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To investigate the pivotal physiological processes modulating lead (Pb) tolerance capacities of poplars, the saplings of two contrasting poplar species, Populus × canescens with high Pb sensitivity and Populus nigra with relatively low Pb sensitivity, were treated with either 0 or 8 mM Pb for 6 weeks. Lead was absorbed by the roots and accumulated massively in the roots and leaves, leading to overproduction of reactive oxygen species, reduced photosynthesis and biomass in both poplar species. Particularly, the tolerance index of P. × canescens was significantly lower than that of P. nigra. Moreover, the physiological responses including the concentrations of nutrient elements, thiols, organic acids, phytohormones and nonenzymatic antioxidants, and the activities of antioxidative enzymes in the roots and leaves were different between the two poplar species. Notably, the differences in concentrations of nutrient elements, organic acids and phytohormones were remarkable between the two poplar species. A further evaluation of the Pb tolerance-related physiological processes showed that the change of ‘sulfur (S) metabolism’ in the roots was greater, and that of ‘organic acid accumulation’ in the roots and ‘phytohormone regulation’ in the leaves were markedly smaller in P. × canescens than those in P. nigra. These results suggest that there are differences in Pb tolerance capacities between P. × canescens and P. nigra, which is probably associated with their contrasting physiological responses to Pb stress, and that S metabolism, organic acid accumulation and phytohormone regulation are probably the key physiological processes modulating the different Pb tolerance capacities between the two poplar species.

Different root zones have distinct capacities for nitrate (NO3−) uptake in Populus species, but the underlying physiological and microRNA (miRNA) regulatory mechanisms remain largely unknown. To address this question, two root zones of Populus × canescens (Ait.) Smith. with contrasting capacities for NO3− uptake were investigated. The region of 0–40 mm (root zone I) to the root apex displayed net influxes, whereas the region of 40–80 mm (root zone II) exhibited net effluxes. Concentrations of NO3− and ammonium (NH4+) as well as nitrate reductase activity were lower in zone II than in zone I. Forty one upregulated and twenty three downregulated miRNAs, and 576 targets of these miRNAs were identified in zone II in comparison with zone I. Particularly, growth-regulating factor 4 (GRF4), a target of upregulated ptc-miR396g-5p and ptc-miR396f_L + 1R-1, was downregulated in zone II in comparison with zone I, probably contributing to lower NO3− uptake rates and assimilation in zone II. Furthermore, several miRNAs and their targets, members of C2H2 zinc finger family and APETALA2/ethylene-responsive element binding protein family, were found in root zones, which probably play important roles in regulating NO3− uptake. These results indicate that differentially expressed miRNA–target pairs play key roles in regulation of distinct NO3− uptake rates and assimilation in different root zones of poplars.

To investigate physiological and transcriptomic regulation mechanisms underlying the distinct net fluxes of NH4+ and NO3− in different root segments of Populus species under low nitrogen (N) conditions, we used saplings of Populus × canescens supplied with either 500 (normal N) or 50 (low N) μM NH4NO3. The net fluxes of NH4+ and NO3−, the concentrations of NH4+, amino acids and organic acids and the enzymatic activities of nitrite reductase (NiR) and glutamine synthetase (GS) in root segment II (SII, 35–70 mm to the apex) were lower than those in root segment I (SI, 0–35 mm to the apex). The net NH4+ influxes and the concentrations of organic acids were elevated, whereas the concentrations of NH4+ and NO3− and the activities of NiR and GS were reduced in SI and SII in response to low N. A number of genes were significantly differentially expressed in SII vs SI and in both segments grown under low vs normal N conditions, and these genes were mainly involved in the transport of NH4+ and NO3−, N metabolism and adenosine triphosphate synthesis. Moreover, the hub gene coexpression networks were dissected and correlated with N physiological processes in SI and SII under normal and low N conditions. These results suggest that the hub gene coexpression networks play pivotal roles in regulating N uptake and assimilation, amino acid metabolism and the levels of organic acids from the tricarboxylic acid cycle in the two root segments of poplars in acclimation to low N availability.

Although poplar plantations are often established on nitrogen (N)-poor soil, the physiological and molecular mechanisms underlying wood properties of poplars in acclimation to low N availability remain largely unknown. To investigate wood properties of poplars in acclimation to low N, Populus � canescens saplings were exposed to either 50 (low N) or 500 (normal N) �M NH4NO3 for 2 months. Low N resulted in decreased xylem width and cell layers of the xylem (the number of cells counted along the ray parenchyma on the stem cross section), narrower lumina of vessels and fibers, greater thickness of double fiber walls (the walls between two adjacent fiber cells), more hemicellulose and lignin deposition, and reduced cellulose accumulation in poplar wood. Consistently, concentrations of gibberellins involved in cell size determination and the abundance of various metabolites including amino acids, carbohydrates and precursors for cell wall biosynthesis were decreased in low N-supplied wood. In line with these anatomical and physiological changes, a number of mRNAs, long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) were significantly differentially expressed. Competing endogenous RNA regulatory networks were identified in the wood of low N-treated poplars. Overall, these results indicate that miRNAs–lncRNAs–mRNAs networks are involved in regulating wood properties and physiological processes of poplars in acclimation to low N availability.

