Genome-wide analysis of NAC transcription factors and exploration of candidate genes regulating selenium metabolism in Broussonetia papyrifera.

MAIN CONCLUSION: Genome-wide identification revealed 79 BpNAC genes belonging to 16 subfamilies, and their gene structures and evolutionary relationships were characterized. Expression analysis highlighted their importance in plant selenium stress responses. Paper mulberry (Broussonetia papyrifera), a deciduous arboreal plant of the Moraceae family, is distinguished by its leaves, which are abundant in proteins, polysaccharides, and flavonoids, positioning it as a novel feedstock. NAC transcription factors, exclusive to plant species, are crucial in regulating growth, development, and response to biotic and abiotic stress. However, extensive characterization of the NAC family within paper mulberry is lacking. In this study, 79 BpNAC genes were identified from the paper mulberry genome, with an uneven distribution across 13 chromosomes. A comprehensive, genome-wide analysis of BpNACs was performed, including investigating gene structures, promoter regions, and chromosomal locations. Phylogenetic tree analysis, alongside comparisons with Arabidopsis thaliana NACs, allowed for categorizing these genes into 16 subfamilies in alignment with gene structure and motif conservation. Collinearity analysis suggested a significant homologous relationship between the NAC genes of paper mulberry and those in Morus notabilis, Ficus hispida, Antiaris toxicaria, and Cannabis sativa. Integrating transcriptome data and Se content revealed that 12 BpNAC genes were associated with selenium biosynthesis. Subsequent RT-qPCR analysis corroborated the correlation between BpNAC59, BpNAC62 with sodium selenate, and BpNAC55 with sodium selenite. Subcellular localization experiments revealed the nuclear functions of BpNAC59 and BpNAC62. This study highlights the potential BpNAC transcription factors involved in selenium metabolism, providing a foundation for strategically breeding selenium-fortified paper mulberry.

Genome-wide identification of HMT gene family explores BpHMT2 enhancing selenium accumulation and tolerance in Broussonetia papyrifera.

Broussonetia papyrifera, a valuable feed resource, is known for its fast growth, wide adaptability, high protein content and strong selenium enrichment capacity. Selenomethionine (SeMet), the main selenium form in selenium fortification B. papyrifera, is safe for animals and this enhances its nutritional value as a feed resource. However, the molecular mechanisms underlying SeMet synthesis remain unclear. This study identified three homocysteine S-methyltransferase genes from the B. papyrifera genome. The phylogenetic tree demonstrated that BpHMTs were divided into two classes, and BpHMT2 in the Class 2-D subfamily evolved earlier and possesses more fundamental functions. On the basis of the correlation between gene expression levels and selenium content, BpHMT2 was identified as a key candidate gene associated with selenium tolerance. Subcellular localization experiments confirmed the targeting of BpHMT2 in nucleus, cell membrane and chloroplasts. Moreover, three BpHMT2 overexpression Arabidopsis thaliana lines were confirmed to enhance plant selenium tolerance and SeMet accumulation. Overall, our finding provides insights into the molecular mechanisms of selenium metabolism in B. papyrifera, highlighting the potential role of BpHMT2 in SeMet synthesis. This research contributes to our understanding of selenium-enriched feed resources, with increased SeMet content contributing to the improved nutritional value of B. papyrifera as a feed resource.

Synergistic effects of ferulic acid esterase-producing lactic acid bacteria, cellulase and xylanase on the fermentation characteristics, fibre and nitrogen components and microbial community structure of Broussonetia papyrifera during ensiling.

