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Hibiscus hamabo Siebold & Zuccarini is one of the few semi-mangrove plants in the genus Hibiscus that can survive in saline-alkali soil and flooded land, but the mechanism underlying its adaptation to salt soil remains unknown. Here, to uncover this unsolved mystery, we characterized the changes in the accumulation of specific metabolites under salt stress in H. hamabo by integrating physiological, metabolic, and transcriptomic data, and found that osmotic adjustment and abscisic acid (ABA) is highly associated with the salt stress response. Further, a weighted gene co-expression network analysis was performed on the root transcriptome data, which identified three key candidate transcription factors responsive to salt stress. Among them, the expression HhERF9 was significantly upregulated under salt stress and ABA treatment and was involved in regulating the expression of genes related to the salt stress response. Further research indicated that HhERF9 enhances the accumulation of proline and soluble sugars by regulating the expression of genes such as NHX2 and P5CS. These findings provide a reference for improving H. hamabo through targeted genetic engineering and lay a theoretical foundation for its future promotion and cultivation in saline-alkali areas., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interests regarding the publication of this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Carboxymethyl cellulose (CMC) and polylactic acid (PLA) are recognized for their environmental friendliness. By merging them into a composite film, packaging solutions can be designed with good performance. Nonetheless, the inherent interface disparity between CMC and PLA poses a challenge, and there may be layer separation issues. This study introduces a straightforward approach to mitigate this challenge by incorporating tannin acid and ferric chloride in the fabrication of the CMC-PLA. The interlayer compatibility was improved by the in situ formation of a cohesive interface. The resulting CMC/TA-PLA/Fe multilayer film, devoid of any layer separation, exhibits exceptional mechanical strength, with a tensile strength exceeding 70 MPa, a high contact angle of 105°, and superior thermal stability. Furthermore, the CMC/TA-PLA/Fe film demonstrates remarkable efficacy in blocking ultraviolet light, effectively minimizing the discoloration of various wood surfaces exposed to UV aging.

Ilex × attenuata 'Sunny Foster' represents a yellow leaf mutant originating from I. × attenuata 'Foster#2', a popular ornamental woody cultivar. However, the molecular mechanisms underlying this leaf color mutation remain unclear. Using a comprehensive approach encompassing cytological, physiological, and transcriptomic methodologies, notable distinctions were discerned between the mutant specimen and its wild type. The mutant phenotype displayed aberrant chloroplast morphology, diminished chlorophyll content, heightened carotenoid/chlorophyll ratios, and a decelerated rate of plant development. Transcriptome analysis identified differentially expressed genes (DEGs) related to chlorophyll metabolism, carotenoid biosynthesis and photosynthesis. The up-regulation of CHLD and CHLI subunits leads to decreased magnesium chelatase activity, while the up-regulation of COX10 increases heme biosynthesis-both impair chlorophyll synthesis. Conversely, the down-regulation of HEMD hindered chlorophyll synthesis, and the up-regulation of SGR enhanced chlorophyll degradation, resulting in reduced chlorophyll content. Additionally, genes linked to carotenoid biosynthesis, flavonoid metabolism, and photosynthesis were significantly down-regulated. We also identified 311 putative differentially expressed transcription factors, including bHLH s and GLK s. These findings shed light on the molecular mechanisms underlying leaf color mutation in I. × attenuata 'Sunny Foster' and provide a substantial gene reservoir for enhancing leaf color through breeding techniques., Competing Interests: The authors declare that they have no conflicts of interest.

Obtaining high-added value compounds from agricultural waste receives increasing attention, as it can both improve resource utilization efficiency and reduce waste generation. In this study, polysaccharides are extracted from the discarded roots of Abelmoschus manihot (L.) by the high-efficiency ultrasound-assisted extraction (UAE). The optimized condition was determined as solid-liquid ratio SL ratio = 1:20, temperature T = 30 °C and time T = 40 min, achieving an extraction yield of 13.41%. Composition analysis revealed that glucose (Glc, 44.65%), rhamnose (Rha, 26.30%), galacturonic acid (GalA, 12.50%) and galactose (Gal, 9.86%) are the major monosaccharides of the extract. The extract showed a low degree of esterification (DE) value of 40.95%, and its Fourier-transform infrared (FT-IR) spectrum exhibited several characteristic peaks of polysaccharides. Inspired by the wide cosmetic applications of polysaccharides, the skincare effect of the extract was evaluated via the moisture retention, total phenolic content (TPC) quantification, 2,2-Diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging activity, anti-hyaluronidase and anti-elastase activity experiments. The extract solutions demonstrated a 48 h moisture retention rate of 10.75%, which is superior to that of commercially available moisturizer hyaluronic acid (HA). Moreover, both the TPC value of 16.16 mg GAE/g (dw) and DPPH-free radical scavenging activity of 89.20% at the concentration of 2 mg/mL indicated the strong anti-oxidant properties of the extract. Furthermore, the anti-hyaluronidase activity and moderate anti-elastase activity were determined as 72.16% and 42.02%, respectively. In general, in vitro skincare effect experiments suggest moisturizing, anti-oxidant, anti-radical and anti-aging activities of the A. manihot root extract, indicating its potential applications in the cosmetic industry.

