Multi-Generation Ecosystem Selection of Rhizosphere Microbial Communities Associated with Plant Genotype and Biomass in Arabidopsis thaliana.

The role of the microbiome in shaping the host's phenotype has emerged as a critical area of investigation, with implications in ecology, evolution, and host health. The complex and dynamic interactions involving plants and their diverse rhizospheres' microbial communities are influenced by a multitude of factors, including but not limited to soil type, environment, and plant genotype. Understanding the impact of these factors on microbial community assembly is key to yielding host-specific and robust benefits for plants, yet it remains challenging. Here, we conducted an artificial ecosystem selection experiment for eight generations of Arabidopsis thaliana Ler and Cvi to select soil microbiomes associated with a higher or lower biomass of the host. This resulted in divergent microbial communities shaped by a complex interplay between random environmental variations, plant genotypes, and biomass selection pressures. In the initial phases of the experiment, the genotype and the biomass selection treatment had modest but significant impacts. Over time, the plant genotype and biomass treatments gained more influence, explaining ~40% of the variation in the microbial community's composition. Furthermore, a genotype-specific association of plant-growth-promoting rhizobacterial taxa, Labraceae with Ler and Rhizobiaceae with Cvi, was observed under selection for high biomass.

Radical shift in the genetic composition of New England chicory populations.

Human activities have been altering the flora and fauna across the planet. Distributions and the diversity of species, and the phenotypes of individuals in those species are changing. New England with its rapidly changing human demographics is an ideal place to investigate these temporal changes in the habitat. The flora of New England consists of both native and nonnative species. Non-indigenous plant species have been introduced since the first Europeans arrived in North America in the 15th century. Cichorium intybus (chicory), native to Eurasia, was first recorded in North America in 1774. Subsequently, chicory spread and became naturalized throughout much of the continent.In this study, we used chloroplast DNA sequences and 12 microsatellite nuclear markers to assess the temporal genetic changes in New England populations of chicory. We analysed 84 herbarium specimens and 18 contemporary extant populations (228 individuals in total).Three chloroplast DNA haplotypes were detected and all were present in New England prior to 1890; however, Hap3 was rare prior to the 1950s. The nuclear DNA markers showed a major shift in the genetic diversity and composition, with all historical herbarium collections belonging to a single genetic cluster and 16 out of 18 contemporary chicory populations belonging to different genetic clusters. This change occurred regionally and also on a local scale with contemporary populations being very different from herbarium specimens collected previously in the corresponding localities. Synthesis. Our results indicate that the genetic diversity and structure of Cichorium intybus populations have changed substantially since the founding populations in New England. These changes may have contributed to the success of this nonnative species and helped to fuel its rapid expansion and adaptation to the changing landscapes in both New England and the rest of North America.

Population structure, seasonal genotypic differentiation, and clonal diversity of weedy dandelions in three Boston area populations (Taraxacum sp.).

Weedy dandelions have a worldwide distribution and thrive in urban environments despite a lack of sexual reproduction throughout most of its range. North American dandelions, introduced from Eurasia, are believed to be primarily, if not exclusively, apomictic triploids. In some European populations, apomicts co-occur with diploid sexual individuals and hybridizations can create genetically unique apomicts, which may subsequently disperse and establish new populations globally. Using six nuclear microsatellite markers and a cpDNA intergenic spacer, we investigate the impact of this unusual natural history on population structure and diversity in three urban Boston area dandelion populations. Our results show high levels of genetic diversity within populations, spatial population structure, and seasonal genotypic differentiation in flowering times. We find evidence that sexual reproduction and recombination, presumably in Europe, and extensive gene flow drive these patterns of diversity and create the appearance of panmixia despite the lack of evidence for local sexual reproduction.

Analyses of synteny between Arabidopsis thaliana and species in the Asteraceae reveal a complex network of small syntenic segments and major chromosomal rearrangements.

Comparative genomic studies among highly divergent species have been problematic because reduced gene similarities make orthologous gene pairs difficult to identify and because colinearity is expected to be low with greater time since divergence from the last common ancestor. Nevertheless, synteny between divergent taxa in several lineages has been detected over short chromosomal segments. We have examined the level of synteny between the model species Arabidopsis thaliana and species in the Compositae, one of the largest and most diverse plant families. While macrosyntenic patterns covering large segments of the chromosomes are not evident, significant levels of local synteny are detected at a fine scale covering segments of 1-Mb regions of A. thaliana and regions of <5 cM in lettuce and sunflower. These syntenic patches are often not colinear, however, and form a network of regions that have likely evolved by duplications followed by differential gene loss.

Population structure in chicory (Cichorium intybus): A successful U.S. weed since the American revolutionary war.

