Pollination mechanism in Serapias with no pollinaria reconfiguration.

Orchidaceae, one of the most numerous families in the world's flora, have evolved various pollination strategies to favour cross-pollination, such as deceptive pollination and pollinarium reconfiguration. Among the terrestrial orchids of the Mediterranean, only species belonging to the genus Serapias show a strategy defined as shelter imitation. The floral elements form a tubular structure that insects use during their resting phases. The purpose of this article was to clarify the mechanisms that guarantee pollination with particular attention to the morphological interactions between orchids and pollinators and whether pollinaria reconfiguration is necessary in the promotion of cross-pollination in Serapias. Breeding system experiments and hand-pollination treatments indicated that Serapias was highly self-compatible, shows low value of natural fruit set and is pollinator limited. Time-lapse photos showed that the pollinarium had no refolding of the stipe or caudicle after its removal from the flower. The morphology of the flower determined the attack of the pollinarium on the occiput/vertex of insect. When the insect left the flower, the pollinarium was unable to encounter the stigma. When the insect made a second visit to another flower, the pollen masses of the first pollinarium ended up on the stigma and at the same time, the insect picked up a second pollinarium. Our observations and analyses suggested that morphological interactions between flower and pollinator are crucial to the success of pollination and to prevent self-pollination and thus that pollinarium reconfiguration is unnecessary in shelter deceptive orchids, such as Serapias species, for the promotion of cross-pollination. Serapias represent a case of interactions between plant and pollinator; the formation of the tubular shape of the flower is an essential preadaptation for the development of resting site mimicry originating exclusively in Serapias among Mediterranean orchids.

Pollinaria Reconfiguration Mechanism of Widespread Euro-Mediterranean Orchids: The Effects of Increasing Air Temperature.

Orchids are fascinating for many reasons: their reproductive strategies, their pollination systems and the various morphological adaptations they have evolved, including the presence of pollen grains agglomerated into two masses, called pollinia, which form a structure known as a pollinarium. After withdrawal from a flower, the pollinarium undergoes a bending movement such that the pollen masses become correctly orientated to strike the stigma. We evaluated the duration of pollinator visits to inflorescences and the effects of temperature on pollinaria reconfiguration in eight orchid species in order to analyze the effects of increasing air temperature on the changes in bending time, and thus on geitonogamy levels. The impact of temperature on insect behavior was not assessed because our priority was to understand the effects of temperature on the process of pollinaria reconfiguration. All the examined species showed natural reconfiguration times that were 1.7-3.0 times longer than the pollinator residency times. A higher temperature showed a reduction in bending time regardless of the species tested. However, the bending time was never shorter than the residence time of the insects on the flower. Our data showed that high temperatures had a limited effect on the pollinarium reconfiguration time, thus indicating that high temperatures had a limited effect on folding compared to the effect that it had on the viability of the pollen.

Corrigendum: Sicoli G, Passalacqua NG, De Giuseppe AB, Palermo AM, Pellegrino G (2019) A new species of Psathyrella (Psathyrellaceae, Agaricales) from Italy. MycoKeys 52: 89-102. https://doi.org/10.3897/mycokeys.52.31415.

[This corrects the article DOI: 10.3897/mycokeys.52.31415.].

A new species of Psathyrella (Psathyrellaceae, Agaricales) from Italy.

Sporophores of a new Psathyrella species have been reported for the first time as growing at the base of Cladiummariscus culms in the Botanical Garden of the University of Calabria, Rende, Cosenza, southern Italy. The fungus was initially identified as P.thujina (= P.almerensis) by means of both ecology and macro- and microscopic characteristics of the basidiomes, then referred to P.cladii-marisci sp. nov. after extraction, amplification, purification and analysis of the rDNA ITS region. We came to this conclusion after comparing our specimen with the descriptions of the taxa available in the literature for the genus Psathyrella.

Functional differentiation in pollination processes among floral traits in Serapias species (Orchidaceae).

Floral displays, influencing attractiveness to insects, increase the number of pollinator visits and the efficiency of each visit in terms of pollen exchange and thus affect the plant reproductive success. Here, we conducted an in situ manipulation experiment to investigate whether the floral modifications affect reproductive success in natural orchid populations of Serapias lingua and Serapias vomeracea. We estimated male and female reproductive success of three treatment groups, disassembly of floral tube, cutting of lip, and painting of the callus surface, in terms of pollinaria removed/deposited and fruit production. Results revealed that phenotypic modification had opposite effects on reproductive success of two examine species. Indeed, reproductive success was significantly increased by the detached of the petals and sepals, and decreased, due to callus painting and lip removal, in S. lingua. On the contrary, unmanipulated plants of S. vomeracea showed significantly higher value of pollinaria removed and deposited and fruit set than manipulated ones. The differences between S. lingua and S. vomeracea agree to the different pollination strategy of examined species. S. vomeracea shows shelter imitation strategy, and thus, the disassembly of tunnel-like corolla does not allow the insects to use the flower as a refuge, while S. lingua is a sexually deceptive orchid and therefore the opening of the flower made more visible callus (visible at a greater distance) increasing the pollinators attraction. This study provides evidence that pollinators were largely sensitive to the experimental modification of the flower phenotype, which is consistent with the presence of significant selection on individual floral characters. Our experimental investigations of the effects of variation in display on pollinator visitation provide insights into the evolution of floral morphology in orchid with shelter imitation strategy.