BACKGROUND: The high fibre content of whole plants of Broussonetia papyrifera limits its efficient utilization. Ferulic acid esterase (FAE), in combination with xylanase, can effectively cleave the lignin-carbohydrate complex, promoting the function of cellulase. However, little is known about the impact of these additives on silage. To effectively utilize natural woody plant resources, FAE-producing Lactiplantibacillus plantarum RO395, xylanase (XY) and cellulase (CE) were used to investigate the dynamic fermentation characteristics, fibre and nitrogen components and microbial community structure during B. papyrifera ensiling. RESULTS: Broussonetia papyrifera was either not treated (CK) or treated with FAE-producing lactic acid bacteria (LP), CE, XY, LP + CE, LP + XY or LP + CE + XY for 3, 7, 15, 30 or 60 days, respectively. In comparison with those in the CK treatment, the L. plantarum and enzyme treatments (LP + CE, LP + XY and LP + XY + CE), especially the LP + XY + CE treatment, significantly increased the lactic acid concentration and decreased the pH and the contents of acid detergent insoluble protein and NH3 -N (P < 0.05). Enzyme addition improved the degradation efficiency of lignocellulose, and a synergistic effect was observed after enzyme treatment in combination with LP; in addition, the lowest acid detergent fibre, neutral detergent fibre, hemicellulose and cellulose contents were detected after the LP + CE + XY treatment (P < 0.05). Moreover, CE, XY and LP additions significantly improved the microbial community structure, increased the relative abundance of Lactiplantibacillus and Firmicutes, and effectively inhibited undesirable bacterial (Enterobacter) growth during ensiling. CONCLUSION: FAE-producing L. plantarum and the two tested enzymes exhibited synergistic effects on improving the quality of silage, which indicates that this combination can serve as an efficient method for improved B. papyrifera silage utilization. © 2023 Society of Chemical Industry.

Comprehensive Profiling of Paper Mulberry (Broussonetia papyrifera) Crotonylome Reveals the Significance of Lysine Crotonylation in Young Leaves.

Lysine crotonylation is a newly discovered and reversible posttranslational modification involved in various biological processes, especially metabolism regulation. A total of 5159 lysine crotonylation sites in 2272 protein groups were identified. Twenty-seven motifs were found to be the preferred amino acid sequences for crotonylation sites. Functional annotation analyses revealed that most crotonylated proteins play important roles in metabolic processes and photosynthesis. Bioinformatics analysis suggested that lysine crotonylation preferentially targets a variety of important biological processes, including ribosome, glyoxylate and dicarboxylate metabolism, carbon fixation in photosynthetic organisms, proteasome and the TCA cycle, indicating lysine crotonylation is involved in the common mechanism of metabolic regulation. A protein interaction network analysis revealed that diverse interactions are modulated by protein crotonylation. These results suggest that lysine crotonylation is involved in a variety of biological processes. HSP70 is a crucial protein involved in protecting plant cells and tissues from thermal or abiotic stress responses, and HSP70 protein was found to be crotonylated in paper mulberry. This systematic analysis provides the first comprehensive analysis of lysine crotonylation in paper mulberry and provides important resources for further study on the regulatory mechanism and function of the lysine crotonylated proteome.

Genome-Wide Analysis of BpYABs and Function Identification Involving in the Leaf and Silique Development in Transgenic Arabidopsis.

YABs play an important role in the leaf development of the paper mulberry (Broussonetia papyrifera) and of the heterophylly. Thus, we investigated the function of BpYABs. Gene cloning, phylogenetic analysis, motif identification, subcellular localization, transactivation activity assay, qRT-PCR, in situ hybridization, and ectopic expression were used in our study. Six BpYABs were isolated, and four of them had transcriptional activity. BpYAB1, BpYAB3, BpYAB4, and BpYAB5 were localized to the nucleus. BpYAB1 was only expressed in the flower, while BpYAB6 was not expressed in any detected tissues; the four remaining BpYABs were expressed in the bud, leaf and flower, and their expression level decreased with leaf development. Further in situ hybridization showed that BpYAB3 and BpYAB5 were expressed in the vascular tissues and lamina, but neither showed the adaxial-abaxial polarity distribution pattern in the mature leaf lamina. Ectopic expression of BpYAB2, BpYAB3, BpYAB4 and BpYAB5 induced increased expression of AtWOX1 and caused the leaf of Arabidopsis to become smaller and curl downwards. Ectopic expression also led to shorter siliques and smaller seeds, but not for BpYAB5. These results suggest that BpYABs have functional divergency and redundancy in regulating leaf and silique development.

Tolerance capacities of Broussonetia papyrifera to heavy metal(loid)s and its phytoremediation potential of the contaminated soil.

