Intrinsic soil property effects on Cd phytotoxicity to Ligustrum japonicum 'Howardii' expressed as different fractions of Cd in forest soils.

A better comprehensive understanding of the influence of soil/solution properties on cadmium (Cd) phytotoxicity is essential for soil Cd ecological risk assessment. The toxicity of soil spiked Cd to Ligustrum japonicum 'Howardii' seedling growth was conducted by the greenhouse pot experiments using 13 typical forest soils selected from mainland of China. The results showed that the ranges of Cd toxicity thresholds of 10% seedling growth inhibition (EC10) and 50% inhibition (EC50) followed the order: soil pore water Cd (EC10 on average 0.88 mg L-1 with the variation of 54.9 folds and EC50 on average 2.28 mg L-1 with variation of 41.8 folds), DTPA extractable Cd (EC10 on average 5.4 mg kg-1 with 20.9 folds variation and EC50 on average 17.86 mg kg-1 with 6.6 folds variation), total added Cd (EC10 on average 6.55 mg kg-1 with 16.7 folds variation and EC50 on average 22.11 mg kg-1 with 5.1 folds variation), which suggested that whatever the available Cd expressed, its toxicity is largely affected by soil properties. The empirical multiple equations were well developed between different fractions of Cd toxicity thresholds ECx (x = 10 or 50) and soil/solution. The results also showed that the pH inversely correlated with EC10 (r2 = 0.54, P < 0.01) and EC50 (r2 = 0.63, P < 0.001) based on soil pore water, indicating the ECx decreased with more toxicity as pH increased. No single significant soil solution properties were found for ECx in DTPA extractable Cd. For the ECx of DTPA extractable and total Cd, the content of aluminum oxides in soil and soil pH were the two significant factors inversely related with ECx, which explained 68%-79% of the inter-soil variation, respectively. Overall, soil or solution pH was the most important factor controlling Cd toxicity thresholds. Meanwhile, significant negative correlations existed between the soil solution pH and the slopes of parameter (b) of the dose-response curves for different fractions of Cd, implying that the growth of toxic effect enhanced as unit Cd dosage increased in low pH soils. These results will be helpful to evaluate the metal ecological risk in forest soils.

Can arbuscular mycorrhizal fungi from non-invaded montane ecosystems facilitate the growth of alien trees?

It is generally assumed that recruitment and expansion of alien species along elevation gradients are constrained by climate. But, if plants are not fully constrained by climate, their expansion could be facilitated or hindered by other factors such as biotic interactions. Here, we assessed the composition of arbuscular mycorrhizal fungi (AMF) in soils along an elevation gradient (i.e. 900 m, 1600 m, 2200 m and 2700 m a.s.l.) through a fungal DNA meta-barcoding approach. In addition, we studied in the greenhouse the effects of AMF on growth and phosphorous (P) nutrition of seedlings of the alien trees Gleditsia triacanthos, Ligustrum lucidum and Pyracantha angustifolia cultivated in soils from those elevations, spanning the elevation at which they already form monospecific stands (below 1450 m a.s.l.) and higher elevations, above their current range of distribution in montane ecosystems of Central Argentina. For comparison, we also included in the experiment the dominant native tree Lithraea molleoides that historically occurs below 1300 m a.s.l. Arbuscular mycorrhizal fungal community composition showed strong community turnover with increasing elevation. The effects of these AMF communities on plant growth and nutrition differed among native and alien trees. While P nutrition in alien species' seedlings was generally enhanced by AMF along the whole gradient, the native species benefited only from AMF that occur in soils from the elevation corresponding to its current altitudinal range of distribution. These results suggest that AMF might foster upper range expansion of these invasive trees over non-invaded higher elevations.

Effects of lead stress on the growth, physiology, and cellular structure of privet seedlings.

