Isoenzyme diversity in Reynoutria (Polygonaceae) taxa: escape from sterility by hybridization

The genus Reynoutria is represented by four taxa in the Czech Republic – R. japonica var. japonica and compacta, R. sachalinensis and R. × bohemica. Using isoenzyme analysis, we determined the degree of genotype variability in all taxa and compared clones of R. japonica var. japonica from the Czech Republic with those from Great Britain. While the rarely occurring tetraploid variety R. japonica var. compacta possesses low variability, the octoploid female clone of R. japonica var. japonica is genetically uniform in the 93 clones sampled and belongs to the same genotype that is present in the whole Europe. R. japonica var. japonica can be fertilized by the pollen of tetraploid R. sachalinensis and a hexaploid hybrid R. × bohemica is produced. In R. sachalinensis, 16 genotypes were found in the 50 clones sampled. R. × bohemica is genetically the most diverse taxon in the study area, with 33 genotypes recorded among 88 clones sampled.     

Morphological and chromosomal variation in Fallopia section Reynoutria (Polygonaceae) in Korea

We have examined morphological and chromosomal variation inFallopia sect.Reynoutria in Korea to clarify their taxonomic identities and to determine whether their morphological variability is associated with ploidy levels. Principal components analysis (PCA) of individuals from 21 populations, using major distinguishing characters, revealed the presence of four major entiries of sect.Reynoutria in Korea; these includeF. sachalinensis, F. japonica var.japonica, F. forbesii, and the Nonsan population consisting of presumed hybrids. Based on morphology, it is hypothesized that the Nonsan population was probably derived from multiple hybridization events involving the three named taxa. The results also indicate thatF. forbesii is distinct fromF. japonica var.japonica. Polyploidy is more prevalent in sect.Reynoutria than has been previously recognized.Fallopia sachalinensis in Korea occurs as dodecaploids with 2n=132; our count is the first dodecaploid count for the species, and represents the highest chromosome number known in the genus.Fallopia japonica var.japonica occurs as tetraploids (2n=44), hexaploids (2n=66), and octoploids (2n=88), whileF. forbesii occurs as hexaploids (2n=66) and octoploids (2n=88); our counts appear to be the first reported chromosome numbers forF. forbesii. Morphological analysis indicates that there is no apparent correlation between the ploidy levels in these taxa and the morphological characters that we have considered in this study except that the tetraploids ofF. japonica var.japonica tend to have somewhat thicker leaves.     

The potential role of polyploidy and hybridisation in the further evolution of the highly invasive <Emphasis Type="Italic">Fallopia</Emphasis> taxa in Europe

Japanese knotweed s.l. comprises Fallopia japonica, F. sachalinensis, F. × bohemica and any F2s or backcrosses. The parental taxa were introduced from the East to the West as garden ornamentals in the nineteenth century, and soon spread beyond the confines of the garden to become widespread and persistent weeds. Since only female F. japonica var. japonica was introduced, its impressive spread has occurred solely by vegetative means. However, the initial lack of genetic variability has been complemented by an extensive series of hybridisations in the adventive range. We examine the history, spread, reproductive biology and ecological impact of these species in the West. The role and importance of polyploidy and hybridisation in their invasion of the West is discussed, as are the implications of these factors for the potential further evolution of the group.     

How does Reynoutria invasion fit the various theories of invasibility?