Broussonetia papyrifera, is a promising fast-growing woody plant for the phytoremediation of heavy metal(loid) (HM)-contaminated soil. In this study, a greenhouse experiment was conducted to explore the tolerance capacities of B. papyrifera and its phytoremediation potential in the HM-contaminated soil. The results indicated that B. papyrifera could effectively decrease malondialdehyde (MDA) content by enhancing the antioxidant enzyme activities along with the cultivation in the HM-contaminated soil. Significant (p < 0.05) negative relationships were found between MDA content and superoxide dismutase (r = -0.620) and catalase activities (r = -0.702) in B. papyrifera leaves. Fourier Transform Infrared Spectroscopy analysis indicated that the main functional groups in B. papyrifera roots were slightly influenced by HMs, and organic acids, carbohydrates, protein, and amino acids might bind with HMs in plant roots to alleviate the adverse effect of HMs on plants growth. Meanwhile, B. papyrifera had great potential used for the phytoextraction of Cd and Zn in HM-contaminated soil. The maximum total Cd and Zn accumulation amount in B. papyrifera shoots could attach to 2.26 and 66.8 mg·pot-1, respectively. These observations suggested that B. papyrifera has large biomass and high tolerance to HMs, which can be regarded as a promising plant for the eco-remediation of HM-contaminated sites.Novelty statement In this study, a fast-growing woody plant, Broussonetia papyrifera, was used for heavy metal(loid) (HM)-contaminated soil remediation. We found that B. papyrifera can effectively alleviate the adverse effect of HMs on plant growth by enhancing the antioxidant enzyme activities in leaves and binding HMs with organic acids, carbohydrates, protein, and amino acids in roots. Furthermore, the maximum total Cd and Zn accumulation amount in B. papyrifera shoots could attach to 2.26 and 66.8 mg·pot-1, which suggested that B. papyrifera might be regarded as a promising woody plant used for the phytoextraction of Cd and Zn in the contaminated soil.

First comprehensive analysis of lysine succinylation in paper mulberry (Broussonetia papyrifera).

BACKGROUND: Lysine succinylation is a naturally occurring post-translational modification (PTM) that is ubiquitous in organisms. Lysine succinylation plays important roles in regulating protein structure and function as well as cellular metabolism. Global lysine succinylation at the proteomic level has been identified in a variety of species; however, limited information on lysine succinylation in plant species, especially paper mulberry, is available. Paper mulberry is not only an important plant in traditional Chinese medicine, but it is also a tree species with significant economic value. Paper mulberry is found in the temperate and tropical zones of China. The present study analyzed the effects of lysine succinylation on the growth, development, and physiology of paper mulberry. RESULTS: A total of 2097 lysine succinylation sites were identified in 935 proteins associated with the citric acid cycle (TCA cycle), glyoxylic acid and dicarboxylic acid metabolism, ribosomes and oxidative phosphorylation; these pathways play a role in carbon fixation in photosynthetic organisms and may be regulated by lysine succinylation. The modified proteins were distributed in multiple subcellular compartments and were involved in a wide variety of biological processes, such as photosynthesis and the Calvin-Benson cycle. CONCLUSION: Lysine-succinylated proteins may play key regulatory roles in metabolism, primarily in photosynthesis and oxidative phosphorylation, as well as in many other cellular processes. In addition to the large number of succinylated proteins associated with photosynthesis and oxidative phosphorylation, some proteins associated with the TCA cycle are succinylated. Our study can serve as a reference for further proteomics studies of the downstream effects of succinylation on the physiology and biochemistry of paper mulberry.

Identification of the PP2C gene family in paper mulberry (Broussonetia papyrifera) and its roles in the regulation mechanism of the response to cold stress.

OBJECTIVES: To study the possible roles of type-2C protein phosphatases (PP2Cs) which have been confirmed to play roles in the response to diverse abiotic stresses in paper mulberry, we launched a series of genomic and functional studies of BpPP2Cs. RESULTS: Sixty-three PP2C proteins in paper mulberry (Broussonetia papyrifera) were classified into 13 clades. Four BpPP2Cs with kinase domains were verified to be highly conserved in organisms ranging from algae to dicots. Seven pairs of BpPP2C genes were found to be expanding, and 18 BpPP2C genes had orthologues in Arabidopsis. BpPP2Cs showed broad expression in different tissues; the expression levels of 18 BpPP2Cs were changed and the phosphorylation levels of seven BpPP2C proteins increased at low temperature. Cold-response elements were found in the promoter region of 31 BpPP2Cs. Finally, Bp01g0320 was found to act as a hub protein and Bp01g0512 and Bp09g1278 played key roles in the ABA-signaling pathway and MAPK cascades, respectively. CONCLUSION: These results suggest that the PP2C gene family of paper mulberry is evolutionarily conserved and participates the regulation of the response to cold stress, which will play a vital role in further research on phosphatases in paper mulberry.

A Chromosome-Scale Genome Assembly of Paper Mulberry (Broussonetia papyrifera) Provides New Insights into Its Forage and Papermaking Usage.

