Physiological Characteristics and Transcriptomic Dissection in Two Root Segments with Contrasting Net Fluxes of Ammonium and Nitrate of Poplar Under Low Nitrogen Availability.

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.

Dissecting MicroRNA-mRNA Regulatory Networks Underlying Sulfur Assimilation and Cadmium Accumulation in Poplar Leaves.

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.

Competing Endogenous RNA Networks Underlying Anatomical and Physiological Characteristics of Poplar Wood in Acclimation to Low Nitrogen 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.

Physiological and molecular mechanisms of heavy metal accumulation in nonmycorrhizal versus mycorrhizal plants.

Uptake, translocation, detoxification, and sequestration of heavy metals (HMs) are key processes in plants to deal with excess amounts of HM. Under natural conditions, plant roots often establish ecto- and/or arbuscular-mycorrhizae with their fungal partners, thereby altering HM accumulation in host plants. This review considers the progress in understanding the physiological and molecular mechanisms involved in HM accumulation in nonmycorrhizal versus mycorrhizal plants. In nonmycorrhizal plants, HM ions in the cells can be detoxified with the aid of several chelators. Furthermore, HMs can be sequestered in cell walls, vacuoles, and the Golgi apparatus of plants. The uptake and translocation of HMs are mediated by members of ZIPs, NRAMPs, and HMAs, and HM detoxification and sequestration are mainly modulated by members of ABCs and MTPs in nonmycorrhizal plants. Mycorrhizal-induced changes in HM accumulation in plants are mainly due to HM sequestration by fungal partners and improvements in the nutritional and antioxidative status of host plants. Furthermore, mycorrhizal fungi can trigger the differential expression of genes involved in HM accumulation in both partners. Understanding the molecular mechanisms that underlie HM accumulation in mycorrhizal plants is crucial for the utilization of fungi and their host plants to remediate HM-contaminated soils.

Physiological and transcriptional regulation in poplar roots and leaves during acclimation to high temperature and drought.

To elucidate the physiological and transcriptional regulatory mechanisms that underlie the responses of poplars to high temperature (HT) and/or drought in woody plants, we exposed Populus alba × Populus tremula var. glandulosa saplings to ambient temperature (AT) or HT under 80 or 40% field capacities (FC), or no watering. HT increased the foliar total carbon (C) concentrations, and foliar δ(13) C and δ(18) O. HT triggered heat stress signaling via increasing levels of abscisic acid (ABA) and indole-3-acetic acid (IAA) in poplar roots and leaves. After perception of HT, poplars initiated osmotic adjustment by increasing foliar sucrose and root galactose levels. In agreement with the HT-induced heat stress and the changes in the levels of ABA and carbohydrates, we detected increased transcript levels of HSP18 and HSP21, as well as NCED3 in the roots and leaves, and the sugar transporter gene STP14 in the roots. Compared with AT, drought induced greater enhancement of foliar δ(13) C and δ(18) O in poplars at HT. Similarly, drought caused greater stimulation of the ABA and foliar glucose levels in poplars at HT than at AT. Correspondingly, desiccation led to greater increases in the mRNA levels of HSP18, HSP21, NCED3, STP14 and INT1 in poplar roots at HT than at AT. These results suggest that HT has detrimental effects on physiological processes and it induces the transcriptional regulation of key genes involved in heat stress responses, ABA biosynthesis and sugar transport and HT can cause greater changes in drought-induced physiological and transcriptional responses in poplar roots and leaves.

Overexpression of bacterial γ-glutamylcysteine synthetase mediates changes in cadmium influx, allocation and detoxification in poplar.

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.

Exogenous abscisic acid alleviates zinc uptake and accumulation in Populus × canescens exposed to excess zinc.

A greenhouse experiment was conducted to study whether exogenous abscisic acid (ABA) mediates the responses of poplars to excess zinc (Zn). Populus × canescens seedlings were treated with either basal or excess Zn levels and either 0 or 10 µm ABA. Excess Zn led to reduced photosynthetic rates, increased Zn accumulation, induced foliar ABA and salicylic acid (SA), decreased foliar gibberellin (GA3 ) and auxin (IAA), elevated root H2 O2 levels, and increased root ratios of glutathione (GSH) to GSSG and foliar ratios of ascorbate (ASC) to dehydroascorbate (DHA) in poplars. While exogenous ABA decreased foliar Zn concentrations with 7 d treatments, it increased levels of endogenous ABA, GA3 and SA in roots, and resulted in highly increased foliar ASC accumulation and ratios of ASC to DHA. The transcript levels of several genes involved in Zn uptake and detoxification, such as yellow stripe-like family protein 2 (YSL2) and plant cadmium resistance protein 2 (PCR2), were enhanced in poplar roots by excess Zn but repressed by exogenous ABA application. These results suggest that exogenous ABA can decrease Zn concentrations in P. × canescens under excess Zn for 7 d, likely by modulating the transcript levels of key genes involved in Zn uptake and detoxification.

A transcriptomic network underlies microstructural and physiological responses to cadmium in Populus x canescens.

