Localization of LA1.

<p>LA1-Citrine localizes to the nucleus (arrows) and cell membrane in (A) <i>N. benthamiana</i> epidermal cells and (B) <i>N. benthamiana</i> mesophyll cells. Inset shows detail of the mesophyll cell membrane. (C) Deletion of the putative NLS in LA1ΔNLS-Citrine abrogates nuclear localization. Scale bars are 20 uM. (D) Western blotting confirms expression of full length LA1-Citrine (lane 1) and LA1ΔNLS-Citrine (lane 2).</p

Alignment of <i>Mu</i> Terminal Inverted Repeats (TIRs).

<p>ClustalW was used to align all known active and potentially active <i>Mu</i> elements. Strings of four or more consecutive bases that are entirely conserved among all elements are shaded in red. The conserved <i>Taq</i>^αI site is shaded in blue. <i>Mu4, 5,</i> and <i>6</i> are inactive and are not included [reviewed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Walbot1" target="_blank">[5]</a>]. <i>Mu9</i> is MuDR. Only the first 39 bp of the <i>Mu10, 11,</i> and <i>12</i> TIRs have been sequenced <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Dietrich1" target="_blank">[62]</a>. However, the primer used to amplify the TIR ended in a 3′ GTC, allowing for the assumption that the sequence continues as GAC (shown as small case) and that the <i>Taq</i>^αI site remains intact. Of the most recently discovered <i>Mu</i> elements (13–19), only <i>Mu13</i> has been confirmed to actively move and create new mutations <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Tan1" target="_blank">[58]</a>. TIR sequences obtained from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Barker1" target="_blank">[3]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Tan1" target="_blank">[58]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-Dietrich1" target="_blank">[62]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087053#pone.0087053-James1" target="_blank">[68]</a>.</p

Identification of <i>la1</i>.

<p>The <i>la1</i> mutant plants (<b>A</b>) exhibit prostrate growth habit caused by a lack of a negative gravitropic response. Wild type maize plants (<b>B</b>) purposefully grow away from gravity (negative gravitropism). (<b>C</b>) Gene structure of <i>la1</i> with the site of the <i>Mu</i> insertion in the <i>la1</i>-<i>mu1</i> mutant allele and the CACTA family insertion in the <i>la1-cacta</i> allele. Exons are shown as empty boxes and UTRs are shown as filled boxes. The <i>Mu</i> insertion has not been characterized and is not to scale. (<b>D</b>) Amino acid ClustalW alignment of the maize and rice predicted LAZY1 proteins. The pair shares 60% identity. The predicted transmembrane domain is shaded in blue, the predicted NLS domain is shaded in orange.</p

Manhattan plot of <i>la1-mu1 Taq</i>^αI sequencing.

<p>(<b>A</b>) Manhattan plot showing the distribution of reads from <i>la1-mu1</i> genomic DNA mapped throughout the B73 genome. Alternating colors represent each of the ten maize chromosomes. Each x-axis pixel represents a bin of 1 Mb and the logarithmic y-axis denotes the number of reads mapping to each bin. The red line represents the known genetic map position for the <i>la1</i> reference mutation. Each triangle below the plot represents the approximate location of mapped MFS. (<b>B</b>) Expanded Manhattan plot of a 1 Mb interval corresponding to the approximate map position of <i>la1</i>. Same as top with each x-axis pixel representing a bin of 1 kb. Filtered genes in the 1 Mb interval are shown as black rectangles. MFS mapping to this interval are shown as inverted red triangles.</p