Paper mulberry (Broussonetia papyrifera) is a well-known woody tree historically used for Cai Lun papermaking, one of the four great inventions of ancient China. More recently, Paper mulberry has also been used as forage to address the shortage of feedstuff because of its digestible crude fiber and high protein contents. In this study, we obtained a chromosome-scale genome assembly for Paper mulberry using integrated approaches, including Illumina and PacBio sequencing platform as well as Hi-C, optical, and genetic maps. The assembled Paper mulberry genome consists of 386.83 Mb, which is close to the estimated size, and 99.25% (383.93 Mb) of the assembly was assigned to 13 pseudochromosomes. Comparative genomic analysis revealed the expansion and contraction in the flavonoid and lignin biosynthetic gene families, respectively, accounting for the enhanced flavonoid and decreased lignin biosynthesis in Paper mulberry. Moreover, the increased ratio of syringyl-lignin to guaiacyl-lignin in Paper mulberry underscores its suitability for use in medicine, forage, papermaking, and barkcloth making. We also identified the root-associated microbiota of Paper mulberry and found that Pseudomonas and Rhizobia were enriched in its roots and may provide the source of nitrogen for its stems and leaves via symbiotic nitrogen fixation. Collectively, these results suggest that Paper mulberry might have undergone adaptive evolution and recruited nitrogen-fixing microbes to promote growth by enhancing flavonoid production and altering lignin monomer composition. Our study provides significant insights into genetic basis of the usefulness of Paper mulberry in papermaking and barkcloth making, and as forage. These insights will facilitate further domestication and selection as well as industrial utilization of Paper mulberry worldwide.

Nitrogen transfer from one plant to another depends on plant biomass production between conspecific and heterospecific species via a common arbuscular mycorrhizal network.

The formation of a common mycorrhizal network (CMN) between roots of different plant species enables nutrient transfers from one plant to another and their coexistence. However, almost all studies on nutrient transfers between CMN-connected plants have separately, but not simultaneously, been demonstrated under the same experimentation. Both conspecific and heterospecific seedlings of Cinnamomum camphora, Bidens pilosa, and Broussonetia papyrifera native to a karst habitat in southwest China were concurrently grown in a growth microcosm that had seven hollowed compartments (six around one in the center) being covered by 35.0-µm and/or 0.45-µm nylon mesh. The Ci. camphora in the central compartment was supplied with or without Glomus etunicatum and 15N to track N transfers between CMN-connected conspecific and heterospecific seedlings. The results showed as follows: significant greater nitrogen accumulations, biomass productions, 15N content, % Ntransfer, and the Ntransfer amount between receiver plant species ranked as Br. papyrifera≈Bi. pilosa > Ci. camphora under both M+ and M-, and as under M+ than under M- for Ci. camphora but not for both Bi. Pilosa and Br. papyrifera; the CMN transferred more nitrogen (15N content, % Ntransfer, and Ntransfer amount) from the donor Ci. camphora to the heterospecific Br. papyrifera and Bi. pilosa, with a lower percentage of nitrogen derived from transfer (%NDFT). These findings suggest that the CMN may potentially regulate the nitrogen transfer from a donor plant to individual heterospecific receiver plants, where the ratio of nitrogen derived from transfer depends on the biomass strength of the individual plants.

New insight into the molecular basis of cadmium stress responses of wild paper mulberry plant by transcriptome analysis.

