我国玉米新病害—玉米尾孢灰斑病研究

发布时间：2018-06-05 21:38

本文选题：玉米灰斑病 + 玉米尾孢　；参考：《沈阳农业大学》2017年博士论文

【摘要】：玉米(Zea mays L.)是世界上最重要的农作物之一,与小麦和水稻并称世界三大粮食作物。玉米不但是人类的主粮,更是饲料、轻工和医药等工业的重要原料,在国民经济发展中具有重要地位。玉米灰斑病是一种在世界范围普遍发生并造成严重损失的病害,目前已成为我国玉米上的重要病害之一。1991年在我国丹东地区发生了由玉蜀黍尾孢菌(Cercospora zeae-maydis Tehon and Daniels)侵染所致的玉米灰斑病。长期以来国内研究者一致认为我国玉米灰斑病的病原菌都是玉蜀黍尾孢,而笔者于2010年在我国云南发现了一种由玉米尾孢菌(Cercosporazeina CrousU.Braun sp.nov.)侵染所致的玉米灰斑病。该病在云南地区病已造成严重损失,且有向北方地区流行蔓延的趋势。目前,国内外尚未见对该病害系统研究的报道,其发生流行规律不清,缺乏有效的防控措施,具有潜在爆发流行的威胁。对此,本研究采用植物病理学和分子生物学相结合的技术,对该病的病害症状、病原学、病害诊断、病菌种群分布、病菌致病性、种质资源抗病性、寄主抗病机制及抗性QTL定位等方面进行系统研究,以期为该病害的诊断、监测预警和有效防控提供科学依据。主要的研究结果如下:1.首次报道了玉米尾孢(Cercospora zeina Crous＆U.Braun sp.nov.)侵染引起的玉米灰斑病在我国发生。2010～2011年间,详细调查了我国云南地区玉米灰斑病的发生及为害情况,观察记录了病害症状特征,并从云南昆明、大理、德宏、保山等地区采集了具有典型症状的玉米叶片,制成标本。通过单孢分离获得病原菌,对病原菌进行了形态学、特异性引物鉴定和rDNA-ITS序列分析,并结合柯赫氏法则证病,证明云南地区发生的玉米灰斑病为我国首次报道的新病害,其病原菌为玉米尾孢。2.系统研究了玉米尾孢菌与玉蜀黍尾孢菌生物学特性的差异。对两种玉米灰斑病病原尾孢菌的生物学特性进行了比较分析,结果表明:两种尾孢菌生长的适宜培养基、碳源、氮源、温度及其分生孢子萌发的适宜温度和pH值均有明显的差异,说明两种尾孢菌在生物学特性上存在明显的差异。3.研究明确了我国玉米灰斑病病原菌的种群及其地域分布。2010～2016年,利用植物病理学和分子生物学技术,对采自我国13省97市(县)的玉米灰斑病样品进行检测和分析,结果表明:我国的玉米灰斑病菌种群中存在玉蜀黍尾孢(C.zeae-maydis)和玉米尾孢(C.zeina)两个种。玉蜀黍尾孢菌在黑龙江、吉林、辽宁、内蒙古、山西、山东、陕西、河南、四川、湖北和湖南等省区均有分布。而玉米尾孢菌主要在云南、贵州、四川、湖南、湖北、陕西和河南等省区有分布,但有以云南为传播流行中心向东、北方向扩散的趋势。4.对300份玉米种质进行了抗玉米尾孢灰斑病和玉蜀黍尾孢灰斑病评价。建立了玉米尾孢灰斑病的人工接种抗病鉴定技术体系。鉴定了 300份玉米种质对玉米尾孢灰斑病和玉蜀黍尾孢灰斑病抗性,结果表明:对玉米尾孢灰斑病表现高抗、抗和中抗的种质分别有10份、100份和121份,表现感和高感的种质依次有62份和7份;对玉蜀黍尾孢灰斑病表现高抗、抗和中抗的种质分别有4份、33份和78份,表现感和高感的种质依次有137份和48份。300份种质中只有77份对两种灰斑病的抗性表现一致,而149份种质对两种灰斑病的抗性表现截然相反,说明两种灰斑病菌的致病性有很大的差异。76.99%的供试种质对玉米尾孢灰斑病表现为高抗、抗或中抗,而61.67%的供试种质对玉蜀黍尾孢灰斑病表现为感或高感,说明玉米尾孢对大多数种质的致病力比玉蜀黍尾孢弱。5.研究明确了玉米尾孢菌侵染对寄主防御酶系活性的影响。利用抗病自交系De813和感病自交系R151Ht1A,通过人工接种的方法,研究玉米尾孢菌侵染后抗、感病玉米自交系体内防御酶活性的变化,从生理生化的角度揭示玉米对玉米尾孢菌的抗性机制。抗病性不同的玉米自交系在未被玉米尾孢菌侵染时,其防御酶(PAL、POD、PPO、SOD和CAT)活性略有差异,但差异不是很大;而接种玉米尾孢菌后,抗病自交系的各防御酶活性升高得明显比感病自交系的更为迅速,而且抗病自交系的各防御酶活性峰值也比感病自交系的明显要高,并且抗病自交系的防御酶活性能够长时间地维持在较高的水平。说明抗病自交系在受到玉米尾孢菌侵染后生理生化反应更为迅速、持久。6.对抗玉米尾孢灰斑病QTL进行了初步定位。以抗玉米尾孢灰斑病自交系De813和感病自交系R151Ht1A为亲本,组建了 F2分离群体,利用SSR-BSA法筛选与玉米尾孢灰斑病抗性连锁的分子标记,并利用F2分离群体(367株)来检测所得的分子标记与抗性基因的连锁程度,以确定抗性基因在染色体上的位置。结果表明:在玉米1号染色体短臂的bin1.04-1.05处有1个抗玉米尾孢灰斑病的主效QTL,该位点位于标记J91和J96之间遗传图距14.73cM、物理距离约为4Mb的区域内,与两个标记的遗传图距分别为8.03cM和6.7cM,该QTL可以解释53.0498%的表型变异率,加性效应达到-2.7218,显性效应达到-1.07。
[Abstract]:Corn (Zea mays L.) is one of the most important crops in the world, with wheat and rice and the three largest food crop in the world. Corn is not only the main food of human, but also an important raw material in the feed, light industry and medicine industry. It has an important position in the development of the national economy. The disease of heavy loss has become one of the most important diseases on corn in our country at present. In the year.1991, the corn gray spot caused by the infection of Cercospora zeae-maydis Tehon and Daniels in the Dandong area of China. In 2010, a kind of corn grey spot disease caused by Cercosporazeina CrousU.Braun sp.nov. infection was found in Yunnan, China. The disease has caused serious loss in Yunnan area and has the trend of spreading to the northern region. At