Genetic variation and structure in native and invasive Solidago canadensis populations

Solidago canadensis is native to North America, but has become a noxious invasive plant in China. We know only a little about its invasion history and the effects of introductions on its genetic composition. Here, we investigated genetic variation and structure between 15 North American and 13 Chinese populations of S. canadensis using AFLP makers. Four AFLP loci suggested relatively high genetic diversity of this weed and similar genetic variation between the invasive range and the native range. Most genetic variation was within populations across two ranges, but the Chinese range had a higher degree of among‐population variation than the North American range. Multiple tests, including Bayesian assignment, UPGMA analysis, PCoA and analysis of ‘isolation by distance’, showed that the Chinese populations originated from at least two distinct native sources and that secondary introduction or dispersal should be common in China. Also, North American populations were possibly a single genetic group. Overall, S. canadensis in China was probably founded from multiple introductions and then spread through long‐distance dispersal associated with human activities. Genetic variability in the species in the invaded range appears to have favoured establishment and spread and may well provide a challenge to successful control.     

Population structure of Solanum carolinense along the Takano River in Kyoto, Japan as determined by amplified fragment length polymorphism analysis

Areas infested with Solanum carolinense, an introduced perennial weed, have increased quickly in Japan. The genetic structure of a S. carolinense population along the Takano River in Kyoto, Japan was investigated using amplified fragment length polymorphism (AFLP) analysis to reveal how introductions and local spread have contributed to the development of this population. Along a terrace beside the river and an adjacent roadside, we defined a cluster of above‐ground shoots in the population as a subpopulation and 60 subpopulations were identified. Seventeen of these were selected and 165 shoots were analyzed by AFLP analysis using three primer pairs. The AFLP profiles revealed 69 genotypes and the presence of several clones, i.e. genotypes that were present in more than one individual. In total, 19 clones, comprising 2–34 identical genotypes, could be identified in the population. Each of these clones, except one, was allocated within each of the subpopulations. Clustering of the subpopulations was supported by high bootstrap values in all cases. Therefore, introductions from distant regions have mainly contributed to the development of this population, and local spread by seed or vegetative reproduction has rarely been important. The most likely introduction route of S. carolinense to this population was via planting associated with construction works.     

Microsatellite DNA variation within and among invasive populations of Ambrosia artemisiifolia from the southern Pannonian Plain

Although the Pannonian Plain presents one of the areas in Europe most highly infested by Ambrosia artemisiifolia, previously reported population genetic studies of the invasive species have included only a few populations from this region. Our goal was to determine the level of genetic diversity and genetic structure of A. artemisiifolia populations from the southern Pannonian Plain using microsatellite DNA markers. We documented a high level of genetic diversity within the Pannonian populations, as compared with the genetic diversity parameters previously published for the other invasive populations of A. artemisiifolia in Europe. The mean number of alleles per locus (N_A) ranged between 6.8 and 9.4, allelic richness (r) between 6.59 and 8.53 and mean number of rare alleles per locus (N_R) ranged between 3.4 and 6.4 per analysed population. A low level of among‐population differentiation was detected, with percentage of variation between populations of 3.3%, generally low pairwise F_ST values (ranged from ?0.007 to 0.152) and also according to principal coordinate analysis. A Bayesian approach revealed that most individuals did not strongly associate with any single genetic cluster, confirming a lack of genetic structuring in the analysed region. Together with the results of quantification of the level of gene flow (N_m = 5.671), these observations indicated a presence of extensive gene exchange and admixture between the populations. Therefore, A. artemisiifolia in the Pannonian Plain region has the potential for rapid expansion.     

