Engineering Laboratory of Genetic Improvement of Horticultural Crops of Shandong Province, Qingdao Agricultural University, Qingdao 266109, China.
College of Horticulture, Qingdao Agricultural University, Qingdao 266109, China.
Int J Mol Sci. 2022 Jun 5;23(11):6323. doi: 10.3390/ijms23116323.
Glomerella leaf spot (GLS), caused by the fungus , is one of the most devastating apple diseases. Our previous study reported that the GLS resistance locus was defined on the chromosome 15 region. Here, we further found a single-nucleotide polymorphism (SNP) site (SNP) in the GLS resistance that was able to distinguish resistant cultivars (lines) from susceptible ones. On the basis of the SNP site, we cloned a TNL gene from the GLS resistant locus and named it (NCBI Accession Number: ON402514). This gene contains a toll/interleukin-1 receptor transmembrane domain (TIR), nucleotide-binding sites (NBS), and leucine-rich repeat (LRR) domain. Subcellular location indicated that MdTNL1 was expressed in the nucleus and cell membrane. Ectopic overexpression of in caused cell death. We further demonstrated allelic polymorphisms in . It is noteworthy that NBS and LRR domains of the MdTNL1 protein serve as the repository for generating allelic diversity. Quantitative real-time PCR (qRT-PCR) assay revealed that was highly expressed in resistant apple cultivar 'Fuji' after inoculation with whereas susceptible cultivar 'Golden Delicious' exhibited low expression after inoculation. Over-expression of in susceptible apple fruits and leaves improved disease resistance, while in 'Orin' calli, silencing the gene conversely decreased GLS resistance. In conclusion, we identified a GLS associated with SNP and demonstrated that a TIR-NBS-LRR gene positively regulates GLS resistance in apple.
黑星病（GLS）由真菌引起，是苹果最具破坏性的病害之一。我们之前的研究报告称，GLS 抗性基因位于第 15 号染色体区域。在这里，我们进一步在 GLS 抗性基因中发现了一个单核苷酸多态性（SNP）位点，该位点能够区分抗性品种（系）和敏感品种。基于该 SNP 位点，我们从 GLS 抗性基因座克隆了一个 TNL 基因，并将其命名为 （NCBI 登录号：ON402514）。该基因包含一个 Toll/白细胞介素-1 受体跨膜结构域（TIR）、核苷酸结合位点（NBS）和富含亮氨酸重复（LRR）结构域。亚细胞定位表明 MdTNL1 在细胞核和细胞膜中表达。在 中异位过表达 导致细胞死亡。我们进一步证明了 中的等位基因多态性。值得注意的是，MdTNL1 蛋白的 NBS 和 LRR 结构域是产生等位基因多样性的储存库。定量实时 PCR（qRT-PCR）检测显示，接种 后，抗性苹果品种‘富士’中 高度表达，而敏感品种‘金冠’接种后表达水平较低。在易感苹果果实和叶片中过表达 可提高抗病性，而在‘Orin’愈伤组织中，沉默 基因则相反降低了 GLS 抗性。综上所述，我们鉴定了一个与 SNP 相关的 GLS，并证实一个 TIR-NBS-LRR 基因 正向调控苹果的 GLS 抗性。
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