Distribution of exchangeable Ca2+ during the process of Larix decidua Mill. pollination and germination.

The involvement of Ca2+ ions in angiosperms sexual processes is well established, while in gymnosperms, such knowledge remains limited and is still a topic of discussion. In this study, we focused on Larix decidua, using Alizarin-red S staining and the pyroantimonate method to examine the tissue and subcellular distribution of free and loosely bound Ca2+ ions at different stages of the male gametophyte's development and its interaction with the ovule. Our findings show that in larch, both the germination of pollen grains and the growth of pollen tubes occur in an environment rich in Ca2+. These ions play a crucial role in the adhesion of the pollen grain to the stigmatic tip and its subsequent movement to the micropylar canal. There is a significant presence of free and loosely bound Ca2+ ions in both the fluid of the micropylar canal and the extracellular matrix of the nucellus. As the pollen tube extends through the nucellus, we observed a notable accumulation of Ca2+ ions just above the entry to the mature archegonium, a region likely crucial for the male gametophyte's directional growth. Meanwhile, the localized presence of free and loosely bound Ca2+ ions within the egg cell cytoplasm may inhibit the pollen tubes growth and rupture, playing an important role in fertilization.

Comparative analysis of microbial community structure in different times of Panax ginseng Rhizosphere microbiome and soil properties under larch forest.

BACKGROUND: Panax ginseng cultivated under the forest is popular because its shape and effective ingredients are similar to wild ginseng. The growth of P. ginseng in the larch forest is generally better than in the broad-leaved forest, and the incidence rate of diseases is low. Therefore, the selection of forest species is one of the basic factors in the successful cropping of P. ginseng. METHODS: Illumina HiSeq high-throughput sequencing was used to analyze the 16S rRNA/ITS gene sequence of P. ginseng rhizosphere soil under larch forest to study the rhizosphere microbiome's diversity and community composition structure. RESULTS: The species classification and richness of rhizosphere bacterial and fungal communities in the same-aged P. ginseng were similar. Consistent with the soil system of commonly cultivated crops, Proteobacteria, Actinobacteriota, Acidobacteriota, Verrucomicrobiota, Chloroflexi, and Basidiomycota, Ascomycota were the dominant phylum of bacteria and fungi, respectively. Compared with the soil without planting P. ginseng, the diversity of microorganisms and community structure of continuous planting for 2 years, 5 years, and 18 years of P. ginseng rhizosphere soil had little change. The accumulation levels of Ilyonectria, Fusarium, Gibberella, and Cylindrocarpon were not significantly increased with planting P. ginseng and the increased age of cropping P. ginseng. CONCLUSIONS: The results of this study showed that the soil function of the larch forest was good, which provided a theoretical basis for the land selection and soil improvement of cultivating P. ginseng under the larch forest.

[Natural regeneration of Larix principis-rupprechtii plantations with different densities and its influencing factors]. / ä¸åŒå¯†åº¦åŽåŒ—è½å¶æ¾æž—å¤©ç„¶æ›´æ–°åŠå ¶å½±å“å› å­.

In this study, we analyzed the natural regeneration of Larix principis-rupprechtii pure plantations with stand densities of 128, 240, 320, 400, 480, 560, 640 and 720 trees·hm-2 in the Guandi Mountains and its influencing factors. The results showed that the regeneration index first increased and then decreased with the increases of stand density, and that the regeneration performance of stand with medium density (400-560 trees·hm-2) was significantly better than other stands. Light conditions, herbaceous plants and litter of the understory had a dual effect on the regeneration of L. principis-rupprechtii. Excessive light, herbaceous plant cover or over-thick litter was not instrumental to the regeneration. Soil organic matter promoted stand regeneration by providing soft soil texture, adequate water content, low phosphorus but high nitrogen. The effects of the examined factors on the regeneration index were as follows: soil water content (0.798) > total nitrogen (0.621) > litter thickness (-0.597) > soil porosity (0.504) > soil organic matter (0.493) > total phosphorus (-0.404) > transmitted total light (-0.274) > herbaceous plants cover (-0.021). In the plantation management, stand density could be controlled at about 480 trees·hm-2 by thinning or replanting, while litter could be cleared properly to improve soil condition and to promote natural regeneration of L. principis-rupprechtii.

