Plants have been proved as a novel production platform for a wide range of biologically important compounds such as enzymes, therapeutic proteins, antibiotics, and proteins with immunological properties. In this context, plastid genetic engineering can be potentially used to produce recombinant proteins. However, several challenges still remain to be overcome if the full potential of plastid transformation technology is to be realized. They include the development of plastid transformation systems for species other than tobacco, the expression of transgenes in non-green plastids, the increase of protein accumulation and the appearance of pleiotropic effects. In this paper, we discuss the novel tools recently developed to overcome some limitations of chloroplast transformation.
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http://dx.doi.org/10.4161/bioe.21452 | DOI Listing |
Microb Biotechnol
December 2024
State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, China.
Chlamydomonas reinhardtii, a model green alga for expressing foreign proteins, faces challenges in multigene expression and enhancing protein expression level in the chloroplast. To address these challenges, we compared heterologous promoters, terminators and intercistronic expression elements (IEEs). We transformed Chlamydomonas chloroplast with a biolistic approach to introduce vectors containing the NanoLuc expression unit regulated by Chlamydomonas or tobacco promoters and terminators.
View Article and Find Full Text PDFACS Synth Biol
December 2024
Center for Agricultural Synthetic Biology (CASB), University of Tennessee, 2640 Morgan Circle Dr., Knoxville, Tennessee 37996, United States.
Plastids represent promising targets in plant genetic engineering for many biotech applications, ranging from their use as bioreactors for the overproduction of valuable molecules to the installation of transgenes for improving plant traits. For over 30 years, routine methods of plastid transformation have relied on homologous recombination integrating vectors. However, nonintegrating episomal plasmids have recently received more attention as an innovative tool for the plastid genetic engineering of plant cells.
View Article and Find Full Text PDFPlant Cell Rep
November 2024
Center for Agricultural Synthetic Biology (CASB), University of Tennessee, 2640 Morgan Circle Dr., Knoxville, TN, 37996, USA.
This study describes an optimized plastid genetic engineering platform to produce full marker-free transplastomic plants with transgene integrated at homoplasmy in one step in tissue culture. Plastid engineering is attractive for both biotechnology and crop improvement due to natural bio-confinement from maternal inheritance, the absence of transgene positional effects and silencing, the ability to express transgenes in operons, and unparalleled production of heterologous proteins. While plastid engineering has had numerous successes in the production of high-value compounds, no transplastomic plants have been approved for use in agriculture.
View Article and Find Full Text PDFPlant Mol Biol
November 2024
Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India.
Existence of potent in vitro regeneration system is a prerequisite for efficient genetic transformation and functional genomics of crop plants. In this study, two contrasting cultivars differencing in their in vitro regeneration efficiency were identified. Tissue culture friendly cultivar Golden Promise (GP) and tissue culture resistant DWRB91(D91) were selected as contrasting cultivars to investigate the molecular basis of regeneration efficiency through multiomics analysis.
View Article and Find Full Text PDFAnn Bot
November 2024
Department of General Botany, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University in Poznań, Uniwersytetu Poznańskiego 6, 61-614 Poznań, Poland.
Background And Aims: The transformation of sieve elements (SEs) from meristematic cells, equipped with a full complement of organelles, to specialized transport tubes devoid of a nucleus, has long been enigmatic. We hypothesized a strong involvement of various degradation pathways, particularly macroautophagy in this context, emphasizing the importance of autophagic selectivity in the remaining viability of these cells.
Methods: Experiments were performed on pioneer roots of Populus trichocarpa cultivated in rhizotrons under field conditions.
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