present, there is no report on the disease system at home and abroad. In this study, the disease symptoms, etiology, disease diagnosis, pathogen distribution, pathogen pathogenicity, disease resistance, disease resistance of germ plasm resources, host resistance mechanism and resistance QTL location of the disease were introduced in this study. In order to provide scientific basis for the diagnosis, monitoring, early warning and effective prevention and control of the disease, the main research results are as follows: 1. the first report of maize caudal Cyclospora (Cercospora Zeina and u. Sp.nov.) caused by the infection of Corn Gray Spot in China from.2010 to 2011, the grey spot of corn in Yunnan area of China was investigated in detail. The symptoms of the disease were recorded and the symptoms of the disease were recorded. The typical symptoms of maize leaves were collected from Kunming, Dali, Dehong, Baoshan and other regions of Yunnan. The pathogen was obtained by separation of monospora. The pathogenic bacteria were identified by morphology, specific primers and rDNA-ITS sequence analysis, combined with Kirsch's method. It was proved that the corn Grey Spot in Yunnan was the first new disease reported in our country. Its pathogen was the difference between the biological characteristics of caudal caudate.2. system and the biological characteristics of maize caudal cyclosporin. The biological characteristics of the two kinds of the pathogen of the pathogen of the pathogen were compared and analyzed. The results showed that two kinds of caudal cyclosporin were found. The suitable culture medium, carbon source, nitrogen source, temperature and the germination of the conidia were obviously different in the temperature and pH value, indicating that there were obvious differences in biological characteristics between the two species of caudal cauda, and the.3. study clearly defined the population and regional distribution of the pathogen of the pathogen of Chinese corn grey spot from.2010 to 2016, using plant pathology and molecule. Biological techniques were used to detect and analyze the samples of corn gray spots in 97 cities and counties of 13 provinces of China. The results showed that there were two species of Maize (C.zeae-maydis) and maize caudal spore (C.zeina) in the population of Chinese Corn Gray Spot bacteria. The fungus of corn was in Heilongjiang, Jilin, Liaoning, Inner Mongolia, Shanxi, Shandong, Shaanxi, Henan, four. Provinces and regions such as Sichuan, Hubei and Hunan are distributed, and caudal Cercospora is mainly distributed in Yunnan, Guizhou, Sichuan, Hunan, Hubei, Shaanxi and Henan, but the trend of the spread of Yunnan as the spreading center to the East and the north direction.4. has been evaluated for 300 maize germplasms. An artificial inoculation resistance identification technique system was established. The resistance of 300 Maize Germplasms to caudal caudal and corn caudal spotted was identified. The results showed that there were 10, 100 and 121 germplasms with high resistance, 100 and 121, and 62 germplasms with a sense of expression and high sensitivity. 