Regional differences in clonal Japanese knotweed revealed by chemometrics-linked attenuated total reflection Fourier-transform infrared spectroscopy.

BACKGROUND: Japanese knotweed (R. japonica var japonica) is one of the world's 100 worst invasive species, causing crop losses, damage to infrastructure, and erosion of ecosystem services. In the UK, this species is an all-female clone, which spreads by vegetative reproduction. Despite this genetic continuity, Japanese knotweed can colonise a wide variety of environmental habitats. However, little is known about the phenotypic plasticity responsible for the ability of Japanese knotweed to invade and thrive in such diverse habitats. We have used attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy, in which the spectral fingerprint generated allows subtle differences in composition to be clearly visualized, to examine regional differences in clonal Japanese knotweed. RESULTS: We have shown distinct differences in the spectral fingerprint region (1800-900 cm- 1) of Japanese knotweed from three different regions in the UK that were sufficient to successfully identify plants from different geographical regions with high accuracy using support vector machine (SVM) chemometrics. CONCLUSIONS: These differences were not correlated with environmental variations between regions, raising the possibility that epigenetic modifications may contribute to the phenotypic plasticity responsible for the ability of R. japonica to invade and thrive in such diverse habitats.

Molecular Analysis of UV-C Induced Resveratrol Accumulation in Polygonum cuspidatum Leaves.

Resveratrol is one of the most studied plant secondary metabolites owing to its numerous health benefits. It is accumulated in some plants following biotic and abiotic stress pressures, including UV-C irradiation. Polygonum cuspidatum represents the major natural source of concentrated resveratrol but the underlying mechanisms as well as the effects of UV-C irradiation on resveratrol content have not yet been documented. Herein, we found that UV-C irradiation significantly increased by 2.6-fold and 1.6-fold the resveratrol content in irradiated leaf samples followed by a dark incubation for 6 h and 12 h, respectively, compared to the untreated samples. De novo transcriptome sequencing and assembly resulted into 165,013 unigenes with 98 unigenes mapped to the resveratrol biosynthetic pathway. Differential expression analysis showed that P. cuspidatum strongly induced the genes directly involved in the resveratrol synthesis, including phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, 4-coumarate-CoA ligase and stilbene synthase (STS) genes, while strongly decreased the chalcone synthase (CHS) genes after exposure to UV-C. Since CHS and STS share the same substrate, P. cuspidatum tends to preferentially divert the substrate to the resveratrol synthesis pathway under UV-C treatment. We identified several members of the MYB, bHLH and ERF families as potential regulators of the resveratrol biosynthesis genes.

Tolerance of Japanese knotweed s.l. to soil artificial polymetallic pollution: early metabolic responses and performance during vegetative multiplication.

The expansion of invasive Japanese knotweed s.l. is of particular concern because of its aptitudes to rapidly colonize diverse environments, especially anthropized habitats generally characterized by their pollution with heavy metals. Whether the presence of heavy metals impacts the performance traits of this plant is a central question to better understand its invasive properties, though no controlled approach to assess these effects was yet reported. In this aim, we undertook greenhouse experiments where rhizome fragments of Japanese knotweed s.l. (Fallopia japonica and Fallopia × bohemica) were grown during 1 and 3 months, in a soil pot artificially polluted or not with heavy metals added in mixture (Cd, Cr, Pb, Zn). Our results showed that (i) the presence of heavy metals delayed rhizome regeneration and induced lowered plant part weights but did not affect plant height after 3 months; (ii) the effect of metals on the metabolic profiles of belowground part extracts was only detectable after 1 month and not after 3 months of growth, though it was possible to highlight the effect of metals independently of time and genotype for root extracts, and torosachrysone seemed to be the most induced compound; and (iii) the hybrid genotype tested was able to accumulate relatively high concentrations of metals, over or close to the highest reported ones for this plant for Cr, Cd and Zn, whereas Pb was not accumulated. These findings evidence that the presence of heavy metals in soil has a low impact on Fallopia sp. overall performance traits during rhizome regeneration, and has a rather stimulating effect on plant growth depending on pollution level.

