The linear plastid chromosomes of maize: terminal sequences, structures, and implications for DNA replication
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The structure of a chromosomal DNA molecule may influence the way in which it is replicated and inherited. For decades plastid DNA (ptDNA) was believed to be circular, with breakage invoked to explain linear forms found upon extraction from the cell. Recent evidence indicates that ptDNA in vivo consists of linear molecules with discrete termini, although these ends were not characterized. We report the sequences of two terminal regions, End1 and End2, for maize (Zea mays L.) ptDNA. We describe structural features of these terminal regions and similarities found in other plant ptDNAs. The terminal sequences are within inverted repeat regions (leading to four genomic isomers) and adjacent to origins of replication. Conceptually, stem-loop structures may be formed following melting of the double-stranded DNA ends. Exonuclease digestion indicates that the ends in maize are unobstructed, but tobacco (Nicotiana tabacum L.) ends may have a 5′-protein. If the terminal structure of ptDNA molecules influences the retention of ptDNA, the unprotected molecular ends in mature leaves of maize may be more susceptible to degradation in vivo than the protected ends in tobacco. The terminal sequences and cumulative GC skew profiles are nearly identical for maize, wheat (Triticum aestivum L.) and rice (Oryza sativa L.), with less similarity among other plants. The linear structure is now confirmed for maize ptDNA and inferred for other plants and suggests a virus-like recombination-dependent replication mechanism for ptDNA. Plastid transformation vectors containing the terminal sequences may increase the chances of success in generating transplastomic cereals.
KeywordsGC skew Herpes simplex virus Chloroplast DNA Recombination-dependent replication Telomeres
High salt buffer
Herpes simplex virus
Long single copy
Origin binding protein
Origin of replication
Pulsed-field gel electrophoresis
Single-strand binding protein
Short single copy
This research was funded by the Junat Fund (a private charitable fund).
Compliance with ethical standards
Conflict of interest
The authors DO and AB are co-inventers for the patent application: “Plastid Transformation Using Linear DNA Vectors”; application nos. PCT/US2013/030775, WO2014/065857A1. The linear vectors described in this patent are based on the terminal ptDNA sequences reported in this manuscript.
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