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Discovery and Validation of Host Resistance Factors for Spring Black Stem and Leaf Spot Disease in Medicago truncatula
Discovery and Validation of Host Resistance Factors for Spring Black Stem and Leaf Spot Di...
Discovery and Validation of Host Resistance Factors for Spring Black Stem and Leaf Spot Disease in Medicago truncatula

Detailed Information

자료유형  
 학위논문 서양
최종처리일시  
20260202103210
ISBN  
9798286441754
DDC  
581
저자명  
Botkin, Jacob R.
서명/저자  
Discovery and Validation of Host Resistance Factors for Spring Black Stem and Leaf Spot Disease in Medicago truncatula
발행사항  
[Sl] : University of Minnesota, 2025
발행사항  
Ann Arbor : ProQuest Dissertations & Theses, 2025
형태사항  
215 p
주기사항  
Source: Dissertations Abstracts International, Volume: 86-12, Section: B.
주기사항  
Advisor: Curtin, Shaun J.
학위논문주기  
Thesis (Ph.D.)--University of Minnesota, 2025.
초록/해제  
요약The model legume, Medicago truncatula, is a self-fertile diploid relative of Medicago sativa (alfalfa). M. truncatula is a valuable resource for understanding legume physiology and plant-microbe interactions. Ascochyta blights cause yield loss in major legume crops. The necrotrophic fungus Ascochyta medicaginicola is the causal agent of spring black stem and leaf spot (SBS) disease, which is one of the most devastating foliar diseases of alfalfa affecting yield, quality, and stand survival in this perennial forage crop. However, host resistance is poorly understood. Quantitative trait loci (QTL) have been described for a variety of agronomic traits to identify targets for plant improvement. In M. truncatula, recessive QTL rnpm1 and rnpm2 were identified in populations generated using M. truncatula genotype HM078 as the resistant parent. In Chapter 1, we generated a genome of HM078 to investigate factors enabling host resistance. We identified 14 candidate genes for disease resistance based on structural variation in QTL regions, with a focus on loss-of-function events in HM078. Next, in Chapter 2 we performed an analysis of the host transcriptome in response to A. medicaginicola infection over time. A total of 192 and 2,908 differentially expressed genes (DEGs) were observed in the resistant (HM078) and susceptible (A17) genotype, respectively. We performed a functional analysis and identified 22 candidate genes for disease resistance based on a variety of factors, such as uniquely upregulated genes in HM078. In Chapter 3, we sought to validate top candidate genes in QTL regions with CRISPR/Cas9-mediated knockouts. We targeted a TIR-NBS-NLR plant disease resistance gene, MtTCAR1, and MtPHO2A in rnpm1, as well as a MtCPR1-like F-box family gene and MtPAM16 in rnpm2. Bi-allelic knockouts of MtTCAR1, MtPHO2A, and MtPAM16 did not affect disease resistance in M. truncatula accession R108. MtCPR1-like mutant plants exhibited a reduction in mean pathogen biomass, increased chlorosis, and variable constitutive expression of pathogenesis-related genes. Interestingly, the constitutive overexpression of an RNA-seq-derived candidate, MtKCS12, enhanced SBS disease resistance significantly, resulting in a ~75% reduction in mean pathogen biomass compared to the null segregant in a detached leaf assay. This study provides insights into omics strategies for investigating disease resistance, as well as gene-editing and transgenic approaches for crop improvement.
일반주제명  
Plant pathology
일반주제명  
Plant sciences
키워드  
Medicago truncatula
키워드  
Ascochyta medicaginicola
키워드  
Quantitative trait loci
기타저자  
University of Minnesota Plant Pathology
기본자료저록  
Dissertations Abstracts International. 86-12B.
전자적 위치 및 접속  
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MARC

