Transcriptome analysis of aphids exposed to glandular trichomes in tomato reveals stress and starvation related responses.

Understanding the responses of insect herbivores to plant chemical defences is pivotal for the management of crops and pests. However, the mechanisms of interaction are not entirely understood. In this study, we compared the whole transcriptome gene expression of the aphid Macrosiphum euphorbiae grown on two different varieties of tomato that differ in their inducible chemical defences. We used two isogenic lines of tomato with a shared genetic background that only differ in the presence of type IV glandular trichomes and their associated acylsucrose excretions. This works also reports a de novo transcriptome of the aphid M. euphorbiae. Subsequently, we identified a unique and distinct gene expression profile for the first time corresponding to aphid´s exposure to type IV glandular trichomes and acylsugars. The analysis of the aphid transcriptome shows that tomato glandular trichomes and their associated secretions are highly efficient in triggering stress-related responses in the aphid, and demonstrating that their role in plant defence goes beyond the physical impediment of herbivore activity. Some of the differentially expressed genes were associated with carbohydrate, lipid and xenobiotic metabolisms, immune system, oxidative stress response and hormone biosynthesis pathways. Also, the observed responses are compatible with a starvation syndrome. The transcriptome analysis puts forward a wide range of genes involved in the synthesis and regulation of detoxification enzymes that reveal important underlying mechanisms in the interaction of the aphid with its host plant and provides a valuable genomic resource for future study of biological processes at the molecular level using this aphid.

Tomato Domestication Affects Potential Functional Molecular Pathways of Root-Associated Soil Bacteria.

While it has been well evidenced that plant domestication affects the structure of the root-associated microbiome, there is a poor understanding of how domestication-mediated differences between rhizosphere microorganisms functionally affect microbial ecosystem services. In this study, we explore how domestication influenced functional assembly patterns of bacterial communities in the root-associated soil of 27 tomato accessions through a transect of evolution, from plant ancestors to landraces to modern cultivars. Based on molecular analysis, functional profiles were predicted and co-occurrence networks were constructed based on the identification of co-presences of functional units in the tomato root-associated microbiome. The results revealed differences in eight metabolic pathway categories and highlighted the influence of the host genotype on the potential functions of soil bacterial communities. In general, wild tomatoes differed from modern cultivars and tomato landraces which showed similar values, although all ancestral functional characteristics have been conserved across time. We also found that certain functional groups tended to be more evolutionarily conserved in bacterial communities associated with tomato landraces than those of modern varieties. We hypothesize that the capacity of soil bacteria to provide ecosystem services is affected by agronomic practices linked to the domestication process, particularly those related to the preservation of soil organic matter.

Effect of plant origin and phenological stage on the allelopathic activity of the invasive species Oxalis pes-caprae.

PREMISE: Invasion processes involve several mechanisms, some of which have received little attention. Allelopathy has been invoked as an invasion driver according to the novel weapon hypothesis (NWH), and polyploidization can also be an important factor for invasion success. However, very few studies have addressed both topics together. We analyzed the allelopathic effect of the polyploid Oxalis pes-caprae, an invader in mediterranean-climate areas worldwide, from different origins and ploidy levels (native: South Africa; invaded: Chile, Australia, California, and the Mediterranean Basin) on Lactuca sativa as a model species. METHODS: We measured seed germination, initial plant height, and aboveground and belowground biomass of Lactuca grown in field soil mixed with Oxalis material and with or without activated carbon in a common garden experiment. We performed the experiment twice, when Oxalis was vigorous and when senescent. RESULTS: Vigorous plants of Oxalis tended to reduce Lactuca germination, but both vigorous and senescent Oxalis plants increased Lactuca biomass, probably due to an increase in nutrients provided by plant material. The highest increase in Lactuca traits occurred with Oxalis plants from the Iberian Peninsula. Allelopathy only happened when Oxalis was senescent and was especially strong with plants from Chile. CONCLUSIONS: Although we did not find broad evidence for the NWH considering all areas together, we reported differences in the allelopathic potential of Oxalis plants depending on their origin. These results highlight the independent evolution of invasiveness traits in distant introduced ranges and the importance of the invaders' origin when testing hypotheses about invasion drivers.

Characterization of the detrimental effects of type IV glandular trichomes on the aphid Macrosiphum euphorbiae in tomato.

