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Advancing Enabling Technology and Genome Editing in Monocot Crops for Disease Resistance and Sustainability- [electronic resource]
Advancing Enabling Technology and Genome Editing in Monocot Crops for Disease Resistance a...
Advancing Enabling Technology and Genome Editing in Monocot Crops for Disease Resistance and Sustainability- [electronic resource]

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자료유형  
 학위논문파일 국외
최종처리일시  
20240214095903
ISBN  
9798380619004
DDC  
574
저자명  
Poddar, Snigdha.
서명/저자  
Advancing Enabling Technology and Genome Editing in Monocot Crops for Disease Resistance and Sustainability - [electronic resource]
발행사항  
[S.l.]: : University of California, Berkeley., 2021
발행사항  
Ann Arbor : : ProQuest Dissertations & Theses,, 2021
형태사항  
1 online resource(55 p.)
주기사항  
Source: Dissertations Abstracts International, Volume: 85-04, Section: B.
주기사항  
Advisor: Cate, Jamie H. D.;Staskawicz, Brian J.
학위논문주기  
Thesis (Ph.D.)--University of California, Berkeley, 2021.
사용제한주기  
This item must not be sold to any third party vendors.
초록/해제  
요약It is projected that by the year 2050, global agriculture will need to increase the output of staple crops by 60% to feed the world's rapidly growing population amidst threats posed by climate change and crop disease. One strategy to meet this challenge is to develop new genetic diversity in the germplasm to generate robust and resilient lines with beneficial traits. Conventional breeding has played an essential and major role in crop trait improvement. However, those methods can be difficult, laborious, and time-consuming. The development of effective technology for precision and time-saving plant breeding is required. A breakthrough tool to this end has been CRISPR-Cas9. Noteworthy for its simple sequence-specific programmability, the gene editing system has unleashed remarkable potential for plant biotechnology and functional genomics. Here, we explore several aspects and parameters to advance Cas9-mediated gene editing technology in the essential monocot food crops: rice and wheat. Early-stage plant gene-editing projects benefit greatly from a rapid transient pipeline using protoplasts to evaluate the efficacy of gene editing reagents. We describe here a novel and sustainable method for protoplast isolation from rice tissue and demonstrate their use in ribonucleoprotein (RNP) based Cas9 gene editing assays. Next, we focus on applications of Cas9 technology in wheat to engineer disease resistance. Wheat is a critical target organism because the complexity of its large allohexaploid genome has rendered genetic manipulation by classic methods extremely difficult. Using DNA plasmids encoding CRISPR-Cas9 gene editing reagents, we optimize conditions and demonstrate that all three homoeologous copies of genes in allohexaploid wheat can be simultaneously edited within a single generation. We recognize, however, that DNA plasmid gene editing approaches have a number of limitations including random integration into the plant genome and unpredictability of expression. To address this, we finally present significant improvements to DNA-free Cas9-RNP based gene editing in wheat. We show that increased temperature treatment greatly enhances Cas9-mediated editing efficiency and regenerate transgene-free edited wheat plants at a high rate. Lastly, we utilize this DNA-free gene editing method to generate de novo partial resistance to the agronomically important pathogen Parastagonospora nodorum.
일반주제명  
Molecular biology.
일반주제명  
Cellular biology.
일반주제명  
Plant sciences.
일반주제명  
Genetics.
키워드  
Monocot crops
키워드  
CRISPR-Cas9
키워드  
Genetic diversity
키워드  
Plant breeding
키워드  
Ribonucleoprotein
기타저자  
University of California, Berkeley Molecular & Cell Biology
기본자료저록  
Dissertations Abstracts International. 85-04B.
기본자료저록  
Dissertation Abstract International
전자적 위치 및 접속  
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MARC