Actual prospects of using some types of larch growing in Kazakhstan in medicine.

Local plants can save natural resources and be used as a source of biologically active compounds, which can be high-quality, effective, and safe ingredients for pharmacological or chemical industries. Therefore, this study aimed to investigate the properties of two medicinal plants - the fine-scaled larch (Lárix Kaémpferi) and Siberian larch (Lárix Sibirica), which are growing in the Republic of Kazakhstan. We compared the two types of larches according to botanical affiliation and species description. We studied the alcoholic extracts from Lárix Kaémpferi and Lárix Sibirica to determine their physical and chemical properties. The data on the chemical composition of extractive compounds were generalized and systematized. The authenticity of Lárix Kaémpferi and Lárix Sibirica was established by external, anatomical, and diagnostic signs in microscopic examination and qualitative reactions. Specific indicators and their norms for raw materials were identified. This is the standard for both types of larch and determines their quality. We experimented with grinding fineness for studied materials and tested the impurities and moisture content of raw materials, total ash, and ash insoluble in a 10% hydrochloric acid solution. Determination of heavy metals and radionuclides was also considered. The quality specifications were developed based on the standardization of fine-scaled larch and Siberian larch.

Phytochemical and biological studies on rare and endangered plants endemic to China. Part XXV. Structurally diverse triterpenoids and diterpenoids from two endangered Pinaceae plants endemic to the Chinese Qinling Mountains and their bioactivities.

A joint phytochemical investigation on the MeOH extracts of the twigs and needles of two endangered Pinaceae plants endemic to the Chinese Qinling Mountains, Picea neoveitchii (an evergreen spruce) and Larix potaninii var. chinensis (a deciduous larch), led to the isolation and characterization of 34 and 24 structurally diverse terpenoids, respectively. Among them, seven are previously undescribed, including a picane-type [i.e., 14(13 â†’ 12)abeo-12αH-serratane] (neoveitchin A) and a serratane-type (neoveitchin B) triterpenoids, and an abietane-type (neoveitchin C) as well as four labdane-type (potalarxins A-D) diterpenoids. Their structures and absolute configurations were established by extensive spectroscopic methods and/or X-ray diffraction analyses. All isolates were evaluated for their inhibitory activities against the human protein tyrosine phosphatase 1B (PTP1B). Serrat-14-en-3α,21ß-diol, betulinic acid, 3ß-hydroxy-11-ursen-13(28)-olide, ursolic acid, and oleanolic acid were found to have considerable inhibitory effects against PTP1B, with IC50 values ranging from 1.1 to 18.1 µM. The interactions of the bioactive triterpenoids with PTP1B were thereafter performed by employing molecular docking studies. In addition, 7-oxo-dehydroabietic acid (an abietane-type diterpenoid) and mangiferonic acid (a cycloartane-type triterpenoid) inhibited acetyl-coenzyme A carboxylase 1 (ACC1), with IC50 values of 3.4 and 6.6 µM, respectively.

Antimicrobial Activity and Wound-Healing Capacity of Birch, Beech and Larch Bark Extracts.

Bark is a major by-product of woodworking industries. The contents of several wood species are known to harbor antimicrobial, antiviral, anti-inflammatory and wound-healing capacities. The aim of this work was to identify beneficial properties of Austrian larch, birch and beech bark extracts for their potential usage as additives or active ingredients in dermatological applications. Bacterial agar diffusion assay and resazurin-based broth microdilution assay were used to evaluate anti-bacterial activity. To gain more insight into the cellular response to bark extracts, viability-, scratch-assays and ELISAs were performed. Birch and beech extracts showed strong antimicrobial activities against Gram-positive bacteria, including Cutibacterium acnes, Staphylococcus epidermidis and MRSA. Wound closure was enhanced with birch and beech extracts as compared to controls in the scratch-assays. Whereas beneficial properties of birch bark components have previously been described, the similar effects of beech extracts are novel. The combined positive effect on wound-healing and antimicrobial activity has great potential for the treatment of various skin diseases, including acne in future dermal applications.

Different Patterns of mRNA Nuclear Retention during Meiotic Prophase in Larch Microsporocytes.

