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Plant Cell. 2019 Jan 16. pii: tpc.00613.2018. doi: 10.1105/tpc.18.00613. [Epub ahead of print]

Genome-scale Sequence Disruption Following Biolistic Transformation in Rice and Maize.

Author information

1
University of Georgia CITY: Athens STATE: Georgia United States Of America [US].
2
Hamilton College CITY: Clinton STATE: NY United States Of America [US].
3
University of Georgia CITY: Athens STATE: GA United States Of America [US].
4
Bionano Genomics CITY: San Diego STATE: CA United States Of America [US].
5
University of Georgia CITY: Athens STATE: Georgia POSTAL_CODE: 30602 United States Of America [US].
6
University of Georgia CITY: Athens STATE: Georgia POSTAL_CODE: 30602-7271 United States Of America [US] kdawe@uga.edu.

Abstract

Biolistic transformation delivers nucleic acids into plant cells by bombarding the cells with microprojectiles, which are micron-scale, typically gold particles. Despite the wide use of this technique, little is known about its effect on the cell's genome. We biolistically transformed linear 48 kb phage lambda and two different circular plasmids into rice (Oryza sativa) and maize (Zea mays) and analyzed the results by whole genome sequencing and optical mapping. While some transgenic events showed simple insertions, others showed extreme genome damage in the form of chromosome truncations, large deletions, partial trisomy, and evidence of chromothripsis and breakage-fusion bridge cycling. Several transgenic events contained megabase-scale arrays of introduced DNA mixed with genomic fragments assembled by non-homologous or microhomology-mediated joining. Damaged regions of the genome, assayed by the presence of small fragments displaced elsewhere, were often repaired without a trace, presumably by homology-dependent repair (HDR). The results suggest a model whereby successful biolistic transformation relies on a combination of end joining to insert foreign DNA and HDR to repair collateral damage caused by the microprojectiles. The differing levels of genome damage observed among transgenic events may reflect the stage of the cell cycle and the availability of templates for HDR.

PMID:
30651345
DOI:
10.1105/tpc.18.00613
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