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■00520240214095903
■006m          o    d                
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■020    ▼a9798380619004
■035    ▼a(MiAaPQ)AAI28867408
■040    ▼aMiAaPQ▼cMiAaPQ
■0820  ▼a574
■1001  ▼aPoddar,  Snigdha.
■24510▼aAdvancing  Enabling  Technology  and  Genome  Editing  in  Monocot  Crops  for  Disease  Resistance  and  Sustainability▼h[electronic  resource]
■260    ▼a[S.l.]:▼bUniversity  of  California,  Berkeley.  ▼c2021
■260  1▼aAnn  Arbor  :▼bProQuest  Dissertations  &  Theses,  ▼c2021
■300    ▼a1  online  resource(55  p.)
■500    ▼aSource:  Dissertations  Abstracts  International,  Volume:  85-04,  Section:  B.
■500    ▼aAdvisor:  Cate,  Jamie  H.  D.;Staskawicz,  Brian  J.
■5021  ▼aThesis  (Ph.D.)--University  of  California,  Berkeley,  2021.
■506    ▼aThis  item  must  not  be  sold  to  any  third  party  vendors.
■520    ▼aIt  is  projected  that  by  the  year  2050,  global  agriculture  will  need  to  increase  the  output  of  staple  crops  by  60%  to  feed  the  world's  rapidly  growing  population  amidst  threats  posed  by  climate  change  and  crop  disease.  One  strategy  to  meet  this  challenge  is  to  develop  new  genetic  diversity  in  the  germplasm  to  generate  robust  and  resilient  lines  with  beneficial  traits.  Conventional  breeding  has  played  an  essential  and  major  role  in  crop  trait  improvement.  However,  those  methods  can  be  difficult,  laborious,  and  time-consuming.  The  development  of  effective  technology  for  precision  and  time-saving  plant  breeding  is  required.  A  breakthrough  tool  to  this  end  has  been  CRISPR-Cas9.  Noteworthy  for  its  simple  sequence-specific  programmability,  the  gene  editing  system  has  unleashed  remarkable  potential  for  plant  biotechnology  and  functional  genomics.  Here,  we  explore  several  aspects  and  parameters  to  advance  Cas9-mediated  gene  editing  technology  in  the  essential  monocot  food  crops:  rice  and  wheat.  Early-stage  plant  gene-editing  projects  benefit  greatly  from  a  rapid  transient  pipeline  using  protoplasts  to  evaluate  the  efficacy  of  gene  editing  reagents.  We  describe  here  a  novel  and  sustainable  method  for  protoplast  isolation  from  rice  tissue  and  demonstrate  their  use  in  ribonucleoprotein  (RNP)  based  Cas9  gene  editing  assays.  Next,  we  focus  on  applications  of  Cas9  technology  in  wheat  to  engineer  disease  resistance.  Wheat  is  a  critical  target  organism  because  the  complexity  of  its  large  allohexaploid  genome  has  rendered  genetic  manipulation  by  classic  methods  extremely  difficult.  Using  DNA  plasmids  encoding  CRISPR-Cas9  gene  editing  reagents,  we  optimize  conditions  and  demonstrate  that  all  three  homoeologous  copies  of  genes  in  allohexaploid  wheat  can  be  simultaneously  edited  within  a  single  generation.  We  recognize,  however,  that  DNA  plasmid  gene  editing  approaches  have  a  number  of  limitations  including  random  integration  into  the  plant  genome  and  unpredictability  of  expression.  To  address  this,  we  finally  present  significant  improvements  to  DNA-free  Cas9-RNP  based  gene  editing  in  wheat.  We  show  that  increased  temperature  treatment  greatly  enhances  Cas9-mediated  editing  efficiency  and  regenerate  transgene-free  edited  wheat  plants  at  a  high  rate.  Lastly,  we  utilize  this  DNA-free  gene  editing  method  to  generate  de  novo  partial  resistance  to  the  agronomically  important  pathogen  Parastagonospora  nodorum.
■590    ▼aSchool  code:  0028.
■650  4▼aMolecular  biology.
■650  4▼aCellular  biology.
■650  4▼aPlant  sciences.
■650  4▼aGenetics.
■653    ▼aMonocot  crops
■653    ▼aCRISPR-Cas9
■653    ▼aGenetic  diversity
■653    ▼aPlant  breeding
■653    ▼aRibonucleoprotein
■690    ▼a0307
■690    ▼a0379
■690    ▼a0369
■690    ▼a0479
■71020▼aUniversity  of  California,  Berkeley▼bMolecular  &  Cell  Biology.
■7730  ▼tDissertations  Abstracts  International▼g85-04B.
■773    ▼tDissertation  Abstract  International
■790    ▼a0028
■791    ▼aPh.D.
■792    ▼a2021
■793    ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T16931064▼nKERIS▼z이  자료의  원문은  한국교육학술정보원에서  제공합니다.
■980    ▼a202402▼f2024

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