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.

Ectomycorrhizas with Paxillus involutus enhance cadmium uptake and tolerance in Populus × canescens.

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.

Anatomical, physiological and transcriptional responses of two contrasting poplar genotypes to drought and re-watering.

Populus × euramericana (Pe) displays higher stable carbon isotope composition (δ(13)C) and intrinsic water use efficiency (WUEi) than Populus cathayana (Pc) under unlimited water conditions, rendering us to hypothesize that Pe is better acclimated to water deficiency than Pc. To examine this hypothesis, saplings of Pc and Pe were exposed to drought and subsequently re-watered. Pc and Pe exhibited distinct anatomical, physiological and transcriptional responses in acclimation to drought and re-watering, mainly due to stronger responsiveness of transcriptional regulation of genes encoding plasma membrane intrinsic proteins (PIPs), higher starch accumulation, δ(13)C, stable nitrogen isotope composition (δ(15)N) and WUEi , and lower reactive oxygen species (ROS) accumulation and scavenging in Pe. In acclimation to drought, both poplar genotypes demonstrated altered anatomical properties, declined height growth, differential expression of PIPs, activation of ABA signaling pathway, decreased total soluble sugars and starch, increased δ(13)C, δ(15)N and WUEi , and shifted homeostasis of ROS production and scavenging, and these changes can be recovered upon re-watering. These data indicate that Pe is more tolerant to drought than Pc, and that anatomical, physiological and transcriptional acclimation to drought and re-watering is essential for poplars to survive and grow under projected dry climate scenarios in the future.

Net fluxes of ammonium and nitrate in association with H+ fluxes in fine roots of Populus popularis.

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.

Nitrogen metabolism of two contrasting poplar species during acclimation to limiting nitrogen availability.

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.

Performance of coralline hydroxyapatite in sinus floor augmentation: a retrospective study.

OBJECTIVE: The purpose of this study is to retrospectively explore the utilization of coralline hydroxyapatite in maxillary sinus augmentation. METHOD: One hundred and eighteen cases of sinus lift with coralline hydroxyapatite (CHA) were included in this study. In detail, simultaneous implantation was conducted in 78 patients (174 implants) and delayed implantation was done in 40 cases (82 implants) around 6 months after bone transplantation. The clinical features and X-ray radiographs after operation were analyzed to evaluate osseointegration procedures according to a planned medical follow-up. In the delayed group, around 6 months, a bone biopsy was taken just during implant placement in order to evaluate the new formed bone from a histological and histomorphometrical point of view. A further 6 months later, abutment connection was performed, and the patients received prosthetic restoration of the missing teeth. RESULT: Clinically, the incisions healed well. No abnormal reactions were found during follow-up period. All the 174 simultaneous implants were successful after 1-5 years of medical review; Out of 82 delayed implants, 3 were found to be loose. Histologically, all the specimens showed signs of active remodeling, and all the tissues had a large amount of osteocyte at sixth month after sinus augmentation. New bone formed dramatically. Radiologically, the density of CHA gradually reduced since the beginning of the third month, and CHA may be completely resolved at about fifth year. CONCLUSION: CHA is proven an ideal bone graft material for its reliable clinical results and favorable histocompatibility in the treatment of sinus atrophy or other kinds of insufficient bone volume in this region. Moreover, CHA's signal application can achieve desired clinical effect. CLINICAL RELEVANCE: This study shows the clinic application of CHA in maxillary sinus augmentation. Compared with popular mixture of autogenous bone and grafting materials, our results show CHA's signal application can achieve ideal osseointegration interface and satisfying clinic effect.

Homolog of Human placenta-specific gene 8, PcPLAC8-10, enhances cadmium uptake by Populus roots.

Cadmium (Cd) pollution of soil occurs worldwide. Phytoremediation is an effective approach for cleaning up Cd polluted soil. Fast growing Populus species with high Cd uptake capacities are desirable for phytoremediation. Thus, it is important to elucidate the molecular functions of genes involved in Cd uptake by poplars. In this study, PcPLAC8-10, a homolog of Human placenta-specific gene 8 (PLAC8) implicated in Cd transport was functionally characterized in Populus × canescens. PcPLAC8-10 was transcriptionally induced in Cd-treated roots and it encoded a plasma membrane-localized transporter. PcPLAC8-10 exhibited Cd uptake activity when expressed in yeast cells. No difference in growth was observed between wild type (WT) and PcPLAC8-10-overexpressing poplars. PcPLAC8-10-overexpressing poplars exhibited increases in net Cd2+ influxes by 192% and Cd accumulation by 57% in the roots. However, similar reductions in biomass were found in WT and transgenic poplars when exposed to Cd. The complete motif of CCXXXXCPC in PcPLAC8-10 was essential for its Cd transport activity. These results suggest that PcPLAC8-10 is a plasma membrane-localized transporter responsible for Cd uptake in the roots and the complete CCXXXXCPC motif of PcPLAC8-10 plays a key role in its Cd transport activity in poplars.

N-fertilization has different effects on the growth, carbon and nitrogen physiology, and wood properties of slow- and fast-growing Populus species.

