Effects of the invasive plant Mikania micrantha H.B.K. on soil nitrogen availability through allelopathy in South China

Allelopathy has been regarded as a mechanism for successful exotic plant invasion. However, it is not clear if and what effects of allelopathic substances may exert on soil nutrient. The exotic plant Mikania micrantha H.B.K. (M. micrantha) has invaded many forests in south China, and recent studies have suggested it has allelopathic potential for other plants and soil microbial community. Thus, we hypothesized that M. micrantha could influence soil nutrients and N transformation through allelopathy. We measured total C and N, NO₃ ⁻, NH₄ ⁺ and pH of the soil beneath M. micrantha and the adjacent open soil, and then measured the above soil properties after treating soil with 3 concentrations of aqueous extracts of M. micrantha (T₁: 0.005 g ml⁻¹; T₂: 0.025 g ml⁻¹; T₃: 0.100 g ml⁻¹). In addition, a bioassay was conducted to determine the allelopathic potential of the soil beneath M. micrantha. The results showed that M. micrantha significantly affected soil nutrients and N transformation. Soil beneath M. micrantha had inhibitory effects on seed germination and seedling growth of test plant, and had significantly higher C, N, ammonia, net nitrification rate than those of open soil. The plant extracts decreased soil pH, and T₁ decreased it the most, and it increased soil C and N, and T₁ represented the greatest increase in both C and N. The extracts also increased both NO₃ ⁻ and NH₄ ⁺ in soil, whereas no significant difference existed among the 3 extract treatments. Compared to the water control, the soil net mineralization rate was higher under T₁, while lower under T₂ and T₃. However, the extracts increased the soil nitrification rates under all the treatments (T₁, T₂ and T₃). Our results suggest that the water soluble allelochemicals of M. micrantha improve soil nutrient availability, through which the invasive plant M. micrantha may successfully invade and establish in new habitats.     

Fine-scale clonal structure and diversity of invasive plant Mikania micrantha H.B.K. and its plant parasite Cuscuta campestris Yunker

The fine-scale clonal structure in the patch of the invasive plant Mikania micrantha H.B.K. and its plant parasite Cuscuta campestris Yunker were investigated by distinguishing genets using ISSR markers. A high level of clonal diversity (G/N = 0.7, D = 0.9579, E = 0.7778) in M. micrantha and a low level of clonal diversity (G/N = 0.2, D = 0.7632, E = 0.9479) in C. campestris might be due to the different reproductive strategies, different migration rates and different number of founder. Clonal composition of M. micrantha (14 genets of 20 ramets) and C. campestris (4 genets of 20 ramets) were significantly different. There was no relationship between host and parasite genetic distance matrices using Mantel test (r = 0.073, P = 0.150). Spatial autocorrelation analysis of M. micrantha showed that the correlation value was positive and significant at 2 m with an x-intercept of 3.5 and 4.0 m for distance class sizes of 1 and 2 m, respectively. Spatial autocorrelation analysis of C. campestris showed that the correlation value was positive and significant at 2 m and significantly negative at 10 m with an x-intercept of 4.8 and 6.6 m for distance class sizes of 1 and 2 m, respectively.     

The anti-tumor activity of Mikania micrantha aqueous extract in vitro and in vivo

Aqueous extract obtained from Mikania micrantha (MMAE) is commonly used as traditional medicine in some countries. We hypothesized that MMAE may inhibit tumor cell growth, both in an in vitro and in vivo setting. In in vitro experiments, two kinds of human cancer cell lines, K562 and Hela were used to test the anti-tumor activity. Inhibitory concentrations (IC₅₀) were obtained from the inhibition curves fitted by regression analysis, inhibitory rates (%) were calculated by MTT assay, morphological changes were observed by transmission electron microscope (TEM), cell cycles were analyzed by flow cytometry (FCM), and DNA ladders were determined by agarose gel electrophoresis. The in vivo anti-tumor activity was evaluated by calculating the tumor inhibitory rates, thymus index and spleen index of S180-bearing mice. Paraffin-embedded sections were used to test the pathologic changes. The result displayed that the growth of K562 and Hela were enhanced when treated with MMAE at 20 μg/mL after 48 h. Other concentrations of MMAE (50, 100, 200, 400 μg/mL) inhibited the proliferation of both kinds of cells. The IC₅₀ values of K562 and Hela at 48 h were 167.16 and 196.27 μg/mL and at 72 h 98.07 and 131.56 μg/mL, respectively. The effects showed time-dose dependence. MMAE led to damages of organelles and induced apoptosis. These results were confirmed by ladder DNA fragmentation profile. MMAE also increased the percentage of cells in G2/M phase and decreased the percentage of cells undergoing G0/G1 and S phase in in vivo tests using S180 cells. MMAE showed antitummor activity in vivo, with its tumor inhibitory rate ranging from 12.1 to 46.9 %. MMAE also induced necrosis, as shown by pathological examination of Hematoxilin-Eosin stained tumor sections. Meanwhile, compared with the control group, the changes of thymus index and spleen index in MMAE treated group were not obvious. This study suggests that MMAE may be an effective agent for cancer therapy with low toxicity.     