BACKGROUND: Glandular trichomes are essential in plants' defence against pests however, the mechanisms of action are not completely understood. While there is considerable evidence of feeding and movement impairment by trichomes, the effect on other traits is less clear. We combined laboratory and greenhouse experiments with molecular analysis to understand how glandular trichomes affect the behavior, population growth, and the expression of biomarkers involved in detoxification, primary metabolism, and developmental pathways of the aphid Macrosiphum euphorbiae. We used two isogenic tomato lines that differ in the presence of type IV glandular trichomes and production of acylsucroses; i.e.,Solanum lycopersicum cv. 'Moneymaker' and an introgressed line from Solanum pimpinellifolium (with trichomes type IV). RESULTS: Type IV glandular trichomes affected host selection and aphid proliferation with aphids avoiding, and showing impaired multiplication on the genotype with trichomes. The exposure to type IV glandular trichomes resulted in the overexpression of detoxication markers (i.e., Hsp70, Hsp17, Hsp10); the repression of the energetic metabolism (GAPDH), and the activation of the ecdysone pathway; all these, underlying the key adaptations and metabolic trade-offs in aphids exposed to glandular trichomes. CONCLUSION: Our results demonstrate the detrimental effect of glandular trichomes (type IV) on the aphid and put forward their mode of action. Given the prevalence of glandular trichomes in wild and cultivated Solanaceae; and of the investigated molecular biomarkers in insects in general, our results provide relevant mechanisms to understand the effect of trichomes not only on herbivorous insects but also on other trophic levels.

Resistance and Not Plant Fruit Traits Determine Root-Associated Bacterial Community Composition along a Domestication Gradient in Tomato.

Soil bacterial communities are involved in multiple ecosystem services, key in determining plant productivity. Crop domestication and intensive agricultural practices often disrupt species interactions with unknown consequences for rhizosphere microbiomes. This study evaluates whether variation in plant traits along a domestication gradient determines the composition of root-associated bacterial communities; and whether these changes are related to targeted plant traits (e.g., fruit traits) or are side effects of less-often-targeted traits (e.g., resistance) during crop breeding. For this purpose, 18 tomato varieties (wild and modern species) differing in fruit and resistance traits were grown in a field experiment, and their root-associated bacterial communities were characterised. Root-associated bacterial community composition was influenced by plant resistance traits and genotype relatedness. When only considering domesticated tomatoes, the effect of resistance on bacterial OTU composition increases, while the effect due to phylogenetic relatedness decreases. Furthermore, bacterial diversity positively correlated with plant resistance traits. These results suggest that resistance traits not selected during domestication are related to the capacity of tomato varieties to associate with different bacterial groups. Taken together, these results evidence the relationship between plant traits and bacterial communities, pointing out the potential of breeding to affect plant microbiomes.

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.

Associations between sex-organ deployment and morph bias in related heterostylous taxa with different stylar polymorphisms.

PREMISE OF THE STUDY: Populations of heterostylous species are characterized by two or three floral morphs with reciprocal positioning of stigmas and anthers. Theoretical models predict equal morph frequencies (isoplethy) when disassortative mating is prevalent in populations, but biased morph ratios may occur when variation in the expression of heterostyly causes deviations from intermorph mating. METHODS: We explore the role of sex-organ deployment in governing morph ratios in two closely related genera of Boraginaceae, exhibiting striking variation in floral traits associated with the heterostylous syndrome. We sampled 66 populations of six species of Glandora and 39 populations of three species of Lithodora across their distributional range in the Mediterranean. In each population we estimated morph ratios and measured several floral traits. We used phylogenetically corrected and noncorrected regressions to test the hypothesis that differences in sex-organ reciprocity and herkogamy are associated with deviations from isoplethy. KEY RESULTS: Biased morph ratios occurred in 24% of populations, particularly in Lithodora. Populations biased for the long-styled morph (L-morph) were more frequent than the short-styled morph (S-morph). Distylous species were less likely to exhibit biased ratios than species with stigma-height dimorphism. In Lithodora fruticosa, a species lacking reciprocity, decreased herkogamy in the S-morph was associated with increasing L-morph bias, perhaps resulting from self-interference. CONCLUSION: Striking variation in the expression of heterostyly in Glandora and Lithodora is associated with biased morph ratios, which probably result from pollinator-mediated mating asymmetries within populations.

Invasion Fosters Change: Independent Evolutionary Shifts in Reproductive Traits after Oxalis pes-caprae L. Introduction.

Biological invasions offer optimal scenarios to study evolutionary changes under contemporary timescales. After long-distance dispersal, exotic species have to cope with strong mate limitation, and shifts toward uniparental reproduction have been hypothesized to be selectively advantageous. Oxalis pes-caprae is a clonal tristylous species native to South Africa, and invasive in Mediterranean regions worldwide. It reproduces sexually and asexually but the importance of each strategy differs between ranges. Native populations reproduce mostly sexually while in invasive ones asexual reproduction is the prevailing strategy due to the dominance of pentaploid monomorphic populations. Nevertheless, two contrasting scenarios have been observed after introduction: transition toward clonality, and re-acquisition of sexuality fueled by multiple introductions of compatible mates. Here, we aimed to assess evolutionary changes of reproductive traits in O. pes-caprae invasive populations and evaluate whether these traits could be related with invasion success and prevalence of certain forms in the western Mediterranean basin. Sexual and asexual reproduction traits were quantified under optimal conditions in a common garden experiment including native and invasive sexual, predominately asexual, and obligated asexual individuals. Different reproductive, ecological, and genetic constraints created by long-distance dispersal seem to have generated different selective pressures in sexual and asexual traits, with our results supporting evolutionary changes in invasive populations of O. pes-caprae. Native plants had higher sexual fitness, while a transition toward clonality was clear for invasive forms, supporting clonal reproduction as a major trait driving invasion. Differences were also observed among invasive plants, with sexual forms having increased dispersal potential; thus, they are expected to be in advantage in comparison with predominantly asexual and obligated asexual plants, and may become widespread in the future. Historical processes, like the initial introduction of predominantly asexual forms followed by sexual forms more recently, could be in the origin of current distribution patterns of O. pes-caprae in the western Mediterranean. This study shows that invasion processes are very dynamic and that ecological and genetic constraints determined by the invasion process may originate different reproductive strategies that are likely to determine invasion success.