Recent studies show a crucial role of post-transcriptional processes in the regulation of gene expression. Our research has shown that mRNA retention in the nucleus plays a significant role in such regulation. We studied larch microsporocytes during meiotic prophase, characterized by pulsatile transcriptional activity. After each pulse, the transcriptional activity is silenced, but the transcripts synthesized at this time are not exported immediately to the cytoplasm but are retained in the cell nucleus and especially in Cajal bodies, where non-fully-spliced transcripts with retained introns are accumulated. Analysis of the transcriptome of these cells and detailed analysis of the nuclear retention and transport dynamics of several mRNAs revealed two main patterns of nuclear accumulation and transport. The majority of studied transcripts followed the first one, consisting of a more extended retention period and slow release to the cytoplasm. We have shown this in detail for the pre-mRNA and mRNA encoding RNA pol II subunit 10. In this pre-mRNA, a second (retained) intron is posttranscriptionally spliced at a precisely defined time. Fully mature mRNA is then released into the cytoplasm, where the RNA pol II complexes are produced. These proteins are necessary for transcription in the next pulse to occur.mRNAs encoding translation factors and SERRATE followed the second pattern, in which the retention period was shorter and transcripts were rapidly transferred to the cytoplasm. The presence of such a mechanism in various cell types from a diverse range of organisms suggests that it is an evolutionarily conserved mechanism of gene regulation.

Valorization of Larix decidua Mill. bark by functionalizing bioextract onto chitosan films for sustainable active food packaging.

The present study explored the use of chitosan films functionalized with antioxidants extracted from Larix decidua Mill. bark for active packaging. The pristine chitosan and extract-incorporated chitosan films were evaluated for their structural, physico-mechanical, thermal, viscoelastic and antioxidant properties using advanced characterization techniques. The infrared spectroscopy revealed hydrogen bonding between the extract polyphenolic antioxidants and chitosan, whereas the surface microscopy studies indicated good compatibility between them. The addition of bark extract caused a significant increase in color parameters and solubility with reduction in swelling and elongation at break of the films. The thermal analysis indicated a drop in thermal stability of chitosan films modified with the extract. The dynamic mechanical analysis confirmed the extract-polymer interactions and the viscoelastic nature of the films. The incorporation of bark extract caused remarkable enhancement in the antioxidant activity of chitosan films. Overall, larch bark extract-functionalized chitosan films demonstrated promising potential for food packaging.

Lactobacilli Supplemented with Larch Arabinogalactan and Colostrum Stimulates an Immune Response towards Peripheral NK Activation and Gut Tolerance.

Probiotics possibly affect local and systemic immune reactions and maintain the intestinal immune homeostasis in healthy individuals and patients with diseases such as irritable bowel syndrome (IBS). In this single-center, blinded trial, we enrolled 40 individuals (20 patients with IBS and 20 healthy individuals) whose blood and fecal samples were collected before and after a 21-day administration of a product comprising Lactobacillus spp., larch arabinogalactan, and colostrum. The percentage of HLA-DR+ natural killer (NK) cells was higher in healthy individuals (p = 0.03) than in patients with IBS after product supplementation. In the fecal samples of patients with IBS, we observed a decline in IL-6, IFN-γ, TNF-α, and secretory IgA levels and, simultaneously, an increase in IL-10 and IL-17A levels after supplementation, although non-significant, whereas, in healthy individuals, we observed a significant decline in IL-6 and IFN-γ levels after supplementation (p < 0.001). Nevertheless, we observed a clinical improvement of symptoms in 65-75% of patients with IBS and the complete resolution of the initial symptoms in five of the 20 patients. We also observed a possible prophylactic effect by the inducing system antiviral impact accompanied by a trend for local immune tolerance in the gut in healthy individuals, where it is the desirable state.

Magnetic molecularly imprinted specific solid-phase extraction for determination of dihydroquercetin from Larix griffithiana using HPLC.

