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Dissertation
DESIGN AND APPLICATION OF THE FIRST PLASTID-SPECIFIC ARTIFICIAL MICRORNA LIBRARY TO EXPLORE THE NUCLEAR-ENCODED GENETIC SPACE OF PLANT CHLOROPLASTS
Washington State University
Doctor of Philosophy (PhD), Washington State University
01/2021
DOI:
https://doi.org/10.7273/000003164
Handle:
https://hdl.handle.net/2376/121756
Abstract
Oxygenic photosynthesis is essential to proper functioning of the Earth’s current ecosystem; it is the source of our atmospheric oxygen and the primary method of carbon fixation. In plants, these key metabolic processes, along with other key physiological activities, are housed in a cellular organelle called the chloroplast. Despite this central role in plant metabolism, most proteins that localize to the chloroplast remain uncharacterized. A high-throughput, comprehensive method for investigating plastid-localized proteins would be useful in expanding the plant science knowledge base. Described here is the construction and utilization of a tool that enables scientific findings with regards to plastid physiology in a rapid and wide-ranging fashion. This work details the construction of the first plastid-specific artificial microRNA (pami) library, including an update of the binary expression vector pGreen for use as the scaffold for library construction. This library contains 7,408 unique amiRNA expression constructs that target the 5,889 nuclear-encoded genes whose products localize to plastids. Mutant plant lines generated using the pami library have successfully reproduced previously published phenotypes, including phenotypes of higher order mutants, in an important validation of library design and function. The potential for new discovery is highlighted by six pami lines whose phenotypes were either previously unidentified or not thought to be associated with plastid physiology. Taken together, the effectiveness and broad-reaching scope of the pami library makes it an important contribution to the plant science community.
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Details
- Title
- DESIGN AND APPLICATION OF THE FIRST PLASTID-SPECIFIC ARTIFICIAL MICRORNA LIBRARY TO EXPLORE THE NUCLEAR-ENCODED GENETIC SPACE OF PLANT CHLOROPLASTS
- Creators
- Anna Pratt
- Contributors
- Andrew McCubbin (Advisor)Hans Henning Kunz (Advisor)Mechthild Tegeder (Committee Member)Michael Neff (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Biological Sciences, School of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Publisher
- Washington State University
- Number of pages
- 167
- Identifiers
- 99900652004901842
- Language
- English
- Resource Type
- Dissertation