Microsatellite variability of sulfonylurea‐resistant and susceptible populations of Schoenoplectus juncoides (Cyperaceae) in Kinki,Japan

Schoenoplectus juncoides is one of the most harmful weeds found in East Asian paddy fields. Recent emergence of biotypes that are resistant to the herbicide sulfonylurea (SU) has made weed control difficult. To examine the effect of the evolution of this herbicide resistance on genetic diversity within local populations, we investigated microsatellite variability within and among paddy field populations of S. juncoides in Kinki, Japan. In vivo assay of acetolactate synthase activity and root elongation assay in the presence of SU revealed that of 21 populations, five were sulfonylurea‐susceptible (SU‐S) and eight were completely sulfonylurea‐resistant (SU‐R). The remaining eight populations were a mixture of SU‐S and SU‐R individuals. The average gene diversity for SU‐R populations (H_S = 0.168) was lower than those for SU‐S (H_S = 0.256) and mixed (H_S = 0.209) populations, but the difference was not significant. This indicates that positive selection for SU‐R phenotype did not cause a genome‐wide reduction in genetic diversity. Genetic differentiation among S. juncoides populations was higher than that observed for most weed species studied previously. Although populations in neighbouring paddy fields showed a high level of differentiation, Bayesian clustering analyses suggested that some level of gene flow occurs among them and that the genetic exchange or colonisation between neighbouring populations could contribute to the geographical expansion of the resistant allele.     

Genetic diversity within and between sulfonylurea‐resistant and susceptible populations of Schoenoplectus juncoides in Japan

To reveal the effects of herbicide selection on genetic diversity in the outcrossing weed species Schoenoplectus juncoides, six sulfonylurea‐resistant (SU‐R) and eight sulfonylurea‐susceptible (SU‐S) populations were analysed using 40 polymorphic inter‐simple sequence repeat loci. The plants were collected from three widely separated regions: the Tohoku, Kanto and Kyushu districts of Japan. Genetic diversity values (Nei's gene diversity, h) within each SU‐S population ranged from h = 0.125 to h = 0.235. The average genetic diversity within the SU‐S populations was H_S = 0.161, and the total genetic diversity was H_T = 0.271. Although the H_S of the SU‐R populations (0.051) was lower than that of the SU‐S populations, the H_T of the SU‐R populations (0.202) was comparable with that of the SU‐S populations. Most of the genetic variation was found within the region for both the SU‐S and SU‐R populations (88% of the genetic variation respectively). Two of the SU‐R populations showed relatively high genetic diversity (h = 0.117 and 0.161), which were comparable with those of the SU‐S populations. In contrast, the genetic diversity within four SU‐R populations was much lower (from h = 0 to 0.018) than in the SU‐S populations. The results suggest that selection by sulfonylurea herbicides has decreased genetic diversity within some SU‐R populations of S. juncoides. The different level of genetic diversity in the SU‐R populations is most likely due to different levels of inbreeding in the populations.     

Inducing competition: intensive grassland seeding to control Ambrosia artemisiifolia

The invasion of Ambrosia artemisiifolia across European countries has been favoured by its ecological amplitude and by its ability to colonise and dominate disturbed/ruderal areas that have lost competition from native species. We supposed that a strong competition for habitat resources may inhibit A. artemisiifolia growth, generating a negative feedback to its establishment. Based on this hypothesis, in this study, we undertook a 1‐year field experiment to assess the effect of mixtures of grassland species on A. artemisiifolia growth and fitness in bare soils. We applied three different treatments within an abandoned quarry area invaded by A. artemisiifolia: (i) spontaneous succession, (ii) hayseed and (iii) a commercial seed mixture. Within plots, we recorded vegetation parameters, A. artemisiifolia abundance and traits. Results obtained after one growing season showed that the commercial seed resulted in the strongest reduction of A. artemisiifolia growth rate in terms of plant height, lateral spread and leaf size. This was ascribed to higher density of plants that play a key role in reducing biomass and fitness of A. artemisiifolia. However, hayseed should be preferred, as it preserves local biodiversity. Seeding mixtures of grassland species can successfully suppress A. artemisiifolia in the first year of establishment on a vegetation‐free soil derived from quarry activities. This study indicated that inducing dominance of different native species in a newly developing plant community should enhance competition for resources, reducing the success of early coloniser non‐native species.     

