F-values obtained from ANOVA to validate the effects of QTL <i>fsqs8.1</i> on FS.

<p>F-values obtained from ANOVA to validate the effects of QTL <i>fsqs8.1</i> on FS.</p

Frequency distribution of the traits across the F₂ population derived from the cross ‘PS’ × PI124112.

<p>(A) FL (Fruit Length), (B) FD (Fruit Diameter) and (C) FS (Fruit Shape). Both parents and F₁ values are marked. In the upper box, 25^th, 50^th and 75^th quartiles are displayed; the sample mean and the 95% confidence interval are represented by a diamond, and the outlier values as dots. The bracket along the edge of the box stands for the part of the graph in which 50% of the observations are gathered together. Mean and standard deviation estimates for a normal distribution are also shown.</p

QTL detected for FL, FD and FS using the merged data from two locations of the F₂ population from the cross ‘PS’ × PI124112.

<p>QTL detected for FL, FD and FS using the merged data from two locations of the F₂ population from the cross ‘PS’ × PI124112.</p

Digenic interactions studied by two-way ANOVA between the FS QTL using the markers significantly linked to them.

<p>(A) <i>fsqs8.1</i> and <i>fsqs2.1</i> (<i>fsqs2.1</i> × <i>fsqs8.1</i>); (B) <i>fsqs12.1</i> (<i>fsqs8.1</i> × <i>fsqs12.1</i>). Alleles at <i>locus flqs2.1</i> are named A and a since this QTL has been previously identified as gene <i>a</i> <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104188#pone.0104188-Prin1" target="_blank">[12]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104188#pone.0104188-Noguera1" target="_blank">[49]</a>. PsPs: homozygous for the allele PS (solid line); PsPi: heterozygous (dashed line); PiPi: homozygous for the allele PI124112 (dotted line).</p

Fruit Shape (FS) polymorphism at each stage of the crossing program to obtain the <i>fsqs8.1</i>-NIL.

<p>Fruits of (A) ‘PS’; (B) PI124112; (C) 2M158-3 (F₁); (D) different F₂ plants to show the transgressive segregation of FS; (E) 3M70-47 (F₂); (F) 5M113-1 × ‘PS’ (BC1); (G) 6M59-13 × ‘PS’ (BC2); (H) 7M36-1 × ‘PS’ (BC3); (I) 8M42-24 × ‘PS’ (BC4); (J) 9M7-15 ∶ (BC4S1); (K) 10M2-30 × ‘PS’ (BC5); (L) 11M27 ∶ (BC5S1), (L.1) 11M27-20: heterozygous at <i>fsqs8.1</i> and (L.2) 11M27-11: homozygous at <i>fsqs8.1</i>; (M) the <i>fsqs8.1</i>-NIL 12M57-3 OP (BC5S2).</p

Mean and Standard Deviation (SD) values of the traits for the parents, the F₁ and the whole F₂.

<p>Mean and Standard Deviation (SD) values of the traits for the parents, the F₁ and the whole F₂.</p

Significant digenic interactions between all the FS QTL.

<p>Significant digenic interactions between all the FS QTL.</p

The placental parameters evaluated by <i>Plasmodium</i> species during infection.

<p>For all placentas, areas of necrosis (B) and intervillous space (C) were measured by overlaying a square grid (A) and counting the number of intersecting points that touched necrotic areas (yellow dots; the white circle indicates an example) or intervillous space areas (blue dots; the black circle indicates an example). The ratios of intervillous space area per necrosis (D) and intervillous space area per placental barrier thickness (E) were calculated. The placentas in the “no plasmodium” group (n = 41; white boxes) appear to have similar necrotic areas and more intervillous space than the placentas in the “P. vivax” group (n = 59; red boxes). The placentas in the “P. falciparum” group (n = 19; grey boxes) exhibited more necrotic areas and less intervillous space. Graphs (B, C, D and E) represent the transformed data. The boxes represent the mean and standard deviation values. The whiskers represent the 5^th and 95^th percentiles. The photograph was taken using a Zeiss Axio Imager M2 light microscope equipped with a Zeiss Axio Cam HRc. Grid overlays and counts were performed using Image J.</p

The syncytial parameters evaluated by <i>Plasmodium</i> species during infection.

<p>Syncytial knotting (A and B) and syncytial rupture (C and D) were evaluated on H&E-stained slides at 100× magnification. Placental barrier thickness (E and F) was evaluated on Masson's trichrome-stained slides at 1000× magnification after overlaying horizontal lines with 5 µm of interspace (see Methods and <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002071#pntd-0002071-t001" target="_blank">Table 1</a>). For all parameters, placentas from the “no plasmodium” group (n = 41; white boxes) had the lowest values, followed by placentas from the “P. vivax” (n = 59; red boxes) and “P. falciparum” (n = 19; grey boxes) groups. Graphs (B, D and F) represent the transformed data. * ANOVA test, P-value≤0,006. The boxes represent the mean and standard deviation values. The whiskers represent the 5^th and 95^th percentiles. Photographs were taken using a Zeiss Axio Imager M2 light microscope equipped with a Zeiss Axio Cam HRc. The grid overlays and counts were conducted using Image J. Arrow heads on A, C and D point to syncytial knots, syncytial rupture and an example of a thickness measurement, respectively.</p

Results and univariate analysis of the placental parameters, evaluated by histology, according to the species of <i>Plasmodium</i> infection during pregnancy^†.

†<p>Women were grouped according to their malaria diagnoses during pregnancy, based on microscopy data. (IQ) 25^th and 75^th percentile. (n) number of women. (%) percentage (N/A) not applicable. For differences between groups all continuous variables were Ln-transformed and one-way ANOVA tests with Bonferroni post-tests were performed. Chi² tests were used to evaluate differences in categorical variables between groups. Refer to the “methods section” for a full description of how each parameter was measured.</p>*<p><i>P. falciparum</i> vs. No Malaria, P = 0.004.</p>§<p><i>P. falciparum</i> vs. <i>P. vivax</i>, P = 0.005.</p>°<p><i>P. falciparum</i> vs. No Malaria, P = 0.048.</p>#<p>χ² test: <i>P.</i> vivax or <i>P.</i> falciparum, P<0.001.</p