Isoflavanones from the allelopathic aqueous root exudate of Desmodium uncinatum

Three isoflavanones, 5,7,2',4'-tetrahydroxy-6-(3-methylbut-2-enyl)isoflavanone (1), 4",5"-dihydro-5,2',4'-trihydroxy-5"-isopropenylfurano-(2",3";7,6)-isoflavanone (2) and 4",5"-dihydro-2'-methoxy-5,4'-dihydroxy-5"-isopropenylfurano-(2",3";7,6)-isoflavanone (3) and a previously known isoflavone 5,7,4'-trihydroxyisoflavone [genistein (4)] were isolated and characterised spectroscopically from the root exudate of the legume Desmodium uncinatum (Jacq.) DC. We propose the names uncinanone A, B, and C for compounds 1, 2 and 3, respectively. Isolated fractions containing uncinanone B (2) induced germination of seeds from the parasitic weed Striga hermonthica (Del.) Benth. and fractions containing uncinanone C (3) moderately inhibited radical growth, the first example of a newly identified potential allelopathic mechanism to prevent S. hermonthica parasitism.     

Changes in Soil Microbial Community Associated with Invasion of the Exotic Weed, Mikania micrantha H.B.K

Invasions of exotic plant species are among the most pervasive and important threats to natural ecosystems. However, the effects of plant invasions on soil processes and soil biota have not been adequately investigated. Changes were studied in soil microbial communities where Mikania micrantha was invading a native forest community in Neilingding Island, Shenzhen, China. The soil microbial community structure (assessed by phospholipid fatty acid [PLFA] profiles) and function (assessed by enzyme activities), as well as soil chemical properties were measured. The results showed that the invasion of M. micrantha into the evergreen broadleaved forests in South China changed most of the characteristics in studied soils. Microbial community structure and function differed significantly among the native, two ecotones, and exotic-derived soils. For PLFA profiles, we observed a significant increase in aerobic bacteria but a decrease in anaerobic bacteria in the M. micrantha monoculture as compared to the native and ecotones. The ratio of cy19:0 to18:1ω7 gradually declined but mono/sat PLFAs increased as M. micrantha became more dominant. Both ratios were significantly related to pH according to regression analysis, therefore, pH was a sensitive indicator reflecting the invaded soil subsystem succession. The microbial community composition clearly separated the native soil from the invaded soils by principal component analysis (PCA) and discriminant analysis (DA). For enzyme activities, 7 of 9 enzymes (β-glucosidase, invertase, protease, urease, acid phosphatase, alkaline phosphatase, and phenol oxidase) showed the similar trend that the activities were highest in the exotic, intermediate in the two ecotones, and lowest in the native community. In most cases, enzyme activities were influenced by soil chemical properties, especially by pH value and soil organic matter. Differences in the structural variables were well correlated to differences in the functional variables as demonstrated by canonical correlation analysis (CCA). It was concluded that M. micrantha invasion had profound effects on the soil subsystem, which must be taken into account when we try to control its invasions.     