A Mathematical Model for Biocontrol of the Invasive Weed Fallopia japonica.

We propose a mathematical model for biocontrol of the invasive weed Fallopia japonica using one of its co-evolved natural enemies, the Japanese sap-sucking psyllid Aphalara itadori. This insect sucks the sap from the stems of the plant thereby weakening it. Its diet is highly specific to F. japonica. We consider a single isolated knotweed stand, the plant's size being described by time-dependent variables for total stem and rhizome biomass. It is the larvae of A. itadori that damage the plant most, so the insect population is described in terms of variables for the numbers of larvae and adults, using a stage-structured modelling approach. The dynamics of the model depends mainly on a parameter h, which measures how long it takes for an insect to handle (digest) one unit of F. japonica stem biomass. If h is too large, then the model does not have a positive equilibrium and the plant biomass and insect numbers both grow together without bound, though at a lower rate than if the insects were absent. If h is sufficiently small, then the model possesses a positive equilibrium which appears to be locally stable. The results based on our model imply that satisfactory long-term control of the knotweed F. japonica using the insect A. itadori is only possible if the insect is able to consume and digest knotweed biomass sufficiently quickly; if it cannot, then the insect can only slow down the growth which is still unbounded.

Vegetative regeneration capacities of five ornamental plant invaders after shredding.

Vegetation management often involves shredding to dispose of cut plant material or to destroy the vegetation itself. In the case of invasive plants, this can represent an environmental risk if the shredded material exhibits vegetative regeneration capacities. We tested the effect of shredding on aboveground and below-ground vegetative material of five ornamental widespread invaders in Western Europe that are likely to be managed by cutting and shredding techniques: Buddleja davidii (butterfly bush, Scrophulariaceae), Fallopia japonica (Japanese knotweed, Polygonaceae), Spiraea × billardii Hérincq (Billard's bridewort, Rosaceae), Solidago gigantea (giant goldenrod, Asteraceae), and Rhus typhina L. (staghorn sumac, Anacardiaceae). We looked at signs of vegetative regeneration and biomass production, and analyzed the data with respect to the season of plant cutting (spring vs summer), the type of plant material (aboveground vs below-ground), and the shredding treatment (shredded vs control). All species were capable of vegetative regeneration, especially the below-ground material. We found differences among species, but the regeneration potential was generally still present after shredding despite a reduction of growth rates. Although it should not be excluded in all cases (e.g., destruction of giant goldenrod and staghorn sumac aboveground material), the use of a shredder to destroy woody alien plant material cannot be considered as a general management option without significant environmental risk.

Plant litter chemistry and microbial priming regulate the accrual, composition and stability of soil carbon in invaded ecosystems.

Soil carbon (C) sequestration, as an ecosystem property, may be strongly influenced by invasive plants capable of depositing disproportionately high quantities of chemically distinct litter that disrupt ecosystem processes. However, a mechanistic understanding of the processes that regulate soil C storage in invaded ecosystems remains surprisingly elusive. Here, we studied the impact of the invasion of two noxious nonnative species, Polygonum cuspidatum, which produces recalcitrant litter, and Pueraria lobata, which produces labile litter, on the quantity, molecular composition, and stability of C in the soils they invade. Compared with an adjacent noninvaded old-field, P. cuspidatum-invaded soils exhibited a 26% increase in C, partially through selective preservation of plant polymers. Despite receiving a 22% higher litter input, P. lobata-invaded Pinus stands exhibited a 28% decrease in soil C and a twofold decrease in plant biomarkers, indicating microbial priming of native soil C. The stability of C exhibited an opposite trend: the proportion of C that was resistant to oxidation was 21% lower in P. cuspidatum-invaded soils and 50% higher in P. lobata-invaded soils. Our results highlight the capacity of invasive plants to feed back to climate change by destabilizing native soil C stocks and indicate that environments that promote the biochemical decomposition of plant litter would enhance the long-term storage of soil C. Further, our study highlights the concurrent influence of dominant plant species on both selective preservation and humification of soil organic matter.

