本文關(guān)鍵詞： 玉米大斑病菌 效應(yīng)分子 交配型基因位點(diǎn) 生物信息學(xué)　出處：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：玉米大斑病(Nothern leaf blight of corn)是我國玉米產(chǎn)區(qū)重要葉部病害之一,由大斑剛毛座腔菌[Setosphaeria turcicaa(Pass.)Leonard et Suggs]侵染引起,其致病力是影響流行的關(guān)鍵因素。目前已有學(xué)者在細(xì)胞壁降解酶、黑色素、營養(yǎng)元素、毒素、及生理分化等因素對(duì)其致病力影響方面進(jìn)行了研究,在效應(yīng)分子和交配型基因位點(diǎn)方面尚無深入研究。本文旨在利用生物信息學(xué)方法預(yù)測玉米大斑病菌候選效應(yīng)分子,為進(jìn)一步研究效應(yīng)分子在病原菌與寄主互作中的作用奠定基礎(chǔ);克隆并分析玉米大斑病菌交配型基因位點(diǎn),明確其在我國主要玉米產(chǎn)區(qū)的分布,探究病原菌有性生殖過程中交配型基因發(fā)揮的作用,為玉米大斑病菌致病機(jī)理的探究及防治提供理論依據(jù)。1.玉米大斑病菌效應(yīng)分子預(yù)測與分析。本文根據(jù)玉米大斑病菌Setosphaeriaa turcicaa 28A全基因組信息,利用生物軟件從玉米大斑病菌中11698條蛋白序列中預(yù)測候選效應(yīng)分子,再通過對(duì)上述蛋白半胱氨酸含量、信號(hào)肽長度及冗余性分析,最終獲得60個(gè)符合條件的候選效應(yīng)分子,其中有24條蛋白序列在數(shù)據(jù)庫中沒有任何與之同源的序列。對(duì)預(yù)測出的60個(gè)候選效應(yīng)因子功能分析結(jié)果表明,3個(gè)效應(yīng)因子具有預(yù)測功能,其余57個(gè)效應(yīng)因子功能未知。2.玉米大斑病菌株分離鑒定。從我國主要玉米產(chǎn)區(qū)黑龍江、吉林、遼寧、四川、云南等省份共采集并分離獲得的99個(gè)菌株中鑒定出73個(gè)玉米大斑病菌(Setosphaeria turcica)菌株,其余為8個(gè)枝孢霉菌(Cladosporium tenuissimum)菌株,6個(gè)玉米生離蠕孢菌(Bipolaris zeicola)菌株,6個(gè)互隔交鏈孢霉菌(Alternaria alternata)菌株,5個(gè)極細(xì)鏈格孢菌(Alternaria tenuissima)菌株,1個(gè)蕓薹鏈格孢菌(Alternariabrassicae)菌株。對(duì)已分離菌株進(jìn)行系統(tǒng)發(fā)育分析,發(fā)現(xiàn)玉米大斑病菌(S.turcica)菌株、互隔交鏈孢霉菌(A.alternata)菌株、玉米生離蠕孢菌(B.zeicolaa)菌株親緣關(guān)系較近,構(gòu)成一個(gè)亞枝;互隔交鏈孢霉菌(A.alternata)菌株,蕓薹鏈格孢菌(A.brassicae)菌株,極細(xì)鏈格孢菌(A.tenuissima)菌株親緣關(guān)系相對(duì)較近,構(gòu)成一個(gè)亞枝;枝孢霉菌(C.tenuissimum)菌株構(gòu)成一個(gè)小分支,親緣關(guān)系與其它真菌相對(duì)較遠(yuǎn)。3.玉米大斑病菌交配型基因位點(diǎn)克隆與生物信息學(xué)分析。本文克隆了玉米大斑病菌的StM471-1/StM4T1-2基因位點(diǎn),全長為12382bp和12284bp,分別包含1個(gè)MAT基因,基因位點(diǎn)側(cè)翼序列分別包含三個(gè)可編碼蛋白和三個(gè)螺旋結(jié)構(gòu),且位于不同交配型基因位點(diǎn)的同一基因相似性均高達(dá)98%以上。兩交配型基因位點(diǎn)核心區(qū)分別編碼379個(gè)和344個(gè)氨基酸,其中StM4T1-1-1編碼保守α結(jié)構(gòu)域,StM4T1-2-1編碼保守HMG盒子結(jié)構(gòu)域,交配型基因通過自身保守結(jié)構(gòu)域在真菌有性生殖中發(fā)揮作用。其側(cè)翼序列包含的GAP、ORF1、Bgl-1蛋白可能在功能上與玉米大斑病菌的有性生殖過程協(xié)同作用。4.玉米大斑病菌交配型鑒定。玉米大斑病菌生殖方式為異宗配合,存在兩種交配型,為Mating-type 1-1(M4T1-1)和Mating-type 1-2(M4T1-2)。供試73個(gè)玉米大斑病菌菌株中,MAT1-1菌株占49.3%;M4T1-2菌株占43.8%,M4T1-1:M4T1-2比率接近1:1;部分菌株中同時(shí)檢測到MAT1-1和 M4T1-2。
[Abstract]:Northern Corn Leaf Blight (Nothern leaf blight of corn) is one of China's important corn leaf disease, the spot bristles seat cavity bacteria [Setosphaeria (Pass.) turcicaa Leonard et Suggs] caused by infection, its pathogenicity is a key factor affecting the epidemic. At present, scholars in the cell wall degrading enzymes, melanin, nutrient elements, toxin, effects and physiological factors on the differentiation of pathogenicity was studied, there is no in-depth study on effect of molecular and mating type loci. This paper aims to use bioinformatics prediction setosphaeriaturcica candidate effectors methodology, which lays a foundation for further study on the molecular effects of pathogen host interaction in the role of; cloning and analysis of setosphaeriaturcica mating type loci, the distribution in the main maize producing areas in China, play the mating type genes of pathogenic bacteria of importance in the process of sexual reproduction, To provide theoretical basis for the