A cytotoxic cardenolide and a saponin from the rhizomes of Tupistra chinensis

A new cardenolide tupichinolide (1) and a new steroidal saponin tupichinin A (2), together with seven known compounds, were isolated from the rhizomes of Tupistra chinensis. Their structures were established using spectroscopic analysis and chemical methods. Compound 1 was the first cardenolide isolated from Tupistra chinensis and exhibited potent cytotoxicity against five human cancer cell lines: HL-60, SMMC-7721, A-549, MCF-7 and SW480.     

Antiproliferative steroidal glycosides from Digitalis ciliata

Two new compounds, a furostanol glycoside (1) and a pregnane glycoside (4), along with eight known compounds, belonging to the classes of spirostane (2,3), pregnane (5–7) and cardenolide (8–10) glycosides, were isolated from the seeds of Digitalis ciliata. Their structures were elucidated by 1D and 2D-NMR experiments as well as ESI-MS analysis. For the first time pregnane glycosides of the diginigenin series have been isolated from D. ciliata. The cytotoxic effects of compounds 1–10 on cell viability of several cancer cell lines, namely human breast cancer (MCF-7), human glioblastoma (T98G), human lung adenocarcinoma (A549), human colon carcinoma (HT-29), and human prostate cancer (PC-3) cell lines were evaluated. Compounds 1, 4, 7 and 8 showed antiproliferative effects against MCF-7, HT-29 and A549 cancer cells with IC₅₀ values ranging from 8.3 to 20 μM. The effects of compounds 1–10 on cell proliferation were evaluated on these three cancer cell lines by cell cycle analysis of DNA content using flow cytometry. Compounds 7, 8 and 10 induced significant changes in G₂/M cell cycle phase of all analyzed cells. The obtained results indicate that compounds 7, 8 and 10 are cytostatic compounds effective in reducing cell proliferation by inducing accumulation of the cells in the G₂/M phase of the cell cycle.     

Isoangustone A induces apoptosis in SW480 human colorectal adenocarcinoma cells by disrupting mitochondrial functions

Licorice and its components have been reported to posses various anti-tumor activities, but its active ingredients and underlying mechanisms are not well understood yet. In the present study, a group of representative licorice-derived compounds that could be detected in rat plasma or urine were screened for anti-tumor activity. Among these compounds, isoangustone A (IAA) was found to promptly inhibit the viability of SW480 human colorectal adenocarcinoma cells in a time- and concentration-dependent manner. Further analyses indicate that IAA activated caspase-dependent pro-apoptotic signaling and induced significant apoptosis, while had little effect on cell cycle. IAA strongly inhibited Akt phosphorylation within 5 min; however, overexpression of constitutively activated Akt could not rescue IAA-mediated inhibition, indicating that inhibition of Akt was not involved in IAA-induced apoptosis. Further examinations show that IAA induced dissipation of mitochondria membrane potential and release of cytochrome C within 1 h, accompanied by swelling of mitochondrial matrix and disrupting of mitochondrial outer membrane, and followed by decreasing of cellular ATP. The above results suggest that IAA induced apoptosis in colorectal cancer cells principally by inducing mitochondrial outer membrane permeabilization, and deserves further investigations as a novel anti-colorectal cancer agent.     

Xylogenesis of compression and opposite wood in mountain pine at a Mediterranean treeline

Key message

Comparisons between compression and opposite wood formation in prostrating Pinus mugo indicate that the secondary meristem can produce more tracheids with thicker walls by also increasing the number of contemporaneously differentiating cells, rather than only increasing the duration or the rate of cell formation.

Context

Although cambium tissues within a stem experience the same climatic conditions, the resulting wood structure and properties can strongly differ. Assessing how meristem differently regulates wood formation to achieve different anatomical properties can help understanding the mechanisms of response and their plasticity.

