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AbstractIn the early twentieth century, Wilhelm Johannsen's breeding experiments on pure lines of beans provided empirical support for his groundbreaking distinction between phenotype and genotype, the foundation stone of classical genetics. In contrast with the controversial history of the genotype concept, the notion of phenotype has remained essentially unrevised since then. The application of the Johannsenian concept of phenotype to modularly built, nonunitary plants, however, needs reexamination. In the first part of this article it is shown that Johannsen's appealing solution for dealing with the multiplicity of nonidentical organs produced by plant individuals (representing individual plant phenotypes by arithmetic means), which has persisted to this day, reflected his intellectual commitment to nineteenth-century typological thinking. Revisitation of Johannsen's results using current statistical tools upholds his major conclusion about the nature of heredity but at the same time falsifies two important ancillary conclusions of his experiments-namely, the alleged homogeneity of pure lines (genotypes) regarding seed weight variability and the lack of transgenerational effects of within-line (within-genotype) seed weight variation. The canonical notion of individual plant phenotypes as arithmetic means should therefore be superseded by a concept of phenotype as a dual property, consisting of central tendency and variability components of organ trait distribution. Phenotype duality offers a unifying framework applicable to all nonunitary organisms.

The long-known, widely documented inverse relationship between body size and environmental temperature ("temperature-size rule") has recently led to predictions of body size decline following current climatic warming ("size shrinking effect"). For keystone pollinators such as wild bees, body shrinking in response to warming can have significant effects on pollination processes but there is still little direct evidence of the phenomenon because adequate tests require controlling for confounding factors linked to climate change (e.g., habitat change). This paper assesses the shrinking effect in a community of solitary bees from well-preserved habitats in the core of a large nature reserve experiencing climatic warming without disturbances or habitat changes. Long-term variation in mean body mass was evaluated using data from 1704 individual bees (137 species, 27 genera, 6 families) sampled over 1990-2023. Climate warmed at a fast rate during this period, annual mean of daily maximum temperature increasing 0.069°C/year on average during 2000-2020. Changes in bee body mass verified expectations from the size shrinking effect. The mean individual body mass of the community of solitary bees declined significantly, irrespective of whether the analysis referred to the full species sample or only to the subset of species that were sampled in both the old (1990-1997) and recent (2022-2023) periods. On average, body mass declined ~0.7%·year -1 , or an estimated average cumulative reduction of ~20 mg per individual bee from 1990 to 2023. Proportional size reduction was greatest among large-bodied species, ranging from around -0.6%·year -1 for the smallest species to -0.9%·year -1 for the largest ones. Declining rate was steeper for cavity-nesting than ground-nesting species. The pollination and mating systems of bee-pollinated plants in the study region are probably undergoing significant alterations as a consequence of supra-annual decline in bee body mass., (© 2023 The Authors. Ecology published by Wiley Periodicals LLC on behalf of The Ecological Society of America.)

The interspecific range of epigenetic variation and the degree to which differences between angiosperm species are related to geography, evolutionary history, ecological settings or species-specific traits, remain essentially unexplored. Genome-wide global DNA cytosine methylation is a tractable 'epiphenotypic' feature suitable for exploring these relationships. Global cytosine methylation was estimated in 279 species from two distant, ecologically disparate geographical regions: Mediterranean Spain and tropical México. At each region, four distinct plant communities were analyzed. Global methylation spanned a 10-fold range among species (4.8-42.2%). Interspecific differences were related to evolutionary trajectories, as denoted by a strong phylogenetic signal. Genomes of tropical species were on average less methylated than those of Mediterranean ones. Woody plants have genomes with lower methylation than perennial herbs, and genomes of widespread species were less methylated than those of species with restricted geographical distribution. The eight communities studied exhibited broad and overlapping interspecific variances in global cytosine methylation and only two of them differed in average methylation. Altogether, our broad taxonomic survey supported global methylation as a plant 'epiphenotypic' trait largely associated with species evolutionary history, genome size, range size and woodiness. Additional studies are required for better understanding the environmental components underlying local and geographical variation., (© 2019 The Authors. New Phytologist © 2019 New Phytologist Trust.)

Floral nectars become easily colonized by microbes, most often species of the ascomycetous yeast genus Metschnikowia. Although it is known that nectar composition can vary tremendously among plant species, most probably corresponding to the nutritional requirements of their main pollinators, far less is known about how variation in nectar chemistry affects intraspecific variation in nectarivorous yeasts. Because variation in nectar traits probably affects growth and abundance of nectar yeasts, nectar yeasts can be expected to display large phenotypic variation in order to cope with varying nectar conditions. To test this hypothesis, we related variation in the phenotypic landscape of a vast collection of nectar-living yeast isolates from two Metschnikowia species (M. reukaufii and M. gruessii) to nectar chemical traits using non-linear redundancy analyses. Nectar yeasts were collected from 19 plant species from different plant families to include as much variation in nectar chemical traits as possible. As expected, nectar yeasts displayed large variation in phenotypic traits, particularly in traits related to growth performance in carbon sources and inhibitors, which was significantly related to the host plant from which they were isolated. Total sugar concentration and relative fructose content significantly explained the observed variation in the phenotypic profile of the investigated yeast species, indicating that sugar concentration and composition are the key traits that affect phenotypic variation in nectarivorous yeasts., (© FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Genetic diversity and genotypic diversity of wild populations of the floricolous yeast Metschnikowia reukaufii exhibit a strong host-mediated component, with genotypes being nonrandomly distributed among flowers of different plant species. To unravel the causal mechanism of this pattern of host-mediated genetic diversity, this paper examines experimentally whether floral nectars of different host plants differ in their quality as a growing substrate for M. reukaufii and also whether genetically distinct yeast strains differ in their relative ability to thrive in nectars of different species (host × genotype interaction). Genetically distinct M. reukaufii strains were grown in natural nectar of different hosts under controlled conditions. Population growth varied widely among nectar hosts, revealing that different host plants provided microhabitats of different quality for M. reukaufii. Different M. reukaufii strains responded in different ways to interspecific nectar variation, and variable growth responses were significantly associated with genetic differences between strains, thus leading to a significant host × genotype interaction. Results of this study provide support for the diversifying selection hypothesis as the underlying mechanism preserving high genetic diversity in wild M. reukaufii populations and also suggest that consequences of functional plant-pollinator diversity may surpass the domain of the mutualistic organisms to implicate associated microorganisms., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)

