Photosystem II of Ligustrum lucidum in response to different levels of manganese exposure.

The toxic effect of excessive manganese (Mn) on photosystem II (PSII) of woody species remains largely unexplored. In this study, five Mn concentrations (0, 12, 24, 36, and 48 mM) were used, and the toxicity of Mn on PSII behavior in leaves of Ligustrum lucidum was investigated using in vivo chlorophyll fluorescence transients. Results showed that excessive Mn levels induced positive L- and K- bands. Variable fluorescence at 2 ms (VJ) and 30 ms (VI), absorption flux (ABS/RC), trapped energy flux (TRo/RC), and dissipated energy flux (DIo/RC) increased in Mn-treated leaves, whereas the performance index (PIABS), electron transport flux (ETo/RC), maximum quantum yield (φPo), quantum yield of electron transport (φEo), and probability that an electron moves further than QA- (ψo) decreased. Also, excessive Mn significantly decreased the net photosynthesis rate and increased intercellular CO2 concentration. The results indicated that Mn blocked the electron transfer from the donor side to the acceptor side in PSII, which might be associated with the accumulation of QA-, hence limiting the net photosynthetic rate.

Response of gas-exchange characteristics and chlorophyll fluorescence to acute sulfur dioxide exposure in landscape plants.

To explore the toxicity and action mechanism of acute sulfur dioxide (SO2) on urban landscape plants, a simulated SO2 stress environment by using fumigation chamber involving increasing SO2 concentration (0, 25, 50, 100, 200 mg m-3) was carried out among three species. After 72 h of exposure, SO2-induced oxidative damage indicated by electrolyte leakage increased with higher dose of SO2. Meanwhile, SO2 decreased the contents of chlorophyll a, chlorophyll b and carotenoid and increased the contents of sulfur. Net photosynthetic rate (Pn) decreased as a result of stomatal closure when SO2 dose was lower than 50 mg m-3, out of this range, non-stomatal limitation play a dominant role in the decline of Pn. Simultaneous measurements of chlorophyll fluorescence imaging (CFI) also revealed that the maximal quantum efficiency of PSII photochemistry in dark-adapted state (Fv/Fm) and the realized operating efficiency of PSII photochemistry (Fq'/Fm') was reduced by SO2 in a dose-dependent manner. In addition, the maximum quantum efficiency of PSII photochemistry in light-adapted state (Fv'/Fm') and the PSII efficiency factor (Fq'/Fv') decreased when exposure to SO2. These results implied that acute SO2 exposure induced photoinhibition of PSII reaction centers in landscape plants. Our study also indicated that different urban landscape plant species resist differently to SO2: Euonymus kiautschovicus > Ligustrum vicaryi > Syringa oblata according to gas-exchange characteristics and chlorophyll fluorescence responses.

Leaf physiological and anatomical responses of Lantana and Ligustrum species under different water availability.

Understanding the plant characteristics that support tolerance to water stress is important in choosing plants in arid or semi-arid environments, such as the Mediterranean. In particular, leaf characteristics can affect the response of plants to water stress. In order to understand how plants with different leaf features can overcome water stress, four water regimes were adopted on two species that are widespread in the Mediterranean environment, Lantana camara and Ligustrum lucidum. The four treatments were: control (C), in which the pot substrate moisture was maintained close to water container capacity (WCC), light deficit irrigation (LDI) irrigated at 75% of WCC, moderate deficit irrigation (MDI) at 50% of WCC, and severe deficit irrigation (SDI) at 25% of WCC. To better understand the action mechanisms, the trial was repeated twice (from January to May, and from May to September). Morphological, anatomical and physiological data were measured to identify the action mechanisms. Water deficit significantly decreased the biomass accumulation in both species during the experimental growth period. In Lantana, significant variations in total leaf area and leaf number were registered between C and SDI, while in Ligustrum, the differences were significant only for total leaf area. The water deficit treatments reduced the leaf thickness especially in Ligustrum. In both species, photosynthesis reduction was related to stomatal closure. Ligustrum showed a higher variability among treatments indicating a faster and more efficient response to water limitations compared to Lantana, as also demonstrated by the lower biomass reduction in the most severe water stress treatment.

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.

Co-occurring nonnative woody shrubs have additive and non-additive soil legacies.