prediction and analysis of.1. of setosphaeriaturcica effector molecules to explore prevention and treatment of setosphaeriaturcica pathogenesis. According to setosphaeriaturcica Setosphaeriaa turcicaa 28A genome information prediction candidate effectors from 11698 protein sequences of setosphaeriaturcica in using biological software, and through the protein the content analysis of signal peptide length and redundancy, eventually won the 60 eligible candidate effectors, of which 24 protein sequence has no homologous sequences in the database. The results show that the predicted 60 candidate effector function analysis, the 3 effect factor has the forecast function, the remaining 57 effect factor of unknown function.2. setosphaeriaturcica isolate. The main maize producing areas in Heilongjiang, Jilin, Liaoning, Sichuan, Yunnan and other provinces were collected and separated Identified 73 setosphaeriaturcica 99 strains obtained in (Setosphaeria turcica) for the rest of the 8 strains of Cladosporium (Cladosporium tenuissimum) strains, 6 maize leaves Helminthosporium (Bipolaris zeicola) strains, 6 Alternaria fungus (Alternaria alternata) strains, 5 very fine Alternaria (Alternaria tenuissima) 1 strains of Alternaria brassicae (Alternariabrassicae) strains. On isolated strains for phylogenetic analysis, found that corn leaf blight (S.turcica) strains of Alternaria fungi (A.alternata) strains of maize leaves Helminthosporium (B.zeicolaa) near the strain relationship the relationship, constitute a sub branch; Alternaria fungi (A.alternata) strains of Alternaria brassicae (A.brassicae) strains from Alternaria tenuissima (A.tenuissima) strains relatively close genetic relationship, constitute a sub branch; Cladosporium (C.tenuissimum) strains. A small branch, the phylogenetic relationship with other fungi relatively distant.3. setosphaeriaturcica mating type loci cloning and bioinformatics analysis. The cloned StM471-1/StM4T1-2 gene locus setosphaeriaturcica, was 12382bp and 12284bp respectively, including 1 MAT gene loci flanking sequences respectively contains three can encoding protein and three helix, the same gene in different mating type loci and the similarity were as high as 98%. Two mating type loci were core region encoding 379 and 344 amino acids, including StM4T1-1-1 encoding StM4T1-2-1 encoding alpha conserved domain, a conserved HMG box domain by mating type genes the conserved domain in the sexual reproduction of fungi play a role. The flanking sequences included GAP, ORF1, Bgl-1 protein may be the process of sexual reproduction of bacteria and exserohilumturcicum in function coordination Mating type identification of pathogen.4. maize. Setosphaeriaturcica reproduction is heterothallic, there are two kinds of mating type, Mating-type 1-1 (M4T1-1) and Mating-type 1-2 (M4T1-2). 73 tested isolates, MAT1-1 strains accounted for 49.3%; M4T1-2 was 43.8%, the ratio of M4T1-1:M4T1-2 to 1:1; some strains detected for MAT1-1 and M4T1-2.

【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S435.131.4

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