Single-site laparoscopic percutaneous extraperitoneal ligation (SLPEL) for pediatric inguinal hernia gained popularity all over the world. However, complications associated with extraperitoneal knotting were not rare in the classical SLPEL(C-SLPEL) procedure. In order to overcome disadvantages, we herein developed the modified SLPEL (M-SLPEL) procedure, intact circuit ligation of the peritoneum around the internal ring using a homemade hernia needle with a single abdominal wall centesis. To evaluated the effectiveness of the M-SLPEL procedure to decrease adverse events associated with ligation knotting, a comparative study was carried out. A total of 3219 patients from multiple centers were divided into two groups according to the operative procedures: M-SLPEL group and C-SLPEL group. All patients were followed up. Data describing the clinical characteristics, operative time, postoperative hospital stay, and complications was collected and retrospectively analyzed. With equivalent operative time, postoperative hospital stay, there was statistically significant difference between two groups in terms of the overall complications incidence (2.6% in C-SLPEL Vs 0.11% in M-SLPEL, P=0.03), including pain in inguinal area, knot foreign body reaction, palpable knot, recurrence. Together, these findings suggest that the M-SLPEL procedure is an effective and safe approach, with unique advantage in reducing adverse events in the inguinal region.

The process of cadmium (Cd) accumulation and detoxification under different sulfur levels remains largely unknown in woody plants. To investigate the physiological and transcriptomic regulation mechanisms of poplars in response to different sulfate (S) supply levels and Cd exposure, we exposed Populus deltoides saplings to one of the low, moderate and high S levels together with either 0 or 50 µM Cd. Cd accumulation was decreased in low S-treated poplar leaves, and it tended to be increased in high S-supplied leaves under the Cd exposure condition. Sulfur nutrition was deficient in low S-supplied poplars, and it was improved in high S-treated leaves. Cd exposure resulted in lower sulfur level in the leaves supplied with moderate S, it exacerbated a Cd-induced sulfur decrease in low S-treated leaves and it caused a higher sulfur concentration in high S-supplied leaves. In line with the physiological changes, a number of mRNAs and microRNAs (miRNAs) involved in Cd accumulation and sulfur assimilation were identified and the miRNA–mRNA networks were dissected. In the networks, miR395 and miR399 members were identified as hub miRNAs and their targets were ATP sulfurylase 3 (ATPS3) and phosphate 2 (PHO2), respectively. These results suggest that Cd accumulation and sulfur assimilation are constrained by low and enhanced by high S supply, and Cd toxicity is aggravated by low and relieved by high S in poplar leaves, and that miRNA–mRNA regulatory networks play pivotal roles in sulfur-mediated Cd accumulation and detoxification in Cd-exposed poplars.

A series of novel 6-(trifluoromethyl)-1,2,3,4-tetrahydropyrimidine-5-carboxylate derivatives were obtained in good yields from ethyl 4,4,4-trifluoro-3-oxobutanoate, urea, and aryl aldehyde via Biginelli multicomponent reaction. All the corresponding products 4a–4p were examined against four human cancer cell lines (A549, HepG2, COLO205 and DU145) and compounds 4e, 4i, and 4 m which showed promising anticancer activity have been identified.

To investigate the physiological responses of poplars to amino acids as sole nitrogen (N) sources, Populus × canescens (Ait.) Smith plants were supplied with one of three nitrogen fertilizers (NH4NO3, phenylalanine (Phe) or the mixture of NH4NO3 and Phe) in sand culture. A larger root system, and decreased leaf size and CO2 assimilation rate was observed in Phe- versus NH4NO3-treated poplars. Consistently, a greater root biomass and a decreased shoot growth were detected in Phe-supplied poplars. Decreased enzymatic activities of nitrate reductase (NR), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH) and elevated activities of nitrite reductase (NiR), phenylalanine ammonia lyase (PAL), glutamine synthetase (GS) and asparagine synthase (AS) were found in Phe-treated roots. Accordingly, reduced concentrations of NH4+, NO3- and total N, and enhanced N-use efficiencies (NUEs) were detected in Phe-supplied poplars. Moreover, the transcript levels of putative Phe transporters ANT1 and ANT3 were upregulated, and the mRNA levels of NR, glutamine synthetase 2 (GS2), NADH-dependent glutamate synthase (NADH-GOGAT), GDH and asparagine synthetase 2 (ASN2) were downexpressed in Phe-treated roots and/or leaves. The 15N-labeled Phe was mainly allocated in the roots and only a small amount of 15N-Phe was translocated to poplar aerial parts. These results indicate that poplar roots can acquire Phe as an N source to support plant growth and that Phe-induced NUEs in the poplars are probably associated with NH4+ re-utilization after Phe deamination and the carbon bonus simultaneously obtained during Phe uptake.