In this study, we investigated the effects of different lead (Pb) concentrations (0, 200, 600, 1000, 1400 mg kg-1 soil) on the growth, ion enrichment in the tissues, photosynthetic and physiological characteristics, and cellular structures of privet seedlings. We observed that with the increase in the concentrations of Pb, the growth of privet seedlings was restricted, and the level of Pb ion increased in the roots, stem, and leaves of the seedlings; however, most of the ions were concentrated in the roots. Moreover, a decreasing trend was observed for chlorophyll a, chlorophyll b, total chlorophyll, net photosynthesis (Pn), transpiration rate (Tr), stomatal conductance (Gs), sub-stomatal CO2 concentration (Ci), maximal photochemical efficiency (Fv/Fm), photochemical quenching (qP), and quantum efficiency of photosystem II (ΦPSII). In contrast, the carotene levels, minimum fluorescence (F0), and non-photochemical quenching (qN) showed an increasing trend. Under Pb stress, the chloroplasts were swollen and deformed, and the thylakoid lamellae were gradually expanded, resulting in separation from the cell wall and eventual shrinkage of the nucleus. Using multiple linear regression analysis, we found that the content of Pb in the leaves exerted the maximum effect on the seedling growth. We observed that the decrease in photosynthetic activation energy, increase in pressure because of the excess activation energy, and decrease in the transpiration rate could result in maximum effect on the photosynthetic abilities of the seedlings under Pb stress. Our results should help in better understanding of the effects of heavy metals on plants and in assessing their potential for use in bioremediation.

Fundamental Host Range of Leptoypha hospita (Hemiptera: Tingidae), a Potential Biological Control Agent of Chinese Privet.

Chinese privet, Ligustrum sinense Lour., is an invasive shrub within riparian areas of the southeastern United States. Biological control is considered the most suitable management option for Chinese privet. The potential host range of the lace bug, Leptoypha hospita Drake et Poor, was evaluated on the basis of adult feeding and oviposition, combined oviposition-nymphal development no-choice tests, nymphal development no-choice tests, multiple generation comparison on Forestiera pubescens Nutt. and L. sinense no-choice tests, and multiple-choice tests with 45 plant species in 13 families. No-choice tests showed that the host range of L. hospita was restricted to the tribe Oleeae. In adult feeding and oviposition no-choice tests, the bug fed and oviposited significantly more on Chinese privet than all other test plant species except for three native Forestiera spp., two nonnative Syringa spp., and another exotic Ligustrum sp. Among those, only F. pubescens supported complete development in numbers comparable to Chinese privet. However, when reared for multiple generations lace bugs reared on F. pubescens were smaller and had lower fecundity than those reared on L. sinense, suggesting F. pubescens is not an optimal host. In multiple-choice tests, L. hospita displayed a strong preference for feeding and ovipositing on Chinese privet over other test plant species, with the exception of the closely related nonnative Syringa spp. and its congenic species Ligustrum vulgare. The results of this study suggest that the risk to nontarget plant species in North America is minimal, and L. hospita would be a promising candidate for Chinese privet biological control.

Quantitative Determination of Secoiridoids and Phenylpropanoids in Different Extracts of Ligustrum Vulgare L. Leaves by a Validated HPTLC-Photodensitometry Method.

INTRODUCTION: The genus Ligustrum (Oleaceae) is distributed in Europe and Asia (south China and Korea), where it is used to prevent hypertension, sore throats, inflammation and diabetes. The main groups of compounds in extracts of Ligustrum vulgare are biologically active secoiridoids and phenylpropanoids. OBJECTIVES: The aim of the study was primarily the development and validation of a HPTLC-photodensitometry method for separation and determination of secoiridoids (oleacein, oleuropein) and phenylpropanoids (echinacoside) in different extracts prepared from leaves of L. vulgare. A secondary issue was the quantitative screening of oleacein, oleuropein and echinacoside in extracts from leaves collected at different stages of plant growth (from May to September). METHODS: A HPTLC-photodensitometry method was developed and validated for quantification of oleuropein, oleacein and echinacoside in plant extracts (aqueous and ethanolic extract, decoction, infusion). Silica gel was used as the stationary phase and dichloromethane:methanol:formic acid:water (80:25:1.5:4, v/v/v/v) as the mobile phase. RESULTS: The HPTLC-photodensitometry method developed for quantification of oleacein, oleuropein and echinacoside was specific, accurate and precise. The presence of oleacein was detected in aqueous extracts, whereas oleuropein was present, in particular, in ethanolic extracts, decoctions and infusions. Echinacoside was detected in all the extracts prepared. The content of secoiridoids was variable from May to September, whereas the amount of echinacoside increased in this term. CONCLUSION: The developed and validated HPTLC-photodensitometry method allowed performing fast screening of quantitative profiles of oleacein, oleuropein and echinacoside in preparations of privet leaves.

