我國蔬菜匍柄霉葉斑病的病原學(xué)及對(duì)啶酰菌胺抗藥性研究
發(fā)布時(shí)間：2019-03-02 19:36
【摘要】：茄匍柄霉(Stemphylium solani)是一種重要的植物病原真菌,寄主范圍廣泛。近年來,由該病原菌引起的番茄匍柄霉葉斑病在我國大面積爆發(fā),造成巨大的經(jīng)濟(jì)損失,該病原菌目前尚缺乏系統(tǒng)的研究。本論文以來源于不同地區(qū)的茄匍柄霉為研究對(duì)象,從病原菌的培養(yǎng)性狀、分生孢子形態(tài)、致病性、遺傳多樣性等方面分析茄匍柄霉的種群分化。建立茄匍柄霉的檢測(cè)技術(shù),為蔬菜匍柄霉葉斑病的早期預(yù)警監(jiān)測(cè)提供有效的技術(shù)手段,同時(shí)分析了茄匍柄霉對(duì)啶酰菌胺的抗藥性,為蔬菜匍柄霉防治藥劑的開發(fā)提供了理論依據(jù)。1.2013年9月至2015年4月,在我國山東、北京、廣東、廣西、遼寧、海南等6個(gè)省市調(diào)查匍柄霉葉斑病的危害,發(fā)現(xiàn)引起番茄、菜豆、長葉萵苣匍柄霉葉斑病的優(yōu)勢(shì)菌株為茄匍柄霉(S.solani)。2.依據(jù)菌落形態(tài)將茄匍柄霉(S.solani)劃分為9種類型。茄匍柄霉的菌落形態(tài)與寄主和地理來源沒有相關(guān)性,茄匍柄霉分生孢子形態(tài)差異顯著,與寄主和地理來源顯著相關(guān)。3.發(fā)現(xiàn)我國不同來源的茄匍柄霉存在明顯的致病力分化。根據(jù)茄匍柄霉(S.solani)對(duì)番茄、菜豆和長葉萵苣的致病力測(cè)定結(jié)果,明確茄匍柄霉存在兩種致病型,致病型與菌株的寄主和地理來源密切相關(guān)。PA致病型分為4種亞致病型,PA1亞致病型對(duì)三種不同寄主的致病力較弱。PB致病型分為3種亞致病型,PB3亞致病型對(duì)三種不同寄主的致病力較強(qiáng)。同一寄主和地理來源的茄匍柄霉致病力存在強(qiáng)、中、弱的差異。4.建立了茄匍柄霉侵染番茄過程研究的最佳接種體系,明確了菌絲侵染是茄匍柄霉對(duì)番茄的重要侵染途徑。菌絲在葉片表面萌發(fā),產(chǎn)生多個(gè)芽管,菌絲向四周擴(kuò)散伸長,到氣孔附近形成侵染菌絲,從氣孔侵入后,分化的次生侵染菌絲繼續(xù)擴(kuò)展侵染。5.茄匍柄霉遺傳類群與寄主來源有一定的相關(guān)性,與地理來源無關(guān)。采用系統(tǒng)發(fā)育分析和分子標(biāo)記對(duì)105株不同來源的茄匍柄霉進(jìn)行遺傳多樣性分析,分別將茄匍柄霉分為4個(gè)遺傳類群。來源于番茄的茄匍柄霉具有豐富的遺傳變異,但與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來源于番茄,因番茄的交叉侵染在不同寄主間傳播,病害的流行與番茄種苗的跨區(qū)域調(diào)運(yùn)存在一定的相關(guān)性。6.以甘油醛-3-磷酸脫氫酶基因(Glyceraldehyde-2-phosphate dehydrogenase,gpd)為檢測(cè)靶標(biāo)基因建立了番茄匍柄霉葉斑病菌的實(shí)時(shí)熒光定量PCR(Quantitative Real-time PCR,qPCR)檢測(cè)技術(shù)。檢測(cè)靈敏度為 4.285×102fg·μL-1以下,高于普通PCR檢測(cè)技術(shù)1000倍,可以實(shí)現(xiàn)番茄匍柄霉葉斑病菌的定量檢測(cè),接種6小時(shí),植株未顯癥前即可檢測(cè)出病原菌。7.以gpd為檢測(cè)靶標(biāo)基因建立了番茄匍柄霉葉斑病菌的LAMP檢測(cè)技術(shù)。LAMP檢測(cè)靈敏度達(dá)2.4×103 fg·μL-1,高于普通PCR檢測(cè)技術(shù)100倍,通過鈣黃綠素進(jìn)行顯色反應(yīng),檢測(cè)時(shí)間在30min內(nèi),可用于病害潛伏、未顯癥期的快速檢測(cè)。8.建立了茄匍柄霉對(duì)啶酰菌胺的敏感性基線,我國已有茄匍柄霉對(duì)啶酰菌胺產(chǎn)生抗性,主要包括SdhB-H272Y和ShdC-H134R兩種靶基因位點(diǎn)突變類型產(chǎn)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.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：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.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S436.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 趙麗萍;趙統(tǒng)敏;余文貴;楊瑪麗;王銀磊;;番茄灰葉斑病研究進(jìn)展[J];江蘇農(nóng)業(yè)學(xué)報(bào);2014年06期
2 張慧麗;段維軍;張吉紅;王江嶺;徐瑛;陳先鋒;;環(huán)介導(dǎo)等溫?cái)U(kuò)增技術(shù)檢測(cè)油棕猝倒病菌的研究[J];浙江農(nóng)業(yè)學(xué)報(bào);2013年02期
3 李長松;徐作s，
本文編號(hào)：2433395
本文鏈接：http://sikaile.net/shoufeilunwen/nykjbs/2433395.html
最近更新
教材專著