Leaf and root extracts of the invasive shrub, Lonicera maackii, inhibit seed germination of three herbs with no autotoxic effects

In addition to effects mediated by resource competition, some invasive plants may impact surrounding vegetation by secreting compounds that are directly inhibitory to growth. Lonicera maackii, an invasive Asian shrub of forests and open areas in eastern and midwestern North America, has devastating effects on understory vegetation, some of which persist even after this shrub is removed. In this study, we explored the potential of aqueous extracts of the leaves and roots of this plant to inhibit seed germination of Impatiens capensis, Alliaria petiolata, Arabidopsis thaliana, and L. maackii in Petri dish bioassays. Both L. maackii root and leaf extracts significantly decreased germination in the three herb species. This inhibitory effect generally increased with increasing extract concentration and was more pronounced with application of leaf extract than root extract. However, when the same extracts were applied to seeds of L. maackii itself, germination was delayed in some cases, but was not significantly reduced by the end of the experiment. Germination of L. maackii seeds even reached significantly higher levels in some extract treatments than in no-extract controls. This implies that L. maackii can successfully inhibit the germination of other plants with few autotoxic effects and may even promote the germination of its own seeds.     

Effects ofMikania micrantha H.B.K. on germination and growth of weed species

Laboratory, greenhouse and field studies were conducted to determine the allelopathic potential ofMikania micrantha H.B.K. on the germination and growth of three weeds, namelyAsystasia intrusa Bl.,Chrysopogon aciculatus (Retz.) Trin. andPaspalum conjugatum Berg. The plant height of the test species decreased with increasing amounts of debris when either leaf or root debris of Mikania was present on the soil surface or incorporated into the soil. In general, incorporated debris caused greater reduction in height and seedling fresh weight than debris placed on the soil surface. Paspalum was more sensitive to either root or leaf of Mikania when incorporated into soil than on the soil surface. Leachate of Mikania leaf caused a significant reduction in radicle length and fresh weight of the seedlings of the test species. However, only Paspalum seeds showed significant decrease in germination when exposed to leaf leachate. Full-strength extract of either leaf or root caused a significant decrease in both germination and fresh weight of the test species. At the highest concentration, root extract of Mikania caused a marked reduction of radicle elongation of Paspalum.     

Production of ajmalicine and ajmaline in hairy root cultures of Rauvolfia micrantha Hook f., a rare and endemic medicinal plant

Hairy roots of Rauvolfia micrantha were induced from hypocotyl explants of 2–3 weeks old aseptic seedlings using Agrobacterium rhizogenes ATCC 15834. Hairy roots grown in half-strength Murashige & Skoog (MS) medium with 0.2 mg indole 3-butyric acid l^–1 and 0.1 mg -naphthaleneacetic acid l^–1 produced more ajmaline (0.01 mg g^–1 dry wt) and ajmalicine (0.006 mg g^–1 dry wt) than roots grown in auxin-free medium. Ajmaline (0.003 mg g^–1 dry wt) and ajmalicine (0.0007 mg g^–1 dry wt) were also produced in normal root cultures. This is the first report of production of ajmaline and ajmalicine in hairy root cultures of Rauvolfia micrantha.     

Differential allelopathic expression of bark and seed of Tamarindus indica L.

Allelopathic performance of the bark and seed of Tamarindus indica L. tree was evaluated through bioassay-guided studies using seven common agronomic crops (asparagus, cucumber, lettuce, radish, sesame, tomato and welsh onion) and seven weed species (barnyard grass, Chinese milk vetch, perennial ryegrass, phacelia, timothy grass, white clover and wild ginger) under laboratory conditions. As demonstrated by a sandwich method, the bark of the tamarind tree caused strong growth inhibition (compared to the corresponding controls) in both radicles and hypocotyls of the species tested, and the inhibitory effect was highest in barnyard grass (52–65%) and lowest in welsh onion (19–13%). The crude-water soluble extracts of bark at different concentrations (1, 5 and 10%) (w/v) exhibited a strong growth inhibition in all the plant species tested, and a proportional increase in the percentage of growth inhibition was observed with an increase in the concentrations of the extracts. The magnitude of inhibition in weed species was higher (5–60%) than those of agronomic crop species (3–40%). The growth of all the weed species tested was strongly inhibited (17–56%), while the agronomic crop species showed both inhibited (5–21%) and stimulated (5–27%) growth due to the effect of crude-water soluble exudates of tamarind seed. Among the agronomic crop species tested, lettuce (22–27%) followed by radish (20–25%) and sesame (5–8%) showed stimulatory growth with the crude-water soluble exudates of seed. In the pot culture experiments using four agronomic crops (lettuce, radish, tomato and cucumber) and two weed species (barnyard grass and white clover), spraying of crude-water soluble extracts of tamarind seed-coat at three different concentrations (1, 5 and 10%) (w/v) showed that the growth of lettuce (35–62%) and radish (32–56%) was stimulated, while all other species tested showed growth inhibition (29–61%). When the spraying of crude extracts of seed-coat was turned off, the growth of both lettuce and radish continued to be stimulated (4–7%) and all other previously inhibited species recovered well, the recovery percentage ranging between 78 and 82%. However, when spraying of crude extracts of seed-coat was continued, growth increased (10–14%) in lettuce and radish, and reduced (37–76%) in four other species tested. The inhibitory or stimulatory effects of the crude extracts on agronomic crop and weed species were higher in the radicle than the hypocotyl and reached a peak with 10% (w/v) concentrations. These results clearly demonstrated the differential allelopathic effects (inhibitory and excitatory) of bark and seed of tamarind tree in the species tested. Thus, it is evident that these two organs contain certain biologically active true growth regulator(s) and are either additively or synergistically involved in the plant-specific expression, particularly by the seed-coat.     