To explore whether lead (Pb)-induced defense responses are responsible for the low root-to-shoot Pb translocation, we exposed saplings of the two contrasting poplar species, Populus × canescens with relatively high root-to-shoot Pb translocation and P. nigra with low Pb translocation, to 0 or 8 mM PbCl2. Pb translocation from the roots to aboveground tissues was lower by 57% in P. nigra than that in P. × canescens. Lower Pb concentrations in the roots and aerial tissues, greater root biomass, and lower ROS overproduction in the roots were found in P. nigra than those in P. × canescens treated with Pb. P. nigra roots had higher proportions of cell walls (CWs)-bound Pb and water insoluble Pb compounds, and higher transcript levels of some pivotal genes related to Pb vacuolar sequestration, such as phytochelatin synthetase 1.1 (PCS1.1), ATP-binding cassette transporter C1.1 (ABCC1.1) and ABCC3.1 than P. × canescens roots. Pb exposure induced defense responses including increases in the contents of pectin and hemicellulose, and elevated oxalic acid accumulation, and the transcriptional upregulation of PCS1.1, ABCC1.1 and ABCC3.1 in the roots of P. nigra and P. × canescens. These results suggest that the stronger defense barriers in P. nigra roots are probably associated with the lower Pb translocation from the roots to aerial tissues, and that Pb exposure-induced defense responses can enhance the barriers against Pb translocation in poplar roots.

Sulfur (S) can modulate plant responses to toxic heavy metals, but the underlying physiological and transcriptional regulation mechanisms remain largely unknown. To investigate the effects of S supply on lead (Pb)-induced toxicity in poplars, Populus deltoides monilifera (Aiton) Eckenw. saplings were exposed to 0 or 50 μM Pb together with one of the three S concentrations (0 (low S), 100 (moderate S) or 1500 (high S) μM Na2SO4). Populus deltoides roots absorbed Pb and it was partially translocated to the aerial organs, thereby decreasing the CO2 assimilation rate and leaf growth. Lead accumulation in poplars caused the overproduction of O2- and H2O2 to induce higher levels of total thiols (T-SH) and glutathione (GSH). Lead uptake by the roots and its accumulation in the aerial organs were repressed by low S application, but stimulated by high S supply. Lead-induced O2- and H2O2 production were exacerbated by S limitation, but alleviated by high S supply. Moreover, the concentrations of S-containing antioxidants including T-SH and GSH were reduced in S-deficient poplars, but increased in high S-treated plants, which corresponded well to the changes in the activities of enzymes involved in S assimilation and GSH biosynthesis. The transcript levels of both genes encoding sulfate transporters, i.e., SULTR1.1 and SULTR2.2, were elevated by low S application or high S supply in the roots, and the transcriptional upregulation of both genes was more pronounced under Pb exposure. Furthermore, the mRNA levels of several genes involved in S assimilation and the biosynthesis of GSH and phytochelatins, i.e., ATPS1, ATPS3, GSHS1, GSHS2 and PCS1, were upregulated in poplar roots with high S supply, particularly under Pb exposure. These results indicate that a high S supply can stimulate Pb accumulation and reduce its toxicity in poplars by improving S assimilation and stimulating the biosynthesis of S-containing compounds including T-SH and GSH.

Phosphorus (P) and nitrogen (N) are the two essential macronutrients for tree growth and development. To elucidate the P and N physiology of woody plants during acclimation to P and/or N starvation, we exposed saplings of the slow-growing Populus simonii Carr (Ps) and the fast-growing Populus × euramericana Dode (Pe) to complete nutrients or starvation of P, N or both elements (NP). P. × euramericana had lower P and N concentrations and greater P and N amounts due to higher biomass production, thereby resulting in greater phosphorus use efficiency/N use efficiency (PUE/NUE) compared with Ps. Compared with the roots of Ps, the roots of Pe exhibited higher enzymatic activities in terms of acid phosphatases (APs) and malate dehydrogenase (MDH), which are involved in P mobilization, and nitrate reductase (NR), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH), which participate in N assimilation. The responsiveness of the transcriptional regulation of key genes encoding transporters for phosphate, ammonium and nitrate was stronger in Pe than in Ps. These results suggest that Pe possesses a higher capacity for P/N uptake and assimilation, which promote faster growth compared with Ps. In both poplars, P or NP starvation caused significant decreases in the P concentrations and increases in PUE. Phosphorus deprivation induced the activity levels of APs, phosphoenolpyruvate carboxylase and MDH in both genotypes. Nitrogen or NP deficiency resulted in lower N concentrations, amino acid levels, NR and GOGAT activities, and higher NUE in both poplars. Thus, in Ps and Pe, the mRNA levels of PHT1;5, PHT1;9, PHT2;1, AMT2;1 and NR increased in the roots, while PHT1;9, PHO1;H1, PHO2, AMT1;1 and NRT2;1 increased in the leaves during acclimation to P, N or NP deprivation. These results suggest that both poplars suppress P/N uptake, mobilization and assimilation during acclimation to P, N or NP starvation.

