玫煙色棒束孢的遺傳多態(tài)性
發(fā)布時間：2018-07-29 20:52
【摘要】：玫煙色棒束孢Isaria fumosorosea在全球分布廣泛,寄主多樣,致病力強,是一種已開發(fā)為微生物殺蟲劑的重要昆蟲病原真菌,在生物防治中起著重要的作用。相同種內(nèi)的不同菌株間,通常具有豐富的遺傳多態(tài)性。通過對遺傳多態(tài)性的研究,能從本質(zhì)上揭示物種多樣性的起源、變異和進化。本文將從生物學(xué)特性、致病性以及分子水平上對玫煙色棒束孢的遺傳多態(tài)性進行研究,其研究結(jié)果不僅對高性能菌株的采集分離、生產(chǎn)育種以及生產(chǎn)過程中的質(zhì)量監(jiān)控具有理論和實踐意義,而且還為今后利用該菌進行害蟲的生物防治提供科學(xué)理論依據(jù)。其主要研究結(jié)果如下:1、玫煙色棒束孢的生物學(xué)特性11株所試玫煙色棒束孢菌株在不同光照條件下的宏觀培養(yǎng)形態(tài)特征呈現(xiàn)極大的差異,尤其是在PDA培養(yǎng)基,其菌落特征差別最明顯,其中產(chǎn)孢梗束的菌株為H3。此外,玫煙色棒束孢的分生孢子梗、瓶梗、分生孢子等顯微特征、產(chǎn)孢量以及萌發(fā)率也顯示極大差異,產(chǎn)孢量最多的是菌株H5,萌發(fā)率最高的是菌株H4。2、玫煙色棒束孢對二斑葉螨Tetranychus urticae的致病力玫煙色棒束孢全部所試菌株對二斑葉螨均有致死現(xiàn)象,不同菌株在相同濃度條件下對同一種害蟲表現(xiàn)出不同程度的致死率。其中菌株H4表現(xiàn)出致死時間短、致死率高的特點。產(chǎn)孢量也較大,接近1.08×107個/ml,此外,菌株H8、H9和H11在產(chǎn)孢量與致病率方面也表現(xiàn)不錯,半致死時間也較短,產(chǎn)孢量分別為0.958×107個/ml、0.82×107個/ml和0.762×107個/ml。由此看出,產(chǎn)孢量與致病性間存在一定的正相關(guān)性。3、基因序列分析及其與生物特性和致病性之間的相關(guān)性分析對玫煙色棒束孢進行ITS序列分析,系統(tǒng)發(fā)育分析表明,玫煙色棒束孢全部被試序列都分離為兩個完全獨立的支系,分別為聚在分支I-1中且支持率為98%的菌株和聚在分支II-1內(nèi)且支持率為81%的菌株;為了確定上述兩支系的真正分類地位,對所試玫煙色棒束孢與爪哇棒束孢菌株進行了多態(tài)性研究,獲得7個單倍型,單倍型多樣性Hd=0.605,核苷酸多樣性Pi=0.04,種群遺傳分化指數(shù)Fst=0.78,基因流Nm=0.07,以及2個不同的DNA序列保守區(qū)等。通過系統(tǒng)發(fā)育及多態(tài)性研究結(jié)果表明,將收集自國內(nèi)外以及于Genbank下載的玫煙色棒束孢菌株看成一個種時,它們無論在種的水平上或假定的種群水平上,其遺傳多樣性都較大、分化明顯,它們的差異已超出了種的邊界,分支I-1中的菌株是典型的玫煙色棒束孢,而II-1中的玫煙色棒束孢菌株可能是被誤定的爪哇棒束孢。對玫煙色棒束孢進行多基因序列聚類樹構(gòu)建,并對所試菌株(H1-H11)間進行比較,結(jié)果證明多基因樹間存在一定的分支差異。其中,分支I的菌株來源相同;分支II的多數(shù)成員在PDA培養(yǎng)基上的菌落極厚,菌絲濃密,并且其分生孢子的萌發(fā)率都較低;菌株H2、H5和H7對二斑葉螨的致病力較低。而分支III的菌株菌落主要含孢梗束,除了H6外,其余菌株分生孢子萌發(fā)率較高,這些菌株致病性都較好,且H3與H9致病率相當,以100%支持率聚在一起。由此可見,玫煙色棒束孢對二斑葉螨致病性的高低,在分子系統(tǒng)樹上通過聚類展示了一定的相關(guān)性。分別獨立成一個分支的菌株H1、H4也無論從形態(tài)特征還是孢子萌發(fā)率方面都存在極大差異。
[Abstract]:Isaria fumosorosea, which is widely distributed worldwide, has a wide variety of host and strong pathogenicity, is an important insect pathogenic fungus developed as a microbial insecticide. It plays an important role in biological control. The genetic polymorphism is usually abundant among the different strains in the same species. In essence, it reveals the origin, variation and evolution of species diversity. This article will study the genetic polymorphism of Rosa ospora from biological characteristics, pathogenicity and molecular level. The results of this study not only have theoretical and practical significance for the collection and separation of high performance strains, production breeding and quality monitoring in the process of production. The main research results are as follows: 1, the biological characteristics of the 11 strains of Rosa colospora have great differences in the macro culture morphological characteristics under different illumination conditions, especially in the PDA medium, and their colony characteristics are poor. It is the most obvious that the strains of the pedicel bundle are H3., the microspore peduncle, the peduncle, the conidium, the sporulation and the germination rate of the spores are very different, the most spore yield is strain H5, the highest germination rate is the strain H4.2, and the pathogenicity of the two leaf mite Tetranychus urticae is the pathogenicity of the rose colospora. All the strains were lethal to the two leaf mite, and the strains of different strains showed a different degree of death to the same pest under the same concentration. Among them, the strain H4 showed a short death time and a high mortality rate. The sporulation was also large and close to 1.08 x 107 /ml. In addition, the strain H8, H9 and H11 were produced in the sporulation and the sporulation. The pathogenic rate was also good, and the half lethal time was also short, the sporulation amount was 0.958 x 107 /ml, 0.82 x 107 /ml and 0.762 x 107 /ml., thus it was found that there was a positive correlation between the sporulation and pathogenicity, and the correlation analysis between the gene sequence and the biological characteristics and pathogenicity of the ITS was carried out to ITS. Sequence analysis and phylogenetic analysis showed that all the tested sequences were separated into two completely independent branches, which were isolated in the branch I-1, with a support rate of 98%, and a strain of 81% in the branch II-1 and the support rate was 81%. 7 haplotypes, haplotype diversity Hd=0.605, nucleotide diversity Pi=0.04, population genetic differentiation index Fst=0.78, gene flow Nm=0.07, and 2 different DNA sequences conserved areas were obtained. The results of phylogenetic and polymorphism research show that the collection will be collected from home and abroad and under Genbank. At the level of species and the assumed population level, they have larger genetic diversity and distinct differentiation, and their differences have exceeded the species boundaries. The strain in the branch I-1 is a typical rose colored spore, while the Rosa colospora in II-1 may be mistaken. The multi gene sequence clustering tree was constructed for the mycelia of Romeo, and the comparison between the tested strains (H1-H11) showed that there was a certain branch difference between the multi gene trees. Among them, the strains of the branch I were the same, the majority of the members of the branch II were thick, the mycelium was thicker, and the conidium on the PDA medium. The germination rate of the strains H2, H5 and H7 was lower than that of the strains of H5 and H7, but the strains of the strains of the branch III were mainly pedicel bundles, except for the H6, the conidium germination rate of the other strains was higher, and the pathogenicity of these strains was good, and the pathogenic rate of H3 and H9 was equal, and the rate of support was together with the two spots. The pathogenicity of mites showed a certain correlation by clustering on the molecular phylogeny tree. The strain H1 was separated into one branch independently, and H4 also had great differences in morphological characteristics and spores germination rate.
