甜瓜蔓枯病抗性基因相关的SSR候选分子标记的筛选

发布时间：2018-03-28 22:28

本文选题：甜瓜　切入点：蔓枯病　出处：《新疆大学》2017年硕士论文

【摘要】：甜瓜是一年生蔓性草本植物,在我国各地广泛分布,是深受人们喜爱的重要水果之一。甜瓜蔓枯病是由Didymella bryoniae引起的真菌病害,其田间的爆发常导致甜瓜叶片、茎部、根部、果实受损,因此导致甜瓜减产、品质降低。本研究采用光学显微镜镜检、病原菌致病力分析、真菌ITS区分子测序鉴定结合的方法,对实验中使用的蔓枯病病原菌进行鉴定,确定了该真菌的归属。对实验室保存的14种甜瓜鉴别寄主与感病品种Hami413,通过苗期人工喷雾分生孢子法筛选出蔓枯病抗性甜瓜材料。以筛选出的抗蔓枯病甜瓜品种Iran H与感蔓枯病甜瓜品种Hami413为亲本,构建F2代分离群体,利用BSA法(混合分组分析法)和SSR分子标记技术,筛选出甜瓜抗病池与感病池之间扩增出差异条带的SSR分子标记,为Iran H抗蔓枯病基因的定位奠定基础。主要结果如下:(1)感蔓枯病甜瓜品种Hami413的接种结果表明:该菌株为致病性较强的菌株;且与已报道的D.bryoniae菌株的菌落及分生孢子形态相似;核糖体序列间隔区的测序结果经Blast序列对比,进一步证实该菌株属于D.bryoniae。(2)苗期人工喷雾分生孢子法接种鉴定,筛选出高抗蔓枯病甜瓜材料Iran H,抗性材料在不同的季节抗性表现稳定。对接种后对抗感材料叶片POD的活性指标进行测定,接种后抗病甜瓜叶片中的POD活性显著高于感病甜瓜,可为甜瓜蔓枯病的接种材料鉴定作为参考。(3)以筛选出的抗蔓枯病甜瓜品种Iran H为供体亲本,以感蔓枯病甜瓜品种Hami413为受体亲本,构建BC1及F2代分离群体,分离群体接种结果符合单个隐形基因的遗传规律。(4)以128株F2代分离群体为材料,利用BSA法筛选出甜瓜抗病池与感病池之间扩增出差异条带的10个SSR分子标记。
[Abstract]:Muskmelon is an annual tendril herb, which is widely distributed all over China and is one of the most popular important fruits. Melon vine blight is a fungal disease caused by Didymella bryoniae, and its outbreak in the field often results in the leaves, stems and roots of muskmelon. The fruit was damaged, which resulted in the decrease of yield and quality of muskmelon. In this study, microscopical examination, pathogenicity analysis of pathogenic bacteria and molecular sequencing of fungal ITS region were used to identify the pathogen of vine blight in the experiment. The identification of 14 varieties of muskmelon cultivars, Hami413, which were preserved in laboratory, was used to screen the resistant melon materials by artificial spray conidium method at seedling stage, and the resistant varieties of muskmelon were screened out by the method of artificial spray conidial spores at the seedling stage. Iran H and Hami413 were parents. Using BSA method (mixed grouping analysis) and SSR molecular marker technique, the SSR molecular markers with different bands were amplified between resistant and susceptible pools of muskmelon. The main results were as follows: (1) the inoculation results of Iran H susceptible muskmelon variety Hami413 showed that the strain was highly pathogenic and similar to the reported D.bryoniae strains in colony and conidial morphology. The sequencing results of ribosomal spacer region further confirmed that the strain belonged to D. bryoniae.2) inoculation with artificial spray conidium method at seedling stage. The resistant melon (Iran H) with high resistance to vine blight was selected, and the resistant material showed stable resistance in different seasons. The activity index of POD in resistant melon leaves after inoculation was significantly higher than that in susceptible melon leaves after inoculation, and the activity of POD in resistant melon leaves was significantly higher than that in susceptible melon leaves after inoculation. It can be used as reference for identification of inoculation material of melon vine blight disease. The isolated populations of BC1 and F2 generation were constructed by using Iran H as donor parent and Hami413 as recipient parent. The inoculation results of the isolated populations were in accordance with the genetic rule of a single invisible gene. (4) using 128 F _ 2 segregated populations as materials, 10 SSR molecular markers with differential bands were amplified from resistant and susceptible pools of muskmelon by BSA method.
【学位授予单位】：新疆大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S436.5

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