Xin Jingjing, Liu Ye, Li Huiyun, Chen Sumei, Jiang Jiafu, Song Aiping, Fang Weimin, Chen Fadi
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Landscaping, Ministry of Agriculture and Rural Affairs, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
Hortic Res. 2021 May 1;8(1):101. doi: 10.1038/s41438-021-00534-x.
The Mildew Resistance Locus O (MLO) gene family has been investigated in many species. However, there are few studies on chrysanthemum MLO genes. We report in this study that CmMLO17 in Chrysanthemum morifolium was upregulated after Alternaria alternata infection. Silencing of CmMLO17 by artificial microRNA resulted in reduced susceptibility of chrysanthemum to A. alternata infection. Genes in the abscisic acid (ABA) and Ca signaling pathways were upregulated in the CmMLO17-silenced line R20 compared to the wild-type plants. We speculated that CmMLO17-silenced plants had a faster and stronger defense response that was mediated by the ABA and Ca signaling pathways, resulting in reduced susceptibility of chrysanthemum to A. alternata infection. In addition, a candidate gene, CmKIC, that may interact with CmMLO17 was discovered by the yeast two-hybrid assay. The interaction between CmMLO17 and CmKIC was confirmed using the yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) analysis. CmMLO17 and CmKIC were both located on the plasma membrane, and CmKIC was also located on the nucleus. CmKIC overexpression increased the susceptibility of chrysanthemum to A. alternata, whereas CmKIC silencing resulted in reduced susceptibility. Therefore, CmMLO17 and CmKIC may work together in C. morifolium to support the growth of A. alternata. The results of this study will provide insight into the potential function of MLO and improve the understanding of plant defense responses to necrotrophic pathogens.
已经在许多物种中对抗白粉病基因座O(MLO)基因家族进行了研究。然而,关于菊花MLO基因的研究很少。我们在本研究中报告,菊花中的CmMLO17在链格孢感染后上调。通过人工microRNA沉默CmMLO17导致菊花对链格孢感染的敏感性降低。与野生型植物相比,脱落酸(ABA)和钙信号通路中的基因在CmMLO17沉默系R20中上调。我们推测,CmMLO17沉默的植物具有更快更强的防御反应,该反应由ABA和钙信号通路介导,导致菊花对链格孢感染的敏感性降低。此外,通过酵母双杂交试验发现了一个可能与CmMLO17相互作用的候选基因CmKIC。使用酵母双杂交试验和双分子荧光互补(BiFC)分析证实了CmMLO17与CmKIC之间的相互作用。CmMLO17和CmKIC都位于质膜上,并且CmKIC也位于细胞核上。CmKIC过表达增加了菊花对链格孢的敏感性,而CmKIC沉默导致敏感性降低。因此,CmMLO17和CmKIC可能在菊花中共同作用以支持链格孢的生长。本研究结果将为深入了解MLO的潜在功能提供见解,并增进对植物对坏死营养型病原体防御反应的理解。