Complex patterns in tolerance and resistance to pests and diseases underpin the domestication of tomato.

A frequent hypothesis explaining the high susceptibility of many crops to pests and diseases is that, in the process of domestication, crops have lost defensive genes and traits against pests and diseases. Ecological theory predicts trade-offs whereby resistance and tolerance go at the cost of each other. We used wild relatives, early domesticated varieties, traditional local landraces and cultivars of tomato (Solanum lycopersicum) to test whether resistance and tolerance trade-offs were phylogenetically structured or varied according to degree of domestication. We exposed tomato genotypes to the aphid Macrosiphum euphorbiae, the cotton leafworm Spodoptera littoralis, the root knot nematode Meloidogyne incognita and two common insect-transmitted plant viruses, and reconstructed their phylogenetic relationships using Genotyping-by-Sequencing. We found differences in the performance and effect of pest and diseases but such differences were not related with domestication degree nor genetic relatedness, which probably underlie a complex genetic basis for resistance and indicate that resistance traits appeared at different stages and in unrelated genetic lineages. Still, wild and early domesticated accessions showed greater resistance to aphids and tolerance to caterpillars, nematodes and diseases than modern cultivars. Our findings help to understand how domestication affects plant-pest interactions and underline the importance of tolerance in crop breeding.

First evidence for the joint dispersal of mycorrhizal fungi and plant diaspores by birds.

Seed dispersal allows plants to colonise new sites and escape from pathogens and intraspecific competition, maintaining plant genetic diversity and regulating plant distribution. Conversely, most plant species form mutualistic associations with arbuscular mycorrhizal (AM) fungi in a symbiosis established immediately after seed germination. Because AM fungi are obligate symbionts, using the same dispersal vector as their host should be highly advantageous for their survival, but the co-dispersal of seeds and AM fungal spores has never been confirmed. We aim to clarify the potential role of European birds, essential dispersers for many plant species, as co-dispersers of seeds and AM fungal spores. In total, 63 bird droppings with intact seeds were placed in sterilised soil and maintained for 4 months in a protected environment to avoid contamination. Additionally, 173 bird droppings and 729 gauze swabs used to clean birds' feet were inspected for AM fungal spores. Although no spores were detected by direct observation of these samples, seven Rubus ulmifolius seedlings obtained from four independent droppings of Erithacus rubecula and Sylvia melanocephala were colonised by AM fungi. Our results show that birds can effectively co-disperse viable seeds and AM fungal spores, potentially over long distances, providing a pivotal mechanism to understand the cosmopolitan distribution of AM fungi.

Integrating plant species contribution to mycorrhizal and seed dispersal mutualistic networks.

Mutualistic interactions like those established between plants and mycorrhizal fungi or seed dispersers are key drivers of plant population dynamics and ecosystem functioning; however, these interactions have rarely been explored together. We assembled a tripartite fungi-plant-disperser network in the Gorongosa National Park-Mozambique, to test (1) if diversity and importance of plant mutualists above- and belowground are correlated, and (2) whether biotically and abiotically dispersed plants are associated with distinct arbuscular mycorrhizal fungi (AMF). We quantified seed dispersal by animals for 1 year and characterized the AMF of 26 common plant species. Sixteen plant species were dispersed by 15 animals and colonized by 48 AMF virtual taxa (VT), while the remaining 10 plant species were not dispersed by animals and associated with 34 AMF VT. We found no evidence for a correlation between the number of plant partners above- and belowground or on plant specialization on both types of partners. We also found no evidence for differentiation of AMF communities between biotically and abiotically dispersed plants. Our results suggest that the establishment of plant interactions with seed dispersers and mycorrhizal fungi is largely independent and that both biotically and abiotically dispersed plants seem to associate with similar communities of AMF.

Should I stay or should I go? Mycorrhizal plants are more likely to invest in long-distance seed dispersal than non-mycorrhizal plants.