【學(xué)位授予單位】：貴州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S476.12
本文編號：2153986
[Abstract]:Isaria fumosorosea, which is widely distributed worldwide, has a wide variety of host and strong pathogenicity, is an important insect pathogenic fungus developed as a microbial insecticide. It plays an important role in biological control. The genetic polymorphism is usually abundant among the different strains in the same species. In essence, it reveals the origin, variation and evolution of species diversity. This article will study the genetic polymorphism of Rosa ospora from biological characteristics, pathogenicity and molecular level. The results of this study not only have theoretical and practical significance for the collection and separation of high performance strains, production breeding and quality monitoring in the process of production. The main research results are as follows: 1, the biological characteristics of the 11 strains of Rosa colospora have great differences in the macro culture morphological characteristics under different illumination conditions, especially in the PDA medium, and their colony characteristics are poor. It is the most obvious that the strains of the pedicel bundle are H3., the microspore peduncle, the peduncle, the conidium, the sporulation and the germination rate of the spores are very different, the most spore yield is strain H5, the highest germination rate is the strain H4.2, and the pathogenicity of the two leaf mite Tetranychus urticae is the pathogenicity of the rose colospora. All the strains were lethal to the two leaf mite, and the strains of different strains showed a different degree of death to the same pest under the same concentration. Among them, the strain H4 showed a short death time and a high mortality rate. The sporulation was also large and close to 1.08 x 107 /ml. In addition, the strain H8, H9 and H11 were produced in the sporulation and the sporulation. The pathogenic rate was also good, and the half lethal time was also short, the sporulation amount was 0.958 x 107 /ml, 0.82 x 107 /ml and 0.762 x 107 /ml., thus it was found that there was a positive correlation between the sporulation and pathogenicity, and the correlation analysis between the gene sequence and the biological characteristics and pathogenicity of the ITS was carried out to ITS. Sequence analysis and phylogenetic analysis showed that all the tested sequences were separated into two completely independent branches, which were isolated in the branch I-1, with a support rate of 98%, and a strain of 81% in the branch II-1 and the support rate was 81%. 7 haplotypes, haplotype diversity Hd=0.605, nucleotide diversity Pi=0.04, population genetic differentiation index Fst=0.78, gene flow Nm=0.07, and 2 different DNA sequences conserved areas were obtained. The results of phylogenetic and polymorphism research show that the collection will be collected from home and abroad and under Genbank. At the level of species and the assumed population level, they have larger genetic diversity and distinct differentiation, and their differences have exceeded the species boundaries. The strain in the branch I-1 is a typical rose colored spore, while the Rosa colospora in II-1 may be mistaken. The multi gene sequence clustering tree was constructed for the mycelia of Romeo, and the comparison between the tested strains (H1-H11) showed that there was a certain branch difference between the multi gene trees. Among them, the strains of the branch I were the same, the majority of the members of the branch II were thick, the mycelium was thicker, and the conidium on the PDA medium. The germination rate of the strains H2, H5 and H7 was lower than that of the strains of H5 and H7, but the strains of the strains of the branch III were mainly pedicel bundles, except for the H6, the conidium germination rate of the other strains was higher, and the pathogenicity of these strains was good, and the pathogenic rate of H3 and H9 was equal, and the rate of support was together with the two spots. The pathogenicity of mites showed a certain correlation by clustering on the molecular phylogeny tree. The strain H1 was separated into one branch independently, and H4 also had great differences in morphological characteristics and spores germination rate.
【學(xué)位授予單位】：貴州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S476.12
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