To maximize limited conservation funds and prioritize management projects that are likely to succeed, accurate assessment of invasive nonnative species impacts is essential. A common challenge to prioritization is a limited knowledge of the difference between the impacts of a single nonnative species compared to the impacts of nonnative species when they co-occur, and in particular predicting when impacts of co-occurring nonnative species will be non-additive. Understanding non-additivity is important for management decisions because the management of only one co-occurring invader will not necessarily lead to a predictable reduction in the impact or growth of the other nonnative plant. Nonnative plants are frequently associated with changes in soil biotic and abiotic characteristics, which lead to plant-soil interactions that influence the performance of other species grown in those soils. Whether co-occurring nonnative plants alter soil properties additively or non-additively relative to their effects on soils when they grow in monoculture is rarely addressed. We use a greenhouse plant-soil feedback experiment to test for non-additive soil impacts of two common invasive nonnative woody shrubs, Lonicera maackii and Ligustrum sinense, in deciduous forests of the southeastern United States. We measured the performance of each nonnative shrub, a native herbaceous community, and a nonnative woody vine in soils conditioned by each shrub singly or together in polyculture. Soils conditioned by both nonnative shrubs had non-additive impacts on native and nonnative performance. Root mass of the native herbaceous community was 1.5 times lower and the root mass of the nonnative L. sinense was 1.8 times higher in soils conditioned by both L. maackii and L. sinense than expected based upon growth in soils conditioned by either shrub singly. This result indicates that when these two nonnative shrubs co-occur, their influence on soils disproportionally favors persistence of the nonnative L. sinense relative to this native herbaceous community, and could provide an explanation of why native species abundance is frequently depressed in these communities. Additionally, the difference between native and nonnative performance demonstrates that invasive impact studies focusing on the impact only of single species can be insufficient for determining the impact of co-occurring invasive plant species.

Plant-soil interactions promote co-occurrence of three nonnative woody shrubs.

Ecosystems containing multiple nonnative plant species are common, but mechanisms promoting their co-occurrence are understudied. Plant-soil interactions contribute to the dominance of singleton species in nonnative ranges because many nonnatives experience stronger positive feedbacks relative to co-occurring natives. Plant-soil interactions could impede other nonnatives if an individual nonnative benefits from its soil community to a greater extent than its neighboring nonnatives, as is seen with natives. However, plant-soil interactions could promote nonnative co-occurrence if a nonnative accumulates beneficial soil mutualists that also assist other nonnatives. Here, we use greenhouse and field experiments to ask whether plant-soil interactions (1) promote the codominance of two common nonnative shrubs (Ligustrum sinense and Lonicera maackii) and (2) facilitate the invasion of a less-common nonnative shrub (Rhamnus davurica) in deciduous forests of the southeastern United States. In the greenhouse, we found that two of the nonnatives, L. maackii and R. davurica, performed better in soils conditioned by nonnative shrubs compared to uninvaded forest soils, which. suggests that positive feedbacks among co-occurring nonnative shrubs can promote continued invasion of a site. In both greenhouse and field experiments, we found consistent signals that the codominance of the nonnatives L. sinense and L. maackii may be at least partially explained by the increased growth of L. sinense in L. maackii soils. Overall, significant effects of plant-soil interactions on shrub performance indicate that plant-soil interactions can potentially structure the co-occurrence patterns of these nonnatives.

Excising stem samples underwater at native tension does not induce xylem cavitation.

Xylem resistance to water stress-induced cavitation is an important trait that is associated with drought tolerance of plants. The level of xylem cavitation experienced by a plant is often assessed as the percentage loss in conductivity (PLC) at different water potentials. Such measurements are constructed with samples that are excised underwater at native tensions. However, a recent study concluded that cutting conduits under significant tension induced cavitation, even when samples were held underwater during cutting. This resulted in artificially increased PLC because of what we have termed a 'tension-cutting artefact'. We tested the hypothesized tension-cutting artefact on five species by measuring PLC at native tension compared with after xylem tensions had been relaxed. Our results did not support the tension-cutting artefact hypothesis, as no differences were observed between native and relaxed samples in four of five species. In a fifth species (Laurus nobilis), differences between native and relaxed samples appear to be due to vessel refilling rather than a tension-cutting effect. We avoided the tension-cutting artefact by cutting samples to slightly longer than their measurement length and subsequent trimming of at least 0.5 cm of sample ends prior to measurement.

GABA, ß-alanine and glycine in the digestive juice of privet-specialist insects: convergent adaptive traits against plant iridoids.