Seed dispersal and mycorrhizal associations are key mutualisms for the functioning and regeneration of plant communities; however, these processes have seldom been explored together. We hypothesised that obligatory mycorrhizal plants will be less likely to have long-distance dispersal (LDD) syndromes since the probability of finding suitable mycorrhizal partners is likely to decrease with distance to the mother plant. We contrasted the mycorrhizal status and LDD syndromes for 1960 European plant species, using phylogenetically corrected log-linear models. Contrary to our expectation, having specialised structures for LDD is more frequent in obligate mycorrhizal plants than in non-mycorrhizal plants, revealing that lack of compatible mutualists does not constrain investment in LDD structures in the European Flora. Ectomycorrhizal plants associated with wind-dispersing fungi are also more likely to have specialised structures for wind dispersal. Habitat specificity and narrower niche of non-mycorrhizal plants might explain the smaller investment in specialised structures for seed dispersal.

Arbuscular mycorrhizal fungi communities from tropical Africa reveal strong ecological structure.

Understanding the distribution and diversity of arbuscular mycorrhizal fungi (AMF) and the rules that govern AMF assemblages has been hampered by a lack of data from natural ecosystems. In addition, the current knowledge on AMF diversity is biased towards temperate ecosystems, whereas little is known about other habitats such as dry tropical ecosystems. We explored the diversity and structure of AMF communities in grasslands, savannas, dry forests and miombo in a protected area under dry tropical climate (Gorongosa National Park, Mozambique) using 454 pyrosequencing. In total, 147 AMF virtual taxa (VT) were detected, including 22 VT new to science. We found a high turnover of AMF with ˂ 12% of VT present in all vegetation types. Forested areas supported more diverse AMF communities than savannas and grassland. Miombo woodlands had the highest AMF richness, number of novel VT, and number of exclusive and indicator taxa. Our data reveal a sharp differentiation of AMF communities between forested areas and periodically flooded savannas and grasslands. This marked ecological structure of AMF communities provides the first comprehensive landscape-scale evidence that, at the background of globally low endemism of AMF, local communities are shaped by regional processes including environmental filtering by edaphic properties and natural disturbance.

Refaunation and the reinstatement of the seed-dispersal function in Gorongosa National Park.

Large animals are important seed dispersers; however, they tend to be under a high extinction risk worldwide. There is compelling evidence that the global biodiversity crisis is leading to the deterioration of several ecosystem functions, but there is virtually no information on how large-scale refaunation efforts can reinstate seed dispersal. We evaluated the effectiveness of a 62-km2 wildlife sanctuary, which was established to recover populations of large mammals in Gorongosa National Park (Mozambique), in restoring seed dispersal. We collected animal scats during the dry season of 2014 (June-August) along 5 transects inside and 5 transects outside the sanctuary fence (50 km total) with the same type of plant community, identified animal and plant species in the transects, and quantified the number of seeds in each scat. Based on these data, we built bipartite networks and calculated network and species-level descriptor values, and we compared data collected inside and outside the sanctuary. There were more scats (268 vs. 207) and more scats containing seeds (132 vs. 94) inside than outside the sanctuary. The number of mammal dispersers was also higher inside (17) than outside the sanctuary (11). Similarly, more seeds (2413 vs. 2124) and plant species (33 vs. 26) were dispersed inside than outside the sanctuary. Overall, the seed-dispersal network was less specialized (0.38 vs. 0.44) and there was a greater overlap (0.16 vs. 0.07) inside than outside the sanctuary. Both networks were significantly modular and antinested. The high number and richness of seeds dispersed inside the sanctuary was explained mostly by a higher abundance of dispersers rather than by disperser identity. Our results suggest conservation efforts aimed at recovering populations of large mammals are helping to reestablish not only target mammal species but also their functional roles as seed dispersers in the ecosystem.

Pre-dispersal predation effect on seed packaging strategies and seed viability.