The naturally occurring quercetin flavonoid, dihydroquercetin, is widely distributed in plant tissues and has a variety of biological activities. Herein, a magnetic molecularly imprinted solid-phase extraction was tailor made for selective determination of dihydroquercetin in Larix griffithiana using high-performance liquid chromatography. Amino-functionalized core-shell magnetic nanoparticles were prepared and characterized using scanning electron microscopy, transmission electron microscopy, vibrating sample magnetometry, and infrared spectroscopy. The polymer had an average diameter of 250 ± 2.56 nm and exhibited good stability and adsorption for template molecule, which is enriched by hydrogen bonding interaction. Multiple factors for extraction, including loading, washing, elution solvents, and extraction time, were optimized. The limit of detection was 1.23 µg/g. The precision determined at various concentration of dihydroquercetin was less than 4% and the mean recovery was between 74.64 and 101.80%. It has therefore been shown that this protocol can be used as an alternative extraction to quantify dihydroquercetin in L. griffithiana and purify quercetin flavonoid from other complex matrices.

Characteristics of soil phosphorus fractions under different thinning intensities in Larix principis-rupprechtii plantation and the affecting factors.

Thinning is an important plantation management strategy. Phosphorus (P) is one of the limiting nutrients in forest ecosystems. The impacts of thinning on soil P remain unclear. In this study, we investigated the characteristics of soil P fractions in topsoil (0-10 cm) of Larix principis-rupprechtii plantation in Taiyue Mountain, Shanxi Province under different thinning intensity (control, CK, 0%; low thinning, LT, 15%; moderate thinning, MT, 35%; high thinning, HT, 50%) and the affecting factors. The soil P fractions were measured using Tissen modified Hedley P fractionation method. The results showed that the total inorganic P content in soil was significantly higher in stands subjected to moderate thinning than in control. The soil acid phosphatase activity (APA) and the contents of Resin-Pi, NaHCO3-Pi, NaHCO3-Po, NaOH-Pi, and microbial biomass phosphorus (MBP) were significantly increased in LT and MT compared with CK, but the content of NaOH-Po showed an opposite trend. Thinning had no significant effect on soil total P, total organic P, non-available P and residual-P. Soil moisture, organic matter, MBP and APA were the important factors affecting soil P availability. Our results demonstrated that MT could enhance soil P availability in Larix principis-rupprechtii plantation.

Fire severity effects on soil carbon and nutrients and microbial processes in a Siberian larch forest.

Fire frequency and severity are increasing in tundra and boreal regions as climate warms, which can directly affect climate feedbacks by increasing carbon (C) emissions from combustion of the large soil C pool and indirectly via changes in vegetation, permafrost thaw, hydrology, and nutrient availability. To better understand the direct and indirect effects of changing fire regimes in northern ecosystems, we examined how differences in soil burn severity (i.e., extent of soil organic matter combustion) affect soil C, nitrogen (N), and phosphorus (P) availability and microbial processes over time. We created experimental burns of three fire severities (low, moderate, and high) in a larch forest in the northeastern Siberian Arctic and analyzed soils at 1, 8 days, and 1 year postfire. Labile dissolved C and N increased with increasing soil burn severity immediately (1 day) postfire by up to an order of magnitude, but declined significantly 1 week later; both variables were comparable or lower than unburned soils by 1 year postfire. Soil burn severity had no effect on P in the organic layer, but P increased with increasing severity in mineral soil horizons. Most extracellular enzyme activities decreased by up to 70% with increasing soil burn severity. Increasing soil burn severity reduced soil respiration 1 year postfire by 50%. However, increasing soil burn severity increased net N mineralization rates 1 year postfire, which were 10-fold higher in the highest burn severity. While fires of high severity consumed approximately five times more soil C than those of low severity, soil C pools will also be driven by indirect effects of fire on soil processes. Our data suggest that despite an initial increase in labile C and nutrients with soil burn severity, soil respiration and extracellular activities related to the turnover of organic matter were greatly reduced, which may mitigate future C losses following fire.

The Use of a Molybdenum Polyoxometalated Compound to Increase the Amount of Extractives from Wood Wastes.