Glutathione (GSH) plays an important role in cadmium (Cd) tolerance in woody plants, but the underlying mechanisms remain largely unknown. To elucidate the physiological and transcriptional regulation mechanisms of GSH-mediated Cd tolerance in woody plants, we exposed Populus × canescens (Ait.) Smith saplings to either 0 or 75 μM Cd together with one of three external GSH levels. Glutathione treatments include buthionine sulfoximine (BSO, an inhibitor of GSH biosynthesis), no external GSH and exogenous GSH. External GSH resulted in higher Cd2+ uptake rate in the roots, greater Cd amount in poplars, lower Cd-induced H2O2 levels in the roots, and higher contents of endogenous GSH in Cd-treated roots and leaves. Furthermore, external GSH led to upregulated transcript levels of several genes including zinc/iron regulated transporter related protein 6.2 (ZIP6.2) and natural resistance-associated macrophage protein 1.3 (NRAMP1.3), which probably take part in Cd uptake, glutathione synthetase 2 (GS2) implicated in Cd detoxification, metal tolerance protein 1 (MTP1) and ATP-binding cassette transporter C3 (ABCC3) involved in Cd vacuolar accumulation in the roots, γ-glutamylcysteine synthetase (ECS) and phytochelatin synthetase family protein 1 (PCS1) involved in Cd detoxification, and oligopeptide transporter 7 (OPT7) probably implicated in Cd detoxification in the leaves of Cd-exposed P. × canescens. In contrast, BSO often displayed the opposite effects on Cd-triggered physiological and transcriptional regulation responses in poplars. These results suggest that exogenous GSH can enhance Cd accumulation and alleviate its toxicity in poplars. This is probably attributed to external-GSH-induced higher net Cd2+ influx in the roots, greater Cd accumulation in aerial parts, stronger scavenging of reactive oxygen species, and transcriptional overexpression of several genes involved in Cd uptake, detoxification and accumulation.

High temperature (HT) and drought are both critical factors that constrain tree growth and survival under global climate change, but it is surprising that the transcriptomic reprogramming and physiological relays involved in the response to HT and/or drought remain unknown in woody plants. Thus, Populus simonii saplings were exposed to either ambient temperature or HT combined with sufficient watering or drought. RNA-sequencing analysis showed that a large number of genes were differentially expressed in poplar roots and leaves in response to HT and/or desiccation, but only a small number of these genes were identified as overlapping heat-/drought-responsive genes that are mainly involved in RNA regulation, transport, hormone metabolism, and stress. Furthermore, the overlapping heat-/drought-responsive genes were co-expressed and formed hierarchical genetic regulatory networks under each condition compared. HT-/drought-induced transcriptomic reprogramming is linked to physiological relays in poplar roots and leaves. For instance, HT- and/or drought-induced abscisic acid accumulation and decreases in auxin and other phytohormones corresponded well with the differential expression of a few genes involved in hormone metabolism. These results suggest that overlapping heat-/drought-responsive genes will play key roles in the transcriptional and physiological reconfiguration of poplars to HT and/or drought under future climatic scenarios.

Overexpression of bacterial γ-glutamylcysteine synthetase in the cytosol of Populus tremula × P. alba produces higher glutathione (GSH) concentrations in leaves, thereby indicating the potential for cadmium (Cd) phytoremediation. However, the net Cd(2+) influx in association with H(+) /Ca(2+) , Cd tolerance, and the underlying molecular and physiological mechanisms are uncharacterized in these poplars. We assessed net Cd(2+) influx, Cd tolerance and the transcriptional regulation of several genes involved in Cd(2+) transport and detoxification in wild-type and transgenic poplars. Poplars exhibited highest net Cd(2+) influxes into roots at pH 5.5 and 0.1 mM Ca(2+) . Transgenics had higher Cd(2+) uptake rates and elevated transcript levels of several genes involved in Cd(2+) transport and detoxification compared with wild-type poplars. Transgenics exhibited greater Cd accumulation in the aerial parts than wild-type plants in response to Cd(2+) exposure. Moreover, transgenic poplars had lower concentrations of O2 ˙(-) and H2 O2 ; higher concentrations of total thiols, GSH and oxidized GSH in roots and/or leaves; and stimulated foliar GSH reductase activity compared with wild-type plants. These results indicate that transgenics are more tolerant of 100 μM Cd(2+) than wild-type plants, probably due to the GSH-mediated induction of the transcription of genes involved in Cd(2+) transport and detoxification.

Objective To investigate the effect and analyze the possible mechanism of celecoxib combined with tegafur, gimeracil and oteracil potassium on growth and lymphangiogenesis of subcutaneously transplanted human gastric cancer in nude mice. Methods Human gastric cancer was transplanted subcutaneously successfully in nude mice. When the largest diameter of tumor reached about 5mm, the nude mice were randomly divided into four groups (6 each): control group, celecoxib group, tegafur+gimeracil+oteracil potassium group, and celecoxib combination group. The drugs were administered respectively for 21 days. Then the tumor tissues were collected, tumor weight was measured, and tumor inhibition rate was calculated. The changes in body weight of the nude mice before and after treatment were recorded, and the side effect of drug therapy was assessed. The expression levels of COX-2 and VEGF-C protein were determined by immunohistochemistry, and the density of lymphatic vessel was calculated. Results No obvious side effect was found, and the body weight increased after the treatment (P0.05). Conclusion When given alone, both celecoxib and tegafur+gimeracil+oteracil potassium show obvious antitumor effect, and combination of these two drugs will give rise a synergistical antitumor effect. Celecoxib and celecoxib combined with tegafur gimeracil oteracil potassium may reduce the expression of COX-2, in turn downregulate the expression of VEGF-C, and then inhibit the tumor lymphangiogenesis. DOI: 10.11855/j.issn.0577-7402.2013.12.03