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.

Genome-Wide Identification and Characterization of Long Noncoding RNAs in Populus × canescens Roots Treated With Different Nitrogen Fertilizers.

Nitrate (NO3 -) and ammonium (NH4 +) are the primary forms of inorganic nitrogen acquired by plant roots. LncRNAs, as key regulators of gene expression, are a class of non-coding RNAs larger than 200 bp. However, knowledge about the regulatory role of lncRNAs in response to different nitrogen forms remains limited, particularly in woody plants. Here, we performed strand-specific RNA-sequencing of P. × canescens roots under three different nitrogen fertilization treatments. In total, 324 lncRNAs and 6,112 mRNAs were identified as showing significantly differential expression between the NO3 - and NH4NO3 treatments. Moreover, 333 lncRNAs and 6,007 mRNAs showed significantly differential expression between the NH4 + and NH4NO3 treatments. Further analysis suggested that these lncRNAs and mRNAs have different response mechanisms for different nitrogen forms. In addition, functional annotation of cis and trans target mRNAs of differentially expressed lncRNAs indicated that 60 lncRNAs corresponding to 49 differentially expressed cis and trans target mRNAs were involved in plant nitrogen metabolism and amino acid biosynthesis and metabolism. Furthermore, 42 lncRNAs were identified as putative precursors of 63 miRNAs, and 28 differentially expressed lncRNAs were potential endogenous target mimics targeted by 96 miRNAs. Moreover, ceRNA regulation networks were constructed. MSTRG.6097.1, MSTRG.13550.1, MSTRG.2693.1, and MSTRG.12899.1, as hub lncRNAs in the ceRNA networks, are potential candidate lncRNAs for studying the regulatory mechanism in poplar roots under different nitrogen fertilization treatments. The results provide a basis for obtaining insight into the molecular mechanisms of lncRNA responses to different nitrogen forms in woody plants.

Identification and Functional Prediction of Poplar Root circRNAs Involved in Treatment With Different Forms of Nitrogen.

Circular RNAs (circRNAs) are a class of noncoding RNA molecules with ring structures formed by covalent bonds and are commonly present in organisms, playing an important regulatory role in plant growth and development. However, the mechanism of circRNAs in poplar root responses to different forms of nitrogen (N) is still unclear. In this study, high-throughput sequencing was used to identify and predict the function of circRNAs in the roots of poplar exposed to three N forms [1 mM NO3 - (T1), 0.5 mM NH4NO3 (T2, control) and 1 mM NH4 + (T3)]. A total of 2,193 circRNAs were identified, and 37, 24 and 45 differentially expressed circRNAs (DECs) were screened in the T1-T2, T3-T2 and T1-T3 comparisons, respectively. In addition, 30 DECs could act as miRNA sponges, and several of them could bind miRNA family members that play key roles in response to different N forms, indicating their important functions in response to N and plant growth and development. Furthermore, we generated a competing endogenous RNA (ceRNA) regulatory network in poplar roots treated with three N forms. DECs could participate in responses to N in poplar roots through the ceRNA regulatory network, which mainly included N metabolism, amino acid metabolism and synthesis, response to NO3 - or NH4 + and remobilization of N. Together, these results provide new insights into the potential role of circRNAs in poplar root responses to different N forms.

Sulfur metabolism, organic acid accumulation and phytohormone regulation are crucial physiological processes modulating the different tolerance to Pb stress of two contrasting poplars.

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.

Net cadmium flux and accumulation reveal tissue-specific oxidative stress and detoxification in Populus × canescens.

To characterize the dynamics of Cd²âº flux in the rhizosphere and to study cadmium (Cd) plant-internal partitioning in roots, wood, bark and leaves in relation to energy metabolism, reactive oxygen species (ROS) formation and antioxidants, Populus × canescens plantlets were exposed to either 0 or 50 µM CdSO4 for up to 20 days in the nutrient solution. A strong net Cd²âº influx in root apex was observed after Cd exposure for 24 h, even if net Cd²âº influx decreased gradually in roots. A large amount of Cd was accumulated in roots. Cd ions were uploaded via the xylem to leaves and further transported to the phloem where significant accumulation was detected. Cd accumulation led to decreased photosynthetic carbon assimilation but not to the depletion in soluble carbohydrates. Increased levels of ROS were present in all tissues, except the bark of Cd-exposed poplars. To combat Cd-induced superoxide and hydrogen peroxide, P. × canescens appeared to rely mainly on the formation of soluble phenolics as these compounds showed the highest accumulation in the bark and the lowest in wood. Other potential radical scavengers such as proline, sugar alcohols and antioxidant enzymes showed tissue- and exposure time-specific responses to Cd. These results indicate a complex pattern of internal Cd allocation in P. × canescens resulting in higher ROS stress in wood than in bark and intermediate responses in roots and leaves, probably because of differential capacities of these tissues for the production of protective phenolic compounds.

Lead exposure-induced defense responses result in low lead translocation from the roots to aerial tissues of two contrasting poplar species.

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.