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Fig. 6. RPL promotes the formation of two valve margins by negatively
regulating JAG and FIL activity. The loss-of-function
JAG allele jag-1, and the activation-tagged allele
jag-5D. (A,B,D-F,H,I,K,L) Cross sections of stage 17 fruit from
various mutants stained with Alcian Blue and Saffranin O. (A) jag-5D
fruit. The replum is replaced by lignified layer and separation layer-like
cell types. (B) Example of a strongly affected jag-5D fruit showing
the fusion of the lignified layer to form a `lignified bridge'. Note that the
two valves have detached from the rest of the fruit as a single unit. (C)
SHP2::GUS reporter activity in a stage 13 jag-5D fruit
expands into the replum region. Note that the SHP2::GUS reporter is
not activated as highly in the vascular bundle where FUL is expressed
(white arrowhead). (D) shp1,2 fruit lack separation and lignified
layer development. The cross section is taken near the base of the fruit where
the shp mutant phenotype is strongest. (E) jag-5D shp1,2
fruit near the base. The ectopic development of valve margin tissues is
suppressed, indicating that SHP is necessary for the replumless
phenotype of jag-5D mutants. (F) rpl fruit have patterning
defects similar to jag-5D. (G) In situ hybridization of JAG
in a stage 8 rpl mutant gynoecium. Expression of JAG is not
detectable in the replum region. (H) jag-1 fruit are similar to wild
type. (I) jag-1 rpl fruit showing a rescue of replum development.
Note that replum rescue is variable and this is a strong example. (J)
Expression of FIL in a stage 8 rpl gynoecium expands into
the replum region (black arrowhead). (K) fil mutant fruit. The replum
tends to be larger than in wild type. (L) fil rpl fruit display a
consistent and strong rescue of replum development. LL, lignified layer; SL,
separation layer; r, replum; v, valve; o, ovule. Scale bars: 50 µm (in A
for A,B,D-F,H,I,K,L; in C; and in G for G,J).
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