Although Taxodium hybrid 'Zhongshanshan' 406 (Taxodium mucronatum Tenore × Taxodium distichum; Taxodium 406) is an extremely flooding-tolerant woody plant, the physiological and molecular mechanisms underlying acclimation of its roots to long-term flooding remain largely unknown. Thus, we exposed saplings of Taxodium 406 to either non-flooding (control) or flooding for 2 months. Flooding resulted in reduced root biomass, which is in line with lower concentrations of citrate, α-ketoglutaric acid, fumaric acid, malic acid and adenosine triphosphate (ATP) in Taxodium 406 roots. Flooding led to elevated activities of pyruvate decarboxylase, alcohol dehydrogenase and lactate dehydrogenase, which is consistent with higher lactate concentration in the roots of Taxodium 406. Flooding brought about stimulated activities of superoxide dismutase and catalase and elevated reduced glutathione (GSH) concentration and GSH/oxidized glutathione, which is in agreement with reduced concentrations of O2- and H2O2 in Taxodium 406 roots. The levels of starch, soluble protein, indole-3-acetic acid, gibberellin A4 and jasmonate were decreased, whereas the concentrations of glucose, total non-structural carbohydrates, most amino acids and 1-aminocyclopropane-1-carboxylate (ACC) were improved in the roots of flooding-treated Taxodium 406. Underlying these changes in growth and physiological characteristics, 12,420 mRNAs and 42 miRNAs were significantly differentially expressed, and 886 miRNA-mRNA pairs were identified in the roots of flooding-exposed Taxodium 406. For instance, 1-aminocyclopropane-1-carboxylate synthase 8 (ACS8) was a target of Th-miR162-3p and 1-aminocyclopropane-1-carboxylate oxidase 4 (ACO4) was a target of Th-miR166i, and the downregulation of Th-miR162-3p and Th-miR166i results in the upregulation of ACS8 and ACO4, probably bringing about higher ACC content in flooding-treated roots. Overall, these results indicate that differentially expressed mRNA and miRNAs are involved in regulating tricarboxylic acid cycle, ATP production, fermentation, and metabolism of carbohydrates, amino acids and phytohormones, as well as reactive oxygen species detoxification of Taxodium 406 roots. These processes play pivotal roles in acclimation to flooding stress. These results will improve our understanding of the molecular and physiological bases underlying woody plant flooding acclimation and provide valuable insights into breeding-flooding tolerant trees., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Introduction: Partial or complete submergence of trees can occur in natural wetlands during times of high waters, but the submergence events have increased in severity and frequency over the past decades. Taxodium distichum is well-known for its waterlogging tolerance, but there are also numerous observations of this species becoming partially or complete submerged for longer periods of time. Consequently, the aims of the present study were to characterize underwater net photosynthesis ( P N ) and leaf anatomy of T. distichum with time of submergence., Methods: We completely submerged 6 months old seedling of T. distichum and diagnosed underwater ( P N ), hydrophobicity, gas film thickness, Chlorophyll concentration and needles anatomy at discrete time points during a 30-day submergence event. We also constructed response curves of underwater P N to CO 2 , light and temperature., Results: During the 30-day submergence period, no growth or formation new leaves were observed, and therefore T. distichum shows a quiescence response to submergence. The hydrophobicity of the needles declined during the submergence event resulting in complete loss of gas films. However, the Chlorophyll concentration of the needles also declined significantly, and it was there not possible to identify the main cause of the corresponding significant decline in underwater P N . Nevertheless, even after 30 days of complete submergence, the needles still retained some capacity for underwater photosynthesis under optimal light and CO 2 conditions., Discussion: However, to fully understand the stunning submergence tolerance of T. distichum , we propose that future research concentrate on unravelling the finer details in needle anatomy and biochemistry as these changes occur during submergence., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Guo, Xue, Yin, Pedersen and Hua.)

The conifer Taxodium hybrid 'Zhongshanshan' ( T. hybrid 'Zhongshanshan') is characterized by rapid growth, strong stress resistance, and high ornamental value and has significant potential for use in afforestation, landscaping, and wood production. The main method of propagating T. hybrid 'Zhongshanshan' is tender branch cutting, but the cutting rooting abilities of different T. hybrid 'Zhongshanshan' clones differ significantly. To explore the causes of rooting ability differences at a molecular level, we analyzed the transcriptome data of cutting base and root tissues of T. hybrid 'Zhongshanshan 149' with a rooting rate of less than 5% and T. hybrid 'Zhongshanshan 118' with rooting rate greater than 60%, at the developmental time points in this study. The results indicated that differentially expressed genes between the two clones were mainly associated with copper ion binding, peroxidase, and oxidoreductase activity, response to oxidative stress, phenylpropanoid and flavonoid biosynthesis, and plant hormone signal transduction, among others. The expression pattern of ThAP2 was different throughout the development of the adventitive roots of the two clone cuttings. Therefore, this gene was selected for further study. It was shown that ThAP2 was a nuclear-localized transcription factor and demonstrated a positive feedback effect on rooting in transgenic Nicotiana benthamiana cuttings. Thus, the results of this study explain the molecular mechanism of cutting rooting and provide candidate gene resources for developing genetic breeding strategies for optimizing superior clones of T. hybrid 'Zhongshanshan'.

Main Conclusion: Taxodium 703 leaves activate fermentation, amino acids metabolism and ROS detoxification, and reduce TCA cycle and ABA biosynthesis in acclimation to prolonged partial submergence stress. Taxodium hybrid 'Zhongshanshan 703' (T. mucronatum × T. distichum; Taxodium 703) is a highly flooding-tolerant woody plant. To investigate the physiological and transcriptional regulatory mechanisms underlying its leaves in acclimation to long-term flooding, we exposed cuttings of Taxodium 703 to either non-flooding (control) or partial submergence for 2 months. The leaf tissues above (AL) and below (BL) flooding-water were separately harvested. Partial submergence decreased concentrations of chlorophyll (a + b) and dehydroascorbate (DHA) and lactate dehydrogenase (LDH) activity in AL, and reduced biomass, concentrations of succinic acid, fumaric acid and malic acid, and transcript levels of genes involved in tricarboxylic acid (TCA) cycle in BL. Under partial submergence, concentrations of starch, malondialdehyde and abscisic acid (ABA) decreased, and also mRNA levels of nine-cis-epoxycarotenoid dioxygenases that are involved in ABA biosynthesis in AL and BL of Taxodium 703. Partial submergence increased O 2 - content in AL, and improved concentrations of pyruvate and soluble sugars and activities of LDH and peroxidase in BL. In addition, partial submergence increased concentrations of ethanol, lactate, alanine, γ-aminobutyric acid, total amino acids and ascorbic acid (ASA), and ASA/DHA, activities of alcohol dehydrogenases (ADH) and ascorbate peroxidase, as well as transcript levels of ADH1A, ADH1B and genes involved in alanine biosynthesis and starch degradation in AL and BL of Taxodium 703. Overall, these results suggest that Taxodium 703 leaves activate fermentation, amino acids metabolism and reactive oxygen species detoxification, and maintain a steady supply of sugars, and reduce TCA cycle and ABA biosynthesis in acclimation to prolonged partial submergence stress., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Michelia compressa (Maxim.) Sarg. is one of the important timber trees in Taiwan province, P. R. China. Michelia 'Zhongshanhanxiao' is a group of variants found among the progeny of M. compressa that exhibit higher growth rates compared with normal individuals, with a significantly increased stem diameter and height, as well as enlarged leaves and flowers. However, the molecular mechanisms fostering the growth advantage and morphological variations are unknown and deserve further study. Through analysing the transcriptome, metabolome and physiological processes of leaves, we identified remarkable differences in gene expression and metabolic profiles between Michelia 'Zhongshanhanxiao' and both the maternal M. compressa and its normal progeny. These differences were widely associated with a plant-pathogen interaction, phenylpropanoid biosynthesis, cyanoamino acid metabolism, carbon fixation in photosynthetic organisms and plant hormone signal transduction. Additionally, physiological measurements showed that Michelia 'Zhongshanhanxiao' possesses stronger photosynthetic capacity and higher plant hormone content. These results suggest that the heterosis of Michelia 'Zhongshanhanxiao' is regulated by candidates related to cell division, resistance to pathogens and the accumulation of organic compounds. The findings of this study provide crucial information on the molecular mechanisms underlying the growth advantages conferred by heterosis in trees., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