An increased understanding of intraspecific seed packaging (i.e. seed size/number strategy) variation across different environments may improve current knowledge of the ecological forces that drive seed evolution in plants. In particular, pre-dispersal seed predation may influence seed packaging strategies, triggering a reduction of the resources allocated to undamaged seeds within the preyed fruits. Assessing plant reactions to pre-dispersal seed predation is crucial to a better understanding of predation effects, but the response of plants to arthropod attacks remains unexplored. We have assessed the effect of cone predation on the size and viability of undamaged seeds in populations of Juniperus thurifera with contrasting seed packaging strategies, namely, North African populations with single-large-seeded cones and South European populations with multi-small-seeded cones. Our results show that the incidence of predation was lower on the single-large-seeded African cones than on the multi-small-seeded European ones. Seeds from non-preyed cones were also larger and had a higher germination success than uneaten seeds from preyed cones, but only in populations with multi-seeded cones and in cones attacked by Trisetacus sp., suggesting a differential plastic response to predation. It is possible that pre-dispersal seed predation has been a strong selective pressure in European populations with high cone predation rates, being a process which maintains multi-small-seeded cones and empty seeds as a strategy to save some seeds from predation. Conversely, pre-dispersal predation might not have a strong effect in the African populations with single-large-seeded cones characterized by seed germination and filling rates higher than those in the European populations. Our results indicate that differences in pre-dispersal seed predators and predation levels may affect both selection on and intraspecific variation in seed packaging.

Invasion genetics of the Bermuda buttercup (Oxalis pes-caprae): complex intercontinental patterns of genetic diversity, polyploidy and heterostyly characterize both native and introduced populations.

Genetic diversity in populations of invasive species is influenced by a variety of factors including reproductive systems, ploidy level, stochastic forces associated with colonization and multiple introductions followed by admixture. Here, we compare genetic variation in native and introduced populations of the clonal plant Oxalis pes-caprae to investigate the influence of reproductive mode and ploidy on levels of diversity. This species is a tristylous geophyte native to South Africa. Invasive populations throughout much of the introduced range are composed of a sterile clonal pentaploid short-styled form. We examined morph ratios, ploidy level, reproductive mode and genetic diversity at nuclear microsatellite loci in 10 and 12 populations from South Africa and the Western Mediterranean region, respectively. Flow cytometry confirmed earlier reports of diploids and tetraploids in the native range, with a single population containing pentaploid individuals. Introduced populations were composed mainly of pentaploids, but sexual tetraploids were also found. There was clear genetic differentiation between ploidy levels, but sexual populations from both regions were not significantly different in levels of diversity. Invasive populations of the pentaploid exhibited dramatically reduced levels of diversity but were not genetically uniform. The occurrence of mixed ploidy levels and stylar polymorphism in the introduced range is consistent with multiple introductions to the Western Mediterranean. This inference was supported by variation patterns at microsatellite loci. Our study indicates that some invasive populations of Oxalis pes-caprae are not entirely clonal, as often assumed, and multiple introductions and recombination have the potential to increase genetic variation in the introduced range.

Adaptive plasticity to heterogeneous environments increases capacity for division of labor in the clonal invader Carpobrotus edulis (Aizoaceae).

UNLABELLED: • PREMISE OF THE STUDY: Clonality has been proposed as an important mechanism favoring plant invasions, but few studies have been conducted to determine the role of clonal traits on successful invaders. An interesting trait associated with clonality is the capacity for division of labor. Division of labor requires a negative spatial correlation between the availabilities of two essential resources and ramet specialization for locally abundant resources to increase the overall performance of the clone. We hypothesized that the capacity for division of labor in the clonal invader Carpobrotus edulis will be selected in those clones from patchy environments where this trait could be an advantage.• METHODS: Morphological and physiological division of labor was compared between clones from coastal sand dunes (where nutrients and light show a negative spatial covariance) and from rocky coasts (where nutrients and light are homogenously distributed).• KEY RESULTS: Clones from coastal sand dunes showed a greater capacity than clones from rocky coasts for division of labor. Specialization for abundance was found at the morphological (biomass allocated to roots) and the physiological (photochemical efficiency) level.• CONCLUSIONS: The greater ability for division of labor in the patchy environment where the presence of this trait would be more beneficial demonstrates the existence of local adaptation and suggests that rapid evolution in clonal traits could be contributing to the success of the invader C. edulis. This study is one of the few showing that division of labor is under selection and is the first reporting adaptive division of labor of an aggressive invader.

Ecological networks: delving into the architecture of biodiversity.