[Evaluation of quality coherence of prepared slices of polygoni cuspidati rhizoma commercially available in China based on HPLC fingerprint].

OBJECTIVE: To evaluate the quality coherence of the prepared slices of Polygoni Cuspidati Rhizoma commercially available in China and provide a reference for their quality evaluation. METHODS: The fingerprints were obtained using HPLC method, and analyzed with Chromatographic Fingerprint Similarity Evaluation System (2004A Version) provided by Chinese Pharmacopoeia Commission. The experiment was carried out with an Agilent TC-C18 column (250 mm x 4.6 mm, 5 microm) using acetonitrile-0.05% aqueous solution of phosphoric acid as mobile phase at the flow rate of 1.0 mL/min, detection wavelength of 230 nm, and temperature of 25 degrees C. RESULTS: The fingerprint for Polygoni Cuspidati Rhizoma was established. 21 common characteristic peaks were identified and the similarities of 24 batches of prepared slices of Polygoni Cuspidati Rhizoma were between 0.882 - 0.988. CONCLUSION: The quality of the prepared slices of Polygoni Cuspidati Rhizoma commercially available in China is comparable.

Elucidating the population dynamics of Japanese knotweed using integral projection models.

Plant demographic studies coupled with population modeling are crucial components of invasive plant management because they inform managers when in a plant's life cycle it is most susceptible to control efforts. Providing land managers with appropriate data can be especially challenging when there is limited data on potentially important transitions that occur belowground. For 2 years, we monitored 4 clonal Japanese knotweed (Polygonumcuspidatum) infestations for emergence, survival, shoot height until leaf senescence, dry shoot biomass after senescence, and rhizome connections for 424 shoots. We developed an integral projection model using both final autumn shoot height and shoot biomass as predictors of survival between years, growth from year to year, and number of rhizomes produced by a shoot (fecundity). Numbers of new shoots within an infestation (population growth rate λ) were projected to increase 13-233% in a year, with the greatest increase at the most frequently disturbed site. Elasticity analysis revealed population growth at 3 of the 4 sites was primarily due to ramet survival between years and to year-to-year growth in shoot height and shoot biomass. Population growth at the fourth site, the most disturbed, was due to the large production of new rhizomes and associated shoots. In contrast to previous studies, our excavation revealed that most of the shoots were not interconnected, suggesting rhizome production may be limited by the size or age of the plants, resource availability, disturbance frequency, or other factors. Future integration of plant population models with more data on belowground growth structures will clarify the critical stages in Japanese knotweed life cycle and support land managers in their management decisions.

Quality assessment of Japanese knotweed (Fallopia japonica) grown on Prince Edward Island as a source of resveratrol.

Japanese knotweed (Fallopia japonica , also known as Polygonum cuspidatum) is a common invasive plant species on Prince Edward Island (PEI), Canada, whereas it has been used in Chinese medicine and more recently as a raw material for extracting resveratrol. This paper reports on the quantification of resveratrol, polydatin, emodin, and physcion in roots, stems, and leaves of Japanese knotweed samples from PEI and British Columbia (BC), Canada, and nine provinces of China, by ultraperformance liquid chromatography (UPLC). The results showed that the root contains a much higher level of resveratrol than the stem and leaf, and it is accumulated in its highest level in October. PEI-grown knotweed contains similar levels of resveratrol and polydatin compared to Chinese samples collected in the month of October, but the contents of the other anthraquinones (emodin and physcion) are different. As such, Japanese knotweed grown in PEI could be a commercially viable source of raw material for resveratrol production; however, caution has to be taken in harvesting the right plant species.

