The fate of extrachromosomal DNAs in the progeny of plastid-transformed tobacco plants

Min, Sung Ran; Jung, Seo Hee; Liu, Jang R.; Jeong, Won-Joong

doi:10.1007/s11816-015-0380-5

Cited by 6 publications

(1 citation statement)

References 14 publications

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“…Shuttle vectors utilizing NICE1 were developed and proven capable of replication in Escherichia coli and able to generate transplastomic tobacco through expression of aadA on the episomal construct (Staub and Maliga, 1994, 1995). Other chloroplast transformation constructs, with promoter and terminator sequences homologous to host plastid DNA, have also been shown to generate episomal plasmids that persist at least to the T3 generation (Min et al ., 2015b). As an alternative approach, a previous attempt was made to develop an episomal plastid transformation system in tobacco that was bioinspired by the dinoflagellate plastome organization (Min et al ., 2015a).…”

Section: Introductionmentioning

confidence: 99%

Mini‐synplastomes for plastid genetic engineering

Occhialini

Pfotenhauer

et al. 2021

Plant Biotechnology Journal

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Summary In the age of synthetic biology, plastid engineering requires a nimble platform to introduce novel synthetic circuits in plants. While effective for integrating relatively small constructs into the plastome, plastid engineering via homologous recombination of transgenes is over 30 years old. Here we show the design–build–test of a novel synthetic genome structure that does not disturb the native plastome: the ‘mini‐synplastome’. The mini‐synplastome was inspired by dinoflagellate plastome organization, which is comprised of numerous minicircles residing in the plastid instead of a single organellar genome molecule. The first mini‐synplastome in plants was developed in vitro to meet the following criteria: (i) episomal replication in plastids; (ii) facile cloning; (iii) predictable transgene expression in plastids; (iv) non‐integration of vector sequences into the endogenous plastome; and (v) autonomous persistence in the plant over generations in the absence of exogenous selection pressure. Mini‐synplastomes are anticipated to revolutionize chloroplast biotechnology, enable facile marker‐free plastid engineering, and provide an unparalleled platform for one‐step metabolic engineering in plants.

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