The treatment of wood wastes of Castanea sativa L., Quercus frainetto, Larix decidua, and Paulownia tomentosa S. in autoclave in the presence of micrometric crystals of H3PMo12O40 showed an impressive increase of the amount of extractives. The extractives were mainly constituted of insoluble compounds that were analyzed by using gas chromatography-mass spectrometry (GC-MS) after acetylation. The GC-MS analysis of the chloroform soluble fraction of the extractives obtained from sativa showed the presence of methyl hexadecanoate and octadecanoic acid, that of the extractives of frainetto showed the presence of octadecanal and some long chain hydrocarbons. decidua extracts showed the presence of large amounts of sesamin, while the extractives of P tomentosa revealed the presence of 4-hydroxy-3,5-dimethoxybenzaldehyde, 4-hydroxy-3,5-dimethoxycimmanaldehyde, and relevant amounts of long chain hydrocarbons. The insoluble fraction showed the presence of relevant amounts of several carbohydrates and, in the case of C. sativa, of inositol.

Mating patterns and pollen dispersal in a Japanese larch (Larix kaempferi) clonal seed orchard: a case study.

Pollination dynamics highly determines the genetic quality of seed orchard crops. However, there is less research about the effect of mating patterns on seed productivity of orchard crops. So far, clonal seed orchards have been producing genetically improved seedlings used for most Japanese larch (Larix kaempferi (Lamb.) Carr.) plantations in China. In the present study, a total of 17 highly variable simple sequence repeat (SSR) markers were used for genotyping a progeny trial population consisting of 647 open-pollinated progenies germinated from seeds which were collected from 63 maternal clones with 140 potential paternal clones in a Japanese larch clonal seed orchard in China. Paternity analysis was used in the present case study in order to evaluate the level of paternal gametic contribution, estimate pollen contamination and selfing rates, and investigate pollination patterns, pollen dispersal patterns and the impact of mating patterns on seed productivity of orchard crops. We observed 93.7% of the success rate of the parental assignment, unequal paternal gametic contribution (0-12.4%) with 6.3% of the progenies derived from pollen contamination or unsampled pollen donors, and absence of evidence for selfing. We also found that pollination rate highly depended on the distance between pollen donors and maternal parents, the majority of the identified crossing (65.7%) occurred between clones within a 150-m radius, and large variations in growth performance existed among the paternal half-siblings. Progeny growth performance (diameter at breast (DBH) and height (HGT)) was measured at Age-20 in order to investigate the impact of mating patterns on timber production of orchard crops. As either the paternal or maternal, two clones (i. e., clones Z38 and Z62) were identified to have produced progenies with higher average stem volume breeding values than that of all of the progenies. Specifically, the genetic gains for volume were 3.53% for the two clones as paternal parents, and 8.26% as the maternal parents at Age-20. Thus, both elite clones were ideal candidates for the construction of next-generation clonal seed orchards due to their synchronous reproductive phenology with greater crossing rate and higher genetic gain. These results improved the pedigree information to provide solid evidence of mating patterns for future design and effective management of seed orchards and for the development of viable long-term breeding strategies for other coniferous species.

Reducing copper use in the environment: the use of larixol and larixyl acetate to treat downy mildew caused by Plasmopara viticola in viticulture.

BACKGROUND: Plant extracts might provide sustainable alternatives to copper fungicides, which are still widely used despite their unfavourable ecotoxicological profile. Larch bark extract and its constituents, larixyl acetate and larixol, have been shown to be effective against grapevine downy mildew (Plasmopara viticola) under semi-controlled conditions. The aim of this study was to reduce the gap between innovation and the registration of a marketable product, namely to develop scalable extraction processes and to evaluate and optimise the performance of larch extracts under different conditions. RESULTS: Toxicologically and technically acceptable solvents like ethanol were used to extract the active compounds larixyl acetate and larixol from bark in sufficient amounts and their combined concentration could be increased by up to 39% by purification steps. The combined concentration of larixyl acetate and larixol from larch turpentine could be increased by up to 66%. The Minimal Inhibitory Concentration (MIC100 ) against P. viticola in vitro (6-23 µg mL-1 ) and the Effective Concentration (EC50 ) in planta under semi-controlled conditions (0.2-0.4 mg mL-1 ) were promising compared with other plant extracts. In vineyards, efficacies of larch extracts reached up to 68% in a stand-alone strategy and 84% in low-copper strategies. CONCLUSION: Larch extracts represent valid candidates for copper reduction in organic vineyards, and their development into a sustainable plant protection product might be feasible. © 2017 Society of Chemical Industry.

