells, SA The results show that the fluorescence intensity within the two cells. ABA-induced inward K current significantly reduced; PD98059 treatment complexing agent B PD98059 and eaZ-good recognition (bis (o a amnophenoxy), thane an N, N Dui pay a tetraaeeticacid) jointly deal with the K ten current no less ; PD98059 can be reversed ABA-induced inward K current decreases, the BAPTA treatment has no effect on this process. Various concentration levels of 2.10 6 mol · L afraid the spoon sBZo358o blocked 7 ABA HZo: ten current induced stomatal closure and inward K varies from one 10 \The SBZo358o epidermal strip, and HZOZ treatment: changes in stomatal aperture inhibitor concentration in the 10 - 9 mol.L 'to 10 7mol · L', stomatal opening decreases, the inhibitor concentration 1 D \lower but different amplitudes, compared When SB203580 concentration of 10 6 mol · L, when, with the ABA treated separately, stomatal opening merely reducing part of each concentration level, the non-active SB202474 and ABA , executive O: common processing, stomatal closure does not affect the analysis is consistent with the epidermal strip: SBZo358O processing HZO: inward K current induced by a process of change that is going through a recovery process is reduced to within 15min rebound after the current value and considerable decline before; Coincidentally min, SBZO358O makes ABA-induced inward K current decreases partially rebounded and stabilized. 3.SB203580, SBZOZ190 respectively different blocks induced by ABA and SA HZO: to produce SBZO358o and ABA jointly deal with DCF fluorescence intensity in guard cells did not rise, but this role BAPTA treatment does not affect SB203580; injected into the guard cells. SB203580, an inhibitor of p38 MAPK, extracellular ABA treatment, both with the same fluorescent guard cells based on the fluorescence intensity of contrasting; SB203580 and H20: with injection into the guard cells, and then treated with ABA, after injection of guard cell DCF fluorescence intensity lower than the party of no injection; the SB203580 and SA jointly deal, DCF fluorescence still elevated. SB202190 and SA stomatal or SB202190 injected into guard cells to SA extracellular processing, HZO: fluorescence intensity did not rise; if the former two treatment liquid was further added BAPTA, Rose O: Fluorescence but can be increased. In visible the, SBZO358O and SBZOZ19O different inhibited guard cells induced by ABA, SA HZO: produce. summary, PD98059 induced by ABA HZO: produce ABA and HZO: induced inward K current decreases the blocking effect MEKI / 2 (the mitogen-activated proteinkinase kinasel 2) positive regulator of ABA-induced Mei O: generate while pD98059 effect may be due to the activation of the Vision O: Clear system activity of of HZOZ clear; P38 The protein kinase probe SBZO358O SB20219O the different results in downstream signaling of ABA and SA Description: P38 homologous kinase in the play.: the signaling pathways play a role in the dependence of c expansion, shows HZO : Signal-specific protein kinase not only involved in stress signal transduction, and specifically regulate H20: signaling pathways. MAPK activation defend the cell HZOZ signal generation, enlarge its specific response signals stimulate reactions The important regulatory role.

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