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在病原菌侵染下,植物体是如何通过超敏反应限制病原菌扩增的,到目前为止还不清楚。最近的研究显示自噬起了必要的作用,可见,研究自噬和超敏反应的机制非常重要。本研究通过观察在非寄主病原菌突变体丁香假单胞菌番茄致病变种(Pseudomonas syringae pv.tomatoDC3000,Pst DC3000)的侵染下,拟南芥光合功能的变化及自噬现象出现的情况,以为进一步研究植物抗病机理提供实验基础。以野生型拟南芥为材料,采用光谱分析和叶绿素荧光成像分析手段,研究不同浓度的Pst DC3000侵染对拟南芥离体叶片光合功能的影响;以转基因野生型拟南芥(绿色荧光蛋白标记的自噬基因8a)幼根为材料,应用共聚焦显微镜,研究不同浓度的Pst DC3000诱导拟南芥自噬的情况。实验发现,OD600=0.2的Pst DC3000侵染,可显著诱导拟南芥叶片活性氧的积累和迸发,并会引起拟南芥叶片光合作用效率的下降。同时发现,该病原菌处理2 h,可导致拟南芥根中自噬小体的产生。用2’,7’-二氯二氢荧光素二乙酯(2’,7’-dichlorodihydrofluorescein diacetate,H2DCFDA)标记活性氧,检测了Pst DC3000作用下叶片活性氧产生量的变化,发现,在OD600=0.2的Pst DC3000作用下,90 min内,叶片组织中的活性氧产生了明显的积累。以上结果说明:Pst DC3000侵染叶片,导致了活性氧迸发和光合系统的受损,进而推测这两个结果具有相关性;在植物与病原菌的互作系统中,光合系统的变化规律可以作为典型的指标来加以检测。
It is still unclear how plants inhibit the multiplication of pathogenic bacteria by the hypersensitivity reaction under pathogen infection. Recent studies have shown that autophagy plays a necessary role. It can be seen that the mechanism of studying autophagy and hypersensitivity is very important. In this study, we observed the changes of photosynthetic function and autophagy in Arabidopsis thaliana under the infection of non-host pathogen mutant Pseudomonas syringae pv.tomatoDC3000 and Pst DC3000, Further study of plant disease resistance mechanism to provide the experimental basis. The effects of different concentrations of Pst DC3000 on leaf photosynthetic function of Arabidopsis thaliana were studied by using spectral analysis and chlorophyll fluorescence imaging analysis. The wild-type Arabidopsis thaliana was used to study the effects of transgenic Arabidopsis thaliana (green fluorescent protein Labeled autophase gene 8a) roots were used as materials. Confocal microscopy was used to study the autophagy of Arabidopsis induced by different concentrations of Pst DC3000. It was found that the infection of Pst DC3000 with OD600 of 0.2 significantly induced the accumulation and burst of reactive oxygen species in Arabidopsis thaliana leaves and caused the decrease of photosynthesis efficiency in Arabidopsis leaves. Also found that the pathogen treatment 2 h, can lead to autophagy in Arabidopsis root body production. The reactive oxygen species (ROS) were labeled with 2 ’, 7’-dichlorodihydrofluorescein diacetate (H2DCFDA), and the changes of reactive oxygen species (ROS) production under the action of Pst DC3000 were measured. = 0.2 Pst DC3000 within 90 min, the reactive oxygen species in leaves have a significant accumulation. The above results indicated that the invasion of leaves by Pst DC3000 led to the burst of reactive oxygen species and the damage of photosynthetic system, and then the correlation between the two results was speculated. In the interaction system between plant and pathogen, the variation of photosynthetic system can be regarded as typical The indicators to be tested.