【摘要】：【目的】通過(guò)對(duì)鐮孢菌(Fusarium)ITS、EF-1α和β-tubulin 3個(gè)基因序列比較分析,篩選適合于鐮孢菌種類(lèi)鑒定的基因序列,并以此序列分析山西省植物病原鐮孢菌種群分布及遺傳變異情況�！痉椒ā繌�2013—2015年在山西省11市28縣(區(qū))采集分離的625株鐮孢菌菌株中選取形態(tài)學(xué)清晰的菌株進(jìn)行ITS、EF-1α和β-tubulin基因序列聯(lián)合分析,運(yùn)用Sequencher軟件對(duì)序列進(jìn)行拼接和校對(duì),將測(cè)序結(jié)果與NCBI及FUSARIUM-ID數(shù)據(jù)庫(kù)中所有已公布的序列進(jìn)行BLAST分析,結(jié)合下載概念清晰或標(biāo)準(zhǔn)種序列,運(yùn)用Clustal X和Gel Doc軟件進(jìn)行序列對(duì)齊和編輯,運(yùn)用MEGA、Excel、DNAstar和Taxon Gap軟件分析鐮孢菌種內(nèi)種間遺傳變異情況,從ITS、EF-1α和β-tubulin中篩選適合鐮孢菌種類(lèi)鑒定的基因序列,并以此序列分析山西省鐮孢菌的種群分布特點(diǎn)等�！窘Y(jié)果】在3個(gè)候選基因序列中,EF-1α基因序列是最適用于鐮孢菌種類(lèi)鑒定的基因序列,其種間平均遺傳距離分別是種內(nèi)平均遺傳距離的24倍,種內(nèi)差異小于種間差異的種的數(shù)量最多,達(dá)到供試種的73%,物種鑒定準(zhǔn)確率最強(qiáng),達(dá)到87%�；贓F-1α基因片段的系統(tǒng)發(fā)育分析結(jié)果表明,在供試的27種鐮孢菌中,有22種表現(xiàn)出單系性,相同種的不同菌株以較高支持率聚成獨(dú)立支。在27種鐮孢菌中,F.oxysporum是山西省鐮孢菌的優(yōu)勢(shì)種,分離頻率最高(22.1%),且分布最廣,在23個(gè)縣(區(qū))均有分布;其次是F.solani(13.8%),在14個(gè)縣(區(qū))有分布。從不同地區(qū)鐮孢菌的種群結(jié)構(gòu)及分布來(lái)看,運(yùn)城、臨汾、忻州、長(zhǎng)治、呂梁、晉中和太原均以F.oxysporum為優(yōu)勢(shì)種,朔州以F.lateritium為優(yōu)勢(shì)種,大同以F.solani為優(yōu)勢(shì)種,晉城以F.verticillioides為優(yōu)勢(shì)種,陽(yáng)泉以F.incarnatum為優(yōu)勢(shì)種;其中晉中和忻州的鐮孢菌種類(lèi)最豐富,臨汾次之,朔州最少。從不同寄主鐮孢菌的種群結(jié)構(gòu)及分布來(lái)看,番茄上鐮孢菌的種類(lèi)最多(15種),其次是馬鈴薯(13種)和大豆(12種);番茄、黃瓜、西瓜、馬鈴薯、茄子、西葫蘆和甘藍(lán),均以F.oxysporum為優(yōu)勢(shì)種,大豆和玉米以F.verticillioides為優(yōu)勢(shì)種,小麥以F.avenaceum和F.graminearum為優(yōu)勢(shì)種�！窘Y(jié)論】鐮孢菌種內(nèi)種間存在豐富的遺傳變異,其中EF-1α基因序列的遺傳變異最適用于鐮孢菌種類(lèi)的鑒定,但形態(tài)學(xué)種和系統(tǒng)發(fā)育學(xué)種不完全吻合。F.oxysporum是山西省鐮孢菌的優(yōu)勢(shì)種,不同地區(qū)、不同寄主上鐮孢菌種群存在明顯的遺傳分化;研究結(jié)果可為鐮孢菌的分類(lèi)鑒定、DNA條形碼篩選、檢驗(yàn)檢疫及其綜合防治提供理論依據(jù)。
[Abstract]:[objective] by comparing and analyzing the three gene sequences of (Fusarium) ITS,EF-1 偽 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 偽 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 偽 and 尾-tubulin. [results] among the three candidate gene sequences, EF-1 偽 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 偽 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 偽 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.
【作者單位】： 山西農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;
【基金】：山西省科技攻關(guān)項(xiàng)目(20120311019-3) 山西省科技基礎(chǔ)條件平臺(tái)建設(shè)項(xiàng)目(1105-0104)
【分類(lèi)號(hào)】：S432.4

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