In recent years, the analysis of interaction networks has grown popular as a framework to explore ecological processes and the relationships between community structure and its functioning. The field has rapidly grown from its infancy to a vibrant youth, as reflected in the variety and quality of the discussions held at the first international symposium on Ecological Networks in Coimbra-Portugal (23-25 October 2013). The meeting gathered 170 scientists from 22 countries, who presented data from a broad geographical range, and covering all stages of network analyses, from sampling strategies to effective ways of communicating results, presenting new analytical tools, incorporation of temporal and spatial dynamics, new applications and visualization tools.(1) During the meeting it became evident that while many of the caveats diagnosed in early network studies are successfully being tackled, new challenges arise, attesting to the health of the discipline.

Differential effectiveness of novel and old legume-rhizobia mutualisms: implications for invasion by exotic legumes.

The degree of specialization in the legume-rhizobium mutualism and the variation in the response to different potential symbionts are crucial factors for understanding the process of invasion by exotic legumes and the consequences for the native resident plants and bacteria. The enhanced novel mutualism hypothesis predicts that exotic invasive legumes would take advantage of native rhizobia present in the invaded soils. However, recent studies have shown that exotic legumes might become invasive by using exotic introduced microsymbionts, and that they could be a source of exotic bacteria for native legumes. To unravel the role of novel and old symbioses in the progress of invasion, nodulation and symbiotic effectiveness were analyzed for exotic invasive plants and native co-occurring legumes in a Mediterranean coastal dune ecosystem. Although most of the studied species nodulated with bacteria from distant origins these novel mutualisms were less effective in terms of nodulation, nitrogenase activity and plant growth than the interactions of plants and bacteria from the same origin. The relative effect of exotic bradyrhizobia was strongly positive for exotic invasive legumes and detrimental for native shrubs. We conclude that (1) the studied invasive legumes do not rely on novel mutualisms but rather need the co-introduction of compatible symbionts, and (2) since exotic rhizobia colonize native legumes in invaded areas, the lack of effectiveness of these novel symbiosis demonstrated here suggests that invasion can disrupt native belowground mutualisms and reduce native legumes fitness.

Author Correction: Multilayer networks reveal the spatial structure of seed-dispersal interactions across the Great Rift landscapes.

The original version of this Article contained Figshare links in the Code availability statement that were not functional. The correct Figshare links to MATLAB scripts and R code used in this study are https://doi.org/10.6084/m9.figshare.4955651 and https://doi.org/10.6084/m9.figshare.4836383, respectively. These errors have now been corrected in both the PDF and HTML versions of the Article.

Multilayer networks reveal the spatial structure of seed-dispersal interactions across the Great Rift landscapes.

Species interaction networks are traditionally explored as discrete entities with well-defined spatial borders, an oversimplification likely impairing their applicability. Using a multilayer network approach, explicitly accounting for inter-habitat connectivity, we investigate the spatial structure of seed-dispersal networks across the Gorongosa National Park, Mozambique. We show that the overall seed-dispersal network is composed by spatially explicit communities of dispersers spanning across habitats, functionally linking the landscape mosaic. Inter-habitat connectivity determines spatial structure, which cannot be accurately described with standard monolayer approaches either splitting or merging habitats. Multilayer modularity cannot be predicted by null models randomizing either interactions within each habitat or those linking habitats; however, as habitat connectivity increases, random processes become more important for overall structure. The importance of dispersers for the overall network structure is captured by multilayer versatility but not by standard metrics. Highly versatile species disperse many plant species across multiple habitats, being critical to landscape functional cohesion.

The shift from plant-plant facilitation to competition under severe water deficit is spatially explicit.

The stress-gradient hypothesis predicts a higher frequency of facilitative interactions as resource limitation increases. Under severe resource limitation, it has been suggested that facilitation may revert to competition, and identifying the presence as well as determining the magnitude of this shift is important for predicting the effect of climate change on biodiversity and plant community dynamics. In this study, we perform a meta-analysis to compare temporal differences of species diversity and productivity under a nurse plant (Retama sphaerocarpa) with varying annual rainfall quantity to test the effect of water limitation on facilitation. Furthermore, we assess spatial differences in the herbaceous community under nurse plants in situ during a year with below-average rainfall. We found evidence that severe rainfall deficit reduced species diversity and plant productivity under nurse plants relative to open areas. Our results indicate that the switch from facilitation to competition in response to rainfall quantity is nonlinear. The magnitude of this switch depended on the aspect around the nurse plant. Hotter south aspects under nurse plants resulted in negative effects on beneficiary species, while the north aspect still showed facilitation. Combined, these results emphasize the importance of spatial heterogeneity under nurse plants for mediating species loss under reduced precipitation, as predicted by future climate change scenarios. However, the decreased water availability expected under climate change will likely reduce overall facilitation and limit the role of nurse plants as refugia, amplifying biodiversity loss.

