https://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
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https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2021, Vol. 48 ›› Issue (11): 2133-2145.doi: 10.16420/j.issn.0513-353x.2021-0060

• 研究论文 • 上一篇    下一篇

苹果MdMYB116对白粉病的抗性研究

吉苗苗1, 万叶1, 张一平1, 马海1, 王西平1,2,*(), 高华1,2,*()   

  1. 1西北农林科技大学园艺学院,旱区作物逆境生物学国家重点实验室,陕西杨凌 712100
    2农业农村部西北地区园艺作物生物学与种质创制重点开放实验室,陕西杨凌 712100
  • 收稿日期:2021-05-21 修回日期:2021-07-20 发布日期:2021-12-02
  • 通讯作者: 王西平,高华 E-mail:wangxiping@nwsuaf.edu.cn;gaohua2378@163.com
  • 基金资助:
    陕西省科技重大专项项目(2020zdzx03-06-02-02);国家现代农业产业技术体系建设专项资金项目(CARS-27)

Studies on the Resistance of Apple MdMYB116 to Powdery Mildew

JI Miaomiao1, WAN Ye1, ZHANG Yiping1, MA Hai1, WANG Xiping1,2,*(), GAO Hua1,2,*()   

  1. 1College of Horticulture,State Key Laboratory of Crop Stress Biology in Arid Areas,Northwest A & F University,Yangling,Shaanxi 712100,China
    2Key Laboratory of Horticultural Crop Biology and Germplasm Development in Northwest China,Ministry of Agriculture and Rural Affairs,Yangling,Shaanxi 712100,China
  • Received:2021-05-21 Revised:2021-07-20 Published:2021-12-02
  • Contact: WANG Xiping,GAO Hua E-mail:wangxiping@nwsuaf.edu.cn;gaohua2378@163.com

摘要:

以抗白粉病的‘嘎拉’苹果为试材,克隆得到MdMYB116的cDNA全长序列。该基因的开放阅读框为900 bp,编码299个氨基酸,含有2个保守结构域。系统进化树结果表明,MdMYB116与梨PbMYB108-like的相似性最高,为93.38%。亚细胞定位试验显示MdMYB116定位在细胞核内。MdMYB116接种苹果白粉病菌6 ~ 12 h表达量上升,并达到高峰。酵母转录激活验证得出MdMYB116全长含有自激活活性。将MdMYB116异源转化拟南芥,得到的转基因植株接种白粉病菌处理后,其抗性显著高于野生型和pad4突变体植株。组织化学染色观察发现,与野生型和pad4突变体植株相比,转基因株系活性氧积累水平更高,死细胞数量和胼胝质积累也更多,说明转基因株系存在较严重的过敏致死情况,证明MdMYB116参与植株对白粉病的免疫调节反应。通过分析相关抗病基因的表达结果,说明MdMYB116通过参与SA和/或MeJA的信号途径增强防御反应。

关键词: 苹果, MdMYB116, 白粉病, 功能分析

Abstract:

The full-length cDNA sequence of MdMYB116 was cloned from the powdery mildew resistant‘Gala’apple. The open reading frame of this gene was 900 bp,encoded 299 amino acids and contained two conserved domains. The phylogenetic tree results showed that MdMYB116 shared close relationship with PbMYB108-like in pear,with a identity of 93.38%. Subcellular localization assay evidenced that MdMYB116 was targeted to the nucleus. The expression level of MdMYB116 increased gradually from 6 h to 12 h and it was induced to a peak at 12 h after inoculation with powdery mildew. Yeast transcriptional activation verification revealed that the full length of MdMYB116 exhibited self-activating activity. MdMYB116 was transformed into Arabidopsis thaliana heterologously,and transgenic plant was obtained. After being inoculated with powdery mildew,it was found that the resistance of the transgenic lines was significantly higher than that of the wild type and pad4 mutant plants. Histochemical staining assays displayed that compared with wild type and pad4 mutant plants,the transgenic lines had a higher level of reactive oxygen species(ROS)burst,and more dead cells and calluses accumulation,indicating that the transgenic lines had a serious allergic lethal situation,which proved that MdMYB116 participated in the plants powdery mildew immunomodulatory response. Expression analysis of some disease related genes speculated that MdMYB116 may enhanced defense responses though SA and/or MeJA signal pathways.

Key words: apple, MdMYB116, powdery mildew, functional analysis

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