Demographic analysis of the fitness of Problepsis superans (Lepidoptera: Geometridae) feeding on three ligustrum (Lamiales: Oleaceae) species.

Using the age-stage, two-sex life table, the effects of three ligustrum species, Ligustrum x vicaryi Hort., Ligustrum quihoui Carrière, and Ligustrum lucidum Aiton, on the fitness of Problepsis superans (Butler, 1885) (Lepidoptera: Geometridae) were assayed by considering life table parameters of P. superans at 27 +/- 1 degrees C, 70 +/- 5% relative humidity, and a photoperiod of 16:8 (L: D) h. The means and SEs of population parameters were calculated using the jackknife and bootstrap methods. The total developmental time of larval stage of P. superans on L. x vicaryi was significantly shorter than that on L. x vicaryi and L. quihoui, whereas higher fecundity was observed on L. x vicaryi. The highest value of the finite rate of increase was observed on L. x vicaryi. The intrinsic rate of increase of P. superans on L. x vicaryi, L. quihoui, and L. lucidum, was 0.147 +/- 0.004, 0.130 +/- 0.004, and 0.112 +/- 0.005, respectively, which differed significantly among the three ligustrum species. The net reproductive rate varied from 122.8 +/- 24.7 female offspring on L. lucidum to 242.2 +/- 36.2 female offspring on L. x vicaryi. The lowest mean generation time was observed on L. x vicaryi. The gross reproductive rate of P. superans on the three ligustrum species did not significantly differ. Based on growth and population parameters, the suitability of the three ligustrum species to P. superans is ranked from high to low in the order as L. x vicaryi, L. quihoui, and L. lucidum.

Seasonal variations in metabolite profiling of the fruits of Ligustrum lucidum Ait.

The metabolite profiling of fruits of the herb Ligustrum lucidum Ait collected during different months has been performed using ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC/QTOFMS) and multivariate statistical analysis techniques. The markers such as oleuropein acid, neonuezhenide, specnuezhenide, oleuropein and ligustrosidic acid accountable for such variations were identified through the loadings plot of principal component analysis (PCA), and the tentative identification of the markers is completed by comparing the mass spectra and retention times with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae and MS/MS data with those of the known compounds published. Furthermore, one of the chemical markers, such as specnuezhenide, which is water-soluble, biologically active and also the predominant compound in this crude drug, was quantified by ultra-performance liquid chromatography coupled with a tunable UV detector (UPLC-TUV). The developed UPLC method provides good linearity (r(2)=0.9991), repeatability (RSD=2.96%), intra- and inter-day precisions (RSD=0.21%, 0.96%), with accuracies of 99.18-100.26% and a recovery of specnuezhenide of 97.57%. The fruits of L. lucidum Ait collected from August to December were tested. The results clearly show that the fruits of L. lucidum Ait harvested in October have the highest yields of specnuezhenide. It is also noted that the variations of content of specnuezhenide obtained by both methods have a strong correlation. This suggests that the newly proposed strategy is a reliable and simple method for the rapid discrimination of subtle variations, within the same plant species or strains, due to different seasonal collection times.

Light regulation to chlorophyll synthesis and plastid development of the chlorophyll-less golden-leaf privet.

Ligustrum vicaryi L. is a hybrid of Ligustrum ovalifolium Hassk. var. aureo-marginatum and Ligustrum vulgale L., and displays a chlorophyll-less phenotype. Therefore it is widely used as a horticultural shrub because of its golden-color leaves. Its putative mechanism, light responses, chlorophyll synthesis and plastid development were studied. L. vicaryi has a higher level of 5-aminolevulinic acid (ALA), but lower levels of chlorophylls compared with L. quihoui. The yellowish phenotype of L. vicaryi upper leaves could be attributed to their hampered conversion from chlorophyllide into chlorophyll a. Despite the enhanced ALA level and the decreased thylakoid stacking in plastids, L. vicaryi golden leaves contain normal levels of Lhcb transcripts and photosystem apoproteins. Furthermore, reactive oxygen species (ROS) accumulation is almost the same in L. vicaryi and L. quihoui. The golden leaves often turn green and the contents of chlorophylls increase with decreasing light intensity. Dynamic changes of chlorophyll-synthesis-system under the light transition were also analyzed.

