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Functional analysis of two host genes that were up-regulated upon Fusarium graminearum virus1 infection : Fusarium graminearum virus1에 감염된 붉은 곰팡이에서 발현량이 증가하는 유전자들에 관한 기능분석

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dc.contributor.advisor김국형-
dc.contributor.author박진만-
dc.date.accessioned2017-07-14T06:43:57Z-
dc.date.available2017-07-14T06:43:57Z-
dc.date.issued2015-02-
dc.identifier.other000000026033-
dc.identifier.urihttps://hdl.handle.net/10371/125895-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부, 2015. 2. 김국형.-
dc.description.abstractFusarium graminearum virus 1(FgV1) is a mycovirus that isolated from Fusarium boothii. It has 6.6kb double stranded RNA as a genome, including putative four open reading frames. Along with FgV1, three other mycoviruses have been reported including FgV2, FgV3, and FgV4. Among four reported viruses, FgV1 and FgV2 infection shows hypovirulence such as a reduction in growth rate, conidiation, and virulence to their host fungi. But infection with the other two viruses, FgV3 and FgV4,makes no dramatic morphological change.
Based upon RNA-sequence analyses of the host transcriptome upon combination of those four mycovirus infection, ten genes that might have relationship with mycovirus infection and/or defense responses of the host fungi are selected. These genes were assorted into four groups depending on their changes of expression level upon virus infection. To investigatepossible functional roles of these genes involved inrelationship between host fungi and virus, targetgene deletion mutants were generated. FgV1 infected deletion mutants were constructed using hyphal fusion mediated mycovirus transmission. While deletion mutants did not show significant alterationin colony morphology and radial growth on solid comparing with wild-type strain, viral RNA accumulations in fgsg_05076 and fgsg_10551 deletion mutantswere decreased comparing with those of virus infected wild-type strain. This study can provide experimental support for RNA-sequence analysis and investigation of individual gene functions which related to FgV1 accumulation in host fungi, F. graminearum.
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dc.description.tableofcontentsABSTRACT……………………………………………………………………5
CONTENTS……………………………………………………………………7
LIST OF TABLES……………………………………………………………. 9
LIST OF FIGURES……………………………………………………….. 10

I.INTRODUCTION………………………………………………………… 11
II. MATERIALS AND METHODS…………………………………………15
1. Fungal strains and culture condition………………………………………….. 15
2. Computational analysis………………………………………………………15
3. Construction of target gene deletion and complementation mutants…………… 15
4. Fungal transformation……………………………………………………. 18
5. Genomic DNA extraction………………………………………………….. 19
6. Southern blot hybridization ………………………………………………. 19
7. Total RNA extraction…………………………………………………….... 20
8. Semi quantitative RT-PCR.……………………………………………….. 20





III. RESULTS……………………………………………………………. 27
1. Selection of target gene.....................................................................…………27
2. Prediction of gene function using blast tools………………………………… 27
3. Generation of gene deletion mutants………………………………………… 28
4. Generation of complementation mutants……………………………………. 28
5. Colony morphology test……………………………………………………. 35
6. Vegetative growth of deletion mutants……………………………………….. 35
7. Quantification of FgV1 viral RNA accumulation using semi-quantitative RT-PCR……………………………………………………………………… .40
IV.DISCUSSION……………………………………………………………. 42
V.LITERATURE CITED…………………………………………………. 47
VI. ABSTRACT IN KOREAN……………………………………………. 53
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dc.formatapplication/pdf-
dc.format.extent2054636 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectFusarium graminearum-
dc.subjectFusarium graminearum virus 1 (FgV1)-
dc.subjecthost factor-
dc.subjectgene deletion-
dc.subjectmycelial growth-
dc.subjectviral RNA accumulation.-
dc.subject.ddc630-
dc.titleFunctional analysis of two host genes that were up-regulated upon Fusarium graminearum virus1 infection-
dc.title.alternativeFusarium graminearum virus1에 감염된 붉은 곰팡이에서 발현량이 증가하는 유전자들에 관한 기능분석-
dc.typeThesis-
dc.contributor.AlternativeAuthorJin Man Park-
dc.description.degreeMaster-
dc.citation.pages55-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2015-02-
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