Aims

We monitored the formation of compression (CW) and opposite (OW) wood within the same stems to understand whether achieved differences in wood structure are caused by modifications in the process of cell formation.

Methods

We collected weekly microcores of compression and opposite wood from the curved stem of ten treeline prostrating mountain pines (Pinus mugo Turra ssp. mugo) at the Majella massif in Central Italy.

Results

Results indicate that cambium formed approximately 1.5 times more cells in CW than OW, despite that CW cell differentiation only extended 2 weeks longer and the residence time of CW cells in the wall-thickening phase was only 20% longer. Differences in their formation were thus mainly related to both the rates and the width of the enlarging and wall-thickening zones (i.e., the number of cells simultaneously under differentiation) and less to duration of cell formation.

Conclusion

We conclude that to achieve such a different wood structures, the efficiency of the secondary meristem, in addition of altered rate of cell division and differentiation, can also modify the width of the developing zones. Thus, deciphering what rules this width is important to link environmental conditions with productivity.

     

Changes in the morphology and functions of vacuoles during the death of ray parenchyma cells in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

Changes in the morphology and functions of vacuoles provide useful information about the mechanism of cell death. In the present study, we monitored the morphology and contents of vacuoles during the death of ray parenchyma cells in the conifer Cryptomeria japonica. In differentiating xylem, ray parenchyma cells had large central vacuoles. In sapwood, vacuoles in ray parenchyma cells contained proteins, an indication that one of the main functions of these vacuoles might be protein storage. A dramatic decrease in the protein content of some vacuoles was detected in the intermediate wood before the initiation of vacuole rupture. Although vacuole rupture was detected from the intermediate wood to the outermost heartwood, some vacuoles were obviously enlarged in the inner intermediate wood. Condensed nuclei were first observed after the rupture of these large vacuoles in ray parenchyma cells. It seems plausible that the autolysis of the contents of ray parenchyma cells might be caused by the rupture of the enlarged vacuoles in the inner intermediate wood.     

Impacts of soil conditions and light availability on natural regeneration of Norway spruce <Emphasis Type="Italic">Picea abies</Emphasis> (L.) H. Karst. in low-elevation mountain forests

? Key message

Natural regeneration of P. abies (L.) H. Karst. may reach high densities in lower mountain elevations. The highest densities were found in sites with moderate light availability, with low pH, and not near the riverbank. However, age-height classes differed in the predicted magnitude of response, but were consistent in response directions. Mosses and understory species typical of coniferous forests were positively correlated with regeneration density.

? Context

Norway spruce Picea abies (L.) H. Karst. in Central Europe is at risk under climate change scenarios, particularly in mountain regions. Little is known about the impact of environmental factors on the natural regeneration of P. abies in low-elevation mountain forests.

? Aims

We aimed to assess impacts of distance from the riverbank, soil pH, and light availability on natural P. abies regeneration. We hypothesized that (1) natural P. abies regeneration would depend on light availability and soil pH and (2) there are understory plant species which may indicate the microsites suitable for natural regeneration of P. abies.

? Methods

The study was conducted in the Sto?owe Mountains National Park (SW Poland, 600–800 m a.s.l.). We established 160 study plots (25 m²) for natural regeneration, light availability, soil pH, and understory vegetation assessment.

? Results

The highest densities of natural regeneration of P. abies were observed in sites with moderate light availability (0.1–0.2 of open sky) and low pH (3.5–4.5), and located relatively far from the riverbank. Cover of 22 understory plant species were correlated with natural P. abies regeneration densities, mostly positively.

? Conclusion

Different stages of natural regeneration of P. abies revealed different regeneration niches. Most understory plant species (bryophytes and herbs typical of coniferous forests) do not compete with natural regeneration of P. abies.

     

Growth phenology in <Emphasis Type="Italic">Pinus halepensis</Emphasis> Mill.: apical shoot bud content and shoot elongation

Key message

The chronology of periods of organogenesis and elongation is highlighted in Pinus halepensis.The two first growth units of an annual shoot are preformed inside the bud during the previous year. The following growth units are formed during the spring or summer of the current year.