Can land managers control Japanese knotweed? Lessons from control tests in Belgium.

Japanese knotweed Fallopia japonica is an extremely abundant invasive plant in Belgium and surrounding countries. To date, no eradication method is available for land managers facing the invasion of this rhizomatous plant. We tested different chemical herbicides with two application methods (spraying and stem injection), as well as mechanical treatments, on knotweed clones throughout southern Belgium. The tested control methods were selected to be potentially usable by managers, e.g., using legally accepted rates for herbicides. Stem volume, height and density reduction were assessed after one or two years, depending on the control method. Labor estimations were made for each control method. No tested control method completely eradicated the clones. Stem injection with glyphosate-based herbicide (3.6 kg ha(-1) of acid equivalent glyphosate) caused the most damage, i.e., no sprouting shoots were observed the year following the injection. The following year, though, stunted shoots appeared. Among the mechanical control methods, repeated cuts combined with native tree transplanting most appreciably reduced knotweed development. The most efficient methods we tested could curb knotweed invasion, but are not likely to be effective in eradicating the species. As such, they should be included in a more integrated restoration strategy, together with prevention and public awareness campaigns.

Invasion of diverse habitats by few Japanese knotweed genotypes is correlated with epigenetic differentiation.

The expansion of invasive species challenges our understanding of the process of adaptation. Given that the invasion process often entails population bottlenecks, it is surprising that many invasives appear to thrive even with low levels of sequence-based genetic variation. Using Amplified Fragment Length Polymorphism (AFLP) and methylation sensitive-AFLP (MS-AFLP) markers, we tested the hypothesis that differentiation of invasive Japanese knotweed in response to new habitats is more correlated with epigenetic variation than DNA sequence variation. We found that the relatively little genetic variation present was differentiated among species, with less differentiation among sites within species. In contrast, we found a great deal of epigenetic differentiation among sites within each species and evidence that some epigenetic loci may respond to local microhabitat conditions. Our findings indicate that epigenetic effects could contribute to phenotypic variation in genetically depauperate invasive populations. Deciphering whether differences in methylation patterns are the cause or effect of habitat differentiation will require manipulative studies.

[Fluorescence microscopy and HPLC assay for rapid detection of distribution and content of resveratrol in Polygonum cuspidatum].

OBJECTIVE: To establish fluorescence microscopy combined with HPLC method for rapid detection the distribution and content of resveratrol tissues in different growth stages of Polygonum cuspidatum. METHODS: Used sequential experiment to design conditions of frozen and observe of the section by fluorescence microscopy; Resveratrol was extracted by ultrasonic-assisted extraction and its content was detected by HPLC. RESULTS: The results showed that frozen condition for concentration of gum Arabic was from 20% (dipping time was 5 - 6 h) to 40% (2 - 5 min), the freezer temperature was -5 degrees C, and the thickness was 15 microm. Resveratrol in polygonum cuspidatum was mainly accumulated in the organs, tissues and cells of fiber and cellulose, its content in rhizomes declined as the following sequence: spinal cord > xylem > phloem > periderm; Its content declined in organ as the following sequence: buds > rhizomes > ground stem > leaves; The content of resveratrol in root increased with age. CONCLUSION: The results of fluorescence microscopic observation is in accordance with the HPLC results, indicating that the method is simple, fast and reliable, and provides a fast and reliable detection method for the determination of optimum harvesting period of Polygonum cuspidatum and acquisition of quality.

[Effect of different light intensity on Polygonum cuspidatum callus].