Ectomycorrhizas (EMs), which are symbiotic organs formed between tree roots and certain fungi, can mediate cadmium (Cd) tolerance of host plants, but the underlying physiological and molecular mechanisms are not fully understood. To investigate EMs mediated Cd tolerance in woody plants, Populus × canescens was inoculated with Paxillus involutus (strain MAJ) to establish mycorrhizal roots. Mycorrhizal poplars and non-mycorrhizal controls were exposed to 0 or 50 μM CdSO4 . EMs displayed higher net Cd(2+) influx than non-mycorrhizal roots. Net Cd(2+) influx was coupled with net H(+) efflux and inactivation of plasma membrane (PM) H(+) -ATPases reduced Cd(2+) uptake of EMs less than of non-mycorrhizal roots. Consistent with higher Cd(2+) uptake in EMs, in most cases, transcript levels of genes involved in Cd(2+) uptake, transport and detoxification processes were increased in EMs compared to non-mycorrhizal roots. Higher CO2 assimilation, improved nutrient and carbohydrate status, and alleviated oxidative stress were found in mycorrhizal compared to non-mycorrhizal poplars despite higher Cd(2+) accumulation. These results indicate that mycorrhizas increase Cd(2+) uptake, probably by an enlarged root volume and overexpression of genes involved in Cd(2+) uptake and transport, and concurrently enhance Po. × canescens Cd tolerance by increased detoxification, improved nutrient and carbohydrate status and defence preparedness.

Bark tissue of Populus × canescens can hyperaccumulate cadmium, but microstructural, transcriptomic, and physiological response mechanisms are poorly understood. Histochemical assays, transmission electron microscopic observations, energy-dispersive x-ray microanalysis, and transcriptomic and physiological analyses have been performed to enhance our understanding of cadmium accumulation and detoxification in P. × canescens. Cadmium was allocated to the phloem of the bark, and subcellular cadmium compartmentalization occurred mainly in vacuoles of phloem cells. Transcripts involved in microstructural alteration, changes in nutrition and primary metabolism, and stimulation of stress responses showed significantly differential expression in the bark of P. × canescens exposed to cadmium. About 48% of the differentially regulated transcripts formed a coregulation network in which 43 hub genes played a central role both in cross talk among distinct biological processes and in coordinating the transcriptomic regulation in the bark of P. × canescens in response to cadmium. The cadmium transcriptome in the bark of P. × canescens was mirrored by physiological readouts. Cadmium accumulation led to decreased total nitrogen, phosphorus, and calcium and increased sulfur in the bark. Cadmium inhibited photosynthesis, resulting in decreased carbohydrate levels. Cadmium induced oxidative stress and antioxidants, including free proline, soluble phenolics, ascorbate, and thiol compounds. These results suggest that orchestrated microstructural, transcriptomic, and physiological regulation may sustain cadmium hyperaccumulation in P. × canescens bark and provide new insights into engineering woody plants for phytoremediation.

Drought-induced tree mortality, which rapidly alters forest ecosystem composition, structure, and function, as well as the feedbacks between the biosphere and climate, has occurred worldwide over the past few decades, and is expected to increase pervasively as climate change progresses. The objectives of this review are to (1) highlight the likely ecological consequences of drought-induced tree mortality, (2) synthesize the hypotheses related to drought-induced tree mortality, (3) discuss the implications of current knowledge for modeling tree mortality processes under climate change, and (4) highlight future research needs. First, we emphasize the likely ecological consequences of tree mortality from ecosystem to biome to continental scales. We then document and criticize multiple non-exclusive tree mortality hypotheses (e.g., carbon starvation — carbon supply is less than carbon demand; and hydraulic failure — desiccation from failed water transport) from a more comprehensive ecological perspective. Next, we extend a forest decline concept model, Manion’s framework, by considering new emerging environmental conditions, for a more thorough understanding of the effects of climate change on forest decline. We find that an increase in drought frequency and (or) climate-change-type droughts may trigger increased background tree mortality rates and severe forest dieback events, accelerating species turnover and ecological regime shifts. The contribution of CO2 fertilization, rising temperature within the optimal growth range, and increased nitrogen deposition may defer or reduce this trend in tree mortality, but such contributions will vary between locations, species, and tree sizes. Multiple hypotheses proposed for drought-induced tree mortality are discussed, but coupling carbon and water cycles could help resolve the debate. The absence of a physiological understanding of tree mortality mechanisms limits the predictive ability of current models from stand-level process-based models to dynamic global vegetation models. We thus suggest that long-term observations, experiments, and models should be tightly interwoven during the research process to better forecast future climate changes and evaluate their impacts on forests.