Seasonal variations in carbon, nitrogen and phosphorus concentrations and C:N:P stoichiometry in different organs of a Larix principis-rupprechtii Mayr. plantation in the Qinling Mountains, China.

Understanding how concentrations of elements and their stoichiometry change with plant growth and age is critical for predicting plant community responses to environmental change. We used long-term field experiments to explore how the leaf, stem and root carbon (C), nitrogen (N) and phosphorous (P) concentrations and their stoichiometry changed with growth and stand age in a L. principis-rupprechtii Mayr. plantation from 2012-2015 in the Qinling Mountains, China. Our results showed that the C, N and P concentrations and stoichiometric ratios in different tissues of larch stands were affected by stand age, organ type and sampling month and displayed multiple correlations with increased stand age in different growing seasons. Generally, leaf C and N concentrations were greatest in the fast-growing season, but leaf P concentrations were greatest in the early growing season. However, no clear seasonal tendencies in the stem and root C, N and P concentrations were observed with growth. In contrast to N and P, few differences were found in organ-specific C concentrations. Leaf N:P was greatest in the fast-growing season, while C:N and C:P were greatest in the late-growing season. No clear variations were observed in stem and root C:N, C:P and N:P throughout the entire growing season, but leaf N:P was less than 14, suggesting that the growth of larch stands was limited by N in our study region. Compared to global plant element concentrations and stoichiometry, the leaves of larch stands had higher C, P, C:N and C:P but lower N and N:P, and the roots had greater P and C:N but lower N, C:P and N:P. Our study provides baseline information for describing the changes in nutritional elements with plant growth, which will facilitates plantation forest management and restoration, and makes a valuable contribution to the global data pool on leaf nutrition and stoichiometry.

Larch Arabinogalactan Attenuates Myocardial Injury by Inhibiting Apoptotic Cascades in a Rat Model of Ischemia-Reperfusion.

We reported previously that supplementation with apple pectin, a dietary fiber, reduced myocardial injury in a rat model of ischemia-reperfusion. In this study, we further investigated an arabinogalactan, one of the constituent polysaccharides of pectin, to determine which domains comprising pectin were responsible for the protection. In a rat model of 30-min ischemia followed by 3-h reperfusion, supplementation with larch arabinogalactan (LAG) over 50 mg/kg/day significantly reduced infarct size. Reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and immunoblot analyses showed that intake of LAG blocked the steps involved in apoptotic cascades through downregulation of gelsolin gene expression at the protein (Gelsolin) level, inhibition of p38 phosphorylation in mitogen-activated protein kinase (MAPK) pathways, decreased bax/bcl-2 ratio at the protein (Bax/Bcl-2) level, which was correlated with the ratio at the mRNA level, inhibition of the conversion of Procaspase protein to Caspase-3 protein, and consequently a decrease in apoptotic cells. In addition, the intake of LAG reduced the hif1-α gene expression at the protein (HIF1-α) level. These findings suggest that arabinogalactan is an active component of pectin for reducing myocardial injury by inhibiting apoptosis in postocclusion steps, possibly indicating that arabinogalactan can be developed as a cardioprotectant to prevent myocardial injury.

Self-assembly as the underlying mechanism for exine development in Larix decidua D. C.

MAIN CONCLUSION: Our findings suggest a new approach to pollen ontogenetic investigations, entailing consideration of physical factors, which enable a better understanding of exine developmental processes. The sporopollenin-containing part of the pollen wall-the exine-is one of the most complex cell walls in plants. By tracing each stage of microspore development in Larix decidua with TEM, we aimed to understand the underlying mechanisms of its exine establishment. Our hypothesis is that self-assembly interferes with exine development. Our specific aim is to generate experimental simulations of the exine developmental pattern. The sequence of events leading to exine development includes the appearance of spherical units in the periplasmic space, their rearrangement into radial columns, and the appearance of white-lined endexine lamellae. The final accumulation of sporopollenin proceeds in the post-tetrad period. The sequence of self-assembling micellar mesophases corresponds with that of the developmental events: spherical micelles; columns of spherical micelles; and laminate micelles separated by strata of water and visible as white-lined lamellae in TEM. Several patterns, simulating structures at different stages of exine development in Larix, were obtained from in vitro experiments. Purely physicochemical processes of self-assembly, which are not under direct genetic control, play an important role in exine development and share control with the genome. These findings suggest that a new approach to ontogenetic investigations, entailing consideration of physical factors (e.g., cell tensegrity), is required for a better understanding of developmental processes.