As a flooding-tolerant tree species, Taxodium distichum has been utilized in afforestation projects and proven to have important value in flooding areas. Alcohol dehydrogenase (ADH), which participates in ethanol fermentation, is essential for tolerance to the anaerobic conditions caused by flooding. In a comprehensive analysis of the ADH gene family in T. distichum , TdADHs were cloned on the basis of whole-genome sequencing, and then bioinformatic analysis, subcellular localization, and gene expression level analysis under flooding were conducted. The results show that the putative protein sequences of 15 cloned genes contained seven TdADHs and eight TdADH-like genes (one Class III ADH included) that were divided into five clades. All the sequences had an ADH_N domain, and except for TdADH-likeE2 , all the other genes had an ADH_zinc_N domain. Moreover, the TdADHs in clades A, B, C, and D had a similar motif composition. Additionally, the number of TdADH amino acids ranged from 277 to 403, with an average of 370.13. Subcellular localization showed that, except for TdADH-likeD3 , which was not expressed in the nucleus, the other genes were predominantly expressed in both the nucleus and cytosol. TdADH-likeC2 was significantly upregulated in all three organs (roots, stems, and leaves), and TdADHA3 was also highly upregulated under 24 h flooding treatment; the two genes might play key roles in ethanol fermentation and flooding tolerance. These findings offer a comprehensive understanding of TdADHs and could provide a foundation for the molecular breeding of T. distichum and current research on the molecular mechanisms driving flooding tolerance.

Taxodium hybrid 'Zhongshanshan' has been widely used as a timber tree in river network areas and coastal regions and is mainly propagated by cuttings. However, when trees age, their capacity to form adventitious roots becomes weaker. We successfully enhanced the rooting ability of shoots in T. hybrid 'Zhongshanshan 302' by their rejuvenation based on grafting. We recorded temporal variation in endogenous auxin, abscisic acid (ABA), gibberellins (GAs), trans-zeatin-riboside (TZR), soluble sugar and H 2 O 2 after root induction. Auxin, soluble sugars and H 2 O 2 levels were higher in rejuvenated shoots than in mature shoots, whereas the opposite was true for ABA and GAs. Notably, indole-3-acetic acid (IAA) and GA3 presented higher contents with more obvious differences in T. hybrid 'Zhongshanshan 302' rejuvenated shoots vs. mature shoots compared with other kinds of auxin and GAs. The evident improvement in the rooting ability of rejuvenated shoots after grafting likely resulted from the differential regulation of plant hormones, carbohydrates and redox signaling. In addition to the physiological basis of improved rooting ability by grafting, this study provided a theoretical basis for the optimization of subsequent propagation techniques in T. hybrid 'Zhongshanshan' and potentially other Taxodium spp.

Taxodium hybrid Zhongshanshan has been widely planted in the Yangtze River Basin (YRB) for soil and carbon conservation, with quantities over 50 million. The objective of this study was to determine how T . hybrid Zhongshanshan plantations affected soil physicochemical properties and bacterial community structure in the YRB, and to examine the consistency of changes by afforestation. Soils under T . Zhongshanshan plantations across six sites of the YRB were compared with soils of adjacent non-forested sites. Soil physicochemical properties and bacterial community structure were determined to clarify edaphic driving factors and reveal the effects of afforestation on bacteria. The results indicated that most soil attributes manifested improvements, e.g., total nitrogen in Jiangxi and Shanghai; available phosphorus in Hubei, Chongqing and Yunnan, exhibited the potential to maintain or ameliorate soil quality. A decrease in soil bulk density caused by plantation was also observed at the expense of soil macro-aggregates augment. Afforestation of T . Zhongshanshan plantation has habitually improved Shannon diversity and Chao1 richness, of which dominant phyla were Proteobacteria, Acidobacteria, and Actinobacteria, and increased the relative abundance of the phyla Proteobacteria and Nitrospirae, and the classes Flavobacteriia , Acidobacteria _Gp5, and Bacilli . We concluded that T . Zhongshanshan plantation can be employed to facilitate soil nutrient accumulation in the YRB, but that the degree, rate and direction of changes in soil attributes are sites dependent. It is recommended that afforestation of nutrient-depleted and less productive lands in the YRB should utilize this fast-growing species in combination with proper fertilization.

Kosteletzkya pentacarpos is a promising plant being developed as a salt-tolerant biofuel crop that also the ability to fix heavy metals. Here, high-throughput sequencing technology was used to sequence and assemble the chloroplast genome of K. pentacarpos . The full length of the chloroplast genome is 161,777 bp, comprising a large single-copy region of 90,019 bp, a small single-copy region of 18,978 bp, and a pair of inverted repeats of 26,390 bp. A total of 113 genes were annotated, including 79 protein-coding, 30 transfer RNA, and 4 ribosomal RNA genes. Phylogenetic analysis based on whole chloroplast genome sequences showed that K. pentacarpos has a close relationship with Abelmoschus in Malvaceae. This study increases the available genomic information on K. pentacarpos , and provides a basis for the rational exploitation and utilization of germplasm resources., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

GRAS proteins are widely distributed plant-specific transcription factors. In this study, we identified 59 GRAS proteins (HhGRASs) from the genomic and transcriptomic datasets of Hibiscus hamabo Sieb. et Zucc. These proteins were phylogenetically divided into nine subfamilies. RNA-seq analysis revealed that most HhGRASs were expressed in response to abiotic stresses. Results from quantitative real-time PCR analysis of nine selected HhGRASs suggested that HhGRAS14 was significantly upregulated under multiple abiotic stresses; therefore, this gene was selected for further study. Silencing HhGRAS14 in H. hamabo reduced the tolerance to drought and salt stress, while overexpression in Arabidopsis thaliana significantly increased the tolerance to drought and salt and reduced the sensitivity to abscisic acid (ABA). In summary, we analyzed the GRAS family of proteins in semi-mangrove plants for the first time and identified a gene that responds to drought and salt stress, which provided the basis for a comprehensive analysis of GRAS genes and insight into the abiotic stress response mechanism in H. hamabo., (Copyright © 2022 Elsevier B.V. All rights reserved.)