To investigate how N-fertilization affects the growth, carbon and nitrogen (N) physiology, and wood properties of poplars with contrasting growth characteristics, slow-growing (Populus popularis, Pp) and fast-growing (P. alba×P. glandulosa, Pg) poplar saplings were exposed to different N levels. Above-ground biomass, leaf area, photosynthetic rates (A), instantaneous photosynthetic nitrogen use efficiency (PNUE (i)), chlorophyll and foliar sugar concentrations were higher in Pg than in Pp. Foliar nitrate reductase (NR) activities and root glutamate synthase (GOGAT) activities were higher in Pg than in Pp as were the N amount and NUE of new shoots. Lignin contents and calorific values of Pg wood were less than that of Pp wood. N-fertilization reduced root biomass of Pg more than of Pp, but increased leaf biomass, leaf area, A, and PNUE(i) of Pg more than of Pp. Among 13 genes involved in the transport of ammonium or nitrate or in N assimilation, transcripts showed more pronounced changes to N-fertilization in Pg than in Pp. Increases in NR activities and N contents due to N-fertilization were larger in Pg than in Pp. In both species, N-fertilization resulted in lower calorific values as well as shorter and wider vessel elements/fibres. These results suggest that growth, carbon and N physiology, and wood properties are more sensitive to increasing N availability in fast-growing poplars than in slow-growing ones, which is probably due to prioritized resource allocation to the leaves and accelerated N physiological processes in fast-growing poplars under higher N levels. peerReviewed

Objective To explore the effects of human miR-451 on the proliferation,apoptosis and invasive ability of colon cancer cell line SW620.Methods Four groups were designed in the present experiment,i.e.,cells were transfected with 100nmol/L of miR-451 mimics in miR-451 mimics group,with 100nmol/L of negative control in negative control(NC) group,with Lipo 2000 alone in blank control group,and without transfection of either Lipo2000 or miRNA in control group.The expression of miR-451 was measured by real-time RT-PCR 24h after transfection,and proliferation of cells was examined by using the Cell Counting Kit-8 at respective time of 24,48,72 hours after transfction.The apoptotic rate was assessed by flow cytometry(FCM) and invasive ability was determined by Transwell assay.The effect of miR-451 mimics on the expression of macrophage migration inhibitory factor(MIF) protein was measured by immunofluorescent and Western blot assay.Results Twenty-four hours after transfection,an increase in expression of miR-451 was noted in miR-451 mimics group(67.96±13.33) when compared with NC group,in which the expression of miR-451 was defined as 1(P < 0.01).In comparison with the other groups,miR-451 mimics group showed a significant inhibition of proliferation at 48 and 72 hours after transfection(P < 0.01),but no significant change in apoptotic rate was found at 48 hour after transfection(P > 0.05).Immunofluorescent assay showed that MIF positive rate of miR-451 mimics group(23.9%±14.9%) was decreased when compared with that of NC group(53.5%±14.1%),blank transfection group(45.2%±19.8%) and blank control group(59.3%±22.2%)(F=7.356,P < 0.01).Transwell assay illustrated that the invasive ability in miR-451 mimics group(37.4±6.1 invasive cells per field) was lower than that in negative control group(55.6±3.6 invasive cells per field),blank transfection group(60.8±7.4 invasive cells per field) and blank control group(61.6±8.6 invasive cells per field,P < 0.01).Conclusion Up-regulation of miR-451 expression may inhibit the bioactivity of colon cancer cells,and it may be attributed to the inhibition of MIF expression.

Heavy metal (HM)-accumulating herbaceous and woody plants are employed for phytoremediation. To develop improved strategies for enhancing phytoremediation efficiency, knowledge of the microstructural, physiological and molecular responses underlying HM-accumulation is required. Here we review the progress in understanding the structural, physiological and molecular mechanisms underlying HM uptake, transport, sequestration and detoxification, as well as the regulation of these processes by signal transduction in response to HM exposure. The significance of genetic engineering for enhancing phytoremediation efficiency is also discussed. In herbaceous plants, HMs are taken up by roots and transported into the root cells via transmembrane carriers for nutritional ions. The HMs absorbed by root cells can be further translocated to the xylem vessels and unloaded into the xylem sap, thereby reaching the aerial parts of plants. HMs can be sequestered in the cell walls, vacuoles and the Golgi apparatuses. Plant roots initially perceive HM stress and trigger the signal transduction, thereby mediating changes at the molecular, physiological, and microstructural level. Signaling molecules such as phytohormones, reactive oxygen species (ROS) and nitric oxide (NO), modulate plant responses to HMs via differentially expressed genes, activation of the antioxidative system and coordinated cross talk among different signaling molecules. A number of genes participated in HM uptake, transport, sequestration and detoxification have been functionally characterized and transformed to target plants for enhancing phytoremediation efficiency. Fast growing woody plants hold an advantage over herbaceous plants for phytoremediation in terms of accumulation of high HM-amounts in their large biomass. Presumably, woody plants accumulate HMs using similar mechanisms as herbaceous counterparts, but the processes of HM accumulation and signal transduction can be more complex in woody plants.

