园艺学报 ›› 2020, Vol. 47 ›› Issue (7): 1264-1276.doi: 10.16420/j.issn.0513-353x.2020-0263

• 研究论文 • 上一篇    下一篇


赵凯茜,丁 茜,王跃进*   

  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,农业部西北地区园艺作物生物与种质创制重点实验室,陕西杨凌 712100
  • 出版日期:2020-07-25 发布日期:2020-07-25
  • 基金资助:

Research on Control of Disease Resistance by Stilbene Synthase Genes Derived from Chinese wild Vitis quinquangularis

ZHAO Kaixi,DING Xi,and WANG Yuejin*   

  1. College of Horticulture,Northwest A & F University,State Key Laboratory of Crop Stress Biology in Arid Areas,Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture,Yangling,Shaanxi 712100,China
  • Online:2020-07-25 Published:2020-07-25

摘要: 利用同源克隆法得到抗白粉病的中国野生毛葡萄‘丹凤–2’芪合酶基因VqSTS11和VqSTS23的开放阅读框,并构建过表达载体;通过器官发生途径诱导不抗白粉病的欧洲葡萄‘无核白’分生愈伤组织并采用农杆菌介导法对其进行转化;经过PCR检测和Western blot鉴定,获得了5株VqSTS11超量表达植株和3株VqSTS23超量表达植株;人工接种白粉菌发现,转基因植株中白粉菌菌丝生长速度慢,孢子萌发受到一定的抑制,并且转基因植株中芪合酶基因相对表达量升高,抗病相关基因表达上调,芪类物质含量积累增多。本研究结果进一步证明中国野生毛葡萄‘丹凤–2’芪合酶基因VqSTS11和VqSTS23对白粉菌具有抗性,‘丹凤–2’可以为改良欧洲葡萄品种抗病性提供抗病基因,作为抗病育种的资源。

关键词: 毛葡萄, 欧洲葡萄, 芪合酶, 白藜芦醇, 白粉病, 抗病性, 转基因

Abstract: Two complete open reading frames of VqSTS11 and VqSTS23,were respectively isolated from Vitis quinquangularis‘Danfeng-2’using homologous cloning technique and further for overexpression vector construction. Meristematic bulk(MB)of cultivated Vitis vinifera‘Thompson Seedless’,were induced through organogenesis and further for Agrobacterium-mediated transformation to obtain transgenic plants. After PCR detection and Western blot identification,5 transgenic lines of VqSTS11 and 3 transgenic lines of VqSTS23 were verified,respectively. Transgenic plants and non-transgenic plants were artificially inoculated with powdery mildew. Microscopic observation demonstrated that the hyphae growth in transgenic lines were slower than those in non-transgenic plants and spore germination was inhibited in transgenic plants. The disease resistance of transgenic plants was further analyzed by qRT-PCR and HPLC. The results displayed that not only the expression of STSs in transgenic plants increased,and the expression of disease resistance-related genes was up-regulated,but also stilbene compounds in the transgenic plants were accumulated. The expression product of‘Danfeng-2’VqSTS11 and VqSTS23 could tolerate powdery mildew. Therefore,‘Danfeng-2’could be used to improve the disease tolerance and provide disease- tolerant candidate genes for European grape cultivars,which could be served as a breeding resources for disease-tolerant breeding.

Key words: Vitis quinquangularis, Vitis vinifera, stilbene synthase, resveratrol, powdery mildew, disease resistance, transgenic