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Morphological transitions associated with ovule diversification provide unique opportunities for studies of developmental evolution. Here, we investigate the underlying mechanisms of one such transition, reduction in integument number, which has occurred several times among diverse angiosperms. In particular, reduction in integument number occurred early in the history of the asterids, a large clade comprising approximately one-third of all flowering plants. Unlike the vast majority of other eudicots, nearly all asterids have a single integument, with the only exceptions in the Ericales, a sister group to the other asterids. Impatiens, a genus of the Ericales, includes species with one integument, two integuments, or an apparently intermediate bifid integument. A comparison of the development of representative Impatiens species and analysis of the expression patterns of putative orthologs of the Arabidopsis thaliana ovule development gene INNER NO OUTER (INO) has enabled us to propose a mechanism responsible for morphological transitions between integument types in this group. We attribute transitions between each of the three integument morphologies to congenital fusion via a combination of variation in the location of subdermal growth beneath primordia and the merging of primordia. Evidence of multiple transitions in integument morphology among Impatiens species suggests that control of underlying developmental programs is relatively plastic and that changes in a small number of genes may have been responsible for the transitions. Our expression data also indicate that the role of INO in the outgrowth and abaxial-adaxial polarity of the outer integument has been conserved between two divergent angiosperms, the rosid Arabidopsis and the asterid Impatiens.

The HUELLENLOS (HLL) gene participates in patterning and growth of the Arabidopsis ovule. We have isolated the HLL gene and shown that it encodes a protein homologous to the L14 proteins of eubacterial ribosomes. The Arabidopsis genome also includes a highly similar gene, HUELLENLOS PARALOG (HLP), and genes for both cytosolic (L23) and chloroplast ribosome L14 proteins. Phylogenetic analysis shows that HLL and HLP differ significantly from these other two classes of such proteins. HLL and HLP fusions to green fluorescent protein were localized to mitochondria. Ectopic expression of HLP complemented the hll mutant, indicating that HLP and HLL share redundant functions. We conclude that HLL and HLP encode L14 subunits of mitochondrial ribosomes. HLL mRNA was at significantly higher levels than HLP mRNA in pistils, with the opposite pattern in leaves. This differential expression can explain the confinement of effects of hll mutations to gynoecia and ovules. Our elucidation of the nature of HLL shows that metabolic defects can have specific effects on developmental patterning.

Arabidopsis superman (sup, also referred to as floral mutant10) mutants have previously been shown to have flowers with supernumerary stamens and reduced carpels as a result of ectopic expression of the floral homeotic gene APETALA3 (AP3). Here, we report that sup mutations also cause specific alterations in ovule development. Growth of the outer integument of wild-type ovules occurs almost exclusively on the abaxial side of the ovule, resulting in a bilaterally symmetrical hoodlike structure. In contrast, the outer integument of sup mutant ovules grows equally on all sides of the ovule, resulting in a nearly radially symmetrical tubular shape. Thus, one role of SUP is to suppress growth of the outer integument on the adaxial side of the ovule. Genetic analyses showed that the effects of sup mutations on ovule development are independent of the presence or absence of AP3 activity. Thus, SUP acts through different mechanisms in its early role in ensuring proper determination of carpel identity and in its later role in asymmetric suppression of outer integument growth.

Ovules are complex structures that are present in all seed bearing plants and are contained within the carpels in flowering plants. Ovules are the site of megasporogenesis and megagametogenesis and, following fertilization, develop into seeds. We combined genetic methods with anatomical and morphological analyses to dissect ovule development. Here, we present a detailed description of the morphological development of Arabidopsis ovules and report on the isolation of two chemically induced mutants, bell (bel1) and short integuments (sin1), with altered ovule development. Phenotypic analyses indicated that bel1 mutants initiate a single integument-like structure that develops aberrantly, sin1 mutants initiate two integuments, but growth of the integuments is disrupted such that cell division continues without normal cell elongation. Both mutants can differentiate archesporial cells, but neither forms a normal embryo sac. Genetic analyses indicated that bel1 segregates as a single recessive mutation, and complementation tests showed that the two mutants are not allelic. The phenotypes of the mutants indicate that normal morphological development of the integuments and proper embryo sac formation are interdependent or are governed in part by common pathways. The ovule mutants that we describe in Arabidopsis represent novel genetic tools for the study of this stage of reproductive development.

We have used a differential plaque hybridization screening procedure to isolate cDNA clones for genes that show elevated or exclusive expression in tomato pistils. Clones that showed maximal expression in immature pistils (premeiotic to early meiosis) and mature pistils (at anthesis) were isolated. Of nine clones that were characterized, four were found also to express at some stage of anther development. In situ hybridization experiments showed that expression of the genes we have identified is very tightly regulated both spatially and temporally within the pistil. One gene was identified that is expressed in the pistil only in the transmitting tissue of the style. A second gene was found to express exclusively in two to three cell layers of the ovules for a period of less than eight days.