Management of roadside populations of invasive Ambrosia artemisiifolia by mowing

Ambrosia artemisiifolia (common ragweed) is a highly allergenic alien weed in Europe, which spreads rapidly along roadsides. Road verges are subject to frequent mowing, which further increases the spreading of the plants' seeds. Ambrosia artemisiifolia reacts to cutting by producing new shoots, which are able to develop flowers and ultimately new seeds. An effective mowing regime that would decrease the production of seeds and their dispersal is desirable to control the spread of the plant, but an appropriate way of mowing has yet to be found. In this study, we explored how the reproductive traits of A. artemisiifolia plants in seven spontaneous roadside populations reacted to the application of different mowing regimes over 3 years. The mowing regimes that were applied differed in the timing and frequency of cuttings. We found that the cutting regime, if appropriately timed, can strongly influence the production of male inflorescences (i.e. allergenic pollen), of female flowers (i.e. seeds) and had an impact on the phenological development of the plant. Based on our findings, we suggest that the optimal management of the plant along roadsides must be adjusted to its phenological development. The most effective mowing method of control consists of a first cut shortly before male flowering, to limit the quantities of released pollen, followed by subsequent cuts before the onset of new flowers on the resprouting lateral shoots.     

Do the introductions by botanical gardens facilitate the invasion of Solidago canadensis (Asterceae) in China?

Many invasive plants have long been suspected of firstly being introduced and cultivated by a local botanical garden or nursery and then escaping into the field after adapting to the novel environment. The role of botanical gardens in the spread of invasive plants has not yet been explored experimentally. In this article, we studied the possible roles of two botanical gardens in the spread of invasive Solidago canadensis (Asterceae) in China by analysing genetic relationships of invasive and native (United States) populations with intersimple sequence repeats markers. Our results showed a high genetic variation (mean He = 0.292) and a large proportion of genetic variation (85.6%) residing within populations. Solidago canadensis was possibly introduced firstly into eastern China. The plants from Lushan Botanical Garden showed distant genetic distance from all of the other populations, suggesting that this botanical garden had little effect on the invasion of S. canadensis. Populations from Wuhan Botanical Garden in central China, however, showed close genetic relationships with local populations and populations in west and south‐west China, suggesting gene exchange between these populations. Thus, risk assessment is critical for plant introduction and conservation, as introductions of alien plants by botanical gardens may facilitate plant invasions, while plants conserved in botanical gardens may be at risk by surrounding plant invasions.     

Major emerging alien plants in Austrian crop fields

Globalisation and increasing trade have led to the introduction of alien plants that are highly competitive and difficult to control in agriculture in central Europe. Our study set out to analyse the invasion process and agricultural impact of the six emerging alien plants Abutilon theophrasti, Ambrosia artemisiifolia, Cyperus esculentus, Datura stramonium, Panicum schinzii and Sorghum halepense in Austria, based on a large distribution data set (694 occurrence records in crop fields). We found that all study species have increased in abundance and range, especially after the year 2000. The rate of spread was highest for A. artemisiifolia and lowest for C. esculentus. At present, records of the study species were largely associated with areas of high summer crop concentration in eastern, southern and north‐western Austria. Based on the CORINE land‐cover data set, we found that the agricultural area at risk of being invaded increased over time. At present, it ranges between 45 500 and 168 000 ha (approximately 2.4% and 8.7% of the total agricultural area) depending on the species. The invasion success of the study species is probably associated with frequent human‐mediated dispersal, specific crop‐dominated rotations and herbicide use. Our results suggest that the study species will successfully spread further. We conclude that these emerging alien species will cause substantial impacts on crops in Austria and probably in other countries of central Europe.     

Ophraella communa,the ragweed leaf beetle,has successfully landed in Europe: fortunate coincidence or threat?

We report the occurrence of the North American ragweed leaf beetle Ophraella communa in Europe. During our surveys to monitor populations of the invasive alien plant Ambrosia artemisiifolia in Europe, we found the beetle south of the Alps, in more than 130 sites in southern Switzerland (Ticino) and northern Italy (Lombardia, Piemonte and Emilia‐Romagna). At sites where O. communa was present, up to 100% of the plants were attacked with damage levels high enough to completely defoliate and prevent flowering and seed set of most ragweed plants. That in its first year of discovery, O. communa was already found over a large area of c. 20 000 km² and in all habitat types occupied by A. artemisiifolia reflects its great dispersal potential and wide habitat suitability. This oligophagous beetle is a successful biological control agent against A. artemisiifolia in China, but despite extensive host specificity tests, the risk of attack and the level of damage of sunflower under field conditions remain unclear. The recently launched COST Action on ‘Sustainable management of Ambrosia artemisiifolia in Europe (SMARTER)’ offers an ideal framework to respond quickly to the recent establishment of O. communa in Europe and to collect data that can help determine whether this event should be considered a troublesome introduction or whether it is likely to become the first case of a successful biological control of an invasive weed in continental Europe.     

