山西省植物病原镰孢菌种群分布及遗传变异分析
发布时间：2018-10-16 21:45
【摘要】：【目的】通过对镰孢菌(Fusarium)ITS、EF-1α和β-tubulin 3个基因序列比较分析,筛选适合于镰孢菌种类鉴定的基因序列,并以此序列分析山西省植物病原镰孢菌种群分布及遗传变异情况。【方法】从2013—2015年在山西省11市28县(区)采集分离的625株镰孢菌菌株中选取形态学清晰的菌株进行ITS、EF-1α和β-tubulin基因序列联合分析,运用Sequencher软件对序列进行拼接和校对,将测序结果与NCBI及FUSARIUM-ID数据库中所有已公布的序列进行BLAST分析,结合下载概念清晰或标准种序列,运用Clustal X和Gel Doc软件进行序列对齐和编辑,运用MEGA、Excel、DNAstar和Taxon Gap软件分析镰孢菌种内种间遗传变异情况,从ITS、EF-1α和β-tubulin中筛选适合镰孢菌种类鉴定的基因序列,并以此序列分析山西省镰孢菌的种群分布特点等。【结果】在3个候选基因序列中,EF-1α基因序列是最适用于镰孢菌种类鉴定的基因序列,其种间平均遗传距离分别是种内平均遗传距离的24倍,种内差异小于种间差异的种的数量最多,达到供试种的73%,物种鉴定准确率最强,达到87%。基于EF-1α基因片段的系统发育分析结果表明,在供试的27种镰孢菌中,有22种表现出单系性,相同种的不同菌株以较高支持率聚成独立支。在27种镰孢菌中,F.oxysporum是山西省镰孢菌的优势种,分离频率最高(22.1%),且分布最广,在23个县(区)均有分布;其次是F.solani(13.8%),在14个县(区)有分布。从不同地区镰孢菌的种群结构及分布来看,运城、临汾、忻州、长治、吕梁、晋中和太原均以F.oxysporum为优势种,朔州以F.lateritium为优势种,大同以F.solani为优势种,晋城以F.verticillioides为优势种,阳泉以F.incarnatum为优势种;其中晋中和忻州的镰孢菌种类最丰富,临汾次之,朔州最少。从不同寄主镰孢菌的种群结构及分布来看,番茄上镰孢菌的种类最多(15种),其次是马铃薯(13种)和大豆(12种);番茄、黄瓜、西瓜、马铃薯、茄子、西葫芦和甘蓝,均以F.oxysporum为优势种,大豆和玉米以F.verticillioides为优势种,小麦以F.avenaceum和F.graminearum为优势种。【结论】镰孢菌种内种间存在丰富的遗传变异,其中EF-1α基因序列的遗传变异最适用于镰孢菌种类的鉴定,但形态学种和系统发育学种不完全吻合。F.oxysporum是山西省镰孢菌的优势种,不同地区、不同寄主上镰孢菌种群存在明显的遗传分化;研究结果可为镰孢菌的分类鉴定、DNA条形码筛选、检验检疫及其综合防治提供理论依据。
[Abstract]:[objective] by comparing and analyzing the three gene sequences of (Fusarium) ITS,EF-1 伪 and 尾-tubulin in Fusarium, the gene sequences suitable for identification of Fusarium species were screened. The population distribution and genetic variation of Fusarium graminearum in Shanxi Province were analyzed by using this sequence. [methods] from 625 Fusarium strains collected from 28 counties (districts) of 11 cities in Shanxi Province from 2013-2015, the morphology of Fusarium spp. Was clear. ITS,EF-1 伪 and 尾-tubulin gene sequences were analyzed. The sequence was spliced and proofread by Sequencher software, and the sequence results were analyzed by BLAST with all the published sequences in NCBI and FUSARIUM-ID database. The sequence alignment and editing were carried out by using Clustal X and Gel Doc software, combined with the clear concept of downloading or standard species sequence. MEGA,Excel,DNAstar and Taxon Gap software were used to analyze the genetic variation of Fusarium intraspecies. The gene sequences suitable for identification of Fusarium species were screened from ITS,EF-1 伪 and 尾-tubulin. [results] among the three candidate gene sequences, EF-1 伪 gene sequence is the most suitable gene sequence for identification of Fusarium species. The average genetic distance between species was 24 times of the average genetic distance within the species. The number of species with the difference within the species was the most than that of the interspecific species, reaching 73 for the test species, and the accuracy of species identification was the strongest, reaching 87. Phylogenetic analysis based on EF-1 伪 gene fragments showed that 22 of the 27 Fusarium species tested showed monophyletic characteristics, and different strains of the same species clustered into independent branches with high support. Among the 27 Fusarium species, F.oxysporum is the dominant species of Fusarium in Shanxi Province, with the highest isolation frequency (22.1%) and the widest distribution in 23 counties (districts), followed by F.solani (13.8%) and 14 counties (districts). According to the population structure and distribution of Fusarium in different regions, Yuncheng, Linfen, Xinzhou, Changzhi, Lv Liang, Jinzhong and Taiyuan all have F.oxysporum as dominant species, Shuozhou as dominant species, Datong as dominant species, F.solani as dominant species, Jincheng as dominant species, and Jincheng as dominant species. F.incarnatum is the dominant species in Yangquan, in which Fusarium species are the most abundant in Jinzhong and Xinzhou, followed by Linfen and Shuozhou. According to the population structure and distribution of Fusarium mongolicum, the species of Fusarium graminearum were the most (15 species), followed by potato (13 species) and soybean (12 species), tomato, cucumber, watermelon, potato, eggplant, squash and cabbage. F.oxysporum was the dominant species, F.verticillioides was the dominant species in soybean and maize, and F.avenaceum and F.graminearum were the dominant species in wheat. [conclusion] there is abundant genetic variation among species of Fusarium, and the genetic variation of EF-1 伪 gene is the most suitable for identification of Fusarium species. But the morphological species and phylogenetic species are not completely consistent. F.oxysporum is the dominant species of Fusarium in Shanxi Province, and there are obvious genetic differentiation among different host populations in different regions, the results can be classified and identified by DNA bar code screening. Inspection and quarantine and its comprehensive prevention and treatment provide theoretical basis.