Questions: 1. How does species richness of recipient communities affect Reynoutria invasion? 2. How does Reynoutria invasion change host community structure? 3. Are there any differences in habitat preferences among three closely related Reynoutria taxa? 4. How does the genetic structure of Reynoutria populations change along the course of a river? Location: River Jizera basin, north Bohemia, Czech Republic. Methods: Nine 0.25 km² plots were chosen along the river. Within each plot all main habitat types were determined and sampled using the Braun‐Blanquet scale to determine the invasibility of various communities. The patches invaded by Reynoutria taxa and surrounding Reynoutria‐free vegetation in the same habitat type were sampled as relevé pairs to compare the composition of invaded and non‐invaded vegetation. In addition, to characterize the genetic structure of Reynoutria populations along the river, 30 samples from different clones were collected. Results and conclusions: 1. The species richness of communities has no influence on the success of Reynoutria invasion in the area studied. The combination of environmental conditions and propagule spread is more important to the invasion success than the number of species in the host community. 2. Reynoutria invasion greatly reduces species diversity. 3. R. japonica invaded more habitat types than R. sachalinensis and R.×bohemica. The hybrid R.×bohemica outcompetes the parental taxa at sites where both taxa co‐occur. 4. Isozyme analysis revealed phenotype variability in the hybrid in contrast to the parental taxa. Different hybrid phenotypes are distributed randomly on the middle and lower reaches of the River Jizera; one of them dominates and the other three occur occasionally. This pattern supports the hypothesis that sexual reproduction occasionally occurs within Reynoutria taxa.     

Reynoutria niche modelling and protected area prioritization for restoration and protection from invasion: A Southeastern Europe case study

An important step in managing invasive species is determining the factors responsible for their current and potential distribution, especially when species distribution is climatically determined like in Reynoutria taxa case. The main aim of this paper was to integrate all available distribution data of the Reynoutria taxa in SE Europe and to predict in which habitats and along which corridors its future spread can be expected. Distribution data of the Reynoutria taxa were obtained from extensive field studies in the period from 2006 to 2016, as well as from the literature and herbarium sources. A total of 4081 localities in Serbia and the Kosovo region, Montenegro, Slovenia, Croatia, Bosnia and Herzegovina, Bulgaria and Romania have been recorded, most frequently in riparian and human-made habitats. Ecologically suitable ranges for the Reynoutria taxa were predicted using the MaxEnt program and 19 bioclimatic variables which were derived from the WorldClim data set. Results of this study showed that the most frequent Reynoutria taxa in SE Europe are R. japonica var. japonica and hybrid R. × bohemica. The most suitable geographical areas for R. japonica and R. sachalinensis are mostly located in the north, while for R. × bohemica the central part of SE Europe is suitable. The precipitation of the warmest quarter had a high influence on the potential distribution modelling of these closely related species. Potential distribution modelling revealed R. × bohemica’s high temperature tolerance in different seasons with high durability of drought (up to 60 mm per year less precipitation than parental species). Moreover, this research predicts which areas are most likely to be invaded, and makes suggestions where to focus management efforts for survey and removal of these invaders, especially in protected areas. According to the future climate analysis R. japonica and R. × bohemica could expand their range in riparian habitats up to 30–40%. Consequently, invasion in Slovenia’s and Croatia’s rivers requires urgent preventative measures. This study provides empirical evidence that these plants could become widespread throughout SE Europe within 25 years under the current trend and emphasizes that regionally specific management plans should be implemented to prevent further spread of these taxa.     

Asexual spread versus sexual reproduction and evolution in Japanese Knotweed s.l. sets the stage for the “Battle of the Clones”

The invasion of NW Europe by a single clone of male-sterile Fallopia japonica var. japonica by purely vegetative reproduction has demonstrated that reproduction by seed and the genetic diversity associated with it may not be an essential feature of plant invasions, at least in the short term. What is less well known is that a significant proportion of the Japanese Knotweed s.l. involved is not F. japonica var. japonica, but the hybrid between it and F. sachalinensis–F. × bohemica. This hybrid is able to backcross to either parent with the potential to replace the missing male F. japonica; by the same process, the hybrid is generating the genetic diversity so conspicuously lacking in F. japonica. In terms of understanding the population structure in a particular country, it is important to be able to identify hybrids and putative back-crosses. We bring together a mixture of published and unpublished information to provide a comprehensive section on morphological and anatomical aids to identification. Regeneration of plants from stem and rhizome fragments varies by taxon, as do responses to control techniques, underlining the importance of correct identification of these plants. In this paper we look closely at this group of taxa, with special emphasis on the role played by hybridization, with an examination of the genetic make up of seedlings produced in the wild or by artificial hybridizations, and the implications that this will have on the future directions of the invasion process.     