Nursery‐linked plantation outbreaks and evidence for multiple introductions of the pitch canker pathogen Fusarium circinatum into South Africa

In recent years, Pinus plantation forestry has been significantly hampered by outbreaks of pitch canker caused by the fungus Fusarium circinatum. This study investigated the role of Pinus host, geographic origin and reproductive mode in structuring the F. circinatum populations in plantations. For this purpose, 159 isolates originating from diseased plantation trees in the Western and Eastern Cape Provinces of South Africa were genotyped using 10 microsatellite markers. Analyses of these data revealed 30 multilocus haplotypes and that the populations were distinct based on geographic origin as well as host. However, shared haplotypes were observed between populations, showing that these populations are connected, possibly through the movement of haplotypes. A second aim was to determine whether the genetic variation found in these populations of the fungus could be attributed to outbreaks of the seedling disease caused by this pathogen in Pinus nurseries. To achieve this goal, an additional set of 43 isolates originating from pine seedling nurseries was genotyped and analysed. The results showed that the populations of F. circinatum in plantations most probably originated from the nursery outbreaks that occurred prior to the plantation outbreak. Inferences regarding reproductive mode further showed that sexual reproduction has little impact on the genetic makeup of the F. circinatum populations and that they primarily reproduce asexually. Overall, the results of this study showed that the F. circinatum diversity in South Africa has arisen due to multiple introductions of the pathogen and is not due to sexual reproduction.     

Genotypic diversity and migration of clonal lineages of Botrytis cinerea from chickpea fields of Bangladesh inferred by microsatellite markers

An analysis of allelic diversity at nine microsatellite loci provided an insight into the population structure of Botrytis cinerea from four fields (sampled in 2003 and 2004) that represented important regional locations for chickpea production in Bangladesh. Although three populations were limited by sample size after clone‐correction, a total of 51 alleles were amplified among 146 B. cinerea isolates from Bangladesh, which revealed a high amount of within‐population and overall genetic diversity (H_S = 0·48 and H_T = 0·54, respectively). The percentage of maximal genotypic diversity (G) ranged between populations (G = 23–40), with a total of 69 haplotypes detected (G = 25). Bayesian cluster analysis depicted two major clusters distributed among the four Bangladesh populations, indicating population admixture from two origins that have spread throughout these regions. Genotype flow between regions was detected and indicated the spread of clonal lineages, consistent with relatively low differentiation among the four populations (mean G_ST = 0·1, P < 0·05). These results highlighted the potential threat of host resistance breakdown as a result of considerable genetic diversity, genotype flow and the evolutionary potential of B. cinerea.     

Genetic variation of the invasive Campuloclinium macrocephalum,Asteraceae in South Africa,inferred from molecular markers

Campuloclinium macrocephalum is native to Central and South America, but is a highly invasive weed in South Africa, where it is commonly known as the ‘pompom weed’. It is targeted for biological control, the success of which will depend on host specificity and biotype compatibility to its full genetic diversity in South Africa. We investigated the genetic diversity and phylogeography of 52 specimens from across South Africa, 14 from Argentina and three from Brazil using nuclear ribosomal ITS regions. We further explored the AFLP marker diversity in 54 South African, 25 Argentine and three Brazilian specimens. Maximum parsimony analysis of the ITS sequence data produced an unresolved phylogeny. However, three haplotypes were recognised via network analysis. All South African, one Brazilian and all bar one of the Argentine individuals shared a single haplotype. AFLP analyses generated two genetic clusters with a low net nucleotide distance of 0.115 and further revealed that most plants were a mixture of alleles from these two genetic clusters. Although there was a significant genetic variation among the populations, genetic differentiation and mean heterozygosity were low, suggesting that clonal reproduction may be occurring. The current South African populations may therefore be clonal products with different proportions of genetic admixture introduced more than once. The original point of entry appears to be Gauteng, South Africa. Long‐distance dispersal appears to have played a major role in its spread across South Africa. Candidate Argentine biological control agents should therefore be effective on C. macrocephalum in South Africa.     