[Spatial patterns of and specific correlations between dominant tree species in a karst evergreen and deciduous broadleaved mixed forest in Mulun Karst National Nature Reserve].

In order to understand the biological characteristics and specific correlations of dominant tree species in a karst characteristic evergreen and deciduous broad-leaved mixed forest in Mulun National Nature Reserve of Guangxi, a point pattern analysis was made on the spatial distribution patterns and inter- and intraspecific correlations of four dominant species in a one-hectare plot. Among the four species, Boniodendron minius dominated in tree sublayer I, while Ligustrum japonicum, Sinosideroxylon wightianum, and Rapanea kwangsiensis dominated in tree sublayers II and III. All the four species had a clumped distribution at scale <10 m, a transition from clumped to random distribution at scale 10-25 m, and a random or regular distribution at scale >25 m. The critical scale from clumped to random distribution varied with species. No significant correlations were observed between the B. minius in sublayer I and the dominant species in sublayer II. The correlations of B. minius with the dominant species in sublayers II and III showed greater fluctuation, with significant positive correlation for L. japonicum at scale <50 m, no significant correlation for S. wightianum, and no significant correlation for R. kwangsiensis at scale <20 m but significant negative correlation at scale 20-50 m.

Interactions of water stress and solar irradiance on the physiology and biochemistry of Ligustrum vulgare.

We studied the interactive effects of water stress and solar irradiance on physiological and biochemical traits in Ligustrum vulgare L., with special emphasis on antioxidant enzymes and flavonoids. Water relations, photosynthetic performance, plant growth, activities of antioxidant enzymes and of phenylalanine-ammonia-lyase, and concentrations of nonstructural carbohydrates and phenylpropanoids were measured in plants growing in 12% (shade) or 100% (sun) sunlight and supplied with 100 or 40% of daily evapotranspiration-demand over a 4-week period. The mild water stress treatment caused leaf water potential and relative water content to decline on average by -0.22 MPa and 4.5%, respectively. In response to the water stress treatment, photosynthetic rates decreased more in sun plants than in shade plants, likely because of declines in photosystem II photochemistry, apparent maximum rate of carboxylation and apparent maximum electron transport rate coupled with significant reductions in stomatal conductance. Antioxidant enzymatic activities, which were much greater in sun leaves than in shade leaves under well-watered conditions, increased (particularly the enzymatic activities associated with hydrogen peroxide removal) in response to water stress only in shade leaves. Antioxidant phenylpropanoids, particularly quercetin and luteolin derivatives, markedly increased in response to full sunlight irrespective of water treatment; however, antioxidant phenylpropanoid concentrations increased in response to water stress only in shade leaves. We suggest that: (1) assimilated carbon in sun plants was used largely to support an effective antioxidant system capable of countering water-stress-induced oxidative damage--an example of cross tolerance; and (2) in shade plants, carbon was also diverted from growth to counter oxidative damage driven by the mild water-stress treatment. Both findings are consistent with the nearly exclusive distribution of L. vulgare in well-watered, partially shaded Mediterranean areas.

Mobilisation of nutrients and transport via the xylem sap in a shrub (Ligustrum ovalifolium) during spring growth: N and C compounds and interactions.

In open-field soilless culture there can be great deal of leaching, particularly in rainy springs. Ligneous plants have the capacity to store large quantities of nutrients in perennial organs. Knowledge of the plant's internal nutrient mobilisation during spring to supply growing organs could lead to reduction of fertiliser application. To quantify the fraction of storage mobilisation available for growth of new organs during spring, Ligustrum ovalifolium shrubs were grown for 2 years with or without fertilisation in the second spring. Nitrogen (N) absorption and N and carbon (C) mobilisation from storage were followed during spring growth via the sap quality. A mathematical combination of the sap composition with flow velocity provided the transported quantities of N and C. Nitrogen and C mobilisation towards new shoots took place during all the spring growth from bud break onwards. In unfertilised plants, C was mobilised primarily as sugars (stachyose, mannose and sucrose) and starch. In fertilised plants, the same sugars were transported in the xylem sap, but at lower concentrations. Stachyose concentration was lower in fertilised than in unfertilised plants and decreased during spring growth. Nitrogen was transported in the xylem sap mainly as amino acids in both fertilisation treatments. Glutamine was the predominant form at bud break and during shoot elongation. In fertilised plants, arginine became predominant after shoot elongation, and was related to low C availability. The interactions of N with C are discussed; specifically, insufficient availability of N limits the use of C, more of which is directed to aerial organs by sap flow.

