我国蔬菜匍柄霉叶斑病的病原学及对啶酰菌胺抗药性研究
发布时间：2019-03-02 19:36
【摘要】：茄匍柄霉(Stemphylium solani)是一种重要的植物病原真菌,寄主范围广泛。近年来,由该病原菌引起的番茄匍柄霉叶斑病在我国大面积爆发,造成巨大的经济损失,该病原菌目前尚缺乏系统的研究。本论文以来源于不同地区的茄匍柄霉为研究对象,从病原菌的培养性状、分生孢子形态、致病性、遗传多样性等方面分析茄匍柄霉的种群分化。建立茄匍柄霉的检测技术,为蔬菜匍柄霉叶斑病的早期预警监测提供有效的技术手段,同时分析了茄匍柄霉对啶酰菌胺的抗药性,为蔬菜匍柄霉防治药剂的开发提供了理论依据。1.2013年9月至2015年4月,在我国山东、北京、广东、广西、辽宁、海南等6个省市调查匍柄霉叶斑病的危害,发现引起番茄、菜豆、长叶莴苣匍柄霉叶斑病的优势菌株为茄匍柄霉(S.solani)。2.依据菌落形态将茄匍柄霉(S.solani)划分为9种类型。茄匍柄霉的菌落形态与寄主和地理来源没有相关性,茄匍柄霉分生孢子形态差异显著,与寄主和地理来源显著相关。3.发现我国不同来源的茄匍柄霉存在明显的致病力分化。根据茄匍柄霉(S.solani)对番茄、菜豆和长叶莴苣的致病力测定结果,明确茄匍柄霉存在两种致病型,致病型与菌株的寄主和地理来源密切相关。PA致病型分为4种亚致病型,PA1亚致病型对三种不同寄主的致病力较弱。PB致病型分为3种亚致病型,PB3亚致病型对三种不同寄主的致病力较强。同一寄主和地理来源的茄匍柄霉致病力存在强、中、弱的差异。4.建立了茄匍柄霉侵染番茄过程研究的最佳接种体系,明确了菌丝侵染是茄匍柄霉对番茄的重要侵染途径。菌丝在叶片表面萌发,产生多个芽管,菌丝向四周扩散伸长,到气孔附近形成侵染菌丝,从气孔侵入后,分化的次生侵染菌丝继续扩展侵染。5.茄匍柄霉遗传类群与寄主来源有一定的相关性,与地理来源无关。采用系统发育分析和分子标记对105株不同来源的茄匍柄霉进行遗传多样性分析,分别将茄匍柄霉分为4个遗传类群。来源于番茄的茄匍柄霉具有丰富的遗传变异,但与其他菌株具有一定的相似性。表明其他寄主的茄匍柄霉可能来源于番茄,因番茄的交叉侵染在不同寄主间传播,病害的流行与番茄种苗的跨区域调运存在一定的相关性。6.以甘油醛-3-磷酸脱氢酶基因(Glyceraldehyde-2-phosphate dehydrogenase,gpd)为检测靶标基因建立了番茄匍柄霉叶斑病菌的实时荧光定量PCR(Quantitative Real-time PCR,qPCR)检测技术。检测灵敏度为 4.285×102fg·μL-1以下,高于普通PCR检测技术1000倍,可以实现番茄匍柄霉叶斑病菌的定量检测,接种6小时,植株未显症前即可检测出病原菌。7.以gpd为检测靶标基因建立了番茄匍柄霉叶斑病菌的LAMP检测技术。LAMP检测灵敏度达2.4×103 fg·μL-1,高于普通PCR检测技术100倍,通过钙黄绿素进行显色反应,检测时间在30min内,可用于病害潜伏、未显症期的快速检测。8.建立了茄匍柄霉对啶酰菌胺的敏感性基线,我国已有茄匍柄霉对啶酰菌胺产生抗性,主要包括SdhB-H272Y和ShdC-H134R两种靶基因位点突变类型产生的抗性。
[Abstract]:Stemphyllium solani is an important phytopathogenic fungus and has a wide range of host. In recent years, the bacterial leaf spot of the tomato, which is caused by the pathogenic bacteria, has burst in a large area in China, causing great economic loss, and the pathogen is still lacking in the research of the system. This paper is based on the study of the species, the conidia, the pathogenicity, the genetic diversity and so on, from the culture character, the conidia form, the pathogenicity and the genetic diversity of the pathogenic bacteria. The invention provides a detection technology for establishing the mould of the solanum solani, and provides an effective technical means for early warning and monitoring of the leaf spot of the leaf spot of the vegetable, and meanwhile, the drug resistance of the solanum lysolani to the mycophenamide is analyzed, and the theoretical foundation is provided for the development of the control agent for the control and control of the vegetable stolon, and the method is from September 2013 to April 2015, In our country, six provinces and cities, such as Shandong, Beijing, Guangdong, Guangxi, Liaoning, and Hainan, were investigated to investigate the harm of the leaf spot of the stolon. It was found that the dominant strain which caused the leaf spot of the tomato, the kidney bean and the long-leaf lettuce was the S. solani. The S. solani was divided into 9 types according to the colony morphology. There was no correlation between the colony morphology and the host and the geographical source, and the morphology of the conidia conidia was significantly different from that of the host and the geographical source. It was found that there was a clear differentiation of the pathogenic force of the solanum lysolani from different sources in China. According to the results