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    20 March 2022, Volume 42 Issue 2
    Systematic and Evolutionary
    Correction of Typographical Errors in the Protologue of Twenty-five Taxa (Ferns) in China
    Yun Lin, Qian Sun, Xiaobing Zhang, Dongdong He, Huibing Wu, Li Song, Wenqun Liu
    2022, 42(2):  161-173.  doi:10.7525/j.issn.1673-5102.2022.02.001
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    According to Article 9.2 in the International Code of Nomenclature for algae, fungi, and plants(Shenzhen Code), the typographical errors in the protologues of twenty-five taxa(ferns) in China were corrected respectively, including Gleichenia cantonensis Ching, Hicriopteris omeiensis Ching & P.S. Chiu, Hypolepis yunnanensis Ching, Coniogramme latibasis Ching ex K.H. Shing, Athyrium costulalisorum Ching, Athyrium exindusiatum Ching, Athyrium gongshanense Ching, Athyrium habaense Ching, Athyrium pseudoepirachis Ching, Athyrium submacrocarpum Ching & S.K. Wu, Lunathyrium sichuanense Z.R. Wang var. jinfoshanense Z.R. Wang, Pseudocystopteris remota Ching, Diacalpe medogensis R.C. Ching & S.K. Wu, Arachniodes falcata Ching, Cyrtogonellum caducum Ching, Cyrtogonellum salicifolium Ching & Y.T. Hsieh, Cyrtomium moupingense Ching & K.H. Shing ex K.H. Shing, Cyrtomium tsinglingense Ching & K.H. Shing ex K.H. Shing, Dryopteris grandiosa Ching & P.C. Chiu, Dryopteris occidental-zhejiangensis Ching & P.C. Chiu, Dryopteris subtenuicula Ching & P.C. Chiu, Hemigramma distinctipetiolata Ching, Trogostolon yunnanensis Ching, Colysis liouii Ching and Lepisorus pseudonudus Ching.

    Leaf Anatomical Characteristics and Environmental Adaptability of Seven Sorbus Species at Longcanggou National Forest Park
    Kailin Zhu, Jiabao Li, Xin Chen
    2022, 42(2):  174-183.  doi:10.7525/j.issn.1673-5102.2022.02.002
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    Leaf anatomical structures of seven Sorbus species at Longcanggou National Forest Park in Sichuan Province were observed by paraffin section method, and the relationship between leaf anatomical structure and environment was discussed in this study. The results showed that: the leaves of seven species were all of typical dorsal-ventral pattern; leaf thickness ranged from 108.16-208.21 μm, significant differences(P<0.01) were observed in the studied species; upper epidermis thickness was greater than the lower, furthermore, papillae were observed in the lower epidermis of pinnate-leaved species; palisade parenchyma was composed of 1-2 layers of cells, the ratio of palisade parenchyma to spongy parenchyma in thickness of S. prattii var. aestivalis was 1.93, and the rest of six species were less than 1 respectively; the vascular bundles of the midrib were all heart-shaped, which were typical collateral vascular bundle; the differences of midrib protuberant degree among species were extremly significant(P<0.01). Among the anatomical structures, the plasticity of the upper and lower epidermis was the largest, which had strong potential adaptability in the habitat; the midrib had the least plasticity and the overall structure was relatively stable. Palisade parenchyma, spongy parenchyma and midrib were the most different anatomical structures among the seven species. Correlation analysis between leaf anatomical structures and environmental variables indicated that palisade parenchyma thickness, the ratio of palisade parenchyma to spongy parenchyma and cell tense ratio were positively correlated with annual precipitation, precipitation of warmest quarter and altitude(P<0.05), and were negatively correlated with temperature seasonality(P<0.05); diameter and protuberant degree of midrib were positively correlated with seasonal temperature(P<0.01), and were negatively correlated with annual precipitation, precipitation of warmest quarter and altitude(P<0.05). The adaptability of leaf anatomical characteristics reflect the survival strategies of seven Sorbus species at Longcanggou National Forest Park.

