Additional file 2: Figure S2. of Function and diversity of P0 proteins among cotton leafroll dwarf virus isolates

Amino acid sequence alignment of cotton leafroll dwarf virus P0s. Sequences were aligned with ClustalW2 and conserved amino acids were shaded in black with Mview. The red bar highlights the conserved F-box-like domain. Functional ring structure residues described in Han et al., 2010 are marked with asterisks. (PNG 1391 kb

<i>AtGRP3</i> Is Implicated in Root Size and Aluminum Response Pathways in <i>Arabidopsis</i>

<div><p>AtGRP3 is a glycine-rich protein (GRP) from <i>Arabidopsis thaliana</i> shown to interact with the receptor-like kinase AtWAK1 in yeast, <i>in vitro</i> and <i>in planta</i>. In this work, phenotypic analyses using transgenic plants were performed in order to better characterize this GRP. Plants of two independent knockout alleles of <i>AtGRP3</i> develop longer roots suggesting its involvement in root size determination. Confocal microscopy analysis showed an abnormal cell division and elongation in <i>grp3-1</i> knockout mutants. Moreover, we also show that <i>grp3-1</i> exhibits an enhanced Aluminum (Al) tolerance, a feature also described in <i>AtWAK1</i> overexpressing plants. Together, these results implicate <i>AtGRP3</i> function root size determination during development and in Al stress.</p></div

Relative expression of cell elongation and/or division molecular markers in Col and <i>grp3-1</i>.

<p>Quantitative real time PCR for <b>a</b> <i>COB</i> (At5g60920). <b>b</b> <i>KOR1</i> (At5g49720). <b>c</b> <i>CESA6</i> (At5g64740). <b>d</b> <i>POM1</i> (At1g05850). <b>e</b> <i>DWF1</i> (At3g19820). <b>f</b> <i>BRI1</i> (At4g39400). <b>g</b> <i>CDC48</i> (At3g09840). <b>h</b> <i>CYCB1;2</i> (At5g06150). <i>Error bars</i> indicate standard error. * indicates p≤ 0.05, *** indicates p≤ 0.005 and **** indicates p≤ 0.001.</p

Confocal analysis of root cells.

<p><b>a-c Division pattern of stele cells. * labels stele rows. a</b> Col wild-type. <b>b</b> <i>grp3-1</i> individual presenting normal division pattern. <b>c</b> <i>grp3-1</i> individual presenting abnormal division pattern. <b>d</b> Counting of number of stele cell rows. <b>e</b> Root diameter measurements. <b>f</b> Cell length measurements. <i>Error bars</i> indicate standard error. * indicates p≤ 0.05, ** indicates p≤ 0.01, *** indicates p≤ 0.005 and **** indicates p≤ 0.001.</p

<i>grp3-1</i> loss-of-function mutant analysis.

<p><b>a</b> Relative expression of <i>AtGRP3</i> transcripts analyzed through real-time quantitative PCR of Col and <i>grp3-1</i> mutant. <b>b</b> Summarized data for root length measurements of 2-week-old plants. <i>Error bars</i> indicate standard error. *** indicates p≤ 0.005 and **** indicates p≤ 0.001.</p