Journal: bioRxiv

Article Title: Maternal control of RNA decay safeguards embryo development

doi: 10.1101/2025.05.28.656625

Figure Lengend Snippet: ( A ) Nomarsky images of embryos in whole mount cleared seeds of wild-type (WT) Col-0 and cer7-4 at 2, 3, 5 and 7 days after pollination (DAP). ( B ) Quantification of embryo stages in WT and cer7 alleles. ( C ) Endosperm cellularization in WT and cer7-4 determined by Feulgen staining at 5 and 7 DAP. ( D ) Quantification of seed abortion in hand self-pollinated heterozygous plants at 14 DAP (top panel), representative images of siliques (bottom panel). ( E ) Quantification of seed abortion at 14 DAP in indicated genetic crosses to determine the maternal eYect in cer7 mutants. ( F ) Representative images of siliques quantified in (E). Bar color code for D and E: dark gray denotes sporophytic maternal plant homozygous for cer7 ; light gray denotes WT and heterozygous cer7 sporophytic maternal plant. Pairwise t-test, **P-value < 0.01; * P-value < 0.05; ns, not significant. Scale bar 50 µm (A and B).

Article Snippet: Siliques were dissected at 3 days after pollination (DAP), and approximately 500 seeds per genotype were collected and stored in RNAlater solution (ThermoFisher, AM7021).

Techniques: Staining

Journal: bioRxiv

Article Title: Maternal control of RNA decay safeguards embryo development

doi: 10.1101/2025.05.28.656625

Figure Lengend Snippet: ( A ) Quantification of seed abortion at 14 days after pollination (DAP) in the indicated genetic crosses. Numbers of analyzed seeds are indicated on the right side of the graph. ( B ) Representative images of siliques quantified in (A). ( C ) Scheme depicting the expected PTGS amplification in reciprocal crosses. Dark grey indicates active PTGS amplification; light grey indicates impaired amplification. Top panel: cer7;SGS3 sporophytic maternal plant and progeny amplify PTGS. Bottom panel: cer7;sgs3 maternal plant with PTGS impairment; progeny retains amplification. ( D ) Quantification of seed abortion at 14 DAP in the indicated genetic crosses. Numbers of analyzed seeds are indicated on the right side of the graph. ( E ) Representative images of siliques quantified in (C). Bar color code for B and E: Dark grey indicates PTGS amplification by the cer7 homozygous sporophytic maternal plant; light grey indicates impaired PTGS amplification in the sporophytic maternal plant. ( F ) As in (A), scheme depicting the expected PTGS amplification in selfed cer7;ago1-27/+ plants. The sporophytic maternal plant and the majority of the progeny amplify PTGS (dark grey). A subset of progeny ( ago1-27 homozygotes, ∼25%) is expected to show impaired PTGS amplification, indicated in light grey in the accompanying pie chart. Pairwise t-test, **P-value < 0.01; * P-value < 0.05; ns, not significant.

Article Snippet: Siliques were dissected at 3 days after pollination (DAP), and approximately 500 seeds per genotype were collected and stored in RNAlater solution (ThermoFisher, AM7021).

Techniques: Amplification

Journal: bioRxiv

Article Title: Maternal control of RNA decay safeguards embryo development

doi: 10.1101/2025.05.28.656625

Figure Lengend Snippet: ( A–D ) CER7-GFP expression driven by the constitutive RPS5A promoter (A), the seed coat-specific dVPE promoter (B), the BAN promoter (C), or a combination of both dVPE and BAN promoters (D). Green represents GFP expression, blue shows autofluorescence. Scale bar 50 µm. ( E ) Stems of 6 week old cer7;pRPS5A::CER7-GFP (left) and cer7;pBAN::CER7-GFP;pdVPE::CER7-GFP (right). White arrows indicate glossy stem phenotype. Scale bar 200 µm. ( F-I ) Quantification of seed abortion in mature siliques in cer7 mutants expressing CER7-GFP under the constitutive RPS5A promoter (F), the seed coat-specific inner integument (ii1) BAN promoter (G), the ii2-3 specific dVPE promoter (H) and the combined expression of d VPE and BAN promoters (ii1-3) (I). Representative images of the indicated siliques depicted in the bottom panels. Numbers of analyzed seeds are indicated on the right side of the graph. DAP, days after pollination. L, individual transgenic line. Pairwise t-test, **P-value < 0.01; * P-value < 0.05; ns, not significant.

Article Snippet: Siliques were dissected at 3 days after pollination (DAP), and approximately 500 seeds per genotype were collected and stored in RNAlater solution (ThermoFisher, AM7021).

Techniques: Expressing, Transgenic Assay