Graduate Thesis Or Dissertation
 

Development of genetic resources for Rubus occidentalis from three segregating sources of resistance to Aphid Amphorophora agathonica

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/5m60r0501

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  • Black raspberry (Rubus occidentalis L.) is a US Pacific Northwest specialty crop prized for its unique flavor profile and nutritional attributes. A major obstacle for the raspberry industry is Black raspberry necrosis virus vectored by the large raspberry aphid (Amphorophora agathonica). Market expansion depends on production, which is currently hindered by aphid vectored viruses, such as Black raspberry necrosis virus (BRNV). This work seeks to enable the development of useful genetic markers for each source of resistance, and to allow pyramiding of these resistance loci in new breeding populations. Identifying genes conferring aphid resistance has the potential to facilitate control of the devastating BRNV. Natural, genetic resistance to aphids has been identified from three geographic sources: Maine (US), Michigan (US), and Ontario (CA). These sources are being utilized to breed cultivars with durable aphid resistance. The genetic mechanisms responsible for aphid resistance are unknown. Utilizing differential expression analysis of the three sources of resistance can provide a tool for identifying genes responsible. Three new segregating populations of black raspberry were developed to study the genetic basis of aphid resistance. Segregation of resistance in each of these populations was evaluated phenotypically by aphid inoculation. Expression profiles in 10 resistant and 10 susceptible seedlings were evaluated with Iso-Seq (Full-Length Isoform Sequencing). Illumina sequencing was used to measure the abundance of mRNA transcripts in leaves following aphid inoculation. Samples were collected from five resistant plants and five susceptible plants from each of the three distinct populations, with each plant sampled both before and after aphid inoculation. High quality RNA was extracted from the collected leaf tissue using a modified RNA extraction protocol. Differential analysis of mRNA transcripts was performed to identify genes that are differentially expressed within each population and in response to aphid treatment. The abundance of transcripts associated with several differing functions were altered after aphid inoculation, including genes with predicted functions related to plant pathogen interactions. This work provides gene candidates for marker development based on genes unique to resistant plants and unique to each population.
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  • Intellectual Property (patent, etc.)
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  • 2022-03-27 to 2023-04-28

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