Plant invasions are recognized as major drivers of ecosystem change, yet the precise cause of these invasions remains unknown for many species. Frequency and modes of introductions during the first, transport and colonization, stages of the invasion process as well as phenotypic changes due to plasticity or changing genetic diversity and adaptation during later establishment and expansion stages can all influence the "success" of invasion. Here, we examine some of these factors in, and the origin of, a very successful weed, Cichorium intybus (chicory) which was introduced to North America in the 18th century and which now can be found in all 48 continental U.S. states and much of Canada. We genotyped a Eurasian collection of 11 chicory cultivars, nine native populations and a North American collection of 20 introduced wild populations which span the species range (592 individuals in total). To detect the geographic sources of North American chicory populations and to assess the genetic diversity among cultivars, native, and introduced populations, we used both a sequenced cpDNA region and 12 nuclear simple sequence repeat (SSR), microsatellite loci. Four cpDNA haplotypes were identified and revealed clear geographic subdivisions in the chicory native range and an interspecific hybrid origin of Radicchio group. Nuclear data suggested that domesticated lines deliberately introduced to North America were major contributors to extant weedy populations, although unintended sources such as seed contaminants likely also played important roles. The high private allelic richness and novel genetic groups were detected in some introduced populations, suggesting the potential for local adaptation in natural sites such as deserts and nature reserves. Our findings suggest that the current populations of weedy U.S. chicory have evolved primarily from several sources of domesticated and weedy ancestors and subsequent admixture among escaped lineages.

An Ac-like transposable element family with transcriptionally active Y-linked copies in the white campion, Silene latifolia.

An RFLP genomic subtraction was used to isolate male-specific sequences in the species Silene latifolia. One isolated fragment, SLP2, shares similarity to a portion of the Activator (Ac) transposase from Zea mays and to related proteins from other plant species. Southern blot analysis of male and female S. latifolia genomic DNA shows that SLP2 belongs to a low-copy-number repeat family with two Y-linked copies. Screening of a S. latifolia male genomic library using SLP2 as a probe led to the isolation of five clones, which were partially sequenced. One clone contains two large open reading frames that can be joined into a sequence encoding a putative protein of 682 amino acids by removing a short intron. Database searches and phylogenetic analysis show that this protein belongs to the hAT superfamily of transposases, closest to Tag2 (Arabidopsis thaliana), and contains all of the defined domains critical for the activity of these transposases. PCR with genomic and cDNA templates from S. latifolia male, female, and hermaphrodite individuals revealed that one of the Y-linked copies is transcriptionally active and alternatively spliced. This is the first report of a transcriptionally active transposable element (TE) family in S. latifolia and the first DNA transposon residing on a plant Y chromosome. The potential activity and regulation of this TE family and its use for Y chromosome gene discovery is discussed.

Potato Diversity in the Andean Center of Crop Domestication.

The diversity and population structure of potato landraces (Solanum spp.) within their center of domestication was studied using isozyme surveys of four polymorphic loci. The objective in assessing the distribution of genetic diversity was to assist in planning conservation strategies of crop genetic resources that are threatened by genetic erosion. In situ conservation methods depend on this type of analysis. Research was conducted in the region of Cusco, Peru. Eight fields spread among two microregions were randomly sampled, and 610 tubers were studied from this sample. In addition, 503 tubers were collected from markets in seven different meso-regions (provinces) surrounding the regional center of Cusco. Thirty genotypes were identified in the field sample and 82 in the regional sample. The frequency and distribution of genotypes and alleles are described. A high degree of genotype endemism was found at both the field and regional levels. Genotypes were unevenly distributed, and most of the genotypic diversity was between rather than within populations. At the allele level, however, we found that a very high percentage of the diversity was within rather than between populations. The genotype is the key unit for maintaining the population of potato landraces. Our findings suggest that collections need to be both geographically extensive and intensive. Because farmers are able to maintain most alleles on relatively small portions of their land, in situ conservation is a viable strategy.

Resumen: La diversidad y estructura poblacional de razas locales de papa (Solanum spp.) en su centro de domesticación andino fué estudiado por medio de isoenzimas en cuatro loci polimórficos. La determinación de la distribución de la diversidad genética se hizo con el fin de ayudar en la planificación de las estrategias para la conservación de germoplasma en cultivos amenazados por la erosión genética, teniendo en cuenta que los métodos de conservación in situ dependen de este tipo de análisis. La investigación fue realizada en la región del Cusco, en Perú. Se muestrearon tubérculos al azar en ocho campos distribuidos en dos microregiones, haciendo un total de 610 tubérculos. Por otro lado, 503 tubérculos fueron colectados en los mercados de siete provincias alrededor de la cuidad del Cusco, Perú. Treinta genotipos fueron identificados en las colectas hechas en los campos de los campesinos y 82 en la colección de los mercados, sirviendo para describir la frequencia y distribución de genotipos y alelos en las dos colecciones. La distribución no fué uniforme, encontrándose la mayor parte de la diversidad entre poblaciones, indicando que el genotipo es la unidad clave en el mantenimiento de las razes locales de papas. Nuestro estudio sugiere que las colecciones de germoplasma de papas deben ser geográficamente amplias e intensas. La habilidad de los campensinos en mantener la mayoria de los alelos en sus pequeños terrenos indica que la conservación in situ es un estrategia viable. La diversidad de la papa en el centro andino de domesticacion de cultivos.