Seasonal variations in water relations in current-year leaves of evergreen trees with delayed greening.

We investigated seasonal patterns of water relations in current-year leaves of three evergreen broad-leaved trees (Ilex pedunculosa Miq., Ligustrum japonicum Thunb., and Eurya japonica Thunb.) with delayed greening in a warm-temperate forest in Japan. We used the pressure-volume method to: (1) assess the extent to which seasonal variation in leaf water relations is attributable to leaf development processes in delayed greening leaves versus seasonal variation in environmental variables; and (2) investigate variation in leaf water relations during the transition from the sapling to the adult tree stage. Leaf mass per unit leaf area was generally lowest just after completion of leaf expansion in May (late spring), and increased gradually throughout the year. Osmotic potential at full turgor (Psi(o) (ft)) and leaf water potential at the turgor loss point (Psi(w) (tlp)) were highest in May, and lowest in midwinter in all species. In response to decreasing air temperature, Psi(o) (ft) dropped at the rate of 0.037 MPa degrees C(-1). Dry-mass-based water content of leaves and the symplastic water fraction of total leaf water content gradually decreased throughout the year in all species. These results indicate that reductions in the symplastic water fraction during leaf development contributed to the passive concentration of solutes in cells and the resulting drop in winter Psi(o) (ft). The ratio of solutes to water volume increased in winter in current-year leaves of L. japonicum and E. japonica, indicating that osmotic adjustment (active accumulation of solutes) also contributed to the drop in winter in Psi(o) (ft). Bulk modulus of elasticity in cell walls fluctuated seasonally, but no general trend was found across species. Over the growing season, Psi(o) (ft) and Psi(w) (tlp) were lower in adult trees than in saplings especially in the case of I. pedunculosa, suggesting that adult-tree leaves are more drought and cold tolerant than sapling leaves. The ontogenetic increase in the stress resistance of I. pedunculosa may be related to its characteristic life form because I. pedunculosa grows taller than the other species studied.

Invasion by Ligustrum lucidum (Oleaceae) in NW Argentina: early stage characteristics in different habitat types.

Currently biological invasions are considered one of the world's most serious conservation problems. Ligustrum lucidum is the most abundant exotic tree in secondary forest patches of montane forests of NW Argentina. We studied the determinants of success of the early stages of its life cycle in distinct habitat types, with the hope of identifying vulnerabilities that could be exploited to control the invasion. Seed arrival, germination, seedling recruitment and survival, and sapling growth were studied in edges, gaps and forest interior. Seed arrival was also assessed under perches and in open fields. Germination was studied in forest and grassland patches. L. lucidum seedling survival and sapling growth were compared with the most abundant native species survival and growth. Seed arrival was strongly seasonal with a peak in mid-August. Seed rain did not differ significantly among habitat types, however there was a tendency for edges to receive more seeds when only dispersed seeds were considered. Perches strongly enhanced seed arrival; more than 40 times the number of seeds were dispersed beneath citrus plants (i.e. perches) than found in paired open areas. In the forest, seeds in gaps and edges had higher germination rates, but there was no difference in seedling survival. Fruits under closed canopy exhibited the lowest germination. Germination and survival were low in open areas. Neither seedling recruitment nor sapling growth differed between gaps and forest interior. L. lucidum saplings grew significantly more than saplings of the most common native species, and also showed higher seedling survival. L. lucidum is a prolific fruit producer, is capable of germinating and surviving in a broad range of forest environments, it is relatively shade tolerant and has higher survival and faster growth rate in comparison to the most common native species. All these characteristics highlight its potency as a successful invader, and point to few vulnerabilities that could be targets of control measures.