    Genetic and Breeding
    Rapid Micropropagation of Fraxinus mandshurica Rupr. Based on Liquid and Solid Alternate Culture
    Fenghui Qi, Wenxuan Wang, Lin Liu, Mingshuo Tang, Feixiang Song, Yaguang Zhan
    2022, 42(2):  184-190.  doi:10.7525/j.issn.1673-5102.2022.02.003
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    A rapid and efficient regeneration system of liquid-solid alternate culture was developed for the tissue culture seedlings of Fraxinus mandshurica with lignificated stalk, aged leaves and dormant apical buds, and the axillary buds were germinated in a short time by liquid suspension culture, and the axillary buds were regenerated in vitro in solid culture to obtain the new tissue culture seedlings. On liquid and solid media supplemented of WPM with different plant growth regulators, axillary buds of F. mandshurica were induced to germinate and grow into seedlings. It was found that the axillary bud germination rate of F. mandshurica could reach 100% within 7-15 d after dark culture in WPM liquid medium supplemented with 0.6 mg·L-1 TDZ. After cutting the sprouted buds and inoculating them into WPM solid medium of 0.05 mg·L-1 TDZ and 0.6 mg·L-1 BA, axillary buds could grow into shoots after 1-2 subcultures with an average height of 2.64 cm and a multiplication coefficient of 4.04. After transplanting, the survival rate of rooting seedlings was 90% after 50 d. The establishment of the method could be conducive to the large-scale propagation of F. mandshurica, and the liquid-solid alternate circulation culture, simple, controllable, easy to operate, suitable for different production conditions and reduces the cost.

    Variation of Lignin Content and Association Analysis of FmPAL Nucleotide Polymorphism in Progenies of Interspecific Hybrids of Fraxinus
    Yibo Yin, Jixiang Li, Yingjie Guo, Ziting Lu, Ying Xiao, Hualing Liu, Yaguang Zhan, Fansuo Zeng
    2022, 42(2):  191-199.  doi:10.7525/j.issn.1673-5102.2022.02.004
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    To improve materials for genetic breeding of Fraxinus mandshurica, the correlation among three hybrid combinations of Fraxinus mandshurica × F. rhynchophyllaF. mandshurica × F. sogdiana and F. mandshurica × F. velutina, and the genetic variation of lignin content and FmPAL nucleotide polymorphism in free-pollination offspring of female parent were analyzed respectively. One-year-old branches of Fraxinus mandshurica with 71 species of interspecies hybrid combinations in Maoer Mountain Forest Farm of Northeast Forestry University were used as materials, the genetic variation regularity of lignin was investigated, the differences in lignin content and variation parameters and mean values were analyzed, and joint analysis on the molecular level of lignin and its synthesis-related gene PAL were carried out. The result showed that the lignin content of the hybrid combinations of Fraxinus was extremely significant, and it was significantly different from the female parent free-pollinated offspring of F. mandshurica. The lignin content of most hybrid combinations of F. mandshurica was higher than that of the female parent F. mandshurica. The results showed that the haploid diversity value of the PAL gene of the F. mandshurica population reached 0.999 7, which was a high level. It confirmed that the FmPAL gene had a high variation. There were 3 SNP sites significantly related to the lignin content. There were significant differences in the lignin content of the cross combinations of F. mandshurica. The SNP sites in the FmPAL gene was significantly related to the lignin content.

    Molecular biology
    Comparative Transcriptome Analysis of Three Artemisia Species in Qinghai Tibet Plateau
    Jingya Yu, Mingze Xia, Hao Xu, Faqi Zhang
    2022, 42(2):  200-210.  doi:10.7525/j.issn.1673-5102.2022.02.005
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    Artemisia dalai-lamaeArtemisia frigida and Artemisia hedinii, the common herbs in arid and semi-arid areas of Northwest China, have the effects of wind prevention and sand fixation, preventing grassland degradation, sterilization and anti-inflammatory, and treating a variety of diseases. The transcriptome of three Artemisia species were sequenced by Illumina method and then 54 268 322, 46 434 864 and 43 971 646 Clean reads were obtained, respectively. SNP and alternative splicing analyses were performed on the sequencing results. Differentially expressed genes were screened from the comparison groups of 6 107(ADL vs AF), 4 822(ADL vs AH) and 3 755(AF vs AH) respectively. Go enrichment annotated the differentially expressed genes into three categories: biological process, cell component and molecular function. KEGG enrichment annotated the differentially expressed genes into 198, 198 and 197 pathways respectively. The top ten high expressed genes in each group were analyzed. We screened out 25 differentially expressed genes related to the active components of alkaloids,terpenoids and flavonoid. This study provid a scientific basis for species identification, stress resistance research and resource utilization of Artemisia.