of the determination of the pathogenic force of the tomato, the kidney bean and the long-leaf lettuce by the S. solani, there are two types of disease-causing type, and the pathogenic type is closely related to the host and the geographical source of the strain. The pathogenicity of PA was divided into four subpathogenic types, and the pathogenicity of PA1 was weaker than that of three different hosts. The pathogenic type of PB is divided into three sub-pathogenic types, and the virulence of PB3 is stronger than that of three different hosts. There was a strong, medium and weak difference in the pathogenicity of the same host and the geographical source. The optimal inoculation system of the tomato process was established, and the infection of the hypha was determined to be the most important way to the tomato. The hyphae germinate on the surface of the leaves, and a plurality of bud tubes were produced. The hyphae spread to the periphery and extended to form the infection hyphae near the air holes. After the invasion of the stomata, the secondary infection hyphae of the differentiation continue to expand the infection. There is a certain correlation between the genetic group and the host source of the tomato and the source of the host, which is not related to the geographical source. A phylogenetic analysis and molecular markers were used to analyze the genetic diversity of the five different sources of the solanum lysolani, which was divided into four genetic groups. The tomato-derived tomato has a rich genetic variation, but has a certain similarity with other strains. It is shown that the tomato can be derived from the tomato, and the cross-infection of the tomato can be spread among the different hosts, and the epidemic of the disease and the cross-region transfer of the tomato seedlings have a certain correlation. The real-time quantitative real-time PCR (qPCR) detection technique was established for the detection of the target gene with glyceraldehyde-3-phosphate dehydrogenase (gpd). The detection sensitivity is 4.285-102fg.mu. L-1, and the detection sensitivity is 1000-fold higher than that of the common PCR detection technology, so that the quantitative detection of the tomato stolon-like leaf spot fungus can be realized, the inoculation is carried out for 6 hours, and the pathogenic bacteria can be detected before the plant is not displayed. And the LAMP detection technology of the tomato stolon and the leaf spot of the tomato is established by using the gpd as the detection target gene. The detection sensitivity of LAMP is up to 2.4 to 103 fg. mu; L-1, which is 100 times higher than that of the common PCR detection technology, and the color development reaction is carried out by the calcein, the detection time is within 30 minutes, and the LAMP detection sensitivity can be used for the rapid detection of the disease latent and outstanding symptoms. In this paper, the sensitivity of the solanum lysolani to the amidogenic amine is established, and the resistance of the solanum lysolani to the mycophenamide has been established, which mainly includes the resistance of the two target gene site mutation types of the SdhB-H272Y and the ShdC-H134R.