To understand the characteristics of net NH 4 + and NO 3 - fluxes and their relation with net H + fluxes in Taxodium , net fluxes of NH 4 + , NO 3 - and H + were detected by a scanning ion-selective electrode technique under different forms of fixed nitrogen (N) and experimental conditions. The results showed that higher net NH 4 + and NO 3 - fluxes occurred at 2.1-3.0 mm from the root apex in T. ascendens and T. distichum . Compared to NH 4 + or NO 3 - alone, more stable net NH 4 + and NO 3 - fluxes were found under NH 4 NO 3 supply conditions, of which net NH 4 + flux was promoted at least 1.71 times by NO 3 - , whereas net NO 3 - flux was reduced more than 81.66% by NH 4 + in all plants, which indicated that NH 4 + is preferred by Taxodium plants. T. ascendens and T. mucronatum had the largest net NH 4 + and total N influxes when NH 4 + :NO 3 - was 3:1. 15 N Atom% and activities of N assimilation enzymes were improved by single N fertilization in the roots of T. distichum . In most cases, net H + fluxes were tightly correlated with net NH 4 + and NO 3 - fluxes. Thus, both N forms and proportions could affect N uptake of Taxodium . These findings could provide useful guidance for N management for better productivity of Taxodium plants.

Hibiscus hamabo is a semi-mangrove species with strong tolerance to salt and waterlogging stress. However, the molecular basis and mechanisms that underlie this strong adaptability to harsh environments remain poorly understood. Here, we assembled a high-quality, chromosome-level genome of this semi-mangrove plant and analyzed its transcriptome under different stress treatments to reveal regulatory responses and mechanisms. Our analyses suggested that H. hamabo has undergone two recent successive polyploidy events, a whole-genome duplication followed by a whole-genome triplication, resulting in an unusually large gene number (107 309 genes). Comparison of the H. hamabo genome with that of its close relative Hibiscus cannabinus , which has not experienced a recent WGT, indicated that genes associated with high stress resistance have been preferentially preserved in the H. hamabo genome, suggesting an underlying association between polyploidy and stronger stress resistance. Transcriptomic data indicated that genes in the roots and leaves responded differently to stress. In roots, genes that regulate ion channels involved in biosynthetic and metabolic processes responded quickly to adjust the ion concentration and provide metabolic products to protect root cells, whereas no such rapid response was observed from genes in leaves. Using co-expression networks, potential stress resistance genes were identified for use in future functional investigations. The genome sequence, along with several transcriptome datasets, provide insights into genome evolution and the mechanism of salt and waterlogging tolerance in H. hamabo , suggesting the importance of polyploidization for environmental adaptation., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nanjing Agricultural University.)

NAC transcription factor is one of the largest plant gene families, participating in the regulation of plant biological and abiotic stresses. In this study, 182 NAC proteins (HhNACs) were identified based on genomic datasets of Hibiscus hamabo Sieb. et Zucc ( H. hamabo ). These proteins were divided into 19 subfamilies based on their phylogenetic relationship, motif pattern, and gene structure analysis. Expression analysis with RNA-seq revealed that most HhNACs were expressed in response to drought and salt stress. Research of quantitative real-time PCR analysis of nine selected HhNACs supported the transcriptome data's dependability and suggested that HhNAC54 was significantly upregulated under multiple abiotic stresses. Overexpression of HhNAC54 in Arabidopsis thaliana ( A. thaliana ) significantly increased its tolerance to salt. This study provides a basis for a comprehensive analysis of NAC transcription factor and insight into the abiotic stress response mechanism in H. hamabo .

Hibiscus coccineus is famous for its wide geographical distribution and the showy flowers of scarlet rose mallow. It belongs to the Malvaceae family and has greatly ornamental and ecological value. In this study, high-throughput sequencing and bioinformatics technology were used to assemble the complete chloroplast genome sequence, which will provide more genomic information for studying the genetic diversity and phylogenetic relationship. The full length of chloroplast genome is 160,280 bp, composed of a large single-copy (LSC) region of 89,121 bp, a small single-copy (SSC) region of 18,673 bp, and two inverted repeats (IRs) of 26,243 bp. A total of 113 genes were annotated, including 79 protein-coding genes, 30 tRNA, and four rRNA genes. Phylogenetic tree analysis revealed that the Hibiscus coccineus is closest to Hibiscus mutabilis in the Hibiscus L., Competing Interests: No potential conflict of interest was reported by the authors., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Hibiscus hamabo Sieb. et Zucc is an important semi-mangrove plant with great morphological features and strong salt resistance. In this study, by combining single molecule real time and next-generation sequencing technologies, we explored the transcriptomic changes in the roots of salt stressed H . hamabo . A total of 94,562 unigenes were obtained by clustering the same isoforms using the PacBio RSII platform, and 2269 differentially expressed genes were obtained under salt stress using the Illumina platform. There were 519 differentially expressed genes co-expressed at each treatment time point under salt stress, and these genes were found to be enriched in ion signal transduction and plant hormone signal transduction. We used Arabidopsis thaliana (L.) Heynh. transformation to confirm the function of the HhWRKY79 gene and discovered that overexpression enhanced salt tolerance. The full-length transcripts generated in this study provide a full characterization of the transcriptome of H . hamabo and may be useful in mining new salt stress-related genes specific to this species, while facilitating the understanding of the salt tolerance mechanisms.