    Bioinformatic Analysis and Expression Pattern of LTP Family Genes in Populus davidiana × P. alba var. pyramidalis
    Qian Sun, Yuhang Wu, Yaxuan Zhang, Jingdan Cao, Jingjing Shi, Chao Wang
    2022, 42(2):  211-223.  doi:10.7525/j.issn.1673-5102.2022.02.006
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    To lay a foundation for studying the molecular regulation mechanism of LTP family gene and molecular genetic improvement of Populus davidiana × P. alba var. pyramidalis, the expression pattern of LTP family genes from P. davidiana × P. alba var. pyramidalis was analyzed.The protein properties, multiple sequence alignments and phylogenetic tree analysis of LTP family members were performed to show their structural features. The P. davidiana × P. alba var. pyramidalis seedlings treated with gravity, salt and PEG stress respectively were subjected to quantitive real time PCR analysis. The expression patterns of PdbLTP family genes were carried out respectively. Eight PdbLTP genes and two subfamily PdbGLTP genes were obtained, and their CDS lengths were ranged from 294-396 bp. LTP family were hydrophobic proteins and had eight cysteine conserved structures, without transmembrane region. qRT-PCR analysis showed that the expression patterns of different PdbLTP family genes were different under gravity processing. Expression levels of PdbLTP1PdbLTP3PdbLTP5 and PdbLTP7 genes were up-regulated in tension wood compared to those in the normal wood. The transcripts levels of PdbLTP1PdbLTP2PdbLTP3PdbLTP5 genes were abundant in stem.Expression of PdbLTP family genes was induced by salt stress except for PdbLTP5. PdbLTP1PdbLTP2PdbLTP3 and PdbLTP5 were up-regulated under drought stress. Different members of PdbLTP gene family play important roles in regulation of xylem development and tolerance to abiotic stress in Populus davidiana × P. alba var. pyramidalis.

    Expression and Functional Analysis of FT Homologous Genes in Saffron(Crocus sativus L.)
    Zhen Wang, Liuyan Yang, Weizhong Pei, Xin Li, Zhen Yang, Yongchun Zhang
    2022, 42(2):  224-233.  doi:10.7525/j.issn.1673-5102.2022.02.007
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    As one of the three floral pathway integrators, FT(FLOWERING LOCUS T) and its homologous genes were considered to be important genes which related to flowering. In order to clarify the function of FT homologous genes and flowering mechanism of saffron(Crocus sativus L.), the three reported FT homologous genes, named CsatFT1CsatFT2 and CsatFT3, were isolated and analyzed. The gDNA contained the ORF with length of 835, 1 642 and 1 132 bp, respectively, and they all had 4 exons and 3 introns. cDNA sequences contained the length of 528, 525 and 540 bp ORF, which encoded 175, 174 and 179 amino acids, respectively. The phylogenetic analysis indicated that CsatFT1, CsatFT2 and CsatFT3 showed relatively close genetic distance with the monocotyledon Narcissus tazetta NtFT, Lilium longiflorum LlFT and Allium cepa AcFT1, respectively. qRT-PCR results indicated that the expression levels of CsatFT1CsatFT2 and CsatFT3 were the highest in the leaves, followed by the lateral roots, and almost no expressed in daughter corms and primary roots at the early stage of replacement corm enlargement. At the late stage of replacement corm enlargement, the expression levels of CsatFT1CsatFT2 and CsatFT3 were all high in daughter corms and were almost not detected in terminal buds. During indoor storage stage, the expression levels of CsatFT1CsatFT2 and CsatFT3 were high in the capital, followed by the leaves, and almost undetectable in the petals and anthers. From the phenotypes of transgenic tobacco and transgenic Arabidopsis plants, the results showed that CsatFT1CsatFT2 and CsatFT3 could promote the early flowering of plants.