Low genetic variation of invasive Fallopia spp. in their northernmost European distribution range

Knowledge about the reproduction strategies of invasive species is fundamental for effective control. The invasive Fallopia taxa (Japanese knotweed s.l.) reproduce mainly clonally in Europe, and preventing spread of vegetative fragments is the most important control measure. However, high levels of genetic variation within the hybrid F. × bohemica indicate that hybridization and seed dispersal could be important. In Norway in northern Europe, it is assumed that these taxa do not reproduce sexually due to low temperatures in the autumn when the plants are flowering. The main objective of this study was to examine the genetic variation of invasive Fallopia taxa in selected areas in Norway in order to evaluate whether the taxa may reproduce by seeds in their most northerly distribution range in Europe. Fallopia stands from different localities in Norway were analyzed with respect to prevalence of taxa, and genetic variation within and between taxa was studied using amplified fragment length polymorphism (AFLP). Taxonomic identification based on morphology corresponded with identification based on simple sequence repeats (SSR) and DNA ploidy levels (8× F. japonica, 6× F. × bohemica and 4× F. sachalinensis). No genetic variation within F. japonica was detected. All F. × bohemica samples belonged to a single AFLP genotype, but one sample had a different SSR genotype. Two SSR genotypes of F. sachalinensis were also detected. Extremely low genetic variation within the invasive Fallopia taxa indicates that these taxa do not reproduce sexually in the region, suggesting that control efforts can be focused on preventing clonal spread. Climate warming may increase sexual reproduction of invasive Fallopia taxa in northern regions. The hermaphrodite F. × bohemica is a potential pollen source for the male‐sterile parental species. Targeted eradication of the hybrid can therefore reduce the risk of increased sexual reproduction under future warmer climate.     

Tree regeneration patterns on Mount Usu, northern Japan, since the 1977–78 eruptions

Although the summit of Mount Usu was deforested by the 1977–78 eruptions, vegetative regeneration on the caldera rim was rapid due to the erosion of thick volcanic deposits by snow and rain. To obtain the mechanisms underlying regeneration patterns after the eruptions, we monitored the growth of permanently-marked stems from 1983 to 1990. Regeneration was from resprouting-branches buried in the volcanic deposits on the caldera rim, while on the crater basin, where thick volcanic deposits accumulated, regeneration was from seedlings. The seedling regeneration lagged approximately 3 years behind vegetative regeneration. Stem densities averaged 14,000 ha^-1 in the vegetatively-regenerated community on the caldera rim, and 28,000 ha^-1 in the seedling regeneration on the crater basin. Populus maximowiczii accounted for ca. 75% of total stems on the caldera rim, while P. maximowiczii accounted for ca. 30% on the crater basin where Salix integra and Betula platyphylla var. japonica were also common. In both stands, immigration and mortality rates were very low. The growth of vegetatively regenerated stems expressed as stem height and diameter was significantly faster than that of stems grown from seedlings. Herbivory damage on the terminal shoots of tall stems was restricted on the caldera rim and was restricted for B. platyphylla var. japonica on the crater basin, perhaps due to fast growth supporting herbivore avoidance or low palatability. Height growth was restricted when neighbors established in close proximity, especially in the seedling-regenerated forest. The results suggest that vegetative regeneration is rapid due to three mechanisms: 1) faster plant growth; 2) herbivore avoidance; and 3) decreased interference by neighboring.     

Factors influencing the production of stilbenes by the knotweed, <Emphasis Type="Italic">Reynoutria</Emphasis> × <Emphasis Type="Italic">bohemica</Emphasis>

Background  

Japanese knotweed, Reynoutria japonica, is known for its high growth rate, even on adverse substrates, and for containing organic substances that are beneficial to human health. Its hybrid, Reynoutria × bohemica, was described in the Czech Republic in 1983 and has been widespread ever since. We examined whether Reynoutria × bohemica as a medicinal plant providing stilbenes and emodin, can be cultivated in spoil bank substrates and hence in the coalmine spoil banks changed into arable fields. We designed a pot experiment and a field experiment to assess the effects of various factors on the growth efficiency of Reynoutria × bohemica on clayish substrates and on the production of stilbenes and emodin in this plant.     