7, there were 4, 33 and 78 germplasms with high resistance, 33 and 78, respectively. There were 137 and 48.300 germplasms, only 77 were in the same resistance to two species of gray spots, and 149 germplasms showed the opposite of resistance to two kinds of gray spots, indicating that two spots were found. The test Germplasms of.76.99% showed high resistance, resistance or moderate resistance to maize caudal Caulis speckle, and 61.67% of the tested germplasms showed a sense of sensation or high sensitivity to the blotch of corn caudal caudorum. It showed that the pathogenicity of caudate to most of the germplasm was better than that of maize caudal Cyclospora.5.. The effect of enzyme activity. Using the disease resistant inbred line De813 and the sense disease inbred line R151Ht1A, the changes of the defense enzyme activity in the maize inbred lines after the infection were studied by artificial inoculation, and the resistance mechanism of Maize to maize Caulis was revealed from the physiological and biochemical point of view. The maize inbred lines with different disease resistance were not The activities of the defense enzymes (PAL, POD, PPO, SOD and CAT) were slightly different, but the difference was not very large, but after inoculation of cauda Cercospora, the activity of defense enzymes in the resistant inbred lines increased significantly more rapidly than that of the susceptible inbred lines, and the activity peaks of the defense enzymes in the disease resistant inbred lines were also significantly higher than those of the inbred line. The defense enzyme activity of the disease resistant inbred line can be maintained at a high level for a long time. It indicates that the physiological and biochemical reaction of the disease resistant inbred line is more rapid after the infection of the caudal cauda Cercospora, and the persistent.6. against maize caudal ash spot QTL is preliminarily located. It is resistant to the inbred line De813 and the susceptible inbred line R151Ht1A. For the parents, the F2 separation group was established, and the molecular markers linked with the resistance of maize caudal cauda were screened by SSR-BSA method, and the linkage degree of the molecular markers and resistance genes was detected by F2 Separation Group (367 strains), so as to determine the position of the resistant gene on the chromosome. The results showed that the bin1.04- in the short arm of the maize chromosome was bin1.04-. There are 1 main effective QTL, which are located between the marker J91 and J96, where the genetic map is between the markers and the 14.73cM, the physical distance is about 4Mb, and the two markers are 8.03cM and 6.7cM respectively. This QTL can explain the 53.0498% phenotypic variation rate, the additive effect reaches -2.7218, and the dominant effect reaches -1.07..
【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】：S435.131.4

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