【作者单位】： 山西农业大学农学院;
【基金】：山西省科技攻关项目(20120311019-3) 山西省科技基础条件平台建设项目(1105-0104)
【分类号】：S432.4
本文编号：2275713
[Abstract]:[objective] by comparing and analyzing the three gene sequences of (Fusarium) ITS,EF-1 伪 and 尾-tubulin in Fusarium, the gene sequences suitable for identification of Fusarium species were screened. The population distribution and genetic variation of Fusarium graminearum in Shanxi Province were analyzed by using this sequence. [methods] from 625 Fusarium strains collected from 28 counties (districts) of 11 cities in Shanxi Province from 2013-2015, the morphology of Fusarium spp. Was clear. ITS,EF-1 伪 and 尾-tubulin gene sequences were analyzed. The sequence was spliced and proofread by Sequencher software, and the sequence results were analyzed by BLAST with all the published sequences in NCBI and FUSARIUM-ID database. The sequence alignment and editing were carried out by using Clustal X and Gel Doc software, combined with the clear concept of downloading or standard species sequence. MEGA,Excel,DNAstar and Taxon Gap software were used to analyze the genetic variation of Fusarium intraspecies. The gene sequences suitable for identification of Fusarium species were screened from ITS,EF-1 伪 and 尾-tubulin. [results] among the three candidate gene sequences, EF-1 伪 gene sequence is the most suitable gene sequence for identification of Fusarium species. The average genetic distance between species was 24 times of the average genetic distance within the species. The number of species with the difference within the species was the most than that of the interspecific species, reaching 73 for the test species, and the accuracy of species identification was the strongest, reaching 87. Phylogenetic analysis based on EF-1 伪 gene fragments showed that 22 of the 27 Fusarium species tested showed monophyletic characteristics, and different strains of the same species clustered into independent branches with high support. Among the 27 Fusarium species, F.oxysporum is the dominant species of Fusarium in Shanxi Province, with the highest isolation frequency (22.1%) and the widest distribution in 23 counties (districts), followed by F.solani (13.8%) and 14 counties (districts). According to the population structure and distribution of Fusarium in different regions, Yuncheng, Linfen, Xinzhou, Changzhi, Lv Liang, Jinzhong and Taiyuan all have F.oxysporum as dominant species, Shuozhou as dominant species, Datong as dominant species, F.solani as dominant species, Jincheng as dominant species, and Jincheng as dominant species. F.incarnatum is the dominant species in Yangquan, in which Fusarium species are the most abundant in Jinzhong and Xinzhou, followed by Linfen and Shuozhou. According to the population structure and distribution of Fusarium mongolicum, the species of Fusarium graminearum were the most (15 species), followed by potato (13 species) and soybean (12 species), tomato, cucumber, watermelon, potato, eggplant, squash and cabbage. F.oxysporum was the dominant species, F.verticillioides was the dominant species in soybean and maize, and F.avenaceum and F.graminearum were the dominant species in wheat. [conclusion] there is abundant genetic variation among species of Fusarium, and the genetic variation of EF-1 伪 gene is the most suitable for identification of Fusarium species. But the morphological species and phylogenetic species are not completely consistent. F.oxysporum is the dominant species of Fusarium in Shanxi Province, and there are obvious genetic differentiation among different host populations in different regions, the results can be classified and identified by DNA bar code screening. Inspection and quarantine and its comprehensive prevention and treatment provide theoretical basis.
【作者单位】： 山西农业大学农学院;
【基金】：山西省科技攻关项目(20120311019-3) 山西省科技基础条件平台建设项目(1105-0104)
【分类号】：S432.4
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