Effects of climatic factors on Fallopia japonica s.l. seedling establishment: evidence from laboratory experiments

The highly invasive Fallopia complex (Fallopia japonica s.l., Fallopia japonica var. japonica, Fallopia sachalinensis and the hybrid Fallopia × bohemica) is known to spread primarily vegetatively by rhizomes and stem fragments in its adventive range. Sexual reproduction has rarely been shown in Europe, despite a high production of germinable seeds. To elucidate this contradiction the aim of the present study was to determine which environmental conditions prevent seed germination or seedling establishment. Several laboratory experiments were conducted focusing on water and light supply as well as the influence of frost. According to morphological and cytological analyses of the parental plants paired with morphological analyses of the seedlings, the seeds used for these experiments were assumed to belong predominantly to the hybrid Fallopia × bohemica. Laboratory experiments revealed that seedling establishment is hampered by severe drought or strong late frost, which are, however, rare events in the study region and could not be crucial in explaining the rarity of seedlings in the field. We discuss the possible role of weather conditions that might lead to a loss of seeds or seedlings under field conditions and include a comparison of several other cited studies carried out in the adventive range of Fallopia japonica s.l. that observed either the presence or the absence of Fallopia seedlings in the field.     

The regeneration process in a mixed forest in central Hokkaido,Japan

The regeneration process in a mixed forest was investigated in Nopporo National Forest, Hokkaido. The analysis of age structure in an 80 m×80 m plot revealed that almost all of the species regenerated intermittently. In eleven gaps observed in the plot, the regeneration of a boreal conifer (Abies sachalinensis) was seldom observed. Temperate hardwoods, even climax species (Acer mono, Quercus mongolica var. grosseserrata, Tilia japonica), regenerated vigorously in the gaps. The age structure in ten additional plots scattered all over the forest showed that Abies tended to regenerate synchronously. From previous records, regeneration of Abies could be ascribed to catastrophic storms causing serious windfalls. On the other hand, regeneration of the temperate hardwoods was not synchronous but independent in different places within the forest. They could regenerate not only after those catastrophic storms but also after less severe disturbances which caused the death and fall of one or several trees. It is concluded that the coexistence of boreal coniferous species and temperate deciduous broad-leaved species in mixed forests may be maintained not only by the difference in habitat but also by the balance between the less frequent large disturbances, and the more frequent smaller ones.     

Unexpected genetic diversity of Fallopia japonica from Central Europe revealed after AFLP analysis

Recently much attention has been paid to genetic aspects of invasive success in Japanese knotweed s.l. One hypothesis to explain the invasive spread of these species is a multiple introduction, which leads to a higher level of genetic diversity in the invaded range. Fallopia japonica is considered to be genetically uniform in Europe, introduced as a single female clone. However, there is some evidence suggesting that invasion history and dynamics differ between Western and Central-Eastern Europe. We used AFLP markers to characterize genetic diversity of three Fallopia taxa that occur in Poland: F. japonica, F. sachalinensis and their hybrid Fallopia × bohemica, growing in so-called ‘homogeneous’ populations, consisting of one taxon and ‘heterogeneous’ populations, composed of the three taxa cohabiting together. No polymorphism, resp. an insignificantly low variability was observed in the ‘homogeneous’ populations. In the ‘heterogeneous’ stands polymorphism was detected within each taxa, with one exception that concerns individuals of F. sachalinensis from a riparian habitat. The highest level of polymorphism was found among individuals of F. × bohemica. The most striking result of our study is the observation of polymorphism between individuals of F. japonica. The AFLP data also showed that F. × bohemica is most diverse when occurring in a heterogeneous configuration with F. japonica and F. sachalinensis in the same habitat. Our results are the first evidence of genetic diversity in F. japonica populations in Central Europe and can implicate the possibility of its multiple introduction in this region or the existence of sexual reproduction of this species.     