OBJECTIVE: To study the effect of light intensity (LI) on the callus induction and resveratrol content of Polygonum cuspidatum. METHOD: Taking leaves and stem segment of P. cuspidatum as explants, the callus was inducted and cultured under different LI. The growth status of callus was observed and analyzed. The content of resveratrol was determined by HPLC. RESULT: The formation and growth of the stem segment callus were optimal under low light treatment (1340-1560 lx). The resveratrol content was 18.350 microg x g(-1) in fresh stem segment and 140.074 microg x g(-1) in dry stem segment, which was two times higher than that of wild P. cuspidatum. CONCLUSION: Introduction and status of P. cuspidatum callus were obvious diverse under different LI, the resveratrol content of P. cuspidatum was enhanced under appropriate LI.

[Contents comparison of resveratrol and polydatin in the wild Polygonum cuspidatum plant and its tissue cultures].

OBJECTIVE: To compare the contents of resveratrol and polydatin in some materials of Polygonum cuspidatum from various sources, so to screen and obtain the suitable cultures for the following metabolism regulation study. METHOD: RP-HPLC method was applied to simultaneously assay resveratrol and polydatin in different samples. RESULT: By the modified methods of extraction and determination, large amount of materials were screened. The results indicated that the contents of resveratrol and polydatin in root and rhizome were evidently higher than those in the leave and stems. The content of polydatin in the seedlings cultured indoor for three months was 1.27% and showed a 1.25-time increse than that in the wild plants, while the content of resveratrol (0.401%) approached that in the wild plants. Both of resveratrol and polydatin could be examined from different tissue cultures of P. cuspidatum, such as the sterile seedlings, callus, suspended cells and hairy roots, and the levels of them were closely related to the growth speed, physiological status and developmental phase. Hairy roots had the highest potentiality in several tested cultures and the increase rate of dry weight was 8.29 when cultured in vitro for 30 days, and showed a 8.4-fold and a 192.8-fold increase compared with those of natural roots and suspended cells, respectively. The content of polydatin in the hairy roots was up to 0.037% and that of resveratrol was 0.007%. CONCLUSION: The established analysis method is rapid, simple and accurate, especially adapted to the simultaneous determination of resveratrol and polydatin in massive biological samples. Hairy-root cultures have the superiority among the tested materials of P. cuspidatum and are suitable for the large-scale biomass and consistent production of efficient constituents.

Applicability and limitations of optimal biomass allocation models: a test of two species from fertile and infertile habitats.

BACKGROUND AND AIMS: The practical applicability of optimal biomass allocation models is not clear. Plants may have constraints in the plasticity of their root : leaf ratio that prevent them from regulating their root : leaf ratio in the optimal manner predicted by the models. The aim of this study was to examine the applicability and limitations of optimal biomass allocation models and to test the assumption that regulation of the root : leaf ratio enables maximization of the relative growth rate (RGR). METHODS: Polygonum cuspidatum from an infertile habitat and Chenopodium album from a fertile habitat were grown under a range of nitrogen availabilities. The biomass allocation, leaf nitrogen concentration (LNC), RGR, net assimilation rate (NAR), and leaf area ratio (LAR) of each species were compared with optimal values determined using an optimal biomass allocation model. KEY RESULTS: The root : leaf ratio of C. album was smaller than the optimal ratio in the low-nitrogen treatment, while it was almost optimal in the high-nitrogen treatment. In contrast, the root : leaf ratio of P. cuspidatum was close to the optimum under both high- and low-nitrogen conditions. Owing to the optimal regulation of the root : leaf ratio, C. album in the high-nitrogen treatment and P. cuspidatum in both treatments had LNC and RGR (with its two components, NAR and LAR) close to their optima. However, in the low-nitrogen treatment, the suboptimal root : leaf ratio of C. album led to a smaller LNC than the optimum, which in turn resulted in a smaller NAR than the optimum and RGR than the theoretical maximum RGR. CONCLUSIONS: The applicability of optimal biomass allocation models is fairly high, although constraints in the plasticity of biomass allocation could prevent optimal regulation of the root : leaf ratio in some species. The assumption that regulation of the root : leaf ratio enables maximization of RGR was supported.