Identification of symbiotic nitrogen-fixing bacteria from three African leguminous trees in Gorongosa National Park.

The symbiosis between leguminous plants and symbiotic nitrogen-fixing bacteria is a key component of terrestrial ecosystems. Woody legumes are well represented in tropical African forests but despite their ecological and socio-economic importance, they have been little studied for this symbiosis. In this study, we examined the identity and diversity of symbiotic-nitrogen fixing bacteria associated with Acacia xanthophloea, Faidherbia albida and Albizia versicolor in the Gorongosa National Park (GNP) in Mozambique. To the best of our knowledge, this is the first report on the identity of symbiotic-nitrogen fixing bacteria in this region. 166 isolates were obtained and subjected to molecular identification. BOX-A1R PCR was used to discriminate different bacterial isolates and PCR-sequencing of 16S rDNA, and two housekeeping genes, glnII and recA, was used to identify the obtained bacteria. The gene nifH was also analyzed to assess the symbiotic capacity of the obtained bacteria. All isolates from F. albida and Al. versicolor belonged to the Bradyrhizobium genus whereas isolates from Ac. xanthophloea clustered with Mesorhizobium, Rhizobium or Ensifer strains. Soil chemical analysis revealed significant differences between the soils occupied by the three studied species. Thus, we found a clear delimitation in the rhizobial communities and soils associated with Ac. xanthophloea, F. albida and Al. versicolor, and higher rhizobial diversity for Ac. xanthophloea than previously reported.

Arbuscular mycorrhizal fungi in Mimosa tenuiflora (Willd.) Poir from Brazilian semi-arid.

Many plant species from Brazilian semi-arid present arbuscular mycorrhizal fungi (AMF) in their rhizosphere. These microorganisms play a key role in the establishment, growth, survival of plants and protection against drought, pathogenic fungi and nematodes. This study presents a quantitative analysis of the AMF species associated with Mimosa tenuiflora, an important native plant of the Caatinga flora. AMF diversity, spore abundance and root colonization were estimated in seven sampling locations in the Ceará and Paraíba States, during September of 2012. There were significant differences in soil properties, spore abundance, percentage of root colonization, and AMF diversity among sites. Altogether, 18 AMF species were identified, and spores of the genera Acaulospora, Claroideoglomus, Dentiscutata, Entrophospora, Funneliformis, Gigaspora, Glomus, Racocetra, Rhizoglomus and Scutellospora were observed. AMF species diversity and their spore abundance found in M. tenuiflora rhizosphere shown that this native plant species is an important host plant to AMF communities from Brazilian semi-arid region. We concluded that: (a) during the dry period and in semi-arid conditions, there is a high spore production in M. tenuiflora root zone; and (b) soil properties, as soil pH and available phosphorous, affect AMF species diversity, thus constituting key factors for the similarity/dissimilarity of AMF communities in the M. tenuiflora root zone among sites.

Putative linkages between below- and aboveground mutualisms during alien plant invasions.

Evidence of the fundamental role of below-aboveground links in controlling ecosystem processes is mostly based on studies done with soil herbivores or mutualists and aboveground herbivores. Much less is known about the links between belowground and aboveground mutualisms, which have been studied separately for decades. It has not been until recently that these mutualisms-mycorrhizas and legume-rhizobia on one hand, and pollinators and seed dispersers on the other hand-have been found to influence each other, with potential ecological and evolutionary consequences. Here we review the mechanisms that may link these two-level mutualisms, mostly reported for native plant species, and make predictions about their relevance during alien plant invasions. We propose that alien plants establishing effective mutualisms with belowground microbes might improve their reproductive success through positive interactions between those mutualists and pollinators and seed dispersers. On the other hand, changes in the abundance and diversity of soil mutualists induced by invasion can also interfere with below-aboveground links for native plant species. We conclude that further research on this topic is needed in the field of invasion ecology as it can provide interesting clues on synergistic interactions and invasional meltdowns during alien plant invasions.