The allelopathic potential of alfalfa root medicagenic acid glycosides and their fate in soil environments

Summary The allelopathic effect of alfalfa (Medicago media Pers.) and red clover (Trifolium pratense L.) root saponins on winter wheat seedling growth and the fate of these chemicals in soil environments were studied. Seed germination, seedling and test fungus growth were suppressed by water and by alcohol extracts of alfalfa roots, and by crude saponins of alfalfa roots, indicating that medicagenic acid glycosides are the inhibitor. Powdered alfalfa roots inhibited wheat seedling growth when added to sand. At concentrations as low as 0.25% (w/w) the root system was completely destroyed whereas seedling shoots suffered little damage. Red clover roots caused some wheat growth inhibition when incorporated to sand, but their effect was much lower than in the alfalfa root treatment. Soil textures had a significant influence on the inhibitory effect of alfalfa roots. The inhibition of seedling growth was more pronounced on light than on heavy soils. This was attribted to the higher sorption of inhibitors by heavy soils. Incubation of alfalfa roots mixed into loose sand, coarse sand, loamy sand and clay loam for a period of 0–8 days resulted in decreased toxicity to bothT. viride and wheat seedlings. This decrease occurred more quickly in heavier soils than in loose sand, due to the hydrolysis of glycosides by soil microorganisms. Soil microbes were capable of detoxifying medicagenic acid glycosides by partial hydrolysis of sugar chain to aglycone. These findings illustrate the importance of medicagenic acid glycosides as an inhibitor of wheat seedling growth, and of their fate in different soil environments.     

入侵植物薇甘菊新枝在海南的月生长动态

【背景】薇甘菊是入侵性和危害性极强的外来入侵植物,对生物多样性和农林业生产造成严重影响。观测薇甘菊新枝的月生长动态,了解薇甘菊快速生长扩散的原因,能够为薇甘菊的合理防治提供参考。【方法】采用定点定时观测法,历时2年对海南7个市县薇甘菊新枝的月生长量进行测定。【结果】薇甘菊在海南全年可生长,一新枝月平均生长66 cm,全年累计平均生长795 cm。薇甘菊生长高峰期在6月和8月,其中6月平均生长120 cm,8月平均生长106 cm。薇甘菊最小生长量在1月和12月,平均生长量均为21 cm。薇甘菊在6—10月生长较快,一新枝累计平均生长489 cm,占全年生长量的61.5%;在4—5月和11月生长次之,一新枝累计平均生长196 cm,占全年生长量的24.7%;在1—3月和12月生长缓慢,一新枝累计平均生长110 cm,占全年生长量的13.8%。薇甘菊的花果期在10月至次年3月,一些种群在1—3月再次开花或在3—5月开花。薇甘菊的生长量与温度呈极显著正相关,相关系数为0.91;与降雨量呈显著正相关,相关系数0.56。【结论与意义】薇甘菊是多分枝植物,一新枝年生长量可达795 cm,能迅速生长形成种群。温度是薇甘菊生长的主要因素,降雨在一定程度促进了薇甘菊的生长。     