The privet tree, Ligustrum obtusifolium (Oleaceae), defends its leaves against insects with a strong lysine-decreasing activity that make proteins non-nutritive. This is caused by oleuropein, an iridoid glycoside. We previously found that some privet-specialist caterpillars adapt by secreting glycine in the digestive juice as a neutralizer that prevents the loss of lysine. Here, we extended the survey into 42 lepidopteran and hymenopteran species. The average concentration of glycine in digestive juice for 11 privet-feeding species (40.396 mM) was higher than that for 32 non-privet-feeding species (2.198 mM). The glycine concentrations exceeded 10 mM in 7 out of 11 privet-feeding species. In Macrophya timida (Hymenoptera), it reached 164.8 mM. Three out of the four remaining privet-feeding species had other amino acids instead. Larvae of a privet-specialist butterfly, Artopoetes pryeri (Lycaenidae), had a high concentration (60.812 mM) of GABA. In two other specialists, ß-alanine was found. GABA, ß-alanine, and glycine as well as alanine, amines, and ammonium ion inhibited the lysine decrease, indicating that amino residues are responsible for the inhibition. However, the three amino acids found in the specialists were far more effective (20 mM showed 80% inhibition) than the rest (>140 mM was required for 80% inhibition). Our results show a clear and rare case of the apparent convergent evolution of herbivores' molecular adaptations of feeding on a plant with a chemical defense in a manner that minimizes the cost of adaptation. The novel role of GABA in plant-herbivore interactions shown here is probably the first reported non-neuronal role of animal-derived GABA.

Mesophyll distribution of 'antioxidant' flavonoid glycosides in Ligustrum vulgare leaves under contrasting sunlight irradiance.

BACKGROUND AND AIMS: Flavonoids have the potential to serve as antioxidants in addition to their function of UV screening in photoprotective mechanisms. However, flavonoids have long been reported to accumulate mostly in epidermal cells and surface organs in response to high sunlight. Therefore, how leaf flavonoids actually carry out their antioxidant functions is still a matter of debate. Here, the distribution of flavonoids with effective antioxidant properties, i.e. the orthodihydroxy B-ring-substituted quercetin and luteolin glycosides, was investigated in the mesophyll of Ligustrum vulgare leaves acclimated to contrasting sunlight irradiance. METHODS: In the first experiment, plants were grown at 20 % (shade) or 100% (sun) natural sunlight. Plants were exposed to 100 % sunlight irradiance in the presence or absence of UV wavelengths, in a second experiment. Fluorescence microspectroscopy and multispectral fluorescence microimaging were used in both cross sections and intact leaf pieces to visualize orthodihydroxy B-ring-substituted flavonoids at inter- and intracellular levels. Identification and quantification of individual hydroxycinnamates and flavonoid glycosides were performed via HPLC-DAD. KEY RESULTS: Quercetin and luteolin derivatives accumulated to a great extent in both the epidermal and mesophyll cells in response to high sunlight. Tissue fluorescence signatures and leaf flavonoid concentrations were strongly related. Monohydroxyflavone glycosides, namely luteolin 4'-O-glucoside and two apigenin 7-O-glycosides were unresponsive to changes in sunlight irradiance. Quercetin and luteolin derivatives accumulated in the vacuoles of mesophyll cells in leaves growing under 100 % natural sunlight in the absence of UV wavelengths. CONCLUSIONS: The above findings lead to the hypothesis that flavonoids play a key role in countering light-induced oxidative stress, and not only in avoiding the penetration of short solar wavelengths in the leaf.

Intraspecific differences in drought tolerance and acclimation in hydraulics of Ligustrum vulgare and Viburnum lantana.

An adequate general drought tolerance and the ability to acclimate to changing hydraulic conditions are important features for long-lived woody plants. In this study, we compared hydraulic safety (water potential at 50% loss of conductivity, Psi(50)), hydraulic efficiency (specific conductivity, k(s)), xylem anatomy (mean tracheid diameter, d(mean), mean hydraulic diameter, d(h), conduit wall thickness, t, conduit wall reinforcement, (t/b)(h)(2)) and stomatal conductance, g(s), of forest plants as well as irrigated and drought-treated garden plants of Ligustrum vulgare L. and Viburnum lantana L. Forest plants of L. vulgare and V. lantana were significantly less resistant to drought-induced cavitation (Psi(50) at -2.82 +/- 0.13 MPa and -2.79 +/- 0.17 MPa) than drought-treated garden plants (- 4.58 +/- 0.26 MPa and -3.57 +/- 0.15 MPa). When previously irrigated garden plants were subjected to drought, a significant decrease in d(mean) and d(h) and an increase in t and (t/b)(h)(2) were observed in L. vulgare. In contrast, in V. lantana conduit diameters increased significantly but no change in t and (t/b)(h)(2) was found. Stomatal closure occurred at similar water potentials (Psi(sc)) in forest plants and drought-treated garden plants, leading to higher safety margins (Psi(sc) - Psi(50)) of the latter (L. vulgare 1.63 MPa and V. lantana 0.43 MPa). These plants also showed higher g(s) at moderate Psi, more abrupt stomatal closure and lower cuticular conductivity. Data indicate that the development of drought-tolerant xylem as well as stomatal regulation play an important role in drought acclimation, whereby structural and physiological responses to drought are species-specific and depend on the plant's hydraulic strategy.

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.

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.