Distribution of Parthenium hysterophorus and one of its biological control agents (Coleoptera: Zygogramma bicolorata) in Nepal

Parthenium hysterophorus is a noxious invasive weed of both agricultural and natural ecosystems, spreading aggressively in Nepal. Management of this weed in Nepal has been limited, mainly because of the lack of geo‐referenced data concerning the weed's distribution. We conducted a nationwide survey of P. hysterophorus and its coleopteran biological control agent Zygogramma bicolorata from 2013 to 2016 to determine their spatial distribution. Both were widespread, with the distribution of Z. bicolorata lagging behind the invasion front of P. hysterophorus. The weed was present in 21.2% of the 4838 locations examined, including several isolated satellite populations. The weed was found in the Tarai, Siwalik, Middle Mountains and High Mountains regions, reaching up to 2000 m asl. It has invaded natural and modified ecosystems including all six protected areas in the Tarai and Siwalik regions. Road access appears to be the major pathway for its long‐distance dispersal. Zygogramma bicolorata had spread from the east to the west and was present in 15.4% of the weed occurrence locations, inflicting a low amount of damage. A CLIMEX modelling projection revealed the presence of additional geographic areas in Nepal which are climatically suitable for both P. hysterophorus and Z. bicolorata. Eradication of satellite populations of the weed by physical and chemical measures, and the release of Z. bicolorata into new, but climatically suitable, locations should be prioritised for P. hysterophorus management in Nepal. In conclusion, P. hysterophorus has rapidly become widespread in Nepal and the currently available biological control agent has not been able to prevent further spread of the weed.     

Latent entry and spread of Colletotrichum acutatum (species complex) in strawberry fields

Anthracnose, caused by Colletotrichum acutatum (species complex), has become a troublesome problem in strawberry production worldwide. This paper reports (i) an optimized sampling method combined with a real‐time PCR technique to detect the latent presence of C. acutatum in cold‐stored strawberry plants used as planting material in several European countries, and (ii) a study of the spread of C. acutatum following a point inoculation under field conditions. Screening of different parts of planting material suggested that C. acutatum is most likely to be present on runners and old petioles. In addition, in seven out of nine batches of planting material from different nurseries, latent infection by C. acutatum was detected in at least one of five replicate samples. Field experiments in 2009 and 2010 showed extensive latent within‐field spread of the pathogen on strawberry leaves, with a within‐row dispersal distance up to at least 1·75 m in 1 week. A straw ground cover between the rows did not decrease C. acutatum spread, probably because introduced (and/or subsequent) inoculum was confined to the plant bed (within the row) and was not present between the beds. Moreover, the number of C. acutatum spores on the symptomless leaves, as estimated using a real‐time PCR method, was significantly (P < 0·05) correlated with the incidence of fruit rot at harvest and post‐harvest (r = 0·56–0·66). These results illustrate the importance of detecting latent infections in planting material and strawberry leaves in the field.     

Genetic structure of Verticillium dahliae isolates infecting olive trees in Tunisia using AFLP,pathogenicity and PCR markers

Since 2006, verticillium wilt of olive induced by Verticillium dahliae has caused considerable economic losses in olive orchards in Tunisia. The genetic structure of V. dahliae isolates collected from different olive growing regions was investigated using virulence tests, vegetative compatibility grouping (VCG) and amplified fragment length polymorphism (AFLP) analyses. In total, 42 isolates of V. dahliae from diseased olive trees were tested. Cluster analysis and principal coordinate analysis revealed that geographic origin was the main factor determining the genetic structure of V. dahliae populations and both methods indicated a genetic separation between the central and coastal isolates. Isolates were divided into two major groups: the AFLP‐I group included all isolates from Sidi Bouzid, Kairouan, Kasserine and Sfax (centre of the country) and the AFLP‐II group included isolates from Monastir, Zaghouane, Sousse, Mahdia (coastal region), and two isolates from Sfax. Analysis of the molecular variance (amova ) indicated a significant level of genetic differentiation among (76%) and within (23%) the two populations. Analyses of both the defoliating (D) and non‐defoliating (ND) pathotypes and VCG markers indicated that most of the isolates belong to VCG 2A and 4B/ND pathotype. The disease severity was highly variable among the isolates tested (P < 0·05) with no evidence of association between aggressiveness and geographical origin of the isolates. Overall, results of this study revealed a clear association between the genetic diversity of the isolates and their geographic origin, but not between genetic diversity and virulence patterns.     