Nitrogen (N) starvation and excess have distinct effects on N uptake and metabolism in poplars, but the global transcriptomic changes underlying morphological and physiological acclimation to altered N availability are unknown. We found that N starvation stimulated the fine root length and surface area by 54 and 49%, respectively, decreased the net photosynthetic rate by 15% and reduced the concentrations of NH4+, NO3(-) and total free amino acids in the roots and leaves of Populus simonii Carr. in comparison with normal N supply, whereas N excess had the opposite effect in most cases. Global transcriptome analysis of roots and leaves elucidated the specific molecular responses to N starvation and excess. Under N starvation and excess, gene ontology (GO) terms related to ion transport and response to auxin stimulus were enriched in roots, whereas the GO term for response to abscisic acid stimulus was overrepresented in leaves. Common GO terms for all N treatments in roots and leaves were related to development, N metabolism, response to stress and hormone stimulus. Approximately 30-40% of the differentially expressed genes formed a transcriptomic regulatory network under each condition. These results suggest that global transcriptomic reprogramming plays a key role in the morphological and physiological acclimation of poplar roots and leaves to N starvation and excess.

To investigate N metabolism of two contrasting Populus species in acclimation to low N availability, saplings of slow-growing species (Populus popularis, Pp) and a fast-growing species (Populus alba × Populus glandulosa, Pg) were exposed to 10, 100, or 1000 μM NH4NO3. Despite greater root biomass and fine root surface area in Pp, lower net influxes of NH4(+) and NO3(-) at the root surface were detected in Pp compared to those in Pg, corresponding well to lower NH4(+) and NO3(-) content and total N concentration in Pp roots. Meanwhile, higher stable N isotope composition (δ(15)N) in roots and stronger responsiveness of transcriptional regulation of 18 genes involved in N metabolism were found in roots and leaves of Pp compared to those of Pg. These results indicate that the N metabolism of Pp is more sensitive to decreasing N availability than that of Pg. In both species, low N treatments decreased net influxes of NH4(+) and NO3(-), root NH4(+) and foliar NO3(-) content, root NR activities, total N concentration in roots and leaves, and transcript levels of most ammonium (AMTs) and nitrate (NRTs) transporter genes in leaves and genes involved in N assimilation in roots and leaves. Low N availability increased fine root surface area, foliar starch concentration, δ(15)N in roots and leaves, and transcript abundance of several AMTs (e.g. AMT1;2) and NRTs (e.g. NRT1;2 and NRT2;4B) in roots of both species. These data indicate that poplar species slow down processes of N acquisition and assimilation in acclimation to limiting N supply. peerReviewed

Poplar plants are cultivated as woody crops, which are often fertilized by addition of ammonium (NH4(+)) and/or nitrate (NO3(-)) to improve yields. However, little is known about net NH4(+)/NO3(-) fluxes and their relation with H(+) fluxes in poplar roots. In this study, net NH4(+)/NO3(-) fluxes in association with H(+) fluxes were measured non-invasively using scanning ion-selective electrode technique in fine roots of Populus popularis. Spatial variability of NH4(+) and NO3(-) fluxes was found along root tips of P. popularis. The maximal net uptake of NH4(+) and NO3(-) occurred, respectively, at 10 and 15 mm from poplar root tips. Net NH4(+) uptake was induced by ca. 48 % with provision of NO3(-) together, but net NO3(-) uptake was inhibited by ca. 39 % with the presence of NH4(+) in poplar roots. Furthermore, inactivation of plasma membrane (PM) H(+)-ATPases by orthovanadate markedly inhibited net NH4(+)/NO3(-) uptake and even led to net NH4(+) release with NO3(-) co-provision. Linear correlations were observed between net NH4(+)/NO3(-) and H(+) fluxes in poplar roots except that no correlation was found between net NH4(+) and H(+) fluxes in roots exposed to NH4Cl and 0 mM vanadate. These results indicate that root tips play a key role in NH4(+)/NO3(-) uptake and that net NH4(+)/NO3(-) fluxes and the interaction of net fluxes of both ions are tightly associated with H(+) fluxes in poplar roots.

Ectomycorrhizas (EMs) alleviate stress tolerance of host plants, but the underlying molecular mechanisms are unknown. To elucidate the basis of EM-induced physiological changes and their involvement in stress adaptation, we investigated metabolic and transcriptional profiles in EM and non-EM roots of gray poplar (Populus × canescens) in the presence and absence of osmotic stress imposed by excess salinity. Colonization with the ectomycorrhizal fungus Paxillus involutus increased root cell volumes, a response associated with carbohydrate accumulation. The stress-related hormones abscisic acid and salicylic acid were increased, whereas jasmonic acid and auxin were decreased in EM compared with non-EM roots. Auxin-responsive reporter plants showed that auxin decreased in the vascular system. The phytohormone changes in EMs are in contrast to those in arbuscular mycorrhizas, suggesting that EMs and arbuscular mycorrhizas recruit different signaling pathways to influence plant stress responses. Transcriptome analyses on a whole genome poplar microarray revealed activation of genes related to abiotic and biotic stress responses as well as of genes involved in vesicle trafficking and suppression of auxin-related pathways. Comparative transcriptome analysis indicated EM-related genes whose transcript abundances were independent of salt stress and a set of salt stress-related genes that were common to EM non-salt-stressed and non-EM salt-stressed plants. Salt-exposed EM roots showed stronger accumulation of myoinositol, abscisic acid, and salicylic acid and higher K+-to-Na+ ratio than stressed non-EM roots. In conclusion, EMs activated stress-related genes and signaling pathways, apparently leading to priming of pathways conferring abiotic stress tolerance.