The use of electrochemical fingerprints for plant identification is an emerging application in biosensors. In this work, Taxodium ascendens , T. distichum , T. mucronatum , and 18 of their hybrid progenies were collected for this purpose. This is the first attempt to use electrochemical fingerprinting for the identification of plant hybrid progeny. Electrochemical fingerprinting in the leaves of Taxodium spp. was recorded under two conditions. The results showed that the electrochemical fingerprints of each species and progeny possessed very suitable reproducibility. These electrochemical fingerprints represent the electrochemical behavior of electrochemically active substances in leaf tissues under specific conditions. Since these species and progenies are very closely related to each other, it is challenging to identify them directly using a particular electrochemical fingerprinting. Therefore, electrochemical fingerprints measured under different conditions were used to perform pattern recognition. We can identify different species and progenies by locating the features in different pattern maps. We also performed a phylogenetic study with data from electrochemical fingerprinting. The results proved that the electrochemical classification results and the relationship between them are closely related.

The basic helix-loop-helix (bHLH) family of transcription factors is one of the most significant and biggest in plants. It is involved in the regulation of both growth and development, as well as stress response. Numerous members of the bHLH family have been found and characterized in woody plants in recent years. However, no systematic study of the bHLH gene family has been published for Hibiscus hamabo Sieb. et Zucc. In this research, we identified 162 bHLH proteins (HhbHLHs) from the genomic and transcriptomic datasets of H. hamabo, which were phylogenetically divided into 19 subfamilies. According to a gene structural study, the number of exon-introns in HhbHLHs varied between zero and seventeen. MEME research revealed that the majority of HhbHLH proteins contained three conserved motifs, 1, 4, and 5. The examination of promoter cis -elements revealed that the majority of HhbHLH genes had several cis -elements involved in plant growth and development and abiotic stress responses. In addition, the overexpression of HhbHLH2 increased salt and drought stress tolerance in Arabidopsis .

Taxodium 'Zhongshanshan 401' is an important economic plant with ornamental and ecological values, and has been widely planted in southeastern China. In this study, the complete chloroplast (cp) genome of T . 'Zhongshanshan 401' was sequenced and illustrated to add the more genetic information. The entire cp genome of T . 'Zhongshanshan 401' was 132,037 bp in length with 35.3% overall GC content. The cp genome contained 120 genes, including 83 protein-coding genes, 33 tRNA genes, and four rRNA genes. Fifteen genes contain two exons and two contains three exons. Phylogenetic analysis based on whole cp genome sequences showed that T . 'Zhongshanshan 401' was more closely related to T . mucronatum., Competing Interests: No potential conflict of interest was reported by the authors., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

The Taxodium hybrid 'Zhongshanshan 406' ( T . hybrid 'Zhongshanshan 406') [ Taxodium mucronatum Tenore × Taxodium distichum (L.). Rich] has an outstanding advantage in flooding tolerance and thus has been widely used in wetland afforestation in China. Alcohol dehydrogenase genes ( ADHs ) played key roles in ethanol metabolism to maintain energy supply for plants in low-oxygen conditions. Two ADH genes were isolated and characterized- ThADH1 and ThADH4 (GenBank ID: AWL83216 and AWL83217-basing on the transcriptome data of T . hybrid 'Zhongshanshan 406' grown under waterlogging stress. Then the functions of these two genes were investigated through transient expression and overexpression. The results showed that the ThADH1 and ThADH4 proteins both fall under ADH III subfamily. ThADH1 was localized in the cytoplasm and nucleus, whereas ThADH4 was only localized in the cytoplasm. The expression of the two genes was stimulated by waterlogging and the expression level in roots was significantly higher than those in stems and leaves. The respective overexpression of ThADH1 and ThADH4 in Populus caused the opposite phenotype, while waterlogging tolerance of the two transgenic Populus significantly improved. Collectively, these results indicated that genes ThADH1 and ThADH4 were involved in the tolerance and adaptation to anaerobic conditions in T . hybrid 'Zhongshanshan 406'.

WRKY transcription factors are known to play important roles in the regulation of various aspects of plant growth and development, including germination, stress resistance, and senescence. Nevertheless, there is little information about the WRKY genes in Hibiscus hamabo Sieb. et Zucc., an important semimangrove plant. In this study, HhWRKY genes in H. hamabo were identificated based on Illumina RNA-sequencing and isoform sequencing from salt-treated roots. Then phylogenetic analysis and conserved motif analysis of the WRKY family in H. hamabo and Arabidopsis thaliana were used to classify WRKY genes. Sixteen HhWRKY genes were selected from different groups to detect their expression patterns using real-time quantitative PCR in different organ (root, old leaf, tender leaf, receptacle, petal, or stamen) from 10-year-old H. hamabo plants grown in their natural environment and in seedlings with 8 to 10 true leaves challenged by phytohormone (salicylic acid, methyl jasmonate, or abscisic acid) and abiotic stress (drought, salt, or high temperature). As a result, the identified 78 HhWRKY genes were divided into two major groups and several subgroups based on their structural and phylogenetic features. Most transcripts of the selected 16 HhWRKY genes were more abundant in old than in tender leaves of H. hamabo . HhWRKY genes were regulated in reaction to abiotic stresses and phytohormone treatments and may participate in signaling networks to improve plant stress resistance. Some of HhWRKY genes behaved as would be predicted based on their homology with A. thaliana WRKY genes, but others showed divergent behavior. This systematic analysis lays the foundation for further identification of WRKY gene functions, with the aim of improving woody plants.