【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S436.3
[Abstract]:Stemphyllium solani is an important phytopathogenic fungus and has a wide range of host. In recent years, the bacterial leaf spot of the tomato, which is caused by the pathogenic bacteria, has burst in a large area in China, causing great economic loss, and the pathogen is still lacking in the research of the system. This paper is based on the study of the species, the conidia, the pathogenicity, the genetic diversity and so on, from the culture character, the conidia form, the pathogenicity and the genetic diversity of the pathogenic bacteria. The invention provides a detection technology for establishing the mould of the solanum solani, and provides an effective technical means for early warning and monitoring of the leaf spot of the leaf spot of the vegetable, and meanwhile, the drug resistance of the solanum lysolani to the mycophenamide is analyzed, and the theoretical foundation is provided for the development of the control agent for the control and control of the vegetable stolon, and the method is from September 2013 to April 2015, In our country, six provinces and cities, such as Shandong, Beijing, Guangdong, Guangxi, Liaoning, and Hainan, were investigated to investigate the harm of the leaf spot of the stolon. It was found that the dominant strain which caused the leaf spot of the tomato, the kidney bean and the long-leaf lettuce was the S. solani. The S. solani was divided into 9 types according to the colony morphology. There was no correlation between the colony morphology and the host and the geographical source, and the morphology of the conidia conidia was significantly different from that of the host and the geographical source. It was found that there was a clear differentiation of the pathogenic force of the solanum lysolani from different sources in China. According to the results of the determination of the pathogenic force of the tomato, the kidney bean and the long-leaf lettuce by the S. solani, there are two types of disease-causing type, and the pathogenic type is closely related to the host and the geographical source of the strain. The pathogenicity of PA was divided into four subpathogenic types, and the pathogenicity of PA1 was weaker than that of three different hosts. The pathogenic type of PB is divided into three sub-pathogenic types, and the virulence of PB3 is stronger than that of three different hosts. There was a strong, medium and weak difference in the pathogenicity of the same host and the geographical source. The optimal inoculation system of the tomato process was established, and the infection of the hypha was determined to be the most important way to the tomato. The hyphae germinate on the surface of the leaves, and a plurality of bud tubes were produced. The hyphae spread to the periphery and extended to form the infection hyphae near the air holes. After the invasion of the stomata, the secondary infection hyphae of the differentiation continue to expand the infection. There is a certain correlation between the genetic group and the host source of the tomato and the source of the host, which is not related to the geographical source. A phylogenetic analysis and molecular markers were used to analyze the genetic diversity of the five different sources of the solanum lysolani, which was divided into four genetic groups. The tomato-derived tomato has a rich genetic variation, but has a certain similarity with other strains. It is shown that the tomato can be derived from the tomato, and the cross-infection of the tomato can be spread among the different hosts, and the epidemic of the disease and the cross-region transfer of the tomato seedlings have a certain correlation. The real-time quantitative real-time PCR (qPCR) detection technique was established for the detection of the target gene with glyceraldehyde-3-phosphate dehydrogenase (gpd). The detection sensitivity is 4.285-102fg.mu. L-1, and the detection sensitivity is 1000-fold higher than that of the common PCR detection technology, so that the quantitative detection of the tomato stolon-like leaf spot fungus can be realized, the inoculation is carried out for 6 hours, and the pathogenic bacteria can be detected before the plant is not displayed. And the LAMP detection technology of the tomato stolon and the leaf spot of the tomato is established by using the gpd as the detection target gene. The detection sensitivity of LAMP is up to 2.4 to 103 fg. mu; L-1, which is 100 times higher than that of the common PCR detection technology, and the color development reaction is carried out by the calcein, the detection time is within 30 minutes, and the LAMP detection sensitivity can be used for the rapid detection of the disease latent and outstanding symptoms. In this paper, the sensitivity of the solanum lysolani to the amidogenic amine is established, and the resistance of the solanum lysolani to the mycophenamide has been established, which mainly includes the resistance of the two target gene site mutation types of the SdhB-H272Y and the ShdC-H134R.
【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S436.3
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相关期刊论文 前10条
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