Enhanced Visualization of the Fine Structure of the Stigmatic Surface of Citrus using Pre-fixation Washes

Stigma characteristics and morphology can be useful in taxonomic and phylogenetic studies, indicate relationships in stigma function and receptivity, and be valuable in evaluating pollen–stigma interactions. Problematic is that in some taxa, copious stigmatic exudate can obscure the fine structural details of the stigmatic surface. Such is the case for Citrus, which has a wet stigma type on which abundant exudate inundates surface papillae. The components of stigmatic surface compounds are highly heterogeneous and include carbohydrates, proteins, lipids, glycoproteins, and phenolic compounds. This study evaluated the efficacy of several pre-fixation wash treatments on removing surface exudate to visualize the underlying stigmatic surface. Wash treatments included various buffer solutions, surfactants, dilute acids/bases, and solvents. Stigmas prepared using conventional fixation methods in glutaraldehyde had considerable accumulations of reticulate surface deposits with stigmatic cells obscured. Pre-fixation washes containing solvents such as methanol, chloroform, and ethanol left accumulations of incompletely removed exudate and crystalline deposits. Alkaline water washes produced a crust-like deposit on stigma surfaces. Buffer washes left residues of plaque-like deposits with perforated areas. In contrast, excellent removal of stigmatic exudate was obtained with a pre-fixation wash composed of 0.2 M Tris buffer, pH 7.2, containing 0.2% Triton X-100 surfactant and allowed clear imaging of the stigma and surface papillae morphology. A central sinus and radially arranged openings on the stigmatic surface were clearly visible and shown for the first time using scanning electron microscopy (SEM).

The floral development and anatomy of Carica papaya (Caricaceae)

Floral development and anatomy of Carica papaya L. have been investigated to shed light on (i) the morphology of the flower, (ii) the structural basis for the pollination mechanism, and (iii) the relationships of the Caricaceae. Carica is mostly dioecious with a strong dimorphism between staminate and pistillate flowers. The development of staminate flowers resembles that of pistillate flowers up to the initiation of the stamens. Further development leads to highly diverging morphologies. In staminate flowers a combination of contorted growth and the development of a common stamen-petal tube produces a long floral tube. The gynoecium grows into a central spearlike pistillode. The pistillate flowers have no traces of stamens and initiate five antesepalous carpel primordia. Common basal growth leads to the development of a large ovary with staglike stigmatic lobes and intruding placentae covered with numerous ascending ovules. Floral anatomy of staminate and pistillate flowers is described. The nature of the colleters is discussed. The morphological basis for reward production in C. papaya is clarified, and conflicting views on pollination are discussed. Nectaries of staminate flowers are located on the central rudimentary pistil and not at the base of the stamens, as previously reported. The anthers contain packages of calcium oxalate crystals. Pistillate flowers produce no nectar but have a stigmatic exudate. We compared the floral development and anatomy of Carica with that of Adenia (Passifloraceae) and Moringa (Moringaceae) in the view of a relationship with other glucosinolate-producing families. Although a derivation of the unisexual flowers from bisexual ancestors is probable, Storey's hypothetical derivation of pistillate flowers is not supported by the floral ontogeny and vasculature.Key words: Adenia, Caricaceae, Moringa anatomy, calcium oxalate packages, dioecy, floral structure, nectaries, ontogeny, pollination, systematic relationships.

A scanning electron microscope study of the pollen tube pathway in pistils of Rhododendron

The pollen tube pathway was observed at regular intervals in pistils of four species of Rhododendron, with emphasis on Rhododendron fortunei. Rhododendron is characterized by a nonpapillate wet stigma, angled stylar canal, placentae with central clefts, and many unitegmic anatropous ovules. Receptive stigmas were hand-pollinated with self pollen 1 – 8 days after anthesis. The pollen, which occurs in permanent tetrads, started germinating during the 1st day. After crossing the stigma surface to one of the grooves leading into the stylar canal, pollen tubes grew straight through the style, and continued into the placental clefts from which they emerged onto the placental surface to grow among the ovules. Tubes reached the ovary in 5 – 10 days depending on the species and took several days after entering the upper ovary to reach the base of the placentae. Single tubes (rarely two or more) diverged from the interovular network and grew under the integument (which is close against the placental surface) to enter the slit-like micropylar opening of an ovule. The morphology of the micropylar slit and the direction of pollen tube entry showed variation among ovules. In R. fortunei ovule entries occurred first on the upper half of the placenta, though not at the top, and in ovules closest to the placental cleft. All portions of the pathway, from stigma surface to micropylar opening, are covered by exudate. Stigmatic exudate increased in amount and became more viscous after pollination, burying the pollen grains and tubes, then gradually dried. Exudate was produced in the style and ovary whether or not pollination occurred. Characteristics of the pollen tube pathway in Rhododendron are discussed relative to those in other angiosperm taxa. Key words: Rhododendron, pollen–pistil interactions, fertilization, transmitting tissues, pistil exudates, ovule entries.

Putrescine Increases Effective Pollination Period in Roses

Flowers of two cultivars of Rosa hybrida were treated or not with putrescine before being pollinated from 2 to 8 days after anther emasculation. On both cultivars the 10-3 M putrescine treatment extended the effective pollination period, as shown by the best hip formation rates and mean number of seeds per hip. On one cultivar, the 10-5 M putrescine treatment increased fertilization efficiency (more hips obtained). The effect of putrescine was proportionally more important on the cultivar characterized by the highest stigmatic exudate pH. Putrescine also influenced in vitro pollen germination by increasing the length of emitted pollen tubes (10-3 and 10-5 M-putrescine) and the quantity of germinated pollen grains (10-5 M putrescine).