Background: Chloroplast (cp) genome information would facilitate the development and utilization of Taxodium resources. However, cp genome characteristics of Taxodium were poorly understood., Results: We determined the complete cp genome sequences of T. distichum, T. mucronatum, and T. ascendens. The cp genomes are 131,947 bp to 132,613 bp in length, encode 120 genes with the same order, and lack typical inverted repeat (IR) regions. The longest small IR, a 282 bp trnQ-containing IR, were involved in the formation of isomers. Comparative analysis of the 3 cp genomes showed that 91.57% of the indels resulted in the periodic variation of tandem repeat (TR) motifs and 72.46% single nucleotide polymorphisms (SNPs) located closely to TRs, suggesting a relationship between TRs and mutational dynamics. Eleven hypervariable regions were identified as candidates for DNA barcode development. Hypothetical cp open reading frame 1(Ycf1) was the only one gene that has an indel in coding DNA sequence, and the indel is composed of a long TR. When extended to cupressophytes, ycf1 genes have undergone a universal insertion of TRs accompanied by extreme length expansion. Meanwhile, ycf1 also located in rearrangement endpoints of cupressophyte cp genomes. All these characteristics highlight the important role of repeats in the evolution of cp genomes., Conclusions: This study added new evidence for the role of repeats in the dynamics mechanism of cp genome mutation and rearrangement. Moreover, the information of TRs and hypervariable regions would provide reliable molecular resources for future research focusing on the infrageneric taxa identification, phylogenetic resolution, population structure and biodiversity for the genus Taxodium and Cupressophytes.

Background: Lectin receptor-like protein kinases (LecRLKs) can transform external stimuli into intracellular signals and play important regulatory roles in plant development and response to environmental stressors. However, research on the LecRLK gene family of conifers has seldom been reported., Methods: Putative LecRLK genes were identified in the transcriptome of Taxodium 'Zhongshanshan'. The classification, domain structures, subcellular localization prediction, and expression patterns of LecRLK genes, as well as co-expressed genes, were analyzed using bioinformatics methods. Fifteen representative genes were further selected for qRT-PCR analysis in six tissues and under five different environmental stressor conditions., Results: In total, 297 LecRLK genes were identified, including 155 G-type, 140 L-type, and 2 C-type. According to the classification, G-type and L-type LecRLK genes both can be organized into seven groups. The domain architecture of G-type proteins were more complex compared with that of L- and C-type proteins. Conservative motifs were found in G-type and L-type diverse lectin domains. Prediction and transient expression experiments to determine subcellular localization showed that LecRLKs were mainly concentrated in the cell membrane system, and some members were located at multiple sites at the same time. RNA-seq-based transcriptomics analysis suggested functional redundancy and divergence within each group. Unigenes co-expressed with LecRLKs in the transcriptome were found to be enriched in pathways related to signal transduction and environmental adaptation. Furthermore, qRT-PCR analysis of representative genes showed evidence of functional divergence between different groups., Conclusions: This is the first study to conduct an identification and expression analysis of the LecRLK gene family in Taxodium . These results provide a basis for future studies on the evolution and function of this important gene family in Taxodium ., Competing Interests: Mingzhi Li is employed by Genepioneer Biotechnologies Co. Ltd.

Adventitious root (AR) formation from cuttings is the primary manner for the commercial vegetative propagation of trees. Cuttings is also the main method for the vegetative reproduction of Taxodium 'Zhongshanshan', while knowledge of the molecular mechanisms regulating the processes is limited. Here, we used mRNA sequencing and an isobaric tag for relative and absolute quantitation-based quantitative proteomic (iTRAQ) analysis to measure changes in gene and protein expression levels during AR formation in Taxodium 'Zhongshanshan'. Three comparison groups were established to represent the three developmental stages in the AR formation process. At the transcript level, 4743 genes showed an expression difference in the comparison groups as detected by RNA sequencing. At the protein level, 4005 proteins differed in their relative abundance levels, as indicated by the quantitative proteomic analysis. A comparison of the transcriptome and proteome data revealed regulatory aspects of metabolism during AR formation and development. In summary, hormonal signal transduction is different at different developmental stages during AR formation. Other factors related to carbohydrate and energy metabolism and protein degradation and some transcription factor activity levels, were also correlated with AR formation. Studying the identified genes and proteins will provide further insights into the molecular mechanisms controlling AR formation.

Background: 'Zhongshanshan' is the general designation for the superior interspecific hybrid clones of Taxodium species, which is widely grown for economic and ecological purposes in southern China. Growth is the priority objective in 'Zhongshanshan' tree improvement. A high-density linkage map is vital to efficiently identify key quantitative trait loci (QTLs) that affect growth., Results: In total, 403.16 Gb of data, containing 2016,336 paired-end reads, was obtained after preprocessing. The average sequencing depth was 28.49 in T. distichum var. distichum, 25.18 in T. mucronatum, and 11.12 in each progeny. In total, 524,662 high-quality SLAFs were detected, of which 249,619 were polymorphic, and 6166 of the polymorphic markers met the requirements for use in constructing a genetic map. The final map harbored 6156 SLAF markers on 11 linkage groups, and was 1137.86 cM in length, with an average distance of 0.18 cM between adjacent markers. Separate QTL analyses of traits in different years by CIM detected 7 QTLs. While combining multiple-year data, 13 QTLs were detected by ICIM. 5 QTLs were repeatedly detected by the two methods, and among them, 3 significant QTLs (q6-2, q4-2 and q2-1) were detected in at least two traits. Bioinformatic analysis discoveried a gene annotated as a leucine-rich repeat receptor-like kinase gene within q4-2., Conclusions: This map is the most saturated one constructed in a Taxodiaceae species to date, and would provide useful information for future comparative mapping, genome assembly, and marker-assisted selection.

As a subfamily of the APETALA 2/ethylene response element binding protein (AP2/EREBP) transcription factor superfamily, the ethylene response factor (ERF) is widely involved in the regulation of growth and response to various abiotic stresses in plants, and has been shown to be the main transcription factor regulating transcription of the genes related to hypoxia and waterlogging stress. In this study, three ThERF genes, with significant differences in expression profile in response to flooding stress, were identified from the transcriptomics data acquired from Taxodium hybrid 'Zhongshanshan 406' ( T. mucronatum Tenore × T. distichum (L.) Rich) under waterlogging stress: ThERF15, ThERF39 and ThRAP2.3 (GenBank ID: KY463467, KY463468 and KY463470, respectively).The full-length cDNA of each of the three ERF s was obtained using the RACE (rapid amplification cDNA ends) method, and all three were intron-free. Multiple protein sequence alignments indicated that ThERF15, ThERF39 and ThRAP2.3 proteins all had only one AP2-ERF domain and belonged to the ERF subfamily. A transient gene expression assay demonstrated that ThERF15, ThERF39 and ThRAP2.3 were all localized to the nucleus. Real-time quantitative PCR (qPCR) revealed that the expression of ThERF15, ThERF39 and ThRAP2.3 exhibited significant differences, compared with the control, in response to two levels of flooding treatment (half-flooding or total-submergence) of 'Zhongshanshan 406'. Quantification of ethylene concentration revealed that ethylene was more relevant to the level of expression than the period of flooding treatment. Based on the experimental results above, ThERF15, ThERF39 and ThRAP2.3 were identified as being related to the regulation of downstream flooding- responsive gene expression in 'Zhongshanshan 406'. ThRAP2.3 is most likely to be a key downstream-response ERF gene to respond to the output of the ethylene signal generated by flooding stress., Competing Interests: The authors declare there are no competing interests.