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■1001  ▼aBotkin,  Jacob  R.
■24510▼aDiscovery  and  Validation  of  Host  Resistance  Factors  for  Spring  Black  Stem  and  Leaf  Spot  Disease  in  Medicago  truncatula
■260    ▼a[Sl]▼bUniversity  of  Minnesota▼c2025
■260  1▼aAnn  Arbor▼bProQuest  Dissertations  &  Theses▼c2025
■300    ▼a215  p
■500    ▼aSource:  Dissertations  Abstracts  International,  Volume:  86-12,  Section:  B.
■500    ▼aAdvisor:  Curtin,  Shaun  J.
■5021  ▼aThesis  (Ph.D.)--University  of  Minnesota,  2025.
■520    ▼aThe  model  legume,  Medicago  truncatula,  is  a  self-fertile  diploid  relative  of  Medicago  sativa  (alfalfa).  M.  truncatula  is  a  valuable  resource  for  understanding  legume  physiology  and  plant-microbe  interactions.  Ascochyta  blights  cause  yield  loss  in  major  legume  crops.  The  necrotrophic  fungus  Ascochyta  medicaginicola  is  the  causal  agent  of  spring  black  stem  and  leaf  spot  (SBS)  disease,  which  is  one  of  the  most  devastating  foliar  diseases  of  alfalfa  affecting  yield,  quality,  and  stand  survival  in  this  perennial  forage  crop.  However,  host  resistance  is  poorly  understood.  Quantitative  trait  loci  (QTL)  have  been  described  for  a  variety  of  agronomic  traits  to  identify  targets  for  plant  improvement.  In  M.  truncatula,  recessive  QTL  rnpm1  and  rnpm2  were  identified  in  populations  generated  using  M.  truncatula  genotype  HM078  as  the  resistant  parent.  In  Chapter  1,  we  generated  a  genome  of  HM078  to  investigate  factors  enabling  host  resistance.  We  identified  14  candidate  genes  for  disease  resistance  based  on  structural  variation  in  QTL  regions,  with  a  focus  on  loss-of-function  events  in  HM078.  Next,  in  Chapter  2  we  performed  an  analysis  of  the  host  transcriptome  in  response  to  A.  medicaginicola  infection  over  time.  A  total  of  192  and  2,908  differentially  expressed  genes  (DEGs)  were  observed  in  the  resistant  (HM078)  and  susceptible  (A17)  genotype,  respectively.  We  performed  a  functional  analysis  and  identified  22  candidate  genes  for  disease  resistance  based  on  a  variety  of  factors,  such  as  uniquely  upregulated  genes  in  HM078.  In  Chapter  3,  we  sought  to  validate  top  candidate  genes  in  QTL  regions  with  CRISPR/Cas9-mediated  knockouts.  We  targeted  a  TIR-NBS-NLR  plant  disease  resistance  gene,  MtTCAR1,  and  MtPHO2A  in  rnpm1,  as  well  as  a  MtCPR1-like  F-box  family  gene  and  MtPAM16  in  rnpm2.  Bi-allelic  knockouts  of  MtTCAR1,  MtPHO2A,  and  MtPAM16  did  not  affect  disease  resistance  in  M.  truncatula  accession  R108.  MtCPR1-like  mutant  plants  exhibited  a  reduction  in  mean  pathogen  biomass,  increased  chlorosis,  and  variable  constitutive  expression  of  pathogenesis-related  genes.  Interestingly,  the  constitutive  overexpression  of  an  RNA-seq-derived  candidate,  MtKCS12,  enhanced  SBS  disease  resistance  significantly,  resulting  in  a  ~75%  reduction  in  mean  pathogen  biomass  compared  to  the  null  segregant  in  a  detached  leaf  assay.  This  study  provides  insights  into  omics  strategies  for  investigating  disease  resistance,  as  well  as  gene-editing  and  transgenic  approaches  for  crop  improvement.
■590    ▼aSchool  code:  0130.
■650  4▼aPlant  pathology
■650  4▼aPlant  sciences
■653    ▼aMedicago  truncatula
■653    ▼aAscochyta  medicaginicola
■653    ▼aQuantitative  trait  loci
■690    ▼a0480
■690    ▼a0479
■71020▼aUniversity  of  Minnesota▼bPlant  Pathology.
■7730  ▼tDissertations  Abstracts  International▼g86-12B.
■790    ▼a0130
■791    ▼aPh.D.
■792    ▼a2025
■793    ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17357339▼nKERIS▼z이  자료의  원문은  한국교육학술정보원에서  제공합니다.

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