Dispersal pathways and genetic differentiation among worldwide populations of the invasive weed Centaurea solstitialis L. (Asteraceae).

The natural history of introduced species is often unclear due to a lack of historical records. Even when historical information is readily available, important factors of the invasions such as genetic bottlenecks, hybridization, historical relationships among populations and adaptive changes are left unknown. In this study, we developed a set of nuclear, simple sequence repeat markers and used these to characterize the genetic diversity and population structure among native (Eurasian) and non-native (North and South American) populations of Centaurea solstitialis L., (yellow starthistle). We used these data to test hypotheses about the invasion pathways of the species that were based on historical and geographical records, and we make inferences about historical relationships among populations and demographic processes following invasion. We confirm that the center of diversity and the native range of the species is likely the eastern Mediterranean region in the vicinity of Turkey. From this region, the species likely proceeded to colonize other parts of Europe and Asia via a slow, stepwise range expansion. Spanish populations were the primary source of seed to invade South America via human-mediated events, as was evident from historical records, but populations from the eastern Mediterranean region were also important. North American populations were largely derived from South America, but had secondary contributors. We suggest that the introduction history of non-native populations from disparate parts of the native range have allowed not just one, but multiple opportunities first in South America then again in North America for the creation of novel genotypes via intraspecific hybridization. We propose that multiple intraspecific hybridization events may have created especially potent conditions for the selection of a noxious invader, and may explain differences in genetic patterns among North and South America populations, inferred differences in demographic processes, as well as morphological differences previously reported from common garden experiments.

No accession-specific effect of rhizosphere soil communities on the growth and competition of Arabidopsis thaliana accessions.

Soil communities associated with specific plant species affect individual plants' growth and competitive ability. Limited evidence suggests that unique soil communities can also differentially influence growth and competition at the ecotype level. Previous work with Arabidopsis thaliana has shown that accessions produce distinct and reproducible rhizosphere bacterial communities, with significant differences in both species composition and relative abundance. We tested the hypothesis that soil communities uniquely affect the growth and reproduction of the plant accessions with which they are associated. Specifically, we examined the growth of four accessions when exposed to their own soil communities and the communities generated by each of the other three accessions. To do this we planted focal accessions inside a ring of six plants that created a "background" soil community. We grew focal plants in this design in three separate soil treatments: non-sterile soil, sterilized soil, and "preconditioned" soil. We preconditioned soil by growing accessions in non-sterile soil for six weeks before the start of the experiment. The main experiment was harvested after seven weeks of growth and we recorded height, silique number, and dry weight of each focal plant. Plants grown in the preconditioned soil treatment showed less growth relative to the non-sterile and sterile soil treatments. In addition, plants in the sterile soil grew larger than those in non-sterile soil. However, we saw no interaction between soil treatment and background accession. We conclude that the soil communities have a negative net impact on Arabidopsis thaliana growth, and that the unique soil communities associated with each accession do not differentially affect growth and competition of study species.

Development of microsatellite markers in Cordia bifurcata (Boraginaceae) and cross-species amplification in Cordia inermis and Cordia pringlei.

We developed 16 microsatellite markers in Cordia bifurcata, a Central and South American shrub. The markers show low polymorphism in C. bifurcata, a species suspected of self-fertilization or apomixis. Of four polymorphic loci, three had only two alleles. However, current research indicates that these markers hold value for interpopulational comparisons of C. bifurcata and for analyses of congeners. In Cordia inermis, a dioecious or subdioecious shrub, seven of the markers produced interpretable amplification products of which five showed polymorphism. In Cordia pringlei, a distylous shrub, nine of the markers produced interpretable amplification products of which six showed polymorphism.

Molecular and morphological evidence reveals introgression in swarms of the invasive taxa Fallopia japonica, F. sachalinensis, and F. xbohemica (Polygonaceae) in the United States.

