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拟轮枝镰孢菌EST-SSR信息分析与标记开发
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引用本文:李新凤,王建明,姜晓东,郝晓娟,张祖维,田宏先.拟轮枝镰孢菌EST-SSR信息分析与标记开发.植物保护学报,2018,45(4):819-826
DOI:10.13802/j.cnki.zwbhxb.2018.2016223
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作者单位E-mail
李新凤 山西农业大学农学院, 太谷 030801  
王建明 山西农业大学农学院, 太谷 030801 jm.w@sohu.com 
姜晓东 山西农业大学农学院, 太谷 030801  
郝晓娟 山西农业大学农学院, 太谷 030801  
张祖维 山西农业大学动物科技学院, 太谷 030801  
田宏先 山西省农业科学院高寒区作物研究所, 大同 037000  
中文摘要:为开发可用于拟轮枝镰孢菌Fusarium verticillioides及其近缘种遗传多样性分析的SSR引物,利用生物信息学方法和PCR技术,通过对从NCBI下载的87 086条拟轮枝镰孢菌的EST序列信息进行分析,设计EST-SSR引物,检测其在拟轮枝镰孢菌及其近缘种中的扩增情况,并用筛选出的多态性引物对15株拟轮枝镰孢菌进行SSR遗传多样性分析。结果表明,在EST序列中,共查找到11 952个SSR位点,592种重复基元,SSR出现频率为1.09%,重复基元出现数量最多的为三核苷酸(54.00%),其中(CAA/TTG) n基元出现频率最高。设计的25对EST-SSR引物在拟轮枝镰孢菌种内的有效扩增率和多态率分别为80.00%与32.00%,对5种近缘镰孢菌种的通用率和多态率分别为40.00%和8.00%。遗传多样性分析结果表明,在相似系数为0.664水平下,供试菌株可划分为4个SSR类群,但类群的划分与菌株的地理来源无关;不同菌株间存在明显的遗传分化。表明基于拟轮枝镰孢菌EST序列开发的SSR引物可用于拟轮枝镰孢菌及其近缘种的遗传多样性分析。
中文关键词:拟轮枝镰孢菌  EST-SSR  信息分析  遗传多样性  通用性
 
EST-SSR information analysis and marker development for genetic diversity analysis of Fusarium verticillioides
Author NameAffiliationE-mail
Li Xinfeng College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
Wang Jianming College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China jm.w@sohu.com 
Jiang Xiaodong College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
Hao Xiaojuan College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
Zhang Zuwei College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China  
Tian Hongxian Research Institute of Alpine Crops, Shanxi Academy of Agricultural Sciences, Datong 037000, Shanxi Province, China  
Abstract:To develop EST-SSR markers for genetic diversity analysis of Fusarium verticillioides and related Fusarium spp. strains, the SSRs of Fusarium verticillioides were analyzed and EST-SSR primers were designed based on 87 086 ESTs obtained from NCBI database using bioinformatics method and PCR technology. The genetic diversity of 15 F. verticillioides strains were analyzed with the polymorphic primers screened. The results showed that a total of 11 952 SSRs loci were detected with a frequency of 1.09%, and 592 types of motifs were identified in F. verticillioides ESTs. The most abundant motif was trimucleotide repeats (54.00%). (CAA/TTG)n occurred more often than other repeat motifs. The effective amplification rate of the 25 designed primers in F. verticillioides strains was 80.00%, and 32.00% of these primers were polymorphic. The rates of amplification transferability and polymorphism in five Fusarium spp. strains were 40.00% and 8.00%, respectively. The genetic diversity results indicated that all the strains were clustered into four groups with a similarity coefficient of 0.664, and there was no correlation between SSR grouping and the geographic origins of strains. Obvious genetic diversity existed among the strains. The results indicated that the EST-SSR markers could be used to analyze the genetic diversity of F. verticillioides and related species in Fusarium.
keywords:Fusarium verticillioides  EST-SSR  information analysis  genetic diversity  transferability
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