推迟播期对向日葵黄萎病发生程度影响以及黄萎病菌遗传多样性研究

发布时间：2018-03-13 12:09

本文选题：向日葵黄萎病　切入点：大丽轮枝菌　出处：《内蒙古农业大学》2017年博士论文　论文类型：学位论文

【摘要】：向日葵黄萎病是向日葵上普遍发生的病害,近年来由于向日葵商品种子的频繁运输及向日葵轮作倒茬的困难,使得黄萎病不断加重,成为制约我国向日葵产业发展的重要因素。本研究以我国主要向日葵产区采集分离到的向日葵黄萎病菌为材料,研究推迟播期对向日葵黄萎病发生及向日葵产量的影响;向日葵种皮携带黄萎病菌的可能性;向日葵黄萎病菌的遗传多样性;以及通过R-PCR技术筛选与向日葵黄萎病菌致病力有关的候选基因,得到以下结论:1.在2010-2012年的不同地点的进行的不同播期对向日葵黄萎病发程度的试验中,除2010年在内蒙古巴彦淖尔市五原县,随着播期的推迟黄萎病发病程度呈现先上升后下降的趋势外,其余各点在不同年份的播期实验中,黄萎病发生程度都随向日葵播期推迟而下降,说明推迟向日葵播期可以降低黄萎病的发生程度。同时,不同播期的测产结果表明随着播期推迟,向日葵黄萎病害逐渐减轻,其产量也呈现出逐渐增高的态势。2.向日葵种子带菌的研究结果表明,利用真菌的通用引物对ITS1/4进行PCR扩增时出现的双带型,说明在向日葵种子种皮上可能存在包括黄萎病菌在内的不同类型的其他真菌。而利用黄萎病菌的特异引物VD1/2进行巢氏PCR再次扩增出单一的条带的结果不仅说明黄萎病菌引物VD1/2的特异性,同时也表明向日葵种皮携带有黄萎病菌。同时,结合荧光显微观察我们确定了向日葵种子的种皮能够携带黄萎病菌进行远距离传播。3.本研究利用国际通用的大丽轮枝菌不同亲合组的标准菌株对57株来自我国不同向日葵产区的黄萎菌进行营养亲和群的划分结果表明,供试的57份菌株可以被划分为三个不同的营养亲和群即VCG4B、VCG4A和VCG2B,其中VCG4B为优势亲和群体。亲和群VCG2B和VCG4B中包括的黄萎病菌菌株与其采集的地域分布没有相关性。4.利用国际黄萎病菌生理小种1和2的通用引物对我国向日葵种植区黄萎病菌进行生理小种鉴定结果表明,所有分离到的向日葵黄萎菌都是2号生理小种。对来源于我国不同向日葵种植区的黄萎菌进行交配型鉴定结果表明,向日葵大丽轮枝菌中有MAT1-1和MAT1-2两种交配型,其中MAT1-1型为优势交配型。5.对我国主要向日葵产区的向日葵大丽轮枝菌进行IGS序列系统发育进行分析的结果表明,供试的57份向日葵黄萎病菌菌株被划分为成3个大的聚类群,其中大多数的鉴定为VCG4亲和群的菌株都分布在一个群中,而VCG2B型的菌株却分布在三个不同的聚类群中,说明VCG2B亲和群的菌株在遗传上具有多源性。6.利用R-PCR技术筛选得到一些向日葵黄萎病菌致病相关基因的EST序列,通过对所筛选到的序列进行功能注释,发现编码甘露醇-1-磷酸5-脱氢酶的基因,编码钙调蛋白、糖苷水解酶基因,编码过氧化物酶、STI35蛋白基因可能参与了向日葵大丽轮枝菌的致病过程。
[Abstract]:Sunflower verticillium wilt is a common disease in sunflower. In recent years, because of the frequent transportation of sunflower commercial seeds and the difficulty of rotation of sunflower rotation, Verticillium wilt becomes more and more serious. This study studied the effect of delaying sowing date on the occurrence of sunflower verticillium wilt and the yield of sunflower. The possibility of sunflower seed coat carrying Verticillium wilt, genetic diversity of sunflower Verticillium wilt, and screening candidate genes related to virulence of sunflower Verticillium wilt by R-PCR, The following conclusions are drawn: 1.In the experiment on the incidence of sunflower verticillium wilt in different locations in 2010-2012, except in Wuyuan County of Bayannur City, Inner Mongolia on 2010, With the delay of sowing date, the incidence degree of Verticillium wilt increased first and then decreased. In the experiment of sowing date in different years, the occurrence degree of Verticillium wilt decreased with the delay of sunflower sowing date. The result showed that delaying the sowing date of sunflower could reduce the occurrence degree of Verticillium wilt, and the result of different sowing date showed that the damage of sunflower Verticillium wilt gradually decreased with the delay of sowing date. The results of the study of sunflower seed carrying bacteria showed that the double band type appeared in the PCR amplification of ITS1/4 by using universal primers of fungi. It is suggested that there may be different types of fungi including Verticillium wilt on the seed coat of sunflower. However, the result of using the specific primer VD1/2 of Verticillium wilt to amplify a single band by nested PCR not only shows that the single band is yellow. The specificity of primer VD1/2 of Fusarium wilt, It also shows that sunflower seed coat carries Verticillium wilt. Combined with fluorescence microscopic observation, we determined that the seed coat of sunflower seeds can carry Verticillium wilt for long-distance transmission. 3. In this study, 57 standard strains from different affinity groups of Verticillium dahliae were used in this study. The results of the classification of the vegetative affinity group of wilt fungus in the same sunflower producing area showed that, The 57 strains tested can be divided into three different vegetative affinity groups: VCG4BnVCG4A and VCG2B. among them, VCG4B is the dominant affinity group. The Verticillium wilt strains included in VCG2B and VCG4B have no correlation with the regional distribution. The results of identification of Verticillium wilt strains in sunflower planting area of China by using universal primers of 1 and 2 species of Verticillium wilt showed that: 1 and 2 primers were used to identify Verticillium wilt in sunflower growing area of China. All the sunflower wilt bacteria isolated were 2 physiological races. The results of mating type identification showed that there were two mating types of MAT1-1 and MAT1-2 in the plants of different sunflower growing areas in China. Among them, MAT1-1 type is the dominant mating type. 5. The IGS sequence phylogeny of Vibrio sunflower in main sunflower producing areas in China was analyzed. The results showed that 57 strains of Verticillium wilt strains were divided into three clusters. Most of the strains identified as VCG4 affinity groups were found in one group, while VCG2B type strains were found in three different clusters. The results showed that the strains with VCG2B affinity group had multi-source. 6. the EST sequences of some pathogenic genes of Verticillium wilt were screened by R-PCR, and the function of the selected genes was annotated. It was found that the genes encoding mannitol -1-phosphate 5-dehydrogenase, calmodulin, glucoside hydrolase and peroxidase STI35 may be involved in the pathogenicity of Vibrio dahliensis.
【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S435.655

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