In vitro propagation of Rauwolfia micrantha, a rare medicinal plant

Shoot tip and single node explants from young shoots of 1-year old flowering plants of Rauwolfia micrantha Hook. f. were cultured on Murashige & Skoog (MS) medium variously supplemented with 6-benzyladenine (BA) and -naphthaleneacetic acid (NAA). A combination of 13.2 M BA and 2.68 M NAA induced high frequency (77%) formation of up to 3 shoots from each node in 8 weeks. The regeneration of shoot tips from the field-grown plants and in vitro shoots placed horizontally differed. Repeated subculturing of the shoot tips and single nodes at 6-week intervals for over a year in combination of 4.4 M BA and 0.27 M NAA enabled mass multiplication of shoots without any evidence of decline. Rooting of the excised shoots on medium containing 2.6 M NAA was preceded by callus formation. The rooted plants were removed off the callus, hardened off and 80% established in pots. Micropropagated plants displayed uniform morphological, growth, flowering, fruiting and seed germination characteristics.Abbreviations BA 6-benzyladenie - IAA indole-3-acetic acid - IBA indole-3-butyrie acid - 2-ip 2-isopentenyladenine - MS Murashige & Skoog (1962) - NAA -naphthaleneacetic acid     

Plantlet differentiation in leaf and root cultures of birch (Betula Pendula roth.)

The newly-formed leaves on plantlets differentiated from shoot bud cultures of Betula pendula, when excised and grown on a fresh medium produced callus from the margins or regenerated leafy shoots, roots and plantlets. After 4 weeks, upon transfer to murashige and Skoog (MS) medium supplemented with 3-indoleacetic acid (IAA) + 6-(4-hydroxy-3-methyl-trans-2-enyl)aminopurine (zeatin) + 6-aminopurine (adenine), 15–20 plantlets were produced from each explant. Likewise, the roots also showed meristematic activity at several sites, and produced nodulated callus on MS + α-naphthaleneacetic acid (NAA) + 6-(3-methyl-2-butenyl-amino)purine (2-iP) + adenine, and ultimately differentiated plantlets. Anatomical studies showed that initiation of callus takes place by meristematic activity in epidermal cells of leaves, and cortical cells of roots. Cytological investigations revealed no change in chromosomal complement.     

Allelopathic potential of Bothriochloa laguroides var. laguroides (DC.) Herter (Poaceae: Andropogoneae)

Bothriochloa laguroides var. laguroides (DC.) Herter, a native bluestem of America, has been shown to produce many biologically active compounds. The allelopathic potential of aqueous extracts from roots, stems and leaves was examined. Lettuce seeds (Lactuca sativa) and maize (Zea mays), the common allelopathy bioassay systems, as well as seeds from two native species, wintergreen paspalum (Paspalum guenoarum) and lovegrass (Eragrostis curvula), were germinated in the presence of aqueous extracts. Percent seed germination, root and shoot elongation were measured. After 4 and 7 days root, stem and leaf extracts caused inhibition of root and shoot elongation in all four species tested. Aqueous extracts were generally less inhibitory to seed germination. Aqueous extracts from different parts of B. laguroides var. laguroides show therefore allelopathic effects inhibiting, in particular, growth of competing plants.     

Some morphological and anatomical observations during alleviation of salinity (NaCI) stress on seed germination and seedling growth of barley by polyamines

In this work, the effects of polyamine (cadaverine, putrescine, spermidine, spermine) pretreatments on the germination of barley seeds, seedling growth under saline (NaCI) conditions and on leaf anatomy of the seedlings grown for 20 days in pots with perlite containing different concentrations of NaCI prepared with Hoagland solutions following the germination period of 7 days were studied. The inhibitive effect of salt on seed germination and seedling growth was alleviated in varying degrees, and dramatically, by polyamine pretreatments at the levels of NaCI studied; particularly with lower levels of salt, the control seeds were able to germinate with great difficulty. Also, the successes of these pretreatments continued, even decreased, with higher levels of NaCI such that the control seeds showed no germination. On the other hand, on the various parameters of leaf anatomy of barley seedlings, interactions between pretreatments of polyamine and salinity, with some of these being statistically important, were observed.     