Context

Analysis of annual shoot length growth phenology is crucial to assess the impact of climate change on tree production. Little is known about the basic growth characteristics and the phenology of pines.

Aims

The present study disentangles the roles of shoot organogenesis vs elongation in the annual growth cycle of the polycyclic Aleppo pine.

Methods

Growth of young Pinus halepensis trees was monitored monthly for 1 year. At each monitoring date, the bud content and meristem dimensions of the main stem shoots apices were analyzed.

Results

The two first growth units of an annual shoot are preformed inside the bud during the previous year. The following growth units are formed during the spring or summer of the current year. The gap between a shoot organogenesis and its elongation may vary from 1 month, for the last growth unit, to half a year, for the first growth units.

Conclusion

Our results underline the importance of taking seasonal environmental conditions from both the previous and the current year into account, in order to study the plasticity of annual shoot growth and its response to climate change and variability.

     

Chemical constituents of Kigelia pinnata twigs and their GLUT4 translocation modulatory effect in skeletal muscle cells

Phytochemical investigation of the ethanolic extract of twigs of Kigelia pinnata DC. afforded one new iridoid 7-hydroxy eucommiol (1), and nine known compounds (2-10). The structure of compounds was elucidated by extensive spectroscopic methods, including 1D, 2D NMR experiments and MS analysis. All these compounds were evaluated for GLUT4 translocation modulatory effect in skeletal muscle cells. Four of the tested compounds 1, 5, 6 and 7 showed significant stimulation of GLUT4 translocation to cell surface in skeletal muscle cells without any adverse effect on cell viability. Effect of these four compounds was concentration-dependent and comparable to standard drug rosiglitazone. These findings indicate that constituents of K. pinnata may provide leads for the therapeutics for insulin resistance and diabetes.     

Factors controlling plasticity of leaf morphology in <Emphasis Type="Italic">Robinia pseudoacacia L.</Emphasis> I: height-associated variation in leaf structure

? Context

Physiological ecologists have been fascinated by height- or position-linked differences of leaf morphology within tall trees >25 m, but the exact cause is still debated, i.e., is it due to light or height-induced water stress?

? Aims

The aim of this study was to demonstrate that relatively small trees (<15 m) have leaf morphologies that vary with height and that such variation depends on site-moisture variability.

? Methods

Leaves were collected from Robinia pseudoacacia trees at two sites in China with contrasting moisture variability to gather baseline data on leaf morphology parameters.

? Results

Most measured parameters changed regularly with height. Water potential linearly decreased with height. Leaf area and stomata area decreased with height, while leaf mass per area, carbon isotope composition (δ ¹³C), and stomata density increased with height. Mesophyll and epidermal cell width decreased with height, while leaf thickness and palisade cell length increased with height. All the morphology parameters between two sites were also significantly different.

? Conclusions

Based on the field results, it is concluded that minor variations in water potential at the time of leaf growth influence leaf morphology at both site-level and height-level. Controlled environment experiments will be conducted to confirm this conclusion.

     

New pregnane saponins from Ecdysanthera rosea and their cytotoxicity

Two new pregnane saponins elucidated as ecdysantheroside A (1) and ecdysantheroside B (2) and six known compounds (3-8) based on spectral data (MS, IR, 1D and 2D NMR) were isolated from the stem bark of Ecdysanthera rosea. The cytotoxicity against six cell lines of these compounds was tested by MTT assay. The results revealed that compounds 5 and 7 showed cytotoxicity against all the cell lines. Compound 2 showed cytotoxicity against cells A549, MDA435, HepG2, and HUVEC, while compound 4 showed cytotoxicity against cells A549, CEM, and HUVEC. Compound 6 had cytotoxicity against the others except cell HepG2.     