Footprint of the eastern euroasian past in Italian populations of Cryptotaenia thomasii (Ten.) DC.

BACKGROUND: The knowledge of the genetic architecture of closely related species and/or populations of a single species can be very useful to shed light on the processes that led to their current distributions. The present study provides a preliminary evaluation of the conservation status of the populations of Cryptotaenia thomasii, a very narrow endemic species occurring in southern Italy. Previous studies showed that C. thomasii was phylogenetically closely related to C. flahaultii, endemic species of Caucasus, and to evaluate its conservation status and the genetic variability of plant species the internal transcribed spacers (ITSs) of nuclear ribosomal DNA were sequenced and the SNPs were analyzed. RESULTS: The restriction analysis of nrDNA with the restriction enzymes allowed to detect the presence of two single mutations (SNPs) among the sequences of two species. Our molecular analysis pointed out that C. thomasii and C. flahaultii, in spite of their geographical disjunction, show a sign of an ancient contact as an extreme case of geographical disjunction Italian-Caucasus. CONCLUSION: From the evolutionary history of the species and its distribution pattern one can reconstruct a possible scenario with some hypotheses that explain the hypothetical ancestral area where the two species were in contact. We speculate the two species may have originated from fragmentation of a common ancestor widespread in the Western Palearctic zone and have survived in two separated refuge areas limited by important mountain systems. This hypothesis is supported by our molecular analysis, in fact, the analysis of SNPs showed that some C. thomasii populations retain the signs of an ancient link with C. flahaultii.

Who helps whom? Pollination strategy of Iris tuberosa and its relationship with a sexually deceptive orchid.

Reproductive success of plants may be affected by interactions with co-flowering species either negatively, through competition for pollinators, or positively, by means of a magnet species effect and floral mimicry. In this study, potential interactions between Iris tuberosa, a rewarding species, and Ophrys fusca, a sexually deceptive orchid, were explored in four populations in southern Italy. In each population plots showing different ratios of the examined species were arranged in the field, and in each plot the number of pollinators and fruit set were assessed. In addition, flower size and floral hydrocarbons produced by the two species were analysed. Morphological and scent data pointed out that flower size and aliphatic compounds did not differ significantly between the two species. Interestingly, both species shared tricosane and 11-nonacosene, electrophysiologically active compounds in the shared dominant pollinator Adrena. We have found that fruit production and number of pollinators in I. tuberosa varied significantly among plots, while percentage of capsules and number of pollinators of O. fusca captured showed no significant differences across plots. These results suggested, that the presence of O. fusca contributes differentially to pollinator attraction, and thus, to total reproductive success of I. tuberosa, according to a different ratio of aggregation. These findings suggest that I. tuberosa profits from the greater abundance of insects attracted by the presence of orchid specimens, and that a sexually deceptive orchid may be a magnet species in pollination strategy.

Effects of population structure on pollen flow, clonality rates and reproductive success in fragmented Serapias lingua populations.

BACKGROUND: Fragmentation of habitats by roads, railroads, fields, buildings and other human activities can affect population size, pollination success, sexual and asexual reproduction specially in plants showing pollinator limitation, such as Mediterranean orchids. In this study, we assessed pollen flow, selfing rates, vegetative reproduction and female reproductive success and their correlations with habitat characters in nine fragmented subpopulations of Serapias lingua. To improve understanding of population structure effects on plant biology, we examined genetic differentiation among populations, pollen flow, selfing rates and clonal reproduction using nuclear microsatellite markers. RESULTS: Smaller populations showed a significant heterozygote deficit occurred at all five nuclear microsatellite loci, the coefficient of genetic differentiation among populations was 0.053 and pairwise FST was significantly correlated with the geographical distance between populations. Paternity analysis of seeds showed that most pollen flow occurred within a population and there was a positive correlation between percentage of received pollen and distance between populations. The fruit production rate varied between 5.10 % and 20.30 % and increased with increasing population size, while the percentage of viable seeds (78-85 %) did not differ significantly among populations. The extent of clonality together with the clonal and sexual reproductive strategies varied greatly among the nine populations and correlated with the habitats where they occur. The small, isolated populations tended to have high clonal diversity and low fruit production, whereas the large populations with little disturbance were prone to have reductions in clonal growth and increased sexual reproduction. CONCLUSIONS: We found that clonality offers an advantage in small and isolated populations of S. lingua, where clones may have a greater ability to persist than sexually reproducing individuals.