    Identification of PeNAC121 Gene Promoter and Stress Response Pattern Analysis in Populus euphratica
    Wang Xie, Tianjing Li, Xinyao Li, Fenming Yang, Yin’an Yan, Yongfeng Gao
    2022, 42(2):  234-242.  doi:10.7525/j.issn.1673-5102.2022.02.008
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    To explore the stress response and regulatory mechanism of NAC transcription factor family members in Populus euphratica, the promoter sequence of PeNAC121 gene was cloned from leaves of Populus euphrachiae by PCR and its structural characteristics were analyzed using bioinformatics tools. The PeNAC121 promoter was used to drive the expression of the GUS reporter gene in the triploid Populus tomentosa, and then GUS staining and enzyme activity quantitative analysis were performed in the obtained transgenic plants after different stress treatments. The cloned promoter sequence of PeNAC121 gene was 1 997 bp(located in the upstream of start codon ATG) in length, included a large number of light-responsive elements, and also contained a number of abiotic stress and phytohormone responsive elements, such as low temperature response element LTR, drought response element MBS, abscisic acid(ABA) response element, as well as gibberellin(GA) response elements, respectively. The result of tissue expression pattern showed that the PeNAC121 gene was mainly expressed in stemsof P. euphratica, with low expression in roots and leaves. The results of GUS histochemical staining and enzyme activity test of transgenic plants showed that the PeNAC121 promoter was significantly induced by NaCl, mannitol, ABA and 4 ℃ low temperature respectively. It was speculated from the above results that PeNAC121 gene was closely related to the response to stress. The results showed that the promoter of PeNAC121 gene was an inducible promoter responding to multiple stresses. The study provid a theoretical reference for elucidating the role of PeNAC121 gene in stress response and regulation of P. euphratica.

    Overexpression of Marine Microbial Metagenomic MbCSP Enhanced Drought and Cold Tolerance of Transgenic Arabidopsisthaliana
    Mengjiao Wang, Yuxue Cao, Yongsheng Xu, Fenge Ding, Qiao Su
    2022, 42(2):  243-251.  doi:10.7525/j.issn.1673-5102.2022.02.009
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    Drought and low temperature are important factors affecting the growth and development of crop, genetically modified crop is an effective way to solve this problem. Cold shock proteins(CSPs)are a class of highly conserved nucleic acid binding proteins, which participate in cell physiological activities such as abiotic stress response, and the transfer of CSP gene can enhance the stress tolerance of crop. In this study, MbCSP gene was cloned by anchor PCR with marine microbial metagenomic DNA as template, and ORF was 216bp, encoded 71 amino acids. Homology analysis showed that the amino acid sequence had homology from 60% to 90% with cold shock proteins such as EcCSPG, EcCSPA(Escherichia coli), BsCSPB(Bacillus subtilis) and BcCSPA(Bacillus cereus), respectively. The results of multiple sequence alignment and phylogenetic tree analysis showed that MbCSP protein contained the classical conserved domains of CSP proteins such as RNP1(KGFGFI) and RNP2(VFVHF), and were closely related to the cold shock proteins of EcCSPG(E. coli) and CmCSPA, CmCSPB(compost metagenome). To further explore the function of MbCSP, a plant expression vector pTF101-MbCSP was constructed, and transfered to Arabidopsis thaliana by floral dip method. Transgenic plants were obtained by herbicide screening and PCR. Semi quantitative RT-PCR analysis was carried out, and the positive strain with the highest expression level was selected for subsequent physiological detection. The results showed that the growth of transgenic Arabidopsis plants were significantly better than that of wild-type, and its biomass was significantly higher than that of wild-type, under drought stress and low temperature stress. The relative water content, chlorophyll content and superoxide dismutase activity of transgenic Arabidopsis plants were higher than those of wild-type, while the content of malondialdehyde was lower than that of wild-type. The results showed that overexpression of marine microbial metagenomic MbCSP could improve drought and cold tolerance of transgenic A.thaliana plant, which laid the foundation for cultivating new varieties of transgenic crops.