Modelling wood formation and structure: power and limits of a morphogenetic gradient in controlling xylem cell proliferation and growth

Key message

The emergence of the characteristic tree-ring pattern during xylogenesis is commonly thought to be controlled by a gradient of morphogen (auxin, TDIF peptide...). We show that this hypothesis accounts for several developmental aspects of wood formation, but not for the final anatomical structure.

Context

Wood formation is a dynamic cellular process displaying three generic features: (i) meristematic cell proliferation is restricted to the small cambial zone, preventing exponential xylem radial growth along the growing season; (ii) developmental processes result in a stable zonation of the developing xylem; (iii) the resulting mature wood cells form the typical tree-ring structure made of early and late wood with a gradient of cell sizes, an important trait for wood functioning in trees and for lumber quality. The mechanisms producing these spatial-temporal patterns remain largely unknown. According to the often-cited morphogenetic-gradient hypothesis, a graded concentration profile of a signalling molecule (e.g. auxin, TDIF) controls xylogenesis by providing positional information to differentiating cells.

Aims

We assessed the predictions of the morphogenetic-gradient theory.

Methods

We developed a computational model of wood formation implementing hypotheses on how a morphogen flows through the developing xylem and controls cell division and growth and we tested it against data produced by studies monitoring wood formation in conifers.

Results

We demonstrated that a morphogenetic gradient could indeed control xylem radial growth and wood-forming tissue zonation. However, it failed to explain the pattern of final cell sizes observed in tree-rings. We discussed the features that candidate additional regulatory mechanisms should meet.

     

Deciphering the ozone-induced changes in cellular processes: a prerequisite for ozone risk assessment at the tree and forest levels

Key message

Ozone, one of the major atmospheric pollutants, alters tree growth, mainly by decreasing carbon assimilation and allocation to stems and roots. To date, the mechanisms of O₃ impact at the cellular level have been investigated mainly on young trees grown in controlled or semi-controlled conditions. In the context of climate change, it is necessary to introduce a valuable defence parameter in the models that currently predict O₃ impact on mature trees and the carbon sequestration capacity of forest ecosystems.

Context

Air pollution is an important factor that affects negatively forest ecosystems. Among oxidative air pollutants, ozone is considered as the most toxic in terms of impact on vegetation.

Aims

This paper focuses on the negative impacts of ozone on trees in controlled conditions or in their natural environment. The current knowledge of the responses at cell level is presented and ways to improve their use for ozone risk assessment of forest stands are discussed.

Methods

Information was collected from original papers or reviews, providing an overview of the research conducted over the last 60 years.

Results

The negative effects of ozone on carbon assimilation and tree biomass production were reviewed and discussed, with a focus on effects on cell processes implied in cell defence, including stomatal regulation, detoxification, signalling, and biosynthesis of wood compound.

Conclusion

In the context of increasing significance of O₃ flux approach, this review intends to shed light into the black box of defence processes, which are playing a crucial part within the effective O₃ dose modelling. Today, it is recognized that tropospheric ozone inhibits tree growth and its role on the future carbon sink of the forest ecosystem is discussed along with the combination of other environmental factors like elevated temperature, water, and nitrogen supply, likely to be modified in the context of climate change.

     

Self-thinning in four pine species: an evaluation of potential climate impacts

Key message

Self-thinning lines are species- and climate-specific, and they should be used when assessing the capacity of different forest stands to increase biomass/carbon storage.

Context

The capacity of forests to store carbon can help to mitigate the effects of atmospheric CO₂ rise and climate change. The self-thinning relationship (average size measure ～ stand density) has been used to identify the potential capacity of biomass storage at a given density and to evaluate the effect of stand management on stored carbon. Here, a study that shows how the self-thinning line varies with species and climate is presented.

Aims

Our main objective is thus testing whether species identity and climate affect the self-thinning line and therefore the potential amount of carbon stored in living biomass.