The genetic and evolutionary relationships among floral nectar-dwelling Pseudomonas 'sensu stricto' isolates associated to South African and Mediterranean plants were investigated by multilocus sequence analysis (MLSA) of four core housekeeping genes (rrs, gyrB, rpoB and rpoD). A total of 35 different sequence types were found for the 38 nectar bacterial isolates characterised. Phylogenetic analyses resulted in the identification of three main clades [nectar groups (NGs) 1, 2 and 3] of nectar pseudomonads, which were closely related to five intrageneric groups: Pseudomonas oryzihabitans (NG 1); P. fluorescens, P. lutea and P. syringae (NG 2); and P. rhizosphaerae (NG 3). Linkage disequilibrium analysis pointed to a mostly clonal population structure, even when the analysis was restricted to isolates from the same floristic region or belonging to the same NG. Nevertheless, signatures of recombination were observed for NG 3, which exclusively included isolates retrieved from the floral nectar of insect-pollinated Mediterranean plants. In contrast, the other two NGs comprised both South African and Mediterranean isolates. Analyses relating diversification to floristic region and pollinator type revealed that there has been more unique evolution of the nectar pseudomonads within the Mediterranean region than would be expected by chance. This is the first work analysing the sequence of multiple loci to reveal geno- and ecotypes of nectar bacteria.

The recent upsurge of interest in the role of floral nectar as a habitat for microorganisms has led to some detailed analyses of nectarivorous yeasts. In contrast, very little is known on the occurrence and diversity of nectar-dwelling bacteria, and bacterial-fungal interactions within nectar remain unexplored. In this work, we studied both the culturable bacteria and microfungi found in the floral nectar of wild Mediterranean plants. In general, bacteria and yeasts were found coexisting in nectar more often than would be expected by chance, and such positive association persisted after accounting for phylogenetic nonindependence of the plant species surveyed. Metschnikowia species were confirmed as the main fungal components of nectar communities, and Acinetobacter was identified as the main bacterial taxa. Finally, individual Operational Taxonomic Units (OTUs) were found to co-occur less frequently than predicted by random expectations. There existed, however, some pairwise associations between OTUs that seemed to account for the general pattern of positive bacteria-yeasts coexistence. We conclude that the culturable communities of nectar microorganisms associated with wild Mediterranean plants are nonrandom assemblages of bacterial and yeast species., (© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

This article documents the addition of 153 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Brassica oleracea, Brycon amazonicus, Dimorphandra wilsonii, Eupallasella percnurus, Helleborus foetidus, Ipomoea purpurea, Phrynops geoffroanus, Prochilodus argenteus, Pyura sp., Sylvia atricapilla, Teratosphaeria suttonii, Trialeurodes vaporariorum and Trypanosoma brucei. These loci were cross-tested on the following species: Dimorphandra coccicinea, Dimorphandra cuprea, Dimorphandra gardneriana, Dimorphandra jorgei, Dimorphandra macrostachya, Dimorphandra mollis, Dimorphandra parviflora and Dimorphandra pennigera., (© 2013 Blackwell Publishing Ltd.)

Floral nectar of insect-pollinated plants often contains dense yeast populations, yet little quantitative information exists on patterns and magnitude of species richness of nectar-dwelling yeasts in natural plant communities. This study evaluates yeast species richness at both the plant community and plant species levels in a montane forest area in southern Spain, and also explores possible correlations between the incidence of different yeast species in nectar and their reported tolerance to high sugar concentrations, and between yeast diversity and pollinator composition. Yeast species occurring in a total of 128 field-collected nectar samples from 24 plant species were identified by sequencing the D1/D2 domain of the large subunit rDNA, and rarefaction-based analyses were used to estimate yeast species richness at the plant community and plant species levels, using nectar drops as elemental sampling units. Individual nectar samples were generally characterized by very low species richness (1.2 yeast species/sample, on average), with the ascomycetous Metschnikowia reukaufii and Metschnikowia gruessii accounting altogether for 84.7% of the 216 isolates identified. Other yeasts recorded included species in the genera Aureobasidium, Rhodotorula, Cryptococcus, Sporobolomyces, and Lecythophora. The shapes and slopes of observed richness accumulation curves were quite similar for the nectar drop and plant species approaches, but the two approaches yielded different expected richness estimates. Expected richness was higher for plant species-based than for nectar drop-based analyses, showing that the coverage of nectar yeast species occurring in the region would be improved by sampling additional host plant species. A significant correlation was found between incidence of yeast species in nectar and their reported ability to grow in a medium containing 50% glucose. Neither diversity nor incidence of yeasts was correlated with pollinator composition across plant species.