Aziridines underwent cyclization reaction with carbon disulfide and isothiocyanate in the presence of organophosphine to afford thiazolidinone derivatives in good to high yields. The mechanistic study revealed that organophosphine serves as a catalyst in the reaction.

We investigated the individual and combined effects of elevated CO 2 concentration and fertilization on aboveground growth of three poplar species (Populus alba L. Clone 2AS-11, P. nigra L. Clone Jean Pourtet and P. × euramericana Clone I-214) growing in a short-rotation coppice culture for two growing seasons after coppicing. Free-air carbon dioxide enrichment (FACE) stimulated the number of shoots per stool, leaf area index measured with a fish-eye-type plant canopy analyzer (LAI optical ), and annual leaf production, but did not affect dominant shoot height or canopy productivity index. Comparison of LAI optical with LAI estimates from litter collections and from allometric relationships showed considerable differences. The increase in biomass in response to FACE was caused by an initial stimulation of absolute and relative growth rates, which disappeared after the first growing season following coppicing. An ontogenetic decline in growth in the FACE treatment, together with strong competition inside the dense plantation, may have caused this decrease. Fertilization did not influence aboveground growth, although some FACE responses were more pronounced in fertilized trees. A species effect was observed for most parameters.

• Mycorrhizal fungi and hydrogels (water-absorbing polymers) can improve water availability for trees. The combination of both factors for plant performance under water limitation has not yet been studied. • To investigate the influence of the ectomycorrhizal fungus Paxillus involutus, hydrogel and the combination of both factors, a drought-sensitive poplar, Populus euphratica, was examined in this study. • After 16 weeks of inoculation, no ectomycorrhizas were found. Nevertheless, P. involutusinoculated poplars displayed increased concentrations of soluble sugars and osmolality, leading to an improved water status. Growth was diminished compared with non-inoculated P. euphratica .T he presence of hydrogels in the rooting medium resulted in increased biomass and higher plant water content and decreased the osmolality of plant tissues. Drought markedly decreased water contents in rooting medium and plants, and leaf chlorophyll fluorescence, and stimulated the root growth, concentrations of soluble sugars and osmolality in plants. Under drought conditions, P. euphratica exhibited osmoregulation by accumulation of low-molecular-weight carbohydrates. • These data indicate that adding hydrogels to soils may improve the plant performance. The reasons for improved osmoregulation by fungi and hydrogels were probably related to their stimulating influence on the formation of soluble carbohydrates under drought conditions. Mots-cles : secheresse / hydrogel / champignon mycorhizien / peuplier / hydrates de carbone

The goal of this study was to investigate whether increased nitrogen use efficiency found in Populus nigra L. (Jean Pourtet) under elevated CO2 would correlate with changes in the production of carbon-based secondary compounds (CBSCs). Using Free-Air CO2 Enrichment (FACE) technology, a poplar plantation was exposed to either ambient (about 370 mu mol mol(-1) CO2) or elevated (about 550 mu mol mol(-1) CO2) [CO2] for 5 years. After three growing seasons, the plantation was coppiced and half of the experimental plots were fertilized with nitrogen. CBSCs, total nitrogen and lignin-bound nitrogen were quantified in secondary sprouts in seasons of active growth and dormancy during 2 years after coppicing. Neither elevated CO2 nor nitrogen fertilisation alone or in combination influenced lignin concentrations in wood. Soluble phenolics and soluble proteins in wood decreased slightly in response to elevated CO2. Higher nitrogen supply stimulated formation of CBSCs and increased protein concentrations. Lignin-bound nitrogen in wood ranged from 0.37-1.01 mg N g(-1) dry mass accounting for 17-26% of total nitrogen in wood, thus, forming a sizeable nitrogen fraction resistant to chemical degradation. The concentration of this nitrogen fraction was significantly decreased by elevated CO2, increased in response to nitrogen fertilisation and showed a significant CO2 x fertilisation interaction. Seasonal changes markedly affected the internal nitrogen pools. Soluble proteins in wood were 52-143% higher in the dormant than in the growth season. Positive correlations existed between the biosynthesis of proteins and CBSCs. The limited responses to elevated CO2 and nitrogen fertilisation indicate that growth and defence are well orchestrated in P. nigra and that changes in the balance of both resources-nitrogen and C-have only marginal effects on wood chemistry.