Plant Pathology, Institute of Integrative Biology (IBZ), ETH Zürich, Zürich, Switzerland.
Plant Biotechnol J. 2014 Aug;12(6):728-33. doi: 10.1111/pbi.12177. Epub 2014 Mar 12.
The fire blight susceptible apple cultivar Malus × domestica Borkh. cv. 'Gala' was transformed with the candidate fire blight resistance gene FB_MR5 originating from the crab apple accession Malus × robusta 5 (Mr5). A total of five different transgenic lines were obtained. All transgenic lines were shown to be stably transformed and originate from different transgenic events. The transgenic lines express the FB_MR5 either driven by the constitutive CaMV 35S promoter and the ocs terminator or by its native promoter and terminator sequences. Phenotyping experiments were performed with Mr5-virulent and Mr5-avirulent strains of Erwinia amylovora, the causal agent of fire blight. Significantly less disease symptoms were detected on transgenic lines after inoculation with two different Mr5-avirulent E. amylovora strains, while significantly more shoot necrosis was observed after inoculation with the Mr5-virulent mutant strain ZYRKD3_1. The results of these experiments demonstrated the ability of a single gene isolated from the native gene pool of apple to protect a susceptible cultivar from fire blight. Furthermore, this gene is confirmed to be the resistance determinant of Mr5 as the transformed lines undergo the same gene-for-gene interaction in the host-pathogen relationship Mr5-E. amylovora.
易感火疫病的苹果栽培品种 Malus × domestica Borkh. cv. 'Gala' 被转化为候选火疫病抗性基因 FB_MR5,该基因源自野苹果 accession Malus × robusta 5 (Mr5)。共获得了五个不同的转基因株系。所有的转基因株系均表现出稳定的转化,并且来源于不同的转基因事件。这些转基因株系表达 FB_MR5,其启动子分别为组成型 CaMV 35S 和 ocs 终止子,或者其天然启动子和终止子序列。用 Erwinia amylovora 的 Mr5 毒力和非毒力菌株进行了表型实验,该菌是火疫病的病原体。在接种两种不同的 Mr5 非毒力 E. amylovora 菌株后,转基因株系上检测到的病害症状明显减少,而在接种 Mr5 毒力突变株 ZYRKD3_1 后,观察到的枝条坏死明显增多。这些实验的结果证明了从苹果天然基因库中分离出的单个基因能够保护易感品种免受火疫病的侵害。此外,由于转化株系在 Mr5-E. amylovora 的宿主-病原体关系中发生相同的基因对基因互作,因此该基因被确认为 Mr5 的抗性决定因子。