Generation of homoplastomic transplastomic plants expressing Cr-PTOX1.

<p>Schematic representation of the plastome region of the wildtype (A) and the transplastomic plant line Cr-PTOX1-I (B) analyzed by Southern blot analysis. Cr-PTOX1-I sequences were cloned in vector pHK40 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041722#pone.0041722-Kuroda1" target="_blank">[22]</a>. The positions of the restriction enzyme BglII used to digest the genomic DNA are shown. The dotted lines represent the size of the expected fragments to be released from wildtype as well as Cr-PTOX1-I after restriction. (C) Total genomic DNA from WT as well as Cr-PTOX1-I was digested and hybridized with rrn16/rps12 digoxigenin labelled probe amplified from WT. (D) Maternal inheritance assay of Cr-PTOX1-I transplastomic plant line. Seeds of Cr-PTOX1-I and WT plants were grown on MS plates with or without 500 mg L⁻¹ spectinomycin at room temperature. Abbreviations: P<i>rrn</i> = 16SRNA operon promoter, T<i>rbcL</i> = Rubisco large subunit terminator, P<i>psbA</i> = <i>psbA</i> promoter, T<i>psbA</i> = <i>psbA</i> terminator, WT = wild type, T = transplastomic, ptDNA = Plastid DNA.