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Fig. 2. Phenotype of zou mutant seeds. (A) Seeds from wild-type plant. (B) Shrivelled seeds from self-pollinated zou-2/+ plant (the bulk of seeds were wild-type and non-shrivelled). (C) Dissected silique of self pollinated zou-2/zou-2 ZOU::ZOU-GFP/- primary transformant. The majority of seeds have a wild-type phenotype, owing to complementation by the transgene. Some seeds (asterisk) do not inherit the transgene, owing to segregation, and therefore have zou mutant phenotype. (D) Embryos dissected from mature seeds of wild-type (left) and zou-2 mutants (right). (E-H) Cleared seeds viewed using DIC microscopy. Arrows indicate embryos which are at globular (E), heart (F), late torpedo (G,H) stages. (I,J) Light microscope images of 1µM sections of resin-embedded seeds. Arrow in J indicates the mis-shapen zou embryo, which has adhered to the surrounding cellularized endosperm. (K,L) TEM images of ultra thin sections of osmium stained seeds with torpedo stage embryos. The cuticularized layer (CL) is indicated by arrows. In zou-2 (K), unlike ZOU+ (L), endosperm (en) is appressed to the cuticle of the embryo (em). Scale bars: 200µm in A-H; 100µm in I,J; 500 nm in K,L.

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