| 草地早熟禾PpMLO14基因的克隆及其对禾布氏白粉菌诱导的响应 |
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| 引用本文：张艺凝,高鹏,朱慧森,赵祥,梁银萍.草地早熟禾PpMLO14基因的克隆及其对禾布氏白粉菌诱导的响应.植物保护学报,2025,52(2):340-350 |
| DOI：10.13802/j.cnki.zwbhxb.2025.2025001 |
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| 作者 | 单位 | E-mail | | 张艺凝 | 山西农业大学草业学院, 山西省草地生态保护与乡土草种质创新重点实验室, 太谷 030801 | | | 高鹏 | 山西农业大学草业学院, 山西省草地生态保护与乡土草种质创新重点实验室, 太谷 030801 | | | 朱慧森 | 山西农业大学草业学院, 山西省草地生态保护与乡土草种质创新重点实验室, 太谷 030801 | | | 赵祥 | 山西农业大学草业学院, 山西省草地生态保护与乡土草种质创新重点实验室, 太谷 030801 | | | 梁银萍 | 山西农业大学草业学院, 山西省草地生态保护与乡土草种质创新重点实验室, 太谷 030801 | liangyinping3@163.com |
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| 中文摘要:为明确禾布氏白粉菌早熟禾专化型Blumeria graminis f.sp.poae在侵染草地早熟禾Poa pra‐tensis过程中对抗白粉病相关基因MLO （mildew resistance locus O）表达的诱导作用，采用PCR技术克隆PpMLO14基因，分析该基因编码蛋白的序列结构及系统进化关系，并通过实时荧光定量PCR技术比较草地早熟禾抗、感品种不同组织部位（根、茎、叶片和叶鞘）以及接种禾布氏白粉菌早熟禾专化型后PpMLO14基因的表达情况。结果显示：克隆得到的草地早熟禾PpMLO14基因编码区为1 851 bp，编码599个氨基酸；具有一个高度保守的MLO超家族结构域，是典型的MLO型抗病家族蛋白。草地早熟禾PpMLO14和节节麦Aegilops tauschii AtMLO14聚在一个分支，亲缘关系较近。草地早熟禾PpMLO14基因在不同抗性品种中均在叶片中高表达，该基因在抗病品种叶片、叶鞘和茎中的相对表达量是根中相对表达量的44.65倍、7.18倍和4.11倍；在感病品种叶片、叶鞘和茎中的相对表达量是根中相对表达量的217.51倍、71.73倍和27.52倍。接种禾布氏白粉菌早熟禾专化型后24 h和48 h，抗病品种中PpMLO14基因的相对表达量显著升高，分别是对照的28.61倍和19.93倍，而感病品种中PpMLO14基因的相对表达量在接种后24 h达到峰值且显著高于对照，是对照的202.44倍。表明PpMLO14基因在植物抗病早期就发挥着重要作用，叶片和病菌侵染后24 h分别是其发挥抗病功能的关键部位与时期。 |
| 中文关键词:草地早熟禾 禾布氏白粉菌早熟禾专化型 基因克隆 序列分析 表达分析 |
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| Cloning of PpMLO14 gene from Kentucky bluegrass Poa pratensis and its response induced by powdery mildew fungus Blumeria graminis |
| Author Name | Affiliation | E-mail | | Zhang Yining | Shanxi Provincial Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, College of Prataculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China | | | Gao Peng | Shanxi Provincial Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, College of Prataculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China | | | Zhu Huisen | Shanxi Provincial Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, College of Prataculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China | | | Zhao Xiang | Shanxi Provincial Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, College of Prataculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China | | | Liang Yinping | Shanxi Provincial Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, College of Prataculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China | liangyinping3@163.com |
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| Abstract:To elucidate the induction effect of Blumeria graminis f. sp. poae (Bgp) on the expression of the PpMLO14 gene during the infection of Kentucky bluegrass Poa pratensis, this gene was cloned via PCR, and the structural characteristics and phylogenetic relationships of the protein encoded by PpMLO14 were analyzed. Quantitative real-time PCR (qPCR) was employed to assess the expression level of PpMLO14 gene across different tissues (roots, stems, leaves, and sheaths) and with post-Bgp inoculation in resistant and susceptible varieties. The results revealed that PpMLO14 had a 1 851-bp coding region encoding 599 amino acids with a highly conserved MLO (mildew resistance locus O) domain of the typical MLO-type disease resistance proteins. Phylogenetic analysis showed that P. pratensis PpMLO14 clusters closely with Aegilops tauschii MLO14, suggesting a close genetic relationship. Tissue-specific expression analysis demonstrated that PpMLO14 was predominantly expressed in leaves across both resistant and susceptible varieties. In resistant varieties, its expression in leaves, sheaths, and stems was 44.65-, 7.18-, and 4.11-fold of that in roots, respectively. In susceptible varieties, the corresponding expression levels, sheaths, and stems was 217.51-, 71.73-, and 27.52-fold of that in roots. Following Bgp inoculation, PpMLO14 expression in resistant varieties significantly increased at 24 h and 48 h, reaching 28.61and 19.93-fold of the control, respectively. In susceptible varieties, expression of the PpMLO14 gene reached a peak at 24 h after inoculation and was significantly higher (202.44fold) than that of control. These findings suggested that PpMLO14 played a critical role in early plant defense responses in the leaves at 24 h post-inoculation representing the key position and period to its disease resistance function. |
| keywords:Poa pratensis Blumeria graminis f. sp. poae gene cloning sequence analysis expression analysis |
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