Fallopia japonica (Japanese knotweed, Polygonaceae) is a well-known East Asian perennial that is established throughout the U.S. and Europe. Another congener, F. sachalinensis, and their hybrid, F. ×bohemica, also persist on both continents. Their invasive success is primarily attributed to their ability to spread via clonal growth. However, mounting evidence suggests invasion history and dynamics differ between continents and that sexual reproduction is more common than previously assumed. We used published morphological traits designed to distinguish the three taxa to characterize their distribution in 24 New England towns. We found continuous variation of all five traits, with 84% of our 81 individuals having at least one trait outside parental limits. Hierarchical cluster analysis, along with two chloroplast and one nuclear species-specific markers, suggests the presence of intercrossing, segregating hybrids, and likely introgression between F1 hybrids and F. japonica. Our markers also show the first evidence of bidirectional hybridization between parental taxa in the U.S., emphasizing the complex structure of populations in our region. This study is a first step toward unraveling the evolutionary forces that have made these taxa such aggressive invaders in the U.S. The data may also affect management strategies originally designed for largely monomorphic, clonal populations.

Genetic diversity and clonal vs. sexual reproduction in Fallopia spp. (Polygonaceae).

Although fundamental to the study of invasion mechanisms, the relationship between mode of reproduction and plant invasion is not well understood. Fallopia japonica (Japanese knotweed), a highly aggressive invasive plant in both Europe and North America, serves as a model species for examining this relationship. In Britain, F. japonica var. japonica is a single female clone reproducing solely through vegetative growth or obligate hybridization with other Fallopia spp. In the U.S., however, there is more evidence for sexual reproduction. Here, simple sequence repeat (SSR) markers were developed, and three Massachusetts populations were sampled at regular intervals. The amount of sexual and clonal reproduction in each population was determined based on within-population genetic diversity. Clonal growth was apparent, but the populations together contained 26 genotypes and had evidence of sexual reproduction. One genotype that was present in all populations matched the single aggressive British clone of F. japonica var. japonica. Also, a potentially diagnostic marker for the F. sachalinensis genome provided evidence of inter- and intraspecific sexual reproduction and introgression. These differences observed in U.S. populations compared to European populations have significant implications for management of Fallopia spp. in the U.S. and underscore the importance of regional studies of invasive species.

Universal markers for comparative mapping and phylogenetic analysis in the Asteraceae (Compositae).

The development of universal markers that can be assayed across taxa, but which are polymorphic within taxa, can facilitate both comparative map-based studies and phylogenetic analyses. Here we describe the development of such markers for use in the Asteraceae, which includes the crops lettuce, sunflower, and safflower as well as dozens of locally important crop and weed species. Using alignments of a conserved orthologous set (COS) of ESTs from lettuce and sunflower and genomic sequences of Arabidopsis, we designed a suite of primer pairs that are conserved across species, but which are predicted to flank introns. We then tested 192 such primer pairs in 8 species from across the family. Of these, 163 produced an amplicon in at least 1 taxon, and 125 amplified in at least half of the taxa surveyed. Thirty-nine amplified in all 8 species. Comparisons amongst sequences within the lettuce and sunflower EST databases indicate that the vast majority of these loci will be polymorphic. As a direct test of the utility of these markers outside the lettuce and sunflower subfamilies, we sequenced a subset of ten loci from a panel of cultivated safflower individuals. All 10 loci proved to be single-locus, and nine of the 10 loci were polymorphic with an average of 12.8 SNPs per kb. Taken together, these loci will provide an initial backbone for comparative genetic analyses within the Asteraceae. Moreover, our results indicate that these loci are phylogenetically informative, and hence can be used to resolve evolutionary relationships between taxa within the family as well as within species.

Comparative analysis of NBS domain sequences of NBS-LRR disease resistance genes from sunflower, lettuce, and chicory.

Plant resistance to many types of pathogens and pests can be achieved by the presence of disease resistance (R) genes. The nucleotide binding site-leucine rich repeat (NBS-LRR) class of R-genes is the most commonly isolated class of R-genes and makes up a super-family, which is often arranged in the genome as large multi-gene clusters. The NBS domain of these genes can be targeted by polymerase chain reaction (PCR) amplification using degenerate primers. Previous studies have used PCR derived NBS sequences to investigate both ancient R-gene evolution and recent evolution within specific plant families. However, comparative studies with the Asteraceae family have largely been ignored. In this study, we address recent evolution of NBS sequences within the Asteraceae and extend the comparison to the Arabidopsis thaliana genome. Using multiple sets of primers, NBS fragments were amplified from genomic DNA of three species from the family Asteraceae: Helianthus annuus (sunflower), Lactuca sativa (lettuce), and Cichorium intybus (chicory). Analysis suggests that Asteraceae species share distinct families of R-genes, composed of genes related to both coiled-coil (CC) and toll-interleukin-receptor homology (TIR) domain containing NBS-LRR R-genes. Between the most closely related species, (lettuce and chicory) a striking similarity of CC subfamily composition was identified, while sunflower showed less similarity in structure. These sequences were also compared to the A. thaliana genome. Asteraceae NBS gene subfamilies appear to be distinct from Arabidopsis gene clades. These data suggest that NBS families in the Asteraceae family are ancient, but also that gene duplication and gene loss events occur and change the composition of these gene subfamilies over time.