Methods

The Ecological and Forest Inventory of Catalonia was used to calculate the self-thinning lines of four common coniferous species in Catalonia, NE Iberian Peninsula (Pinus halepensis, Pinus nigra, Pinus sylvestris and Pinus uncinata). Quadratic mean diameter at breast height was chosen as the average size measure. The self-thinning lines were used to predict the potential diameter at a given density and study the effect of environmental variability.

Results

Species-specific self-thinning lines were obtained. The self-thinning exponent was consistent with the predicted values of ?3/2 and ?4/3 for mass-based scaling for all species except P. sylvestris. Species identity and climatic variability within species affected self-thinning line parameters.

Conclusion

Self-thinning lines are species-specific and are affected by climatic conditions. These relationships can be used to refine predictions of the capacity of different forest stands to increase biomass/carbon storage.

     

Variation in pinewood nematode susceptibility among <Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait. provenances from the Iberian Peninsula and France

Key message

Pinus pinaster Ait. susceptibility to pinewood nematode significantly differed among provenances, and the two Atlantic provenances of the Iberian Peninsula being the most affected. However, significant provenance × environment interaction was found. Provenance susceptibility was related to basal diameter, number of branches and oleoresin flow, and some climatic parameters.

Context

The pinewood nematode Bursaphelenchus xylophilus, native to North America, is an important pest affecting pine forests throughout Eurasia. In Europe, it has been detected in Portugal and Spain and is primarily associated with Pinus pinaster, an important Mediterranean tree species.

Aims

We have investigated the differences in susceptibility among several P. pinaster provenances in the Iberian Peninsula and France, as well as their relationship to certain growth traits and physiological parameters.

Methods

Three independent inoculation tests were performed on 3 to 4-year-old trees, followed by assessment of growth traits and physiological variables, along with time course destructive sampling for nematode quantification.

Results

The results showed significant differences among provenances for almost all growth traits, wilting, and mortality, though a significant provenance × environment interaction was also detected. Two Atlantic provenances, Noroeste-Litoral and Leiria, displayed the largest susceptibility to pinewood nematode. Changes in susceptibility to B. xylophilus between experiments were influenced by temperature and seasonality. Autumn precipitation and mean maximum temperature during summer at the original provenance sites could be related to provenance susceptibility.

Conclusion

Noroeste-Litoral and Leiria were the most disease-affected provenances. This study emphasizes the need for further research on how tree growth stage influences susceptibility and on the possibility of cross-breeding among provenances.

     

Radial growth of <Emphasis Type="Italic">Cinnamomum kanehirae</Emphasis> Hayata displays a larger temperature sensitivity in dominant than codominant trees

Key message

The radial wood growth curves of Cinnamomum kanehirae Hayata (an endangered species of subtropical Taiwan) exhibit an S shape. The dominant trees displayed a larger radial growth than the codominant trees, and their growth was more sensitive to air temperature.

Context

Knowledge of wood radial growth is important for evaluating the factors that limit tree growth performance. The relevant experiments have mostly been conducted in cold and temperate ecosystems, but rarely in subtropical ecosystems.

Aims

In this study, we aimed to construct a unified radial growth model for Cinnamomum kanehirae Hayata and to identify its sensitivity to temperature.

Methods

The wood radial increments were quantified for 3 years by either pinning or microcoring. The radial wood growth curves were modelled integratively by semiparametric regression and individually by curve fitting. The effects of tree social class, interannual and environmental factors on radial growth were analysed quantitatively.

Results

A unified S-shaped growth model for C. kanehirae was successfully constructed. By including the social class effect, the model was significantly improved. The maximum radial increment (A) was significantly correlated with the maximum growth rate (μ); both A and μ were significantly higher in dominant than in codominant trees. The time-varying radial growth rate was more sensitive to air temperature in dominant than in codominant trees.

