Somatic embryogenesis requires auxin and establishment of the shoot apical meristem (SAM). WUSCHEL (WUS) is critical for stem cell fate determination in the SAM of higher plants. However, regulation of WUS expression by auxin during somatic embryogenesis is poorly understood. Here, we show that expression of several regulatory genes important in zygotic embryogenesis were up-regulated during somatic embryogenesis of Arabidopsis. Interestingly, WUS expression was induced within the embryonic callus at a time when somatic embryos could not be identified morphologically or molecularly. CorrectWUS expression, regulated by a defined critical level of exogenous auxin, is essential for somatic embryo induction. Furthermore, it was found that auxin gradients were established in specific regions that could then give rise to somatic embryos. The establishment of auxin gradients was correlated with the induced WUS expression. Moreover, the auxin gradients appear to activate PIN1 polar localization within the embryonic callus. Polarized PIN1 is probably responsible for the observed polar auxin transport and auxin accumulation in the SAM and somatic embryo. Suppression of WUS and PIN1 indicated that both genes are necessary for embryo induction through their regulation of downstream gene expression. Our results reveal that establishment of auxin gradients and PIN1-mediated polar auxin transport are essential for WUS induction and somatic embryogenesis. This study sheds new light on how auxin regulates stem cell formation during somatic embryogenesis.

Homeobox genes are master regulatory genes that specify the body plan and control development of many eukaryotic organisms, including plants. We isolated and characterized a cDNA designated ATML1 (for Arabidopsis thaliana meristem L1 layer) that encodes a novel homeodomain protein. The ATML1 protein shares high sequence homology inside and outside of the homeodomain with both the Phalaenopsis O39 and the Arabidopsis GLABRA2 (GL2) homeodomain proteins, which together define a new class of plant homeodomain-containing proteins, designated HD-GL2. The ATML1 gene was first expressed in the apical cell after the first asymmetric division of the zygote and continued to be expressed in all proembryo cells until the eight-cell stage. In the 16-cell proembryo, the ATML1 gene showed a distinct pattern of expression, with its mRNA becoming restricted to the protoderm. In the torpedo stage of embryo development, ATML1 mRNA disappeared altogether but reappeared later only in the L1 layer of the shoot apical meristem in the mature embryo. After germination, this L1 layer-specific pattern of expression was maintained in the vegetative shoot apical meristem, inflorescence, and floral meristems, as well as in the young floral organ primordia. Finally, ATML1 mRNA accumulated in the protoderm of the ovule primordia and integuments and gradually became restricted in its expression to the endothelium surrounding the embryo sac. We propose that ATML1 may be involved in setting up morphogenetic boundaries of positional information necessary for controlling cell specification and pattern formation. In addition, ATML1 provides an early molecular marker for the establishment of both apical-basal and radial patterns during plant embryogenesis.

Pollination regulates a syndrome of developmental responses that contributes to successful sexual reproduction in higher plants. Pollination-regulated developmental events collectively prepare the flower for fertilization and embryogenesis while bringing about the loss of floral organs that have completed their function in pollen dispersal and reception. Components of this process include changes in flower pigmentation, senescence and abscission of floral organs, growth and development of the ovary, and, in certain cases, pollination also triggers ovule and female gametophyte development in anticipation of fertilization. Pollination-regulated development is initiated by the primary pollination event at the stigma surface, but because developmental processes occur in distal floral organs, the activity of interorgan signals that amplify and transmit the primary pollination signal to floral organs is implicated. Interorgan signaling and signal amplification involves the regulation of ethylene biosynthetic gene expression and interorgan transport of hormones and their precursors. The coordination of pollination- regulated flower development including gametophyte, embryo, and ovary development; pollination signaling; the molecular regulation of ethylene biosynthesis; and interorgan communication are presented.

Pollination initiates a syndrome of developmental events that contribute to successful reproduction, including perianth senescence, changes in pigmentation, and ovule differentiation in preparation for impending fertilization. In orchid flowers, initiation of each of these processes in distinct floral organs is strictly and coordinately controlled by pollination, thus providing a unique opportunity to study the signals that coordinate interorgan postpollination development. Because ethylene has been implicated in contributing to regulation of severa1 aspects of postpollination development, we focused on determining the expression of its biosynthetic genes and their possible role in regulation. The abundance of mRNA encoding both 1-aminocyclopropane-l-carboxylic acid (ACC) synthase and ACC oxidase in the stigma, ovary, and labellum was found to be coordinately regulated by emasculation, auxin, and ethylene. Although petals contribute up to 26% of total flower ethylene and accumulate high levels of ACC oxidase mRNA and activity following pollination, no ACC synthase mRNA or activity was detected in this tissue. Together, these results support a model of interorgan regulation of postpollination development that depends on pollination-stimulated accumulation of mRNA encoding ethylene biosynthetic enzymes in a developmentally regulated and tissue-specific manner. This model relies on the translocation of a soluble hormone precursor, ACC, rather than on the translocation of the hormone itself. In this way, ACC serves to actuate the response already initiated by ethylene perceived by other parts of the flower. Thus, ACC may function as a secondary transmissible signal that coordinates postpollination development in diverse floral organs.

The indoleamine melatonin, a well-known animal chemical, has been identified in extracts from several plant species. The function of melatonin in plants is unknown. Two major functions of melatonin in animals are dark signaling and antioxidant protection. Fruit ripening was used as a model physiological process that involves changes in the oxidative status of an organ. Tomato fruits at various stages of ripeness were sampled. Morning glory (Pharbitis nil Choisy, cv. Violet) and tomato (Lycopersicon esculentum Mill., cv. T5 and Castlemart) organs were collected throughout a light/dark cycle to determine whether melatonin levels increased during the night. No consistent evidence was found that melatonin increased significantly in organs of these plants during the night, as it does in many animals. The melatonin content of the fruits generally increased during ripening up to the mature ripe stage and thereafter as the fruit became over ripe.