Fungal effectors play important roles in host-pathogen interactions. Botryosphaeria dothidea is an ascomycetous fungus that is responsible for the diseases of hundreds of woody plant species, including apple ring rot, which seriously affects apples worldwide. However, little is known about the effectors of B. dothidea. In this study, we analyzed the B. dothidea genome and predicted 320 candidate effector genes, 124 of which were successfully amplified and cloned. We investigated the effects of these genes on plant cell death in Nicotiana benthamiana while using a transient expression system. Twenty-four hours after initial inoculation with Agrobacterium tumefaciens cells carrying candidate effectors, the infiltrated leaves were challenged with A. tumefaciens cells carrying the BAX gene. In total, 116 candidate effectors completely inhibited, while one partially inhibited, the programmed cell death (PCD) of N. benthamiana induced by BAX, whereas seven candidate effectors had no effect. We then further tested seven candidate effectors able to suppress BAX-triggered PCD (BT-PCD) and found that they all completely inhibited PCD triggered by the elicitors INF1, MKK1, and NPK1. This result suggests that these effectors were activated in order to suppress pathogen-associated molecular pattern-triggered immunity. The signal peptides of these candidate effectors exhibited secretory activity in yeast (pSUC2 vector). Moreover, the respective deletion of Bdo_11198 and Bdo_12090 significantly reduced the virulence of B. dothidea. These results suggest that these effectors play important roles in the interaction of B. dothidea with its hosts.

译文

:真菌效应子在宿主-病原体相互作用中起重要作用。 Botryosphaeria dothidea是一种子囊菌,负责数百种木本植物的病害,包括严重影响全球苹果的苹果环腐病。然而,关于双歧杆菌的效应子知之甚少。在这项研究中,我们分析了B. dothidea基因组并预测了320个候选效应基因,其中124个被成功扩增和克隆。我们使用瞬时表达系统调查了这些基因对本生烟草中植物细胞死亡的影响。在初始接种携带候选效应子的根癌农杆菌细胞后二十四小时,用携带BAX基因的根癌农杆菌细胞攻击浸润的叶片。总共有116种候选效应物完全抑制了BAX诱导的本氏烟草的程序性细胞死亡(PCD),而部分抑制了BAX诱导的本氏烟草的程序性细胞死亡(PCD),而有7种候选效应物没有任何作用。然后,我们进一步测试了7种能够抑制BAX触发的PCD(BT-PCD)的候选效应子,发现它们都完全抑制了由激发子INF1,MKK1和NPK1触发的PCD。该结果表明,激活这些效应子是为了抑制病原体相关的分子模式触发的免疫力。这些候选效应子的信号肽在酵母(pSUC2载体)中表现出分泌活性。此外,Bdo_11198和Bdo_12090的各自缺失显着降低了十二指肠杆菌的毒力。这些结果表明,这些效应子在B. dothidea与其宿主的相互作用中起重要作用。

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