Search Results (4)

There is little information on the use of pollen in molecular research, despite the increased interest in genome editing by pollen-mediated transformation. This paper presents an essential toolbox of technical procedures and observations for molecular studies on onion (Allium cepa L.) pollen. PCR is a useful tool as an express method to evaluate editing results before pollination. A direct PCR protocol for pollen suspension has been adapted without needing DNA pre-extraction. We showed that the outer layer of lipids known as pollenkitt is a limiting factor for successful PCR on pollen. A simple pre-washing step of pollen suspension was able to eliminate the pollenkitt and enormously affect the PCR results. Additionally, our pollenkitt study helped us develop a simple and effective pollination method using wetted onion pollen grains. Classical manual pollination usually is conducted by intact pollen without wetting. Most existing methods of the editing system delivery into pollen are carried out in a wet medium with consequent drying before pollination, which adversely affects the viability of pollen. The optimal medium for wet pollination was 12% sucrose water solution. Our method of using wetted pollen grains for pollination might be very beneficial for pollen genetic manipulation. Full article

Although pollination is one of the most crucial biological processes that ensures plant reproduction, its mechanisms are poorly understood. Especially in insect-mediated pollination, a pollen undergoes several attachment and detachment cycles when being transferred from anther to insect and from insect to stigma. The influence of the properties of pollen, insect and floral surfaces on the adhesion forces that mediate pollen transfer have been poorly studied. Here, we investigate the adhesive properties of Hypochaeris radicata pollen and their dependence on pollen aging by quantifying the pull-off forces from glass slides using centrifugation and atomic force microscopy. We found that the properties of the pollenkitt—the viscous, lipid liquid on the surface of most pollen grains—influences the forces necessary to detach a pollen from hydrophilic surfaces. Our results show that aged H. radicata pollen form weaker adhesions to hydrophilic glass than fresh ones. On the other hand, when a pollen grain ages in contact with glass, the adhesion between the two surfaces increases over time. This study shows for the first time the pollen aging effect on the pollination mechanism. Full article

Our study attempted to elucidate the significance of floral and pollen traits of the highly nutritious tropical trees Moringa oleifera and Moringa peregrina for their reproductive success under arid conditions. We found that the pollen grains of both species were immersed in a pollenkitt that constituted ~60% of the pollen. Successful pollination was achieved by large bees inserting the pollen into a narrow stylar tube. We found that, upon removal of the pollenkitt, approximately 65% fewer pollen grains penetrated the stylar tube for both species. The pollenkitt protected against heat and desiccation, while removing the pollenkitt resulted in significantly reduced levels of the viability of pollen grains, especially in M. oleifera, and significantly reduced levels of germinability in both species. The stylar tube provided high protection for pollen grains against heat and desiccation even when the pollenkitt was removed. Chemical analysis of pollenkitts of the two species revealed a waxy blend of 21 hydrocarbon compounds, in which n-alkanes constituted >90% of the compounds and their identity corresponded to known plant and animal hydrocarbons, associated with protection against heat and water stress. We concluded that, under arid conditions, the reproductive success of both Moringa species is potentially enhanced by their unique floral and pollen traits. This supports the prospect of cultivating M. oleifera and Moringa peregrina as food crops in arid regions across the globe. Full article

In recent decades, airborne allergens for allergic respiratory diseases have been found to increase significantly by a process of converting coniferous forests into broad-leaved forests in Korea. This study was conducted to evaluate factors, including airborne pollen counts, micromorphology, and flowering phenology, that can affect oak pollen-related allergic symptoms. The catkin of Mongolian oak (Quercus mongolica Fisch. ex Ledeb.) showed the most rapidly blooming catkin on Julian day 104 in flower development. Among six species, the last flowering was observed on Julian day 119 in Korean oak (Quercus dentata Thunb.). The pollen dispersal was persisted for about 32 days from Julian day 104 to Julian day 136. Airborne pollen was observed about 2 weeks after flowering phase H, the senescence phase. Pollen size varied by species, with the largest from Q. mongolica (polar axis length, PL = 31.72 µm, equatorial axis length, EL = 39.05 µm) and the smallest from Jolcham oak (Quercus serrata Murray) (PL = 26.47 µm, EL = 32.32 µm). Regarding pollen wall structure, endexine of Q. dentata was coarsely laminated or fragmented. The endexine thicknesses of Sawtooth oak (Quercus acutissima Carruth.) and Q. serrata were thick and stable, whereas Galcham oak (Quercus aliena Blume), Q. mongolica, and Oriental cork oak (Quercus variabilis Blume) had thinner endexines. The area occupied by pollenkitt of Q. variabilis was significantly larger than that of Q. acutissima. Importantly, Q. variabilis had a distinctly thick 17 kDa protein band, a presumed major allergen. Oak species differ in pollen protein composition, and thus there is a possibility that the allergenic activity of pollen proteins vary depending on oak species. This study highlights the fact that native oak species in Korea differ in flowering pattern of male flowers, pollen morphology, and pollen chemical constituents. These discrepancies in flowering and pollen properties imply variable allergic responses to oak pollen from different species. Full article