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.

Strength through unity: spatial affinity between morphs improves fitness in incompatible heterostylous Melochia (Malvaceae) species.

In heterostylous plants, both stylar polymorphism and incompatibility system favor legitimate pollination among individuals. Weak or partial expression of incompatibility may ensure progeny when mates or pollinators are scarce in unstable habitats, but under these conditions plants with heteromorphic incompatibility would be in disadvantage. In this work we determine how the spatial distribution of morphs and the effect of proximity to the nearest potential mates affect plants' reproductive output in four Melochia species. The general prediction of decreasing reproductive success with an increasing isolation of floral morphs in plants with heteromorphic incompatibility was corroborated only in one species (i.e. M. tomentosa). Meanwhile, the other species exhibit a spatial affinity between morphs (i.e. the number of individuals with the nearest neighbor of the opposite morph exceeds expectations upon a random distribution). For M. savannarum and M. villosa we could not detect any effect of proximity to potential mates on the seed-ovule ratio. This may be due to: (1) existence of pollinators with long flying distances, like butterflies, in the populations and/or, (2) the possible occurrence of resource limitation. Spatial affinity between morphs in populations of heterostylous plants with heteromorphic incompatibility system increases reproductive success and may facilitate colonization of ephemeral habitats.

When did plants become important to leaf-nosed bats? Diversification of feeding habits in the family Phyllostomidae.

A great proportion of bats of the New World family Phyllostomidae feed on fruit, nectar and pollen, and many of them present adaptations to feed also on insects and small vertebrates. So far, attempts to examine the diversification of feeding specialization in this group, and particularly the evolution of nectarivory and frugivory, have provided contradictory results. Here we propose a molecular phylogenetic hypothesis for phyllostomids. On the basis of a matrix of feeding habits that takes into account geographical and seasonal variation, we tested different hypotheses of the evolution of feeding specializations in the group. We find strong support for the evolutionary model of a direct dietary diversification from insectivory. The estimates of divergence times of phyllostomid bats and the reconstruction of ancestral states with a Bayesian approach support the parallel evolution of frugivory in five lineages and of nectarivory in three lineages during the Miocene. On the basis of these findings, and recent dietary studies, we propose that during the evolution of phyllostomids switches to new feeding mechanisms to access to abundant and/or underexploited resources provided selective advantages that favoured the appearance of ecological innovations independently in different lineages of the family. We did not find evidences to support or reject the hypothesis that the insectivorous most recent common ancestor of all phyllostomids was also phytophagous.

A new approach to the quantification of degree of reciprocity in distylous (sensu lato) plant populations.

BACKGROUND AND AIMS: Although evolution of sexual polymorphism has been traditionally analysed using discrete characters, most of these polymorphisms are continuous. This is the case of heterostyly. Heterostyly is a floral polymorphism successfully used as a model to study the evolution of the sexual systems in plants. It involves the reciprocal positioning of anthers and stigmas in flowers of different plants within the same population. Studies of the functioning of heterostyly require the quantification of the degree of reciprocity between morphs of heterostylous species. Some reciprocity indices have been proposed previously, but they show significant limitations that need to be dealt with. This paper analyses these existing indices, and proposes a new index that aims to avoid their main problems (e.g. takes into account population variability and offers a single value per population). METHODS: The new index is based on the comparison of the position of every single sexual organ in the population with each and every organ of the opposite sex. To carry out all the calculations, a macro was programmed with MS Visual Basic in MS Excel. The behaviour of the index is tested using hypothetical data to simulate different situations of dimorphic populations; the index is also tested with some actual populations of different species of the genus Lithodora. RESULTS AND CONCLUSIONS: The index of reciprocity proposed here is a sound alternative to previous indices: it compares stigma-stamen height gaps for all potential crosses in the population, it comprises stigma-stamen distance as well as dispersion, it is not skewed by the more frequent sex, and it can be meaningfully compared between populations and species. It has produced solid results for both hypothetical and natural populations.