[Effects of Different Vegetation Types and Reclamation Years on Soil Bacterial Community Structure in Reclaimed Mine Areas].

Effects of different vegetation types (Ulmus pumila, Larix gmelinii, Armeniaca vulgaris, Picea asperata and Robinia pseudoacacia) and reclamation years (15 and 20 years) on soil bacterial community structure in reclaimed Antaibao opencast mine areas were investigated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and clone sequencing. For 20-year reclaimed soils, the significantly highest and lowest bacterial diversity were found in U. pumila and A. vulgaris stand, respectively, whereas no significant differences were found between the other three vegetation types. Under 15-year plantations, soil bacterial diversity index of P. asperata was significantly higher than that of R. pseudoacacia. Soil bacterial diversity index significantly increased in R. pseudoacacia planted soils but decreased in P. asperata treatment with the increase of reclaimed years. No significant change of soil bacterial community structure was observed in the same reclamation years based on the similarity coefficient analysis, cluster analysis and principal component analysis (PCA). Pearson correlation analysis demonstrated that bacterial diversity index was significantly positively correlated with soil pH. Nitrospira, Sphingomonas, Arthrobacter, Brachybacterium, Rhizobium as well as Mesorhizobium, which play important roles in the nitrogen cycle, degradation of polycyclic aromatic hydrocarbons and other organic matter, were identified by clone sequencing of the DGGE bands. Our results indicated that U. pumila and P. asperata were conducive to the recovery of soil bacterial diversity. The most dominant bacterial community from reclaimed mine soil would be beneficial for restoring wasteland contaminated soil and improving soil fertility.

Larix laricina bark, a traditional medicine used by the Cree of Eeyou Istchee: Antioxidant constituents and in vitro permeability across Caco-2 cell monolayers.

ETHNOPHARMACOLOGICAL RELEVANCE: Larix laricina, a native tree of North America, is a highly respected medicinal plant used for generations by Indigenous Peoples across its range, including the Cree of northern Québec who use the bark to treat symptoms of diabetes. This study investigates the antioxidant capacity and bioavailability of active constituents identified in L. laricina bark extracts. MATERIALS AND METHODS: (1) Oxygen radical absorbance capacity (ORAC) assay was employed to test antioxidant capacity of organic extracts (80% ethanol) from bark of L. laricina as well as fractions, isolated compounds, and media samples collected during permeability assays. (2) Caco-2 cell monolayer cultures were used to determine the permeability of identified antioxidants, which were quantified in basolateral media samples using liquid chromatography - tandem mass spectrometry (HPLC-ESI-MS/MS). RESULTS: Crude ethanolic extract possessed strong antioxidant potential in vitro (7.1±0.3 Trolox equivalents (TE) µM/mg). Among the 16 L. laricina fractions obtained by chromatographic separation, fraction 10 (F10) showed the highest antioxidant capacity (21.8±1.7µm TE/mg). Among other identified antioxidants, the stilbene rhaponticin (isolated from F10) was the most potent (24.6±1.1µm TE/mg). Caco-2 transport studies revealed that none of the identified compounds were detectable in basolateral samples after 2-h treatment with crude extract. In monolayers treated with F10 (60% rhaponticin), small quantities of rhaponticin were increasingly detected over time in basolateral samples with an apparent permeability coefficient (Papp) of 1.86×10-8cm/s (0-60min). To model potential effects on blood redox status, we evaluated the antioxidant capacity of collected basolateral samples and observed enhanced activity over time after exposure to both extract and F10 (75µg/mL) relative to control. CONCLUSIONS: By profiling the antioxidant constituents of L. laricina bark, we identified rhaponticin as the most potent oxygen radical scavenger and observed low permeability in Caco-2 cell monolayers but an increase in basolateral antioxidant capacity.