Landscape dynamics and habitat selection by the alien invasive <Emphasis Type="Italic">Fallopia</Emphasis> (Polygonaceae) in Belgium

Habitat patch colonization dynamics and distribution patterns were analysed at a landscape scale in four invasive Fallopia (Polygonaceae) species. Fallopia sachalinensis and F. aubertii were uncommon and population expansion was not evident during the three consecutive years of study. The two most widespread species, F. japonica and F. × bohemica displayed similar habitat selection patterns with ruderal and natural/semi-natural forests favoured. The highest densities of F. japonica and F. × bohemica individuals were at the edge of preferred habitat patches with different patterns of edge selection. Linear network played an important role in species invasion, with 71% of all F. japonica and F. × bohemica occurring within a 10 m buffer of total linear networks (roads, railways, and rivers). However, the buffer represented only 14.5% of the total landscape surface. The rate of population increase was higher for F. japonica (75.8% and 35.2%, in 2002 and 2003, respectively) than for F. × bohemica (63.6% and 0% in 2002 and 2003, respectively) and was largely the result of intra-patch dynamics with low inter-patch colonization. The total surface area occupied by Fallopia clones in the landscape grew by 34.7% over 2 years of the study, with comparable area growth means for F. japonica and F. × bohemica (34.9% and 34.7%, respectively). The hypothesis that F. × bohemica exhibits higher invasive dynamics due to both clonal and sexual reproduction was not supported by our results.     

Allozyme variation in Japanese species ofChionographis (Liliaceae)

Allozyme variation was examined in three diploid taxaChionographis japonica var.japonica, var.kurokamiana, andC. koidzumiana and three tetraploid taxaC. japonica var.kurohimensis, ssp.hisauchiana, and ssp.minoensis. Results show thatC. japonica var.kurokamiana is genetically closer toC. koidzumiana than to var.japonica. In the tetraploid taxa, fixed heterozygosities were found at several loci, and this supports the hypothesis that these taxa are allotetraploids. Furthermore, the tetraploid taxa have many unique alleles not found in the diploid taxa. This suggests that sufficient time has passed since the origin of tetraploids for new mutations to have been fixed.     

Variation of growth and functional traits of invasive knotweeds (<Emphasis Type="Italic">Fallopia</Emphasis> spp.) in Belgium

Three invasive Fallopia taxa are present in Belgium: F. japonica (FJ), F. sachalinensis (FS) and their hybrid F. × bohemica (FB). FS is the least invasive of the three taxa. In this study, we compared the taxa, in sites where they co-occur, for differences in functional traits that might influence their competitive ability and invasiveness—shoot height and ramification, leaf size, specific leaf area (SLA) and foliar nitrogen (N) concentration. The three taxa exhibited similar growth kinetics and similar SLA. However, FS differed in its architecture and allocation of leaf area, having less ramified shoots and a steeper gradient of decreasing leaf size along the main shoot. Also, FS had greater foliar N and less efficient N resorption from senescing leaves. These traits values may result in lower competitive ability of FS for light and nitrogen. For the same traits, FB was generally intermediate between FS and FJ, but often closer to the latter. FB was more variable than FS and FJ, possibly due to larger genetic variation. SLA and ramification varied greatly amongst sites for all taxa, due in part to plastic response to contrasting light regimes. Variation in functional traits values may in part explain the variation in invasiveness amongst the members of the Fallopia complex in Belgium.     

Herbivore resistance of invasive <Emphasis Type="Italic">Fallopia</Emphasis> species and their hybrids

Hybridization has been proposed as a mechanism by which exotic plants can increase their invasiveness. By generating novel recombinants, hybridization may result in phenotypes that are better adapted to the new environment than their parental species. We experimentally assessed the resistance of five exotic Fallopia taxa, F. japonica var. japonica, F. sachalinensis and F. baldschuanica, the two hybrids F. × bohemica and F. × conollyana, and the common European plants Rumex obtusifolius and Taraxacum officinale to four native European herbivores, the slug Arion lusitanicus, the moth Noctua pronuba, the grasshopper Metrioptera roeselii and the beetle Gastrophysa viridula. Leaf area consumed and relative growth rate of the herbivores differed significantly between the Fallopia taxa and the native species, as well as among the Fallopia taxa, and was partly influenced by interspecific variation in leaf morphology and physiology. Fallopia japonica, the most abundant Fallopia taxon in Europe, showed the highest level of resistance against all herbivores tested. The level of resistance of the hybrids compared to that of their parental species varied depending on hybrid taxon and herbivore species. Genotypes of the hybrid F. × bohemica varied significantly in herbivore resistance, but no evidence was found that hybridization has generated novel recombinants that are inherently better defended against resident herbivores than their parental species, thereby increasing the hybrid’s invasion success. In general, exotic Fallopia taxa showed higher levels of herbivore resistance than the two native plant species, suggesting that both parental and hybrid Fallopia taxa largely escape from herbivory in Europe.     

