H. F. Linskens

Distribution of unesterified and esterified pectins in cell walls of pollen tubes of flowering plants

Immunocytochemical localization of polygalacturonic acid (pectin) and methyl-esterified pectin in the walls of pollen tubes of 20 species of flowering plants grown in vitro was investigated by using monoclonal antibodies (MAbs) JIM5 and JIM7 and by means of confocal laser scanning microscopy (CLSM). In general, periodic annular deposits of pectins were found coating the tube wall in species possessing solid styles, and a more uniform pectin sheath in tube walls in species having hollow styles or no styles. We hypothesize that the periodic ring-like structure of the pectin sheath reinforces pollen tubes for passing through the transmitting tract in the style. Esterified pectin which prevents Ca2+-induced gelification of pectate is located predominantly at the apex. This implies that pectin esterification is related to tip wall “loosening” that is required for cell wall expansion during tip growth of pollen tubes. The occurrence of unesterified pectins in other areas of pollen tube walls suggests that de-esterification of pectin following tip expansion leads to a more rigid form of pectin that contributes to the construction of the pollen tube wall.

Pollen viability and pollen vigor

SummaryInvestigations were carried out to correlate pollen viability, assessed on the basis of a fluorochromatic reaction (FCR) test, with pollen vigor, assessed on the basis of the time taken for in vitro germination in pollen grains subjected to high humidity (>95% RH) and temperature (38 °C) or storage stress of Nicotiana tabacum, Agave sp., Tradescantia virginiana, and Iris sp. Both high RH and temperature, as well as storage stresses, affected pollen vigor before affecting pollen viability. The results are discussed in the light of available data on the viability and vigor of stressed pollen and of aged seeds. The need for consideration of pollen vigor, particularly in stored pollen, the inadequacy of the methods presently used, and some of the methods suitable to assess pollen vigor are elaborated.

Enforced growth-rate fluctuation causes pectin ring formation in the cell wall of Lilium longiflorum pollen tubes

Normally growing lily (Lilium longiflorum Thunb.) pollen tubes cultured in standard sucrose medium display a relatively steady tip-growth pattern and a rather even pectin sheath in the cell wall. In an attempt to better understand pulsatory growth, observed in some species, e.g., Petunia, and its possible role in causing the formation of thickened cell wall rings, we have imposed marked fluctuations in the growth-rate of lily pollen tubes. The appropriate growth-perturbing conditions were achieved by modulating the medium osmolarity or by applying caffeine, a non-turgor inhibitor, in a specially designed incubation chamber with a controlled medium flow. The relatively non-esterified pectin deposition in the wall of the growth-interrupted pollen tubes was detected by immunofluorescence microscopy using a monoclonal antibody, JIM 5. The observations show that the periods of slow or inhibited growth correspond to the times when the thickened walls are deposited. Since the growth fluctuations were induced by both turgor- and non-turgor-related means, the proposed endogenous regulatory role of turgor pressure is questioned. Other factors, such as the tip-focused Ca2+ gradient which was demonstrated by ratiometric ion imaging, and the alteration in the extensibility of the cell wall, which correlated with pectin esterification/de-esterification, emerge as candidates for the regulation of growth fluctuations.

Responses of tobacco pollen to high humidity and heat stress: viability and germinability in vitro and in vivo

SummaryResponses of pollen grains of Nicotiana tabacum to high humidity (95% RH, 4 h) and temperature (38°/45° C, 4 h) stresses were investigated. Pollen grains were subjected to only RH or only temperature, or to both of these stresses. Their viability was assessed on the basis of the fluorochromatic reaction (FCR) test, and vigour was assessed on the basis of the time taken for in vitro germination as well as on the emergence of pollen tubes through the cut end of semi-vivo implanted styles. None of the stress conditions affected pollen viability and high RH or high temperature stress did not individually affect pollen vigour. However, pollen vigour was markedly affected when both the stresses were given together. Pollen grains subjected to high RH at 38° C took a longer time to germinate in vitro and the pollen tubes emerged later from the cut end of the semi-vivo styles; division of the generative cell was also delayed. Pollen grains subjected to high RH at 45° C failed to germinate in vitro, but did germinate on the stigma. Many pollen tubes subjected to this treatment showed abnormalities, and the growth of pollen tubes in the pistil was much slower than that observed in other treatments. Pollen samples subjected to all of the stress conditions were able to induce fruit and seed set. The implications of these results on the relationship between the FCR test and viability, and between viability and vigour, especially in stressed pollen, are discussed.

Pollen-allergy as an ecological phenomenon: A review

Allergy is the specific reaction of the human body to foreign molecules: The organism has to be able to recognize the molecules and to become sensitized. Those molecules which cause an allergic reaction can be found in the normal human environment and represent, besides molecules which are consumed with the food, those substances derived from the surface of foreign organisms, such as dander, feces, fungal spores, pollen grains. In extreme cases the result can be death. The reacting organism needs a first encounter with the allergen in order to become sensitized. The second and following encounters result in the allergic reaction of different kinds and on different parts and organs of the body (atopic reaction, atopy). Molecules acting as allergens come from organisms, called allergen producers. The allergens are produced in low concentrations, but the sensitized counterpart reacts nevertheless. One has to distinguish between the allergenic molecules as such, and the allergen carriers (JORDE & LINSKENS, 1979), that are the organisms or parts of the organisms (tissues, diaspores, mites, dander), which carry the allergens attached to their cells and/or surfaces. We get the following scheme:

Entrapment of long-distance transported pollen grains by various moss species in coastal Victoria Land, Antarctica

SummaryIn northern Victoria Land (continental Antarctica, between 72° and 76°S, 162° and 169°E), 18 moss samples have been collected and analysed for the presence of pollen. In turfs and cushions of 8 different moss species, at least 27 pollen taxa could be identified. The pinus-type pollen and those of grasses were very common. More than 60% of the total grains were damaged or could not be identified. There is evidence that the Antarctic continent could act as a sink for wind-transported pollen from sub-Antarctic islands or from plants (native or cultivated) in South America, Australia, New Zealand and South Africa. However, the pollen concentration in air (⩽ 1 pollen grain/100 m3) and its entrapment rate on moss (about 0.12 grain/cm2/year) result in a very low pollen density in these plants.

Localization of ubiquitin in anthers and pistils of Nicotiana

Ubiquitin-conjugated compounds were localized in anthers and pistils of Nicotiana alata by immuno-cytochemistry. In young anthers, antibodies to ubiquitin bound to callose cell walls surrounding pollen mother cells and to organelles in the endothecium. At the freespore stage, antibodies bound to circular-cell cluster cells subtending the stomium and to organelles and cell walls of endothecial cells. Near anther dehiscence, locular material was labeled. In pistils, cell walls of stylar transmitting tissue were labeled in a beaded pattern. Antibodies bound to a thin layer surrounding ovules, to the lining of embryo sacs, to cytoplasm of eggs and synergids, and to starch grains in central cells. Sites of localization were tissue- and time-specific, suggesting a regulatory role for ubiquitin in development of reproductive structures in flowering plants.

DNA Fingerprinting of Italian Grape Varieties : A Test of Reliability in RAPDs.

Random Ampliefied Polymorphic DNA (RAPD) fingerprinting is shown to be a useful system for identifying varieties of Italian grape varieties. Replicate accessions give the same fingerprints, indicating that contaminants on leaf surfaces do not lead to errors. Three 10 base random primers Operon A-03, A-05, and A07 were particularly useful and each was usually capable of identifying all varieties sampled.