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Elucidating the complex interplay between host factors and soybean mosaic virus (SMV): Study on the GmPAP 2.1 and HSP70 proteins that play antagonistic roles in the development of SMV infection. : 콩 모자이크 바이러스와 기주 인자 간 상호작용 기작의 구명 : 콩 모자이크 바이러스의 감염에 대한 GmPAP 2.1과 HSP 70 단백질의 서로 다른 역할

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dc.contributor.advisorKim, Kook-Hyung-
dc.contributor.author크리스틴위드야사리-
dc.date.accessioned2022-06-16T06:45:16Z-
dc.date.available2024-04-18T05:54:01Z-
dc.date.issued2022-
dc.identifier.other000000169491-
dc.identifier.urihttps://hdl.handle.net/10371/181291-
dc.identifier.urihttps://dcollection.snu.ac.kr/common/orgView/000000169491ko_KR
dc.description학위논문(박사) -- 서울대학교대학원 : 농업생명과학대학 농생명공학부, 2022.2. Kim, Kook-Hyung.-
dc.description.abstractInteraction between plant pathogens and their host plants is a complex interplay that involves numerous host factors. These interactions, however, will determine the fate of either successful infection in plants or the development of resistance response. Hence, understanding the interaction between invading virus and host factors may give us deep insight into the virus infection processes. In this study we disclosure interaction between soybean mosaic virus (SMV) and two soybean host factors that lead to two antagonistic developments of SMV infection. We found that overexpression of GmPAP2.1 from L29 conferred SMV resistance to a susceptible cultivar, Lee 74. We determined that GmPAP2.1 interacted with the SMV protein P1 in the chloroplasts, resulting in the up-regulation of the ICS1 gene, which in turn promoted the pathogen-induced salicylic acid (SA) pathway. On the other hand, SMV infections in Nicotiana benthamiana provoke upregulation of genes that encodes BIPs, members of the HSP70 family protein. Subcellular co-localization and yeast two hybrids analyses demonstrated an interaction between NIb protein and BIP 2 in the ER. Transient knock-down of this gene impairs SMV infection, suggesting the importance of BIP 2 protein in the SMV infection in N.benthamiana. Furthermore, I also observed the upregulation of a gene that encodes BIP 2 protein in Glycine max. Together, my study provides new information about host factors that play antagonistic roles in the SMV infection, and subsequently, these finding may extent the possibility to assist management of SMV incidence in the future through modification of host factor that involves in the SMV infection.-
dc.description.abstract식물 병원균과 그들의 기주 간의 상호작용은 다양한 기주인자들이 연관되어 있는 복합적인 과정이다. 이러한 상호작용들에 의해 병원체의 성공적인 기주 식물 내의 감염이나 반대로 기주 식물의 저항성 반응이 일어난다. 따라서, 바이러스와 기주인자 간의 상호작용에 관한 연구는 바이러스 감염 과정을 이해하고 나아가 바이러스에 대한 방제를 위한 방법을 개발하는데 더 많은 통찰력을 줄 수 있다. 본 연구에서는, 콩 모자이크 바이러스(SMV)와 상호작용하는 두 가지 콩 기주인자인 GmPAP 2.1 와 BIP2 들의 기능을 구명하고, 이들과 바이러스 간의 서로 다른 기작의 상호작용을 밝히고자 하였다. 우선, purple acid phosphatase, GmPAP 2.1 유전자는 SMV에 저항성을 보이는 cultivar 에서 그 발현이 확인되었고, SMV에 감수성을 보이는 cultivar에 GmPAP 2.1 유전자의 과발현을 유도하면 기주 식물에 저항성을 부여하는 것으로 확인되었다. 이와 같은 저항성 반응은 GmPAP2.1와 SMV의 한 단백질인 P1과의 상호작용에 의한 것으로 보여지며, 이 두 단백질간의 상호작용은 살리실산(SA) 관련 유전자들의 발현량 증가를 야기하는 것을 확인하였다. 살리실산 관련유전자들의 발현의 증가는 SA 의 축적과 SAR 의 활성을 유도하였다. 반면에, Nicotiana benthamiana 식물에서의 SMV의 감염은 HSP70 family protein 중의 하나인, BIP2 유전자의 발현량 증가를 야기한다. 해당 유전자를 담배에서 knock-down 할 경우, SMV 의 감염이 일어나지 잘 일어나지 않음을 확인하였다. 세포 내에서의 위치와 Yeast two hybrid 실험을 통해 SMV의 NIb 단백질과 BIP2 단백질이 공통적으로 ER에 존재하며, 이러한 현상은 BIP2 단백질이 SMV 감염을 증진시키기 위해 NIb와의 상호작용을 통하여 viral replication complex를 재구성할 것임을 시사한다. 콩에서도 SMV 감염시 HS70 protein을 암호화하는 유전자의 발현이 증가하는 것으로 보아, 콩-SMV 간의 pathosystem에서도 해당 유전자가 비슷한 역할을 할 것으로 기대된다. 따라서, 본 연구는 SMV 감염 시 서로 반대되는 역할을 하는 기주 인자들을 새롭게 밝혀 내었으며, 이 두 기주인자들에 관한 연구는 SMV에 대한 콩의 병 저항성 연구와 방제에 관련한 도움이 되는 정보를 제공하게 될 것이다.-
dc.description.tableofcontentsCONTENTS
ABSTRACT i
CONTENTS iii
LIST OF TABLES viii
LIST OF FIGURES viii