    Phytochemistry
    Separation and Purification of Three Active Components from Dracocephalum tanguticum Maxim. by High-Speed Counter-current Chromatography Combined with Semi-Preparative High Performance Liquid Chromatography
    Jiaben Zhou, Caidan Rezeng, Maoxian Re, Cairang Limao
    2022, 42(2):  252-258.  doi:10.7525/j.issn.1673-5102.2022.02.010
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    An efficient method was established for the separation and preparation of chlorogenic acid, pedaliin-6″-acetate and rosmarinic acid from Dracocephalum tanguticum Maxim.. The chlorogenic acid, pedaliin-6″-acetate and rosmarinic acid was enriched by semi-preparative high performance liquid chromatography(SP-HPLC) from ethyl acetate extract, and then separated and purification by high-speed counter-current chromatography(HSCCC) and preparative high performance liquid chromatography(Pre-HPLC). A total of 54 mg of chlorogenic acid, 150 mg of pedaliin-6″- acetate and 200 mg of rosmarnic acid were obtained from ethyl acetate extract with purities of 96.9%, 97.9% and 95.1% by HPLC analysis, respectively. Their structures were identified by nuclear magnetic resonance spectrum. The result show that this method is suitable for the isolation and purification of chlorogenic acid, pedaliin-6″-acetate and rosmarnic acid from D. tanguticum. It also avoids the disadvantages of traditional separation methods such as many operations, large reagent consumption and non-recyclability, provides a reference basis for the study of separating of active components and preparation reference substance from D.tanguticum.

    Optimization on Extraction of Total Phenols by Response Surface from Saxifraga stolonifera and Its Correlation in Inhibiting the Activity of PLA2 in Deinagkistrodon acutus Venom
    Tongtong Sun, Zhongjuan Wang, Xiujuan Zhang, Shihan Yang, Xiaoqiang Chen, Ying Zhang
    2022, 42(2):  259-267.  doi:10.7525/j.issn.1673-5102.2022.02.011
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    To reveal the relationship between total phenols(TP) of Saxifraga stolonifera and inhibition of the activity of phospholipase A2(PIR) in Deinagkistrodon acutus venom, we used the single factor and Box-Behnken methods to optimize the extraction conditions of TP of Saxifraga stolonifera and PIR. The effect of various experimental factors on the yield of TP and PIR in the extraction of S. stolonifera by ultrasonic assisted extraction were investigated respectively, and the correlation between the content of TP and PIR were analyzed. The results showed that when the particle size was 60 mesh, the material to liquid ratio was 1∶50, the ultrasonic power was 250 W, and the ultrasonic time was 1.8 h, the content of TP in the aqueous extract of S. stolonifera and its inhibitory rate(PIR) of the PLA2 activity in D. acutus venom could reach the optimal ultrasonic-assisted extraction process conditions. The content of TP was (86.90±0.46) mg·g-1, and the PIR was(40.91±0.21)%. The results of correlation analysis showed that the content of TP and the PIR were extremely significant correlation(P<0.01), indicating that the TP might be the material basis of S. stolonifera to inhibit the PLA2 activity in D. acutus venom .

    Physiology and Ecology
    Development-specific Investigation of Leaf Primary Metabolism of Acanthopanax senticosus and Acanthopanax sessiliflorus
    Ying Song, Kexin Wu, Wenda Shao, Yuli Liu, Jia Liu, Yang Liu, Zhonghua Tang
    2022, 42(2):  268-277.  doi:10.7525/j.issn.1673-5102.2022.02.012
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    Metabolomics analysis of Acanthopanax senticosus(Rupr. et Maxim.)Harms and Acanthopanax sessiliflorus(Rupr. et Maxim.)Seem leaves was performed using GC-MS technology. With analysis of PCA and OPLS-DA, the leaves of A. senticosus and A. sessiliflorus were divided into three developmental periods: growth period, exuberant period, and autumn period. A total of 53 differential compounds such as sugars, amino acids, organic acids, fatty acids and polyols were filtered from the leaves of A. senticosus at three periods. A total of 51 differential compounds such as sugars, amino acids, organic acids, fatty acids and polyols were filtered from the leaves of A. sessiliflorus at three periods. Further analysis showed that the leaves of A. senticosus and A. sessiliflorus had the most differential compounds in the exuberant period and autumn period. Among these differential compounds, the leaves of A. senticosus mainly included sugars and organic acids. The leaves of A. sessiliflorus also included fatty acids and polyamine compounds except for sugars and organic acids. The results initially reveal the accumulation patterns of primary metabolites in the two medicinal plants from the same genus at different periods, and provid a theoretical basis for the leaf utilization of the two plants.

