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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Molecular identification of Phytophthora resistant genes in soybean

Liyang Chen (8744436) 29 July 2021 (has links)
<p>Phytophthora root and stem rot (PRSR), caused by oomycete <i>Phytophthora sojae</i>, is the most severe soil-borne disease of soybean (<i>Glycine max</i> (L.) Merr.) worldwide. The disease can be effectively managed by introducing resistance to <i>P. sojae</i> (<i>Rps</i>) genes into soybean cultivars by breeding, which requires continuous efforts on identification of resistance resources from soybean germplasm. Previously, two resistance genes, <i>Rps2-cas</i> (former name <i>Rps2-das</i>) and <i>Rps14 </i>(former name<i> Rps1-f</i>), were mapped by linkage analysis from soybean landraces, PI 594549 C and PI 340029, respectively. The resistance underlying PI 594592 also need further characterization given its broad resistance spectrum. In this study, <i>Rps-2cas</i> and <i>Rps14</i> were further mapped, and <i>Rps2-b</i>, was identified and initial mapped from PI 594592. Thus, this thesis research was divided into three parts for three <i>Rps</i> genes.</p><p>The first part mainly focuses advances on <i>Rps2-cas</i>. Marker-assisted spectrum analysis was performed for <i>Rps-2cas</i> to confirm its potential in disease management. A high-quality genome assembly of PI 594549 C was generated, and KASP markers were developed based on comparison between new reference and Williams 82 reference genome. The gene was further mapped to a 32.67-kb region on PI 594549 C reference genome harboring three expressed NLRs by 24 recombinants screened from a large F<sub>4</sub> population. Comparative genomics analysis suggests the only intact NBS-LRR gene in the fine mapping region is the best candidate gene for <i>Rps2cas</i>, and its function was validated by stable transformation. Evidences from other high-quality assembly genomes suggest <i>Rps2-cas</i> originated from an ancient unequal crossing over event.</p> <p>In the second part, <i>Rps14</i> was further mapped using 21 recombinants identified from a F<sub>3 </sub>population consisting of 473 plants. In commonly used Williams 82 reference genome, the assembly of fine mapping region was incomplete, and <i>Rps14</i> region showed drastic variation in size and copy number of NLRs in 23 high-quality genome assemblies, suggesting the complexity of <i>Rps14</i> region and high-quality reference sequence of donor line is required for isolation of <i>Rps14</i> candidate genes. Marker assisted resistance test showed <i>Rps14</i> had wider resistance spectrum to different <i>P. sojae </i>isolates comparing to other <i>Rps</i> genes on chromosome 3, and phylogenic analysis further supported the potential of <i>Rps14</i> to be a novel resistance gene. </p> <p>For the third part, an F<sub>2 </sub>population derived from a cross between PI 594592 and Williams was tested by <i>P. sojae</i> race 1. The 3:1 and 1:2:1 Mendelian segregation ratios were observed in F<sub>2 </sub>individuals and F<sub>2:3 </sub>families, respectively, suggesting a single dominant <i>Rps</i> gene in PI 594592. The gene was initially mapped to the distal end chromosome 16 overlapped with <i>Rps2</i>, and the gene was tentatively named as <i>Rps2-b</i>. Polymorphic SSR markers and InDel markers designed based on re-sequencing data of PI 594592 and Williams was used to genotyping all the F<sub>2:3 </sub>families, and a linkage map was constructed for <i>Rps2-b</i>. <i>Rps2-b</i> was mapped to a 461.8-kb region flanked by SSR marker Satt431 and InDel marker InDel3668 according to the reference genome (Wm82. a2). Marker-assisted resistance test showed <i>Rps2-b</i> hold a wide resistance spectrum. </p>

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