Plastid transformation biotechnology: increasing efficiency and expanding application
Description
TitlePlastid transformation biotechnology: increasing efficiency and expanding application
Date Created2021
Other Date2021-10 (degree)
Extent1 online resource (ix, 97 pages) : illustrations
DescriptionPlastid transformation biotechnology holds great potential for many agricultural and pharmaceutical applications, including the development of insect tolerance by expressing dsRNAs, high-level production of bioencapsulated vaccines and antibodies, producing novel metabolic pathways and improving photosynthetic efficiency. However, the plastid transformation technology has yet to become a developed tool in most crop plants.
I was part of the group that developed a system for plastid transformation in Arabidopsis thaliana, a Brassicaceae species, that is potentially applicable to other Brassica species including rapa (turnip, napa cabbage, bok choy), oleracea (broccoli, kale, cauliflower, and cabbage), and napus (oilseed), which are major constituents of the vegetables and vegetable-based products consumed globally. Efficient plastid transformation in Arabidopsis is enabled by eliminating a duplicate fatty acid biosynthetic pathway in chloroplasts. Moreover, the use of a robust tissue culture system increases the number of events acquired from each experiment, enhancing the overall efficiency of the Arabidopsis plastid transformation system.
Infertility in regenerated Arabidopsis transplastomic shoots disables routine use of the plastid transformation system, but a relative, Brassica napus, holds great potential to overcome this bottleneck. To expand plastid transformation here, I generated the tissue culture protocol and tested the plastid transformation efficiency in the newly developed acc2-knockout lines using new plastid transformation vectors for efficient screening of transformed events. Plastid transformation technology in B. napus has a host of applications, such as the increasing seed oil content in the Westar cultivar, the pedigree line that produces canola vegetable oil.
Plastid transformation technology has the potential to greatly impact the pharmaceutical industry. Most therapeutic protein drugs are currently produced on bacterial and yeast platforms, the same basic platforms used since the 1970s. These systems require extensive purification processes and cold storage for transportation, which drive up the costs beyond what is affordable by most people worldwide. The high-level expression platform presented by plastid transformation can overcome many of the manufacturing hurdles incurred by the antiquated systems, while bioencapsulation by the chloroplast presents a system of storage and bioavailability that obviates costly purification processes. Furthermore, a plant production platform for therapeutic drugs can be made available wherever there is need, simply by providing the seeds, and scaled up quickly with straightforward agricultural practices. My project in producing recombinant muscle regulators in tobacco chloroplasts queries the potential of chloroplast transformation for its effectiveness as a competitive therapeutic protein production platform and an alternative to needle-based delivery.
NotePh.D.
NoteIncludes bibliographical references
Genretheses
LanguageEnglish
CollectionSchool of Graduate Studies Electronic Theses and Dissertations
Organization NameRutgers, The State University of New Jersey
RightsThe author owns the copyright to this work.