抗苹果斑点落叶病基因Mal d 1的克隆及功能鉴定

发布时间：2018-01-25 06:00

本文关键词： 苹果 Mal d 表达分析转基因斑点落叶病菌　出处：《园艺学报》2017年02期 　论文类型：期刊论文

【摘要】：以苹果叶片总RNA为模板,通过克隆获得抗苹果斑点落叶病基因Mal d 1。Mal d 1开放阅读框长度为480 bp,编码159个氨基酸残基,包含2个外显子和1个内含子。蛋白结构分析显示,Mal d 1蛋白包含bet v 1-like结构域。荧光定量PCR分析结果表明,Mal d 1在苹果叶、花、果皮、果肉中均有表达,但各器官中表达水平存在差异;在不同发育时期的果皮中都有表达,表达水平随时间呈现先上升后下降的趋势;在抗性品种叶片中表达水平较高;在叶片人工接种病菌条件下表达量随时间的延长明显高于对照组。利用PRI101-AN载体和农杆菌介导的遗传转化方法转化苹果愈伤组织,转基因愈伤组织的抗病鉴定结果显示,Mal d 1过量表达增强愈伤组织对苹果斑点落叶病的抗性。
[Abstract]:With the total RNA of apple leaves as the template for resistance to Alternaria leaf spot of apple Mal gene D 1.Mal d 1 and an ORF of 480 BP by cloning, encoding 159 amino acid residues, including 2 exons and 1 introns. Analysis showed that the protein structure, Mal d 1 Bet v 1-like domain containing protein. Quantitative PCR analysis showed that Mal d 1 in apple leaves, flowers, fruit, the flesh is expressed, but the expression levels were different in different organs; at different developmental stages in the pericarp were expressed, expression levels over time showed a downward trend after rising; high expression level in leaves of resistant varieties; extended amount with time was significantly higher than the control group expressed in the leaves inoculated with the pathogen. Under the condition of transformation of Apple callus by genetic transformation method of PRI101-AN vector and Agrobacterium mediated resistance, the identification results of transgenic callus showed that Mal The overexpression of D 1 enhanced the resistance of callus to Apple spotted deciduous disease.

【作者单位】：中国农业科学院果树研究所农业部园艺作物种质资源利用重点实验室;
【基金】：国家现代农业产业技术体系建设专项资金项目(CARS-28) 国家科技支撑计划项目(2013BAD02B01)
【分类号】：S436.611.1
【正文快照】： 苹果斑点落叶病(Apple Alternaria Blotch)又称褐纹病,最早发现于美国(Roberts,1924),是一种在世界范围内发生,在亚洲产区危害最为严重的病害之一(Miyamoto et al.,2009;Li et al.,2011)。目前全球50%的苹果生产受到该病威胁(Zhao et al.,2008)。近年来,该病在中国北方苹果主

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