BACKGROUND: Heavy metal contamination is becoming a limitation to the utilization of soil and the distribution of vegetation. In particular, cadmium (Cd) pollution has had a serious impact on the food chain. Broussonetia papyrifera is a widely distributed pioneer tree species of heavy metal contaminated areas with important economic value. However, little is known about the genomic background of the Cd-tolerance mechanism in B. papyrifera. RESULTS: The CdCl2 responsive physiology was evaluated and proved to be involved in antioxidase activity and active oxygen species (ROS) accumulation. The leaf and root transcriptomes derived from B. papyrifera grown under normal and CdCl2 stress conditions were systematically investigated using the Illumina HiSeq method. A total of 180,678,660 bp (27.1 GB) clean reads were assembled into 589,487 high-quality unigenes, of which 256,025 (43.43% of the total) and 250,251 (42.45% of the total) were aligned in Gene Ontology (GO) and Protein family (Pfam), respectively. A total of 24,414 differentially expressed genes (DEGs) were GO-annotated into 53, 23, 55, and 60 terms from the transcriptomes of root and leaf tissues under Cd stress and control conditions. A total of 117,547 Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology (KO)-annotated DEGs were enriched in at least 47 KEGG pathway terms among the four comparisons. Many genes encoding important transcription factors (e.g., auxin/indole-3-acetic acid (AUX/IAA), basic helix-loop-helix (bHLH), DNA-binding one zinc finger (Dof), and MYB) and proteins involved in plant-pathogen interactions, phenylpropanoid biosynthesis, plant hormone signal transduction, oxidative phosphorylation, carbon fixation, peroxisomes, flavonoid biosynthesis, and glutathione metabolism, among others, were substantially upregulated under CdCl2 stress. CONCLUSIONS: These genes represent important candidates for studying Cd-response mechanisms and molecular biology of B. papyrifera and related species. Our findings provide a genomic sequence resource for functional genetic assignments in B. papyrifera, which will help elucidate the molecular mechanisms of its Cd-stress responses and facilitate the bioremediation of heavy metal contaminated areas via breeding of new stress-tolerant cultivars.

De Novo assembly of expressed transcripts and global transcriptomic analysis from seedlings of the paper mulberry (Broussonetia kazinoki x Broussonetia papyifera).

The paper mulberry is one of the multifunctional tree species in agroforestry systems and is also commonly utilized in traditional medicine in China and other Asian countries. However, little is known about its molecular genetics, which hinders research on and exploitation of this valuable resource. To discern the correlation between gene expression and the essential properties of the paper mulberry, we performed a transcriptomics analysis, assembling a total of 37,725 unigenes from 54,638,676 reads generated by RNA-seq. Among these, 22,692 unigenes showed greater than 60% similarity with genes from other species. The lengths of 13,566 annotated unigenes were longer than 1,000 bp. Functional clustering analysis with COG (Cluster of Orthologous Groups) revealed that 17,184 unigenes are primarily involved in transcription, translation, signal transduction, carbohydrate metabolism, secondary metabolism, and energy metabolism. GO (Gene Ontology) annotation suggests enrichment of genes encoding antioxidant activity, transporter activity, biosynthesis, metabolism and stress response, with a total of 30,659 unigenes falling in these categories. KEGG (Kyoto Encyclopedia of Genes and Genomes) metabolic pathway analysis showed that 7,199 unigenes are associated with 119 metabolic pathways. In addition to the basic metabolism, these genes are enriched for plant pathogen interaction, flavonoid metabolism and other secondary metabolic processes. Furthermore, differences in the transcriptomes of leaf, stem and root tissues were analyzed and 7,233 specifically expressed unigenes were identified. This global expression analysis provided novel insights about the molecular mechanisms of the biosynthesis of flavonoid, lignin and cellulose, as well as on the response to biotic and abiotic stresses including the remediation of contaminated soil by the paper mulberry.

Functional analysis of BpDREB2 gene involved in salt and drought response from a woody plant Broussonetia papyrifera.

The dehydration-responsive element binding proteins (DREBs) are important transcription factors in the regulation of plant responses to abiotic stresses. In this study, BpDREB2, an AP2/DREB-type transcription factor gene, was cloned from a woody plant, Broussonetia papyrifera by RACE-PCR. Sequence analyses revealed that BpDREB2 protein has three characteristic domains, including an AP2/EREBP, a nuclear localization signal and an acidic activation domain. Yeast one-hybrid assays showed that BpDREB2 protein specifically binds to the DRE sequence and activates the expression of reporter genes in yeast. These results suggested that BpDREB2 protein could function as a transcription factor of DREB family. The expression of BpDREB2 gene was remarkably induced by dehydration and high-salt treatments, but no significant change was observed under ABA or low-temperature conditions. Importantly, transgenic expression of BpDREB2 gene in Arabidopsis significantly enhanced its tolerance to salt and freezing without causing growth retardation. Taken together, these results suggested that BpDREB2 is a novel member of the AP2/EREBP trans-acting factor family which could enhance salt stress tolerance of plants and has the potential application in the improvement of crops and economical tree species.

Enhanced expression of vacuolar H+-ATPase subunit E in the roots is associated with the adaptation of Broussonetia papyrifera to salt stress.