Tideland reclamation has drastic effects on coastal ecosystem involved in soil microorganisms. However, the knowledge regarding temporal variations of microbial community along reclamation chronosequence and their environmental variable predictor is still poorly known. Using Illumina sequencing, we qualified bacterial community composition in soils collected from one tideland and four reclamation stages, i.e. 2-year, 7-year, 19-year and 39-year in Jiangsu, Eastern China. Across all samples, the dominant groups were Proteobacteria, Bacteroidete, Acidobacteria, Planctomycetes and Chloroflexi. Reclamation activity and its histories greatly altered bacterial community structure, and only 0.28% of phylotypes were shared by five soils. Specially, some typical marine bacteria (Gaetulibacter, Alcanivorax …) disappeared in reclamation soils, while other groups (Niabella, Flavisolibacter…) were gradually eminent. Generally, bacterial diversity and richness increased with reclamation histories. Bacterial community was correlated with most of soil physico-chemical properties. Amongst, mean weight diameter of soil aggregates (MWD) was detected as a primary factor predicting bacterial community composition. Together, our results indicated that effects of reclamation on bacterial community varied with diked histories, and MWD was a major factor predicting bacterial community during progressive reclamation. These findings offer predicting case study for understanding the impact of reclamation and its histories on microbial community in a coastal ecosystem.

Among the GRAS family of transcription factors, SHORT ROOT (SHR) and SCARECROW (SCR) are key regulators of the formation of root tissues. In this study, we isolated and characterized two genes encoding SHR proteins and one gene encoding an SCR protein: ThSHR1 (Accession Number MF045148), ThSHR2 (Accession Number MF045149) and ThSCR (Accession Number MF045152) in the adventitious roots of Taxodium hybrid 'Zhongshanshan'. Gene structure analysis indicated that ThSHR1 , ThSHR2 and ThSCR are all intron free. Multiple protein sequence alignments showed that each of the corresponding proteins, ThSHR1, ThSHR2 and ThSCR, contained five well-conserved domains: leucine heptad repeat I (LHRI), the VHIID motif, leucine heptad repeat II (LHR II), the PFYRE motif, and the SAW motif. The phylogenetic analysis indicated that ThSCR was positioned in the SCR clade with the SCR proteins from eight other species, while ThSHR1 and ThSHR2 were positioned in the SHR clade with the SHR proteins from six other species. Temporal expression patterns of these genes were profiled during the process of adventitious root development on stem cuttings. Whereas expression of both ThSHR2 and ThSCR increased up to primary root formation before declining, that of ThSHR1 increased steadily throughout adventitious root formation. Subcellular localization studies in transgenic poplar protoplasts revealed that ThSHR1, ThSHR2 and ThSCR were localized in the nucleus. Collectively, these results suggest that the three genes encode Taxodium GRAS family transcription factors, and the findings contribute to improving our understanding of the expression and function of SHR and SCR during adventitious root production, which may then be manipulated to achieve high rates of asexual propagation of valuable tree species., Competing Interests: The research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Taxodium is a genus renowned for its fast growth, good form and tolerance of flooding, salt, alkalinity, disease and strong winds. In this study, a genetic linkage map was constructed using sequence-related amplified polymorphism (SRAP) and simple sequence repeat (SSR) markers based on an F1 population containing 148 individuals generated from a cross between T. 'Zhongshanshan 302' and T. mucronatum. The map has a total length of 976.5 cM, with a mean distance of 7.0 cM between markers, and contains 34 linkage groups with 179 markers (171 SRAPs and 8 SSRs). Quantitative trait loci (QTLs) affecting growth traits, such as seedling height, basal diameter and crown width, were detected based on the constructed linkage map. Four significant QTLs were identified, three of which, namely qtSH-1 for seedling height, qtBD-1 for basal diameter and qtCW-1 for crown width, were located at 2.659 cM of LG7 with logarithm odds values of 3.72, 3.49 and 3.93, respectively, and explained 24.9, 27.0 and 21.7 % of the total variation of the three grown traits, respectively. Another QTL for crown width (qtCW-2) was detected at 1.0 cM on LG13, with a logarithm of odds value of 3.15, and explained 31.7 % of the total variation of crown width. This is the first report on the construction of a genetic linkage map and QTL analysis in Taxodium, laying the groundwork for the construction of a high-density genetic map and QTL mapping in the genus Taxodium.