The current status of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiiidae) in Turkey and baseline toxicity of some insecticides

Tomato is one of the most important vegetable crops in Turkey, with national production of over 10 million tonnes in 2010. Tuta absoluta (Meyrick) is an important pest of tomato, and was first recorded in Urla District of Izmir Province in the Aegean region of Turkey in August 2009. It has since spread rapidly to the other regions of Turkey and become the main pest of tomato. Since its dispersal, chemical control has been the main method of control. Intensive use of insecticides has led to the development of resistance in T. absoluta. In this study, the baseline toxicity (LC₅₀ values) of some insecticides was determined using a leaf‐dip bioassay method in Antalya and Ankara populations of T. absoluta.     

Microsatellite and mating type markers reveal unexpected patterns of genetic diversity in the pine root‐infecting fungus Grosmannia alacris

Grosmannia alacris is a fungus commonly associated with root‐infesting bark beetles occurring on Pinus spp. The fungus has been recorded in South Africa, the USA, France, Portugal and Spain and importantly, has been associated with pine root diseases in South Africa and the USA. Nothing is known regarding the population genetics or origin of G. alacris, although its association with root‐infesting beetles native to Europe suggests that it is an invasive alien in South Africa. In this study, microsatellite markers together with newly developed mating type markers were used to characterize a total of 170 isolates of G. alacris from South Africa and the USA. The results showed that the genotypic diversity of the South African population of G. alacris was very high when compared to the USA populations. Two mating types were also present in South African isolates and the MAT1‐1/MAT1‐2 ratio did not differ from 1:1 (χ² = 1·39, P = 0·24). This suggests that sexual reproduction most probably occurs in the fungus in South Africa, although a sexual state has never been seen in nature. In contrast, the large collection of USA isolates harboured only a single mating type. The results suggest that multiple introductions, followed by random mating, have influenced the population structure in South Africa. In contrast, limited introductions of probably a single mating type (MAT1‐2) may best explain the clonality of USA populations.     

A quantitative genetic examination of non‐target‐site resistance applied to Avena species

Quantitative genetics tools can be used to assess whether using herbicides at low doses drive selection on standing genetic variation in populations leading to non‐target‐site resistance (NTSR). These tools are particularly important for estimating the number of genes involved and the potential speed of evolution. A short cut to answering questions about the evolution of NTSR may be to measure heritability. The heritability index (H) provides a measure of the potential to develop NTSR and can be simply calculated from classical dose–response experiments. This measure and the associated experimental designs are discussed with two applied examples on Avena spp. (A. fatua and A. sterilis). In these examples, H values ranged from 0.24 to 0.73, which means that selection for NTSR is highly probable in cases with high H value. We suggest that structuring plants into genetic groups (e.g. families or populations) can contribute to our understanding of the evolutionary potential of populations and plant species to evolve resistance, without increasing experimental cost and time.     

Fine‐tuned ability to predict future competitive environment in Ambrosia artemisiifolia seeds

Competition from native species is a key mechanism for biotic resistance to invasion. Accelerated germination to pre‐empt resources or delayed germination and induced dormancy until the next growing season are two alternative strategies for annual invasive plants to avoid the drawbacks of competition at the seed stage. In Ambrosia artemisiifolia, both of these tactics could theoretically increase its long‐term fitness. However, their relative importance has never been tested. We studied the germination pattern of A. artemisiifolia seeds in various competitive environments by experimentally modifying the life stage (seed, seedling, adult), density (low, high) and also the identity (intraspecific and 3 interspecific competitors) of neighbours in controlled conditions. When facing competition of seeds and seedlings at high densities or of particular identity, A. artemisiifolia accelerated its germination. In contrast, A. artemisiifolia followed a competition avoidance strategy in the presence of established adult heterospecific neighbours by delaying germination and reducing the germination fraction through induction of secondary dormancy. By testing the seedlings' performance in the same competition situations as those of seeds, we showed that the germination responses were beneficial in the case of heterospecific, but not of conspecific neighbours.