Interacting effects of grass height and herbivores on the establishment of an encroaching savanna shrub

Shrub encroachment is a widely observed problem in Southern African savannas. Although the effects of herbivory and grass height on woody species recruitment have been studied individually, little information exists about how these factors interact. In this study seeds and seedlings of the encroaching shrub Dichrostachys cinerea were planted in clipped and unclipped grass plots, with and without large herbivores present. Seed germination, seedling survival and seedling predation were monitored for 8 months. Germination started earlier in plots where herbivores were excluded. Overall, the earlier the seeds germinated, the longer the seedlings survived. Clipping positively affected the number of germinated seeds, seedling growth and survival but effects varied among herbivore exclusion treatments and sites. Invertebrates caused the majority of the seedling damage. We conclude the recruitment of D. cinerea is influenced by the interplay of grass height and herbivory. In this study, the presence of large herbivores early in the wet season, and the absence of simulated grazing later on, affected the regeneration of D. cinerea negatively. However, differences in effects among sites suggest that the mechanisms found here may work differently in other habitats.      

Effects of leaf litter on woody seedlings in xeric successional communities

We investigated the effect of leaf litter on the establishment of Eucalyptus incrassata, a mallee eucalypt. It has been suggested that litter accumulation may hinder seedling establishment, and that the removal of litter may be one of the mechanisms through which fire enhances recruitment. We conducted factorial experiments testing the effects of three kinds of leaf litter on E. incrassata seeds and seedlings at three contiguous sites with different land use histories. One site was an uncleared E. incrassata open mallee woodland (Mallee site), one a cleared area that had been ungrazed for about five years (Pasture site) and the third an area of mallee rolled some 40 years ago and permitted to regenerate (Regrowth site). Litter had no effect on emergence of planted E. incrassata seeds, but emergence differed between sites. Overall, the percentage of seeds that germinated and emerged was substantial (mean 35.2% ± 25.9%). Seedling shoot biomass did not differ between sites or litter treatments. Although seedlings grown in Pasture litter suffered higher mortality rates, overall mortality rates were low (mean 13.2% ± 15.5%), suggesting that leaf litter has little effect on recruitment rates during winter and spring. We conclude that leaf litter does not affect emergence or growth in young E. incrassata seedlings during winter and spring, when most establishment occurs. Our results emphasize the difficulty in predicting litter effects on recruitment.     

Differential effects of salinity on leaf elongation kinetics of three grass species

This study focuses on the inhibitory effect of salinity on the leaf extension of three different grass species: Hordeum jubatum L., Hordeum vulgare L. and Zea mays L. Leaf elongation rates (LER) were measured on the third leaf of the plants. NaCl was added to the hydroponic solution (0, 40, 80 and 120 mM) and changes in LER were measured over time with a displacement transducer. Salinity inhibited LER immediately in all three species, and a new, but lower steady-state LER was reached within 5 h. The decrease in LER was proportional to the salinity level. Differences in salt tolerance (% of control LER) were evident between genotypes within 5 h after salinization, but the relative salt tolerance of the plant at this stage was not necessarily indicative of the long-term salt tolerance of the species. In general, H. jubatum was more tolerant than maize, which was more tolerant than barley to these short-term salinity stresses. In contrast, barley is more salt tolerant than maize over the long term. The mechanisms of inhibition of LER by salinity, as tested by the applied-tension technique, varied with the species examined, affecting either the apparent yield threshold, the hydraulic conductance of the whole plant or both. The cell wall extensibility was not significantly affected by salinity in the three species tested in this study.     

Tillage and N fertilization effects on maize root growth and root:shoot ratio

Two methods for estimating the size of the maize (Zea mays l.) root system from soil cores taken in the field were compared. The spatially weighed block method of estimation accounted for variation in root density by using 18 samples per plant which varied in distance from plant and soil depth. This method was compared to an estimation which averaged all of the 18 samples together. Both methods gave surprisingly similar estimates for total root growth. Increased root growth in the surface soil layers, due to tillage and N fertilization, did not impact on the estimation of total root growth. Total root length remained unchanged or increased with N fertilization, while root weight remained the same or decreased. Root mass per length decreased with N fertilization. The estimated size of the root system was used to calculate root:shoot weight ratios. The largest root:shoot ratio was found in the vegetative stage and decreased throughout the rest of the season. In this field experiment, the estimated size of the root system at 8 weeks after planting was not significantly different from the size at silking or harvest. Nitrogen fertilization significantly decreased the root:shoot weight ratio. However, tillage did not significantly change the ratio.