CHAPTER 1. SOYBEAN RESISTANCE TO SOYBEAN MOSAIC VIRUS 1
ABSTRACT 2
INTRODUCTION 3
BIOLOGICAL PROPERTIES AND TRANSMISSION OF SMV 6
I. SMV genome and gene function 6
II. Biological and molecular properties of SMV transmission 8
RESISTANCE GENES (R-GENES): SOYBEAN RESPONSE TO SMV INFECTION 11
I. NLR Gene Family-Mediated Resistance to SMV 11
II. Rsv Genes 12
III. Rsc Genes 21
INDEPENDENT HOST FACTORS INVOLVED IN SOYBEAN-SMV INTERACTION 25
CONCLUSIONS AND FUTURE PERSPECTIVES 32
LITERATURE CITED 35

CHAPTER 2. OVEREXPRESSION OF A PURPLE ACID PHOSPHATASE GMPAP2.1 CONFERS SOYBEAN MOSAIC VIRUS RESISTANT IN A SUSCEPTIBLE SOYBEAN CULTIVAR 53
ABSTRACT 54
INTRODUCTION 55
MATERIALS AND METHODS 59
I. Plants growth and virus strains 59
II. Construction of expression clones 59
III. In vitro transcription and translation 60
IV. Plant inoculation and visual assessment 61
V. Quantitative RT PCR (RT-qPCR) 62
VI. Sequence analysis of GmPAP2.1 homologs 63
VII. Protein domain search and phylogenetic analysis of GmPAP2.1 63
VIII. Subcellular localization analysis in epidermal cells of N. benthamiana 64
IX. Nuclear localization assay 65
X. Genes knock-down constructs 65
XI. Inoculation of the knock-down constructs and challenge inoculation of pSMV-G5H::GFP::GmPAP2.1 or pSMV-G5H::GFP 66
XII. Yeast two-hybrid and beta-galactosidase assays 67
XIII. Construction of expression clone for transient expression in N. benthamiana 67
XIV. Co-immunoprecipitation (Co-IP) assay 68
XV. Measurement of SA level 69
XVI. Statistical analysis 70
RESULTS 75
I. Characterization of GmPAP2.1 75
II. Analyzes of nucleotide sequence and the predicted structure of GmPAP2.1 from cultivar L29 and other cultivars 82
III. Overexpression of GmPAP2.1 from L29 conferred viral resistance to a susceptible cultivar Lee 74 86
IV. Addition of GmPAP2.1 right downstream of the SMV-P1 protein does not affect P1 protease activity 91
V. GmPAP2.1 interacts with the SMV-P1 protein in the chloroplast 93
VI. The C-terminal region of GmPAP2.1 is crucial for resistance against SMV infection 99
VII. GmPAP2.1s from another cultivar than L29 does not confer resistance against SMV 103
VIII. GmPAP2.1-derived resistance may not necessary for Rsv3 mediated resistance 106
IX. Co-expression of GmPAP2.1 with SMV induces accumulation of SA level 110
DISCUSSION 116
LITERATURE CITED 126

CHAPTER 3. A HEAT SHOCK PROTEIN 70 FAMILY MEMBER, BIP2, IS NECESSARY FOR SOYBEAN MOSAIC VIRUS INFECTION IN NICOTIANA BENTHAMIANA 137
ABSTRACT 138
INTRODUCTION 139
MATERIALS AND METHODS 143
I. Plants growth condition and virus inoculation 143
II. Quantitative RT PCR (RT-qPCR) 143
III. Gene knock-down construct and agroinfiltration 144
IV. Assay of viral infection following knock-down of NbHSP70 family proteins 145
V. Construction of expression clone for transient expression in N. benthamiana 145
VI. Subcellular localization analysis in epidermal cells of N. benthamiana 146
VII. Yeast two-hybrid and beta-galactosidase assays 147
VIII. Phylogenetic analysis 147
IX. Statistical analysis 148
RESULTS 151
I. SMV strain G5H and G7H are causes infection in N. benthamiana plant. 151
II. SMV infection in N. benthamiana induces transcription of HSP70 family proteins. 153
III. Knock-down of HSP70 family proteins impairs SMV infection in N. benthamiana. 157
IV. NbBIP2 interacts with SMV RdRP in the endoplasmic reticulum. 161
V. NbBIP2 homologs in Glycine max were highly regulated by SMV infection. 164
DISCUSSION 169
LITERATURE CITED 175
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dc.format.extentxi, 186-
dc.language.isoeng-
dc.publisher서울대학교 대학원-
dc.subjectAgriculture, Plant pathology, Plant virus, Plant virus-Host interaction, Resistance response-
dc.subject.ddc630-
dc.titleElucidating the complex interplay between host factors and soybean mosaic virus (SMV): Study on the GmPAP 2.1 and HSP70 proteins that play antagonistic roles in the development of SMV infection.-
dc.title.alternative콩 모자이크 바이러스와 기주 인자 간 상호작용 기작의 구명 : 콩 모자이크 바이러스의 감염에 대한 GmPAP 2.1과 HSP 70 단백질의 서로 다른 역할-
dc.typeThesis-
dc.typeDissertation-
dc.contributor.AlternativeAuthorKristin Widyasari-
dc.contributor.department농업생명과학대학 농생명공학부-
dc.description.degree박사-
dc.date.awarded2022-02-
dc.identifier.uciI804:11032-000000169491-
dc.identifier.holdings000000000047▲000000000054▲000000169491▲-
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