Pollen production is one plant characteristic that is considered to be altered by changes in environmental conditions. In this study, we investigated pollen production of the three anemophilous species Betula pendula , Plantago lanceolata , and Dactylis glomerata along an urbanization gradient in Ingolstadt, Germany. We compared pollen production with the potential influencing factors urbanization, air temperature, and the air pollutants nitrogen dioxide (NO 2 ) and ozone (O 3 ). While we measured air temperature in the field, we computed concentration levels of NO 2 and O 3 from a land use regression model. The results showed that average pollen production (in million pollen grains) was 1.2 ± 1.0 per catkin of Betula pendula , 5.0 ± 2.4 per inflorescence of Plantago lanceolata , and 0.7 ± 0.5 per spikelet of Dactylis glomerata. Pollen production was higher in rural compared to urban locations on average for B. pendula (+ 73%) and P. lanceolata (+ 31%), while the opposite was the case for D. glomerata (− 14%). We found that there was substantial heterogeneity across the three species with respect to the association of pollen production and environmental influences. Pollen production decreased for all species with increasing temperature and urbanization, while for increasing pollutant concentrations, decreases were observed for B. pendula , P. lanceolata , and increases for D. glomerata . Additionally, pollen production was found to be highly variable across species and within species—even at small spatial distances. Experiments should be conducted to further explore plant responses to altering environmental conditions.

Key message: Paternity analyses show that effective pollination of ash (Fraxinus excelsior L.) in a seed orchard and a floodplain forest affected by ash dieback is more likely to be facilitated by healthier males. Thereby, natural selection can have a positive effect on the health of future generations. Context: Ongoing ash dieback and increasing fragmentation of ash populations may result in reduced pollen flow, which can reduce pollination success of future generations of ash trees. Therefore, it is essential to further improve our understanding of gene flow patterns, especially with respect to ash dieback. Aims: In this study, paternity analyses were conducted in a seed orchard and a floodplain forest in Germany in 2018 to explain the relationship between pollination success and the health status of ash trees and distances of effective pollen transport. Methods: Cambium samples (i.e., from twigs and stumps) were collected from 251 ash trees (putative father and mother trees) for genotyping, and the health status of each tree was documented using a scoring system to evaluate vitality. Additionally, seeds were harvested from 12 mother trees per site. Genetic analyses using nuclear microsatellites were performed to determine paternal trees. Paternities were assigned based on the likelihood model implemented in the Cervus 3.0.7 software. Results: Our results showed that the average pollination distance was 76 m in the seed orchard and 166 m in the floodplain forest. In general, pollination success decreased substantially with increasing distance to the mother tree. Despite the dense tree cover in the floodplain forest, pollen were transported over long distances (greater than 550 m), suggesting that non‑local sources also play a role in pollination. This is supported by the foreign pollen input identified in the seed orchard (66.5%). Self‑pollination was detected only to a very small extent, and thus had no major influence on reproduction. In addition, both healthy and slightly diseased father trees showed similar mating success. However, this was not the case for the severely diseased ash trees (more than 50% of crown damage) because only a few offspring could be assigned to them. Nevertheless, in contrast to the floodplain forest, there was no significant correlation between damage classes and pollination success in the seed orchard. Conclusion: Long‑distance pollen transport contributes to the connectivity of ash trees in the landscape. Additionally, both healthy and slightly diseased fathers have a greater contribution to pollination, thus potentially improving the health of the next generation of ash trees. Moreover, gene flow between stepping stone populations is necessary to ensure the positive impact on the genetic diversity of ash populations in the future.

High-altitude environments are highly susceptible to the effects of climate change. Thus, it is crucial to examine and understand the behaviour of specific plant traits along altitudinal gradients, which offer a real-life laboratory for analysing future impacts of climate change. The available information on how pollen production varies at different altitudes in mountainous areas is limited. In this study, we investigated pollen production of 17 birch (Betula pubescens Ehrh.) individuals along an altitudinal gradient in the European Alps. We sampled catkins at nine locations in the years 2020-2021 and monitored air temperatures. We investigated how birch pollen, flowers and inflorescences are produced in relation to thermal factors at various elevations. We found that mean pollen production of Betula pubescens Ehrh. varied between 0.4 and 8.3 million pollen grains per catkin. We did not observe any significant relationships between the studied reproductive metrics and altitude. However, minimum temperature of the previous summer was found to be significantly correlated to pollen (rs = 0.504, p = 0.039), flower (rs = 0.613, p = 0.009) and catkin (rs = 0.642, p = 0.005) production per volume unit of crown. Therefore, we suggest that temperature variability even at such small scales is very important for studying the response related to pollen production.

Intraspecific genetic variation might limit the relevance of environmental factors on plant traits. For example, the interaction between genetics and (a-)biotic factors regulating pollen production are still poorly understood. In this study, we investigated pollen production of 28 birch (Betula pendula Roth) individuals in the years 2019-2021. We sampled catkins of eleven groups of genetically identical trees, which were partially topped, but of the same age and located in a seed plantation in southern Germany characterized by similar microclimatic conditions. Furthermore, we monitored environmental factors such as air temperature, characterized air quality (NO2, NOx and O3), and assessed potential solar radiation. We especially checked for differences between years as well as between and within clones and assessed the synchronicity of years with high/low pollen production. We present a robust mean for the pollen production of Betula pendula (1.66 million pollen grains per catkin). Our findings show temporal (H(2) = 46.29, p < 0.001) and clonal variations (H(4) = 21.44, p < 0.001) in pollen production. We conclude that synchronized high or low pollen production is not utterly site-specific and, in addition, not strictly dependent on genotypes. We suggest that appropriate clone selection based on application (seed plantation, urban planting) might be advantageous and encourage a long-term monitoring.

Fragmented ash populations due to ash dieback may lead to a limited gene flow and pollination success. Therefore, the viability of ash pollen plays a major role for the survival of the species. The extent to which the long-distance transport of pollen affects pollen viability was investigated with experiments in a climate chamber using ash pollen samples from a seed orchard in Emmendingen, Germany. Furthermore, experiments with a volumetric pollen trap were conducted. A suitable storage temperature for ash pollen was determined by using four viability tests; TTC test, pollen germination, Alexander’s stain and Acetocarmine. An optimization of the germination medium was performed. We found a strong influence of prevailing temperatures on pollen viability, which decreased faster under warmer conditions. At moderate temperatures, viable pollen could still be observed after 28 days. Thus, a possible successful pollination can also be associated to long-range transported pollen. Storage experiments showed that pollen viability could be maintained longer at temperatures of −20 °C and −80 °C than at 4 °C. In particular, the TTC test has proven to be suitable for determining viability. Therefore, properly stored pollen can be used for breeding programs to support the survival of Fraxinus excelsior.