Conclusion

Semiparametric models revealed an S-shaped growth curve of C. kanehirae and confirmed the higher temperature sensitivity of dominant trees compared to codominant trees in humid subtropical areas.

     

Two new amide alkaloids with anti-leukaemia activities from Aconitum taipeicum

Two new amide alkaloids, named 3-isopropyl-tetrahydropyrrolo [1,2-a] pyrimidine-2,4(1H,3H)-dione (1) and 1-acetyl-2,3,6-triisopropyl-tetrahydropyrimidin-4(1H)-one (2), were isolated from the roots of Aconitum taipeicum. Their chemical structures were characterized on the basis of MS, 1D- and 2D NMR. The anti-leukaemia activities of the compounds were also tested. The results indicated that the compounds exhibited more significant cell growth inhibitory activities against HL-60 cells than adriamycin, with the IC₅₀ of 1.1 ± 0.03 μg/mL and 1.6 ± 0.07 μg/mL respectively. In addition, two compounds showed anti-tumor activities against K562 cells as well.     

Radial growth changes following hemlock woolly adelgid infestation of eastern hemlock

? Context

Hemlock woolly adelgid (Adelges tsugae) is an invasive insect that is defoliating and killing eastern hemlock (Tsuga canadensis) in the USA.

? Aims

We quantified changes in tree-ring growth rates and wood anatomy for living trees infested with hemlock woolly adelgid across six sites from Massachusetts (42°41′N) to Georgia (34°53′N) to identify growth responses of eastern hemlock that had survived infestation.

? Methods

Annual ring widths from infested eastern hemlocks were cross-dated and measured. Growth rates before and after infestation were compared. Two infested trees from Virginia were cut, and thin sections were prepared to identify changes in cell properties.

? Results

At three sites, trees experienced a significant decrease in radial growth after hemlock woolly adelgid arrival; however, the other three sites showed no change or increase in growth. Latewood produced after hemlock woolly adelgid infestation had significantly smaller cells with reduced cell wall thickness compared to latewood prior to infestation.

? Conclusion

At half the sites where hemlock woolly adelgid infested eastern hemlock trees were sampled, radial growth increased or remained unchanged. This unexpected response may be due to reduced competition due to mortality of other eastern hemlocks or physiological compensatory responses of increased photosynthetic rate and increased water use efficiency experienced by eastern hemlock infested with hemlock woolly adelgid.     

Genetic variation and genotype by environment interaction in the susceptibility of <Emphasis Type="Italic">Abies nordmanniana</Emphasis> (Steven) Spach to the fungus <Emphasis Type="Italic">Neonectria neomacrospora</Emphasis> (Booth &amp; Samuels) Mantiri &amp; Samuels

Key message

Pronounced clonal variation and moderate to high broad-sense heritability estimates of susceptibility to Neonectria neomacrospora were found in Abies nordmanniana in three sites. Significant genotype by environment (G × E) interaction was detected across sites.

Context

Nordmann fir, a widely used Christmas tree species in Europe, has, since 2011, been increasingly damaged by a canker disease caused by Neonectria neomacrospora.

Aims

The objective was to study the genetic variation and genotype by environment interaction in the susceptibility of Nordmann fir to N. neomacrospora.

Methods

Damage caused by N. neomacrospora was evaluated using a visual scale in three Nordmann fir clonal seed orchards in Denmark, partly containing the same clones.

Results

Damage due to N. neomacrospora was substantial at all three sites, and no clone was completely resistant to N. neomacrospora, but a large genetic variation in the susceptibility was detected among clones. Estimates of single-site individual broad-sense heritability for susceptibility varied between 0.38 and 0.47. The average type-B genetic correlation for damage score across sites was 0.34.

Conclusion

Genetic variation was very pronounced, and significant G × E interactions were detected for susceptibility. Further investigations of narrow-sense heritability, expression of the trait in younger material, and identification of the cause of G × E for N. neomacrospora susceptibility in Nordmann fir across different sites are recommended.