    Regulation Mechanism of Endogenous Hormones in Adventitious Roots Formation of Platycladus orientalis ‘Beverleyensis’
    Guobin Liu, Ting Liao, Ye Wang, Liqin Guo, Jinzhe Zhao, Yanwu Yao, Jun Cao
    2022, 42(2):  278-288.  doi:10.7525/j.issn.1673-5102.2022.02.013
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    Platycladus orientalis ‘Beverleyensis’ is an important ornamental tree species. Plant endogenous hormones(i.e., indole-3-acetic acid(IAA), zeatin(ZT), abscisic acid(ABA) and jasmonic(JA)) play an important role in regulating the adventitious roots regeneration process, but the dynamic changes of endogenous hormones at different developmental stages and their effects on adventitious roots are still unclear. Using semi-woody branches as materials, the dynamic changes of four endogenous hormones were detected by high performance liquid chromatography- mass spectrum(LC-MS). The results showed that the root primordium originated from callus, medullary ray, xylem, vascular cambium, secondary phloem, cortex, and the junction of medullary ray and cambium respectively, which belonged to multiple locus occurrence mode and the multiple rooting type. Hormonal dynamics results showed that the content of IAA and ZT decreased, ABA and JA increased with the formation of callus; the content of IAA and ZT increased slowly, ABA and JA decreased with the differentiation of root primordium; IAA, ZT, JA gradually increased, and ABA maintained at a low level during adventitious root formation and elongation. Hormone balance analysis results showed that the IAA/ABA ratio and IAA/JA ratio both decreased, and the increasing of IAA/ZT ratio was beneficial to the formation of callus, and on the contrary, it was beneficial to the induction and differentiation of root primordia, while the IAA/ABA ratio increased, IAA/ZT and IAA/ JA maintained at a low level was conducive to the formation and elongation of adventitious roots. The results provide a basis for revealing the regulation effect of different endogenous hormones on the development of adventitious roots of P. orientalis ‘Beverleyensis’.

    Dynamic Changes of Total Carbon Release During Mixed Decomposition of Leaf Litter of Pinus massoniana and Native Broad-leaved Tree Species
    Xun Li, Yan Zhang, Simeng Song, Yang Zhou, Jian Zhang
    2022, 42(2):  309-320.  doi:10.7525/j.issn.1673-5102.2022.02.016
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    In order to improve the structure of low-efficiency forest in hilly, the total carbon(C) release of Pinus massoniana and native broad-leaved tree species in the process of mixed decomposition of litter was investigated. The litters of Pinus massonianaSassafras tzumuCinnamomum camphora and Toona sinensis that widely distributed in South Chinawere divided into 35 treatments according to different tree species combination and mixed proportion, and then field decomposition experiments were carried out to explore the best combination and mixing ratio of C release. The results showed that the C release of leaf litter of T. sinensis was the fastest, followed by S. tzumu and C. camphora litter, and the slowest in P. massoniana litter among the four tree species, In 31 mixed litter leaves treatments, the non-additive effect of C release increased at first and then decreased with the extension of decomposition time. Compared with other seasons, the non-additive effect of C release rate decreased in autumn. Among the combinations of P. massoniana with a broad tree, the synergistic effect of PC73 and PC64 was stronger in the treatment with broadleaf accounting for more than 30% in the combination of P. massoniana and C. camphora leaf litter; Among the combination of P. massoniana with two or three broad trees, the treatment with broad leaf accounting for more than 30% and containing T. sinensis litter: PST613 and 712, PCT631 and 613, PSCT7111 and 6121 showed stronger synergistic effect.