Genetic diversity and differentiation of Juniperus thurifera in Spain and Morocco as determined by SSR.

Juniperus thurifera L. is an important tree endemic to the western Mediterranean basin that it is able to grow in semi-arid climates. It nowadays exhibits a disjunct distribution pattern, occurring in North Africa, Spain, France and the Italian Alps. The Strait of Gibraltar has acted as an efficient barrier against gene flow between African and European populations, which are considered different subspecies by some authors. We aimed at describing the intraspecific genetic diversity of J. thurifera in populations from the Iberian Peninsula and Morocco and the phylogeographical relationships among these populations. The ploidy level of J. thurifera was examined and eleven nuclear microsatellites (nSSRs) developed for J. thurifera were assessed for genotyping this species. Six nSSRs were polymorphic and subsequently used to assess the genetic diversity and structure of the studied populations. Genotyping of the tetraploid J. thurifera using nuclear microsatellites supports the separation of Moroccan and Spanish populations into two genetically differentiated groups that correspond to the proposed subspecies africana and thurifera. High values of within population genetic diversity were found, that accounted for 90% of the total genetic variance, while population structure was weak. The estimators of genetic diversity were higher in populations of Spain than in populations of Morocco pointing for a possible loss of genetic diversity during the spread of this species to Africa from Europe.

Genetic diversity of root nodulating bacteria associated with Retama sphaerocarpa in sites with different soil and environmental conditions.

The genetic diversity of root nodulating bacteria isolated from Retama sphaerocarpa was studied using BOX-A1R PCR and phylogenetic analysis of the 16S rRNA region, as well as the housekeeping genes atpD, glnII and recA. A total of 193 isolates were obtained from eight different sites with different soil and environmental conditions in the Iberian Peninsula. These isolates corresponded to 31 different strains that successfully nodulated R. sphaerocarpa seedlings in reinoculation trials. About one-third of the strains clustered with B. canariense or B. cytisi within Bradyrhizobium group I. The remaining strains clustered with B. elkanii/B. pachyrhizi within Bradyrhizobium group II or in separate clades that could represent new lineages. Based on the 16S rRNA and combined atpD+glnII+recA sequences, two to three lineages of root nodulating bacteria were found at each sampling site, except for Collado Garcia where five species were detected. B. canariense and B. elkanii/B. pachyrhizi were the most abundant species, whereas the least abundant were those related to B. retamae and a putative new lineage. B. canariense was found only in soils with neutral and acid pH, whereas B. retamae was the dominant species in alkaline soils.

Disparate origins of Bradyrhizobium symbionts for invasive populations of Cytisus scoparius (Leguminosae) in North America.

To identify the geographic origin of nodule bacteria associated with invasion of the European legume Cytisus scoparius in the United States, isolates from 15 sites in six states were compared to > 200 Bradyrhizobium strains from indigenous legumes in the U.S., Mexico, Europe (six countries), Morocco, and Australia. Portions of five housekeeping loci (2849 bp) were sequenced, along with the nifD locus in the symbiosis island (SI) portion of the Bradyrhizobium chromosome. Bayesian phylogenetic analysis showed that North American C. scoparius symbionts had highly heterogeneous ancestry. Some were grouped into three distinct clades of European C. scoparius symbionts. One isolate had both housekeeping and SI genes belonging to a Bradyrhizobium clade from native legumes in western North America. Two other clades had mosaic ancestry: sequences for nifD as well as two other SI genes (nifH, nodC) were highly similar or identical to a C. scoparius strain from Spain, while their housekeeping loci belonged to American Bradyrhizobium clades. Thus, it appears that bacteria ancestrally associated with other North American legumes have evolved to utilize C. scoparius, by acquiring SI-region genes from European C. scoparius symbionts. Inoculation assays indicated that North American isolates were as competent as European strains in promoting plant growth, consistent with the findings on symbiont ancestry.