Vacuolar H(+)-ATPase (V-H(+)-ATPase) may play a pivotal role in maintenance of ion homeostasis inside plant cells. In the present study, the expression of V-H(+)-ATPase genes was analyzed in the roots and leaves of a woody plant, Broussonetia papyrifera, which was stressed with 50, 100 and 150 mM NaCl. Moreover, the expression and distribution of the subunit E protein were investigated by Western blot and immunocytochemistry. These showed that treatment of B. papyrifera with NaCl distinctly changed the hydrolytic activity of V-H(+)-ATPase in the roots and leaves. Salinity induced a dramatic increase in V-H(+)-ATPase hydrolytic activity in the roots. However, only slight changes in V-H(+)-ATPase hydrolytic activity were observed in the leaves. In contrast, increased H(+) pumping activity of V-H(+)-ATPase was observed in both the roots and leaves. In addition, NaCl treatment led to an increase in H(+)-pyrophosphatase (V-H(+)-PPase) activity in the roots. Moreover, NaCl treatment triggered the enhancement of mRNA levels for subunits A, E and c of V-H(+)-ATPase in the roots, whereas only subunit c mRNA was observed to increase in the leaves. By Western blot and immunocytological analysis, subunit E was shown to be augmented in response to salinity stress in the roots. These findings provide evidence that under salt stress, increased V-H(+)-ATPase activity in the roots was positively correlated with higher transcript and protein levels of V-H(+)-ATPase subunit E. Altogether, our results suggest an essential role for V-H(+)-ATPase subunit E in the response of plants to salinity stress.

[Absorption and allocation characteristics of K+, Ca2+, Na+ and Cl- in different organs of Broussonetia papyrifera seedlings under NaCl stress].

One-year-old Broussonetia papyrifera seedlings were subjected to 0.4, 1, 2, 3, and 4 g x kg(-1) of soil NaCl stress, and their biomass accumulation, leaf plasma membrane permeability, and the absorption, allocation and translocation of K+, Ca2+, Na+, and Cl-, as well as the symptoms of salt injury, were studied and investigated. The leaf plasma membrane permeability increased with the increase of soil NaCl concentration and of the duration of soil NaCl stress, and the seedling's root/shoot ratio also increased with increasing soil NaCl concentration. When the soil NaCl concentration exceeded 3 g x kg(-1), leaf plasma membrane permeability and seedling' s biomass accumulation were affected significantly. The Na+ and Cl- concentrations in different organs of seedlings increased with increasing soil NaCl concentration while the K+ and Ca2+ concentrations were in adverse, and the ion contents in leaves were always much higher than those in other organs, illustrating that soil NaCl stress affected the K+ and Ca2+ absorbing capability of roots, and inhibited the selective translocation of K+ and Ca2+ to aboveground parts. As a result, the K+ and Ca2+ concentrations in leaves and stems decreased. The study showed that B. papyrifera could effectively resist the injury of osmotic stress from soil salt via absorbing and accumulating Na+ and Cl-, but excessive accumulation of Na+ and Cl- could induce salt toxicity. As a non-halophyte species with relatively strong salt resistance, the aboveground parts of B. papyrifera did not have significant salt-exclusion effect.

[Responses of delta13 C values of plant leaves to environmental gradients along environmental gradient factors in rocky desertified area of a typical karst Ggorge].

We analyzed the responses of delta13 C values of plant leaves to environmental factors (namely, soil water storage, air relative humidity, light intensity, depths of soil, soil organic content, average temperature and soil water content) and the correlations between them, by measuring delta13 C values of leaves for 11 plants species from 4 typical communities with different karst rocky desertification backgrounds in a typical karst catchments basin, Huajiang Gorge. It is revealed that, the delta13 C values and water use efficient of most species decrease with the increasing of water supply; but a few species exhibit an opposite trend and several others exhibit no change in delta13 C values or water use efficiency when these environmental factors varied. Moreover, the correlation analysis indicates that the soil water storage is the leading factor for Pistacia weinmannifolia, Mallotus repandus and Alchornea trewioides, while the depths of soil is essential factor for Nephrolepis cordifolia and Mallotus japonicus var. floccosus, and the light intensity is leading factor for N. cordifolia, Alangium chinense, Broussonetia papyrifera. However, the leading factor for some species like Rapanea kwangsiensis, Sapium rotundifolium and Cipadessa cinerascens are yet not clear, which mean their delta13 C values are affected by more comprehensive factors. Hence it could be concluded that high delta13 C values of leaves could indicate the adaptability of plants for low water regime, high light and low resource environments.