Yousaf Ayesha, Baldi Paolo, Piazza Stefano, Gualandri Valeria, Komjanc Matteo, Dalla Costa Lorenza, Patocchi Andrea, Malnoy Mickael
Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige, Italy.
Department of Agricultural, Food, Environmental and Animal Sciences, Università Degli Studi di Udine, Udine, Italy.
Plant J. 2025 Jan;121(2):e17214. doi: 10.1111/tpj.17214. Epub 2024 Dec 18.
To enhance the breeding of new scab-resistant apple cultivars, a comprehensive understanding of the mechanisms governing major scab resistance genes is essential. Rvi12_Cd5 was previously identified as the best candidate gene for the Rvi12 scab resistance of the crab apple "Hansen's baccata #2" by gene prediction and in silico analysis. In the present study, Rvi12_Cd5 was used to transform the scab-susceptible apple cultivar "Gala Galaxy." Two constructs were prepared: the first carrying Rvi12_Cd5 under the control of a 35S promoter and E9 terminator, and the second carrying Rvi12_Cd5 under the control of its native promoter and terminator. All the transgenic lines were analyzed for T-DNA integration, copy number, and expression of Rvi12_Cd5 and phenotypically evaluated for scab resistance. The "Gala Galaxy" lines carrying the 35S promoter expressed Rvi12_Cd5 at a high level, showing partial to high resistance against a mixed inoculum of Venturia inaequalis, with symptoms ranging from class 0 to 3b on the Chevalier scale. The transgenic lines carrying the native promoter showed a lower expression of Rvi12_Cd5 compared with the 35S lines. Nevertheless, the low expression was sufficient to induce a resistance level comparable to that of the transgenic lines carrying the 35S promoter. These results indicate that Rvi12_Cd5 confers scab resistance to a susceptible apple cultivar and that even a low level of gene transcript can trigger a plant response to V. inaequalis infection. After HcrVf2 and Vr2-C, Rvi12_Cd5 is the third major apple scab resistance gene being functionally proven.
为加强新的抗黑星病苹果品种的培育,全面了解控制主要抗黑星病基因的机制至关重要。通过基因预测和电子分析,Rvi12_Cd5先前被确定为海棠“汉森海棠#2”Rvi12抗黑星病的最佳候选基因。在本研究中,Rvi12_Cd5被用于转化感黑星病的苹果品种“嘎啦银河”。制备了两种构建体:第一种在35S启动子和E9终止子的控制下携带Rvi12_Cd5,第二种在其天然启动子和终止子的控制下携带Rvi12_Cd5。对所有转基因株系进行了T-DNA整合、拷贝数以及Rvi12_Cd5表达分析,并对抗黑星病进行了表型评估。携带35S启动子的“嘎啦银河”株系高水平表达Rvi12_Cd5,对苹果黑星病菌混合接种物表现出部分至高抗性,在骑士分级标准下症状为0至3b级。与携带35S启动子的株系相比,携带天然启动子的转基因株系Rvi12_Cd5表达较低。然而,低表达足以诱导出与携带35S启动子的转基因株系相当的抗性水平。这些结果表明,Rvi12_Cd5赋予感病苹果品种抗黑星病能力,即使低水平的基因转录也能触发植物对苹果黑星病菌感染的反应。继HcrVf2和Vr2-C之后,Rvi12_Cd5是第三个功能得到验证的主要苹果抗黑星病基因。