Taxodium 'Zhongshanshan' is an interspecies hybrid of Taxodium distichum and Taxodium mucronatum, and has been widely planted in southeastern China. It has great ecological and economic potential. However, the scant genomic resources in genus Taxodium have greatly hindered further exploration of its underlying salinity-tolerance mechanism. To understand the genetic basis of its salt tolerance, high-throughput sequencing of mRNA (RNA-Seq) was used to analyze transcriptome changes of 'Zhongshanshan 405' clone roots treated with NaCl stress. After de novo assembly, 70,312 unigenes were achieved, and 41,059 of them were annotated. 9038 differentially expressed genes (DEGs) were identified among the treatments, and 7959 DEGs were found between salt-stressed roots and control, with 489 up-regulated and 570 down-regulated shared by all of the treatments. Genes related to transport, signal transductions as well as undescribed transcripts were among those DEGs in response to salt stress. Gene ontology classification analysis revealed that salt stress-related categories including 'oxidoreductase activity', 'metal ion binding', and 'membrane' were highly enriched among these DEGs. Moreover, the gene expression pattern of 12 unigenes revealed by quantitative real-time polymerase chain reaction (qRT-PCR) confirmed the RNA-Seq data. Our study not only provided the large-scale assessment of transcriptome resources of Taxodium but also guidelines for probing the molecular mechanism underlying 'Zhongshanshan' salt tolerance., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Background: Taxodium is renowned for its strong tolerance to waterlogging stress, thus it has great ecological and economic potential. However, the scant genomic resources in genus Taxodium have greatly hindered further exploration of its underlying flood-tolerance mechanism. Taxodium 'Zhongshansa' is an interspecies hybrid of T. distichum and T. mucronatum, and has been widely planted in southeastern China. To understand the genetic basis of its flood tolerance, we analyzed the transcriptomes of Taxodium 'Zhongshansa' roots and shoots in response to short-term waterlogging., Results: RNA-seq was used to analyze genome-wide transcriptome changes of Taxodium 'Zhongshansa 406' clone root and shoot treated with 1 h of soil-waterlogging stress. After de novo assembly, 108,692 unigenes were achieved, and 70,260 (64.64%) of them were annotated. There were 2090 differentially expressed genes (DEGs) found in roots and 394 in shoots, with 174 shared by both of them, indicating that the aerial parts were also affected. Under waterlogging stress, the primary reaction of hypoxic-treated root was to activate the antioxidative defense system to prevent cells experiencing reactive oxygen species (ROS) poisoning. As respiration was inhibited and ATP decreased, another quick coping mechanism was repressing the energy-consuming biosynthetic processes through the whole plant. The glycolysis and fermentation pathway was activated to maintain ATP production in the hypoxic root. Constantly, the demand for carbohydrates increased, and carbohydrate metabolism were accumulated in the root as well as the shoot, possibly indicating that systemic communications between waterlogged and non-waterlogged tissues facilated survival. Amino acid metabolism was also greatly influenced, with down-regulation of genes involvedin serine degradation and up-regulation of aspartic acid degradation. Additionally, a non-symbiotic hemoglobin class 1 gene was up-regulated, which may also help the ATP production. Moreover, the gene expression pattern of 5 unigenes involving in the glycolysis pathway revealed by qRT-PCR confirmed the RNA-Seq data., Conclusions: We conclude that ROS detoxification and energy maintenance were the primary coping mechanisms of 'Zhongshansa' in surviving oxygen deficiency, which may be responsible for its remarkable waterlogging tolerance. Our study not only provided the first large-scale assessment of genomic resources of Taxodium but also guidelines for probing the molecular mechanism underlying 'Zhongshansa' waterlogging tolerance.

The variations of Cd accumulation in three rootstalk crop species (radish, carrot and potato) were investigated by using twelve cultivars grown in acidic Ferralsols and neutral Cambisols under two Cd treatments (0.3 and 0.6 mg kg(-1)) in a pot experiment The result showed that the total Cd uptake was significantly affected by genotype, soil type and interaction between them, suggesting the importance of selecting proper cultivars for phytoextraction in a given soil type. Among the cultivars tested, potato cultivar Luyin No.1 in Ferralsols and radish cultivar Zhedachang in Cambisols exhibited the highest Cd phytoextraction efficiency in aerial parts (4.45% and 0.59%, respectively) under 0.6 mg kg-1 Cd treatment. Furthermore, the Cd concentrations in their edible parts were below the National Food Hygiene Standard of China (0.1 mg kg(-1), fresh weight). Therefore, phytomanagement of slightly Cd-contaminated soils using rootstalk crops for safe food production combined with long-term phytoextraction was feasible, and potato cultivar Luyin No.1 for Ferralsols and radish cultivar Zhedachang for Cambisols were promising candidates for this approach.

Lead (Pb) contamination of soil poses severe health risks to humans through vegetable consumption. The variations of Pb concentration in different parts of rootstalk vegetables (radish, carrot and potato) were investigated by using twelve cultivars grown in acidic Ferralsols and neutral Cambisols under two Pb treatments (125 mg kg(-1) and 250 mg kg(-1) for Ferralsols; 150 mg kg(-1) and 300 mg kg(-1) for Cambisols) in a pot experiment. The Pb concentration in edible parts was higher in Ferralsols under two Pb treatments, with range from 0.28 to 4.14, 0.42-10.66 mg kg(-1) (fresh weight) respectively, and all of them exceeded the food safety standard (0.1 mg kg(-1)) recommended by the Codex Alimentarius Commission of FAO and WHO. The Pb concentration in edible parts was significantly affected by genotype, soil type and the interaction between these two factors. The variation of Pb concentration in different cultivars was partially governed by Pb absorption and the transfer of Pb from aerial to edible part. The results revealed that caution should be paid to the cultivation of rootstalk vegetables in Pb-contaminated Ferralsols without any agronomic management to reduce Pb availability and plant uptake. For Cambisols with slight to moderate Pb contamination, growing potato cultivar Shandong No.1 and Chongqing No.1 was effective in reducing the risk of Pb entering human food chain. The results suggest the possibility of developing cultivar- and soil-specific planting and monitoring guidelines for the cultivation of rootstalk vegetables on slight to moderate Pb-contaminated soils., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This paper measured the concentrations of metallic elements, N and S in soils and Sabina chinensis cv. pyramidalis leaves along the urban-rural roadways (urban trunk roads, belt highway, and suburban roads) of Nanjing city, with botanical garden as un-traffic impacted reference site, and discussed the transfer of test elements among atmosphere, plant and soil, as well as the impact of traffic-related pollution. The results showed that the concentrations of Al, Fe, Mo, Zn, Cd, Pb, As and Cr in Sabina chinensis leaves were significantly higher along urban-rural roadways than at reference site, being most remarkable for Al, Fe, Mo and Zn. The positive correlations between soil and Sabina chinensis leaf Fe, Zn and As concentrations in road environment indicated that Sabina chinensis might be a good indicator plant of soil Fe, Zn and As. The correlations between soil and plant leaf Al, Mo, Cu, Pb, Cd and Cr concentrations were not remarkable, and thus, their high concentrations in Sabina chinensis leaf in road environment might be caused by air pollution. The concentrations of N and S in Sabina chinensis leaf were higher along urban trunk roads and belt highway than at reference site, while those along suburban roads were similar to the reference. There was a negative correlation between soil and plant leaf N, and a positive correlation between soil and plant leaf S. The N content in roadside soil was lower, while the S content was higher than the reference, indicating that the higher Sabina chinensis leaf N content in road environment might be caused by the NO(x) from exhaust gas emission, and the higher leaf S content should be come from soil.