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Exploring cell-type and single cell specific responses in plants to environmental stimuli
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Bar, Cantug (2020) Exploring cell-type and single cell specific responses in plants to environmental stimuli. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3678093
Abstract
Organisms are in constant interaction with their environment where they respond to abiotic and biotic changes, and in turn affect their surroundings. During these interactions, they undergo molecular, physiological and developmental changes in order to adapt to the dynamic circumstances. As our understanding of organism environment interactions has grown it was found that multicellular organisms not only have cell-type specific responses, but these responses are not uniform amongst cells of that cell type types. Studying environmental responses at a whole organism level is thus a challenge, whereby using mixed cell samples can mask the detection of specific effects. In this thesis, the aim was therefore to investigate cell-type and single-cell specific responses towards environmental stress conditions in plants.
In Chapter 3, materials for researching nitrogen responses in root tissue types of the model legume plant Medicago truncatula were generated. Vector constructs were assembled using promoter sequences with known epidermis, cortex or pericycle tissue-specific expression activity to drive reporter proteins. Tissue specific expression for two out of six constructs was confirmed using transient transformation, paving the way for stable transformation of M. truncatula to study tissue-specific expression in legume root systems. In Chapter 4, cell-specific variability in perception of the bacterial pathogen Pseudomonas syringae in Arabidopsis thaliana was investigated using an inducible reporter line. High variability in reporter protein expression was observed amongst genetically identical leaf spongy mesophyll cells. This variation was found to be highly correlated with nuclear size, which has been shown previously to be an indication of ploidy number. Transcriptomic work in this chapter led to the conclusion that cells exhibiting high reporter protein expression possessed an overall higher transcription capacity. In Chapter 5, potential methods of exploiting the cellular variability identified in Chapter 4 were investigated, with the purpose of regenerating a whole plant from a single cell exhibiting higher expression capabilities. To that end, fluorescent activated cell sorting for isolating live protoplasts was optimised, increasing the efficiency from 0.01% to 30%. Low-density protoplast regeneration trials proved challenging, but this method development paves the way for future work.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QH Natural history Q Science > QK Botany |
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Library of Congress Subject Headings (LCSH): | Ecology, Ecophysiology, Plants -- Effect of stress on -- Genetic aspects, Medicago -- Effect of nitrogen on, Genetic vectors, Plant gene expression, Symbiosis | ||||
Official Date: | January 2020 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Gifford, Miriam L. ; Jackson, Stephen D. | ||||
Format of File: | |||||
Extent: | x, 179 leaves : illustrations | ||||
Language: | eng |
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