Nitrogen translocation via rhizome systems in monoclonal stands ofReynoutria japonica in an oligotrophic desert on Mt Fuji: Field experiments

Mechanisms which enableReynoutria japonica, a dominant pioneer herb, to be successful in maintaining large stands in an oligotrophic volcanic desert on Mt Fuji were investigated with special reference to its nitrogen acquisition.Reynoutria japonica forms circular stands, each of which comprises only one genet. As a stand develops outwards, the number of aerial shoots per unit area decreases in the center. Shoots grow vigorously in the peripheral area where the available nitrogen from soil and precipitation (about 2.4 g m⁻² year⁻²) was much less than total nitrogen in the shoots (6.1–9.1 g m⁻²). Leaf nitrogen content per unit mass was also greater in the leaves of the peripheral shoots. When rhizomes extending radially from the center to the periphery were severed, the dry mass of shoots in the periphery diminished by 75% on a ground area basis. In the periphery, leaf nitrogen content also reduced significantly and no flowers were produced. When fertilizer was applied to the peripheral shoots with severed rhizomes, neither growth, survival nor flower production of the shoots was significantly smaller than the control levels. In these shoots, it is also found that the nitrogen content in the youngest leaves decreased for about 1 month and then increased to above that in the control leaves. These results suggest that (i) nitrogen accumulated in the central part is translocated to peripheral shoots via rhizomes, and that long-distance translocation enables the stands to develop outwards, and (ii) aerial shoots in the periphery utilize the nitrogen translocated by rhizomes in the beginning of the growth season, whereas once the shoots have established, they begin to take up nitrogen with their own roots. Since the peripheral shoots are in sunnier environments than the shoots inside the stand, the acropetal nitrogen translocation via rhizomes will raise the production efficiency of a whole stand.     

Somatic embryogenesis in Canary Island date palm

Shoot regeneration was obtained from leaves of in vitro cultures of wild pear genotypes. The highest regeneration rates, ranging from 40% to 64%, depending on the genotype, were obtained using leaves wounded by three cuts transversely to the mid-rib, a Quoirin and Lepoivre macro-salt composition, 250 mg l^-1 cefotaxime and maintaining the explants in darkness for the first 30 days (induction phase), then transferring them to an auxin-free medium in light (expression phase). A concentration of 8.8 μM BA induced the highest number of explants to produce adventitious shoots. TDZ was less effective than BA and induced hyperhydricity in regenerated shoots. The histological studies revealed that the regenerated shoots originated mainly from callus formed by epidermal and sub-epidermal cells and by cells of the vascular tissue. The regenerated shoots were micropropagated, rooted and transplanted to the soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

<Emphasis Type="Italic">In vitro</Emphasis> plant regeneration from the immature seeds of <Emphasis Type="Italic">Cymbidium faberi</Emphasis>

An in vitro plant regeneration protocol of Cymbidium faberi from immature seeds was established. The immature seeds of 50 days old started to form rhizomes 4 months after they were cultured on hormone free medium. The rhizomes multiplied 5 times when subcultured on the medium containing 1.0 mg l^–1 -naphthalene acetic acid (NAA) for 40 days and more than 90% of the rhizomes initiated shoots within 60 days on the media containing 0.5 or 1.0 mg l^–1 NAA plus 2.0 or 5.0 mg l^–1N⁶-benzylaminopurine (BA). Plantlets were regenerated when the shoots were planted on the basal medium amended with 1 g l^–1 activated charcoal for 50 days and the plantlets grew normally after transplanting.