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    20 January 2022, Volume 42 Issue 1
    New findings of the Genus Lonicera L. in Zhejiang Province
    Zhenghai Chen, Wenyuan Xie, Yifei Lu, Shenglong Liu, Xiaofeng Jin
    2022, 42(1):  1-11.  doi:10.7525/j.issn.1673-5102.2022.01.001
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    Some new discoveries in the taxonomy of the genus Lonicera of Caprifoliaceae in Zhejiang Province were reported. The distribution of two species, L. similis in Zhejiang and L. macrantha in China were respectively excluded. L. guillonii was considerably reinstalled as a species with a new synonym L. macrantha var. heterotricha. Furthermore, L. guillonii var. macranthoides was newly combined. A new species L. sinomacrantha and a new form L. hypoglauca f. pulchra were described. The key to the complex of L. macrantha in Zhejiang was compiled.

    Research Report
    Flower Bud Differentiation of Hippeastrum hybridum ‘Merry Christmas’
    Xin Li, Liuyan Yang, Zhen Wang, Yi Sun, Junxu Xu, Yongchun Zhang
    2022, 42(1):  12-20.  doi:10.7525/j.issn.1673-5102.2022.01.002
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    In order to clarify the characteristics of flower bud differentiation of Hippeastrum hybridum and provide theoretical basis for the research of flower development, flowering regulation, cross breeding and systematic classification, anatomical observation and paraffin sectioning techniques were used to study the growth of flower buds, flower organ differentiation, and sex cell differentiation in the H. hybridum variety of ‘Merry Christmas’. The main results were as follows:Each year H. hybridum ‘Merry Christmas’ produced two inflorescence buds, the flower buds in which completed floral organs differentiation in the second year, then bloomed in the third year after low temperature treatment, and of which the second inflorescence aborted occasionally. The floral organ differentiation process included flower primordium differentiation stage, outer perianth primordium differentiation stage, inner perianth primordium differentiation stage, stamen primordium differentiation stage and carpel primordium differentiation stage, the corresponding length of flower buds were about 0.02, 0.05, 0.1, 0.2, 0.3 cm, respectively. The primordia of all floral organs initiated spirally and centripetally. The anthers of H. hybridum were tetrasporangiate, and the anther wall was composed of epidermis, endothecium, middle layer, tapetum from outside to inside. The tapetum belonged to secretory type. The cytokinesis in the microsporocyte meiosis was successive. The microspore tetrads were cruciform and the mature pollen grains were 2-celled. The gynoecium of H. hybridum was tricarpellary and the ovary was inferior with axile placentation and trilocular with two rows of anatropous ovules in each locule. The ovule was bitegmic and crassinucellate. The development of embryo sac was of an Allium type.

    Effects of Carbon Source and Other Factors on the Proliferation of Embryogenic Callus of Elite Clone of Pinus elliottii
    Qiyang Gao, Yulong Huang, Wenbing Guo, Fencheng Zhao, Yang Liu
    2022, 42(1):  21-28.  doi:10.7525/j.issn.1673-5102.2022.01.003
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    To explore the effects of carbon sources and other main factors on the proliferation of embryogenic callus in Pinus elliottii, and provide technical methods and theoretical basis for further improving proliferation efficiency. The embryogenic callus of the elite clone PEE3-13 was used, the effects of different types and concentrations of carbon sources, pH and the organic nitrogen sources, L-glutamine and its substitutes, L-alanine-L-glutamine on its proliferation were studied. The pH and the type of carbon source had significant effects on its proliferation. The autoclaved medium pH ranged from 5.68 to 6.28, it was the most suitable for the embryogenic callus growth. The maximum proliferation rate was achieved when using 30 g·L-1 white sugar was used as carbon source. When 450 mg·L-1 L-glutamine was added, the embryogenic callus had the highest proliferation rate and the best activity, however, L-alanine-L-glutamine could not replace L-glutamine. The optimal medium was 30 g·L-1 white sugar and 450 mg·L-1 L-glutamine and autoclaved pH was ranged from 5.68 to 6.28 and was most suitable for embryogenic callus proliferation, and the maximum proliferation rate was h 851.27%.

    Identification and Expression Analysis of Expansin Genes in Moso Bamboo (Phyllostachys edulis)
    Shuang Ma, Boya Wang, Ying Cao, Shanglian Hu, Zhimin Gao
    2022, 42(1):  29-38.  doi:10.7525/j.issn.1673-5102.2022.01.004
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    In order to fully understand the molecular characteristics and expression patterns of expansin genes in moso bamboo(Phyllostachys edulis), bioinformatics methods were used in this study,and a total of 43 members of the expansin gene family was identified from the genome of moso bamboo, and the members were divided into four subfamilies(EXPA, EXPB, EXLA and EXLB), which contained 18, 17, 7 and 1 members respectively, and distributed on 37 Scaffolds. Except that PeEXPA1 had no introns and PeEXLB1 had 11 introns, other expansin genes have 1 to 5 introns respectively. The amino acid length of the expansin was 91~508 aa respectively, all amino acids had high-frequency codons, and most of the expansins were basic and hydrophilic. Protein structure analysis showed that most of the expansins of moso bamboo accounted for the smallest proportion of β-turns and the largest proportion of β-sheets in the secondary structure. The tertiary structure of most members in each subfamily was similar. The qRT-PCR results of 18 EXPA subfamily members showed that their expression were significantly different in different tissues, and the highest expression values were found in leaves except PeEXPA2 and PeEXPA6, indicating that they might play an important role in the growth of leaves.

    Cloning and Disease Resistance Response Potential of Walnut JrNPR1 Gene
    Kaiheng Ma, Jianing He, Muhong Xie, Bin Ren, Qianxue Huang, Guiyan Yang
    2022, 42(1):  39-46.  doi:10.7525/j.issn.1673-5102.2022.01.005
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    Nonexpressor of pathogenesis-related gene 1(NPR1) played a key role in regulating plant disease resistance. Walnut(Juglans regia) was an important economic oil tree for rural revitalization in China, and its growth and yield were severely restricted by pests and diseases. In order to explore the physiological mechanism of walnut disease resistance and screen disease resistance genes, the JrNPR1 gene was cloned from ‘Xiangling’ walnut, and its basic biological information and disease response were analyzed to predict the function of JrNPR1. The results showed that the ORF of JrNPR1 gene was 1 782 bp, encoded 593 amino acids, and the theoretical isoelectric point was 6.40. Multi-sequence comparison with Morella rubraRicinus communis and other homologous proteins showed that they all had NPR1-like-C conserved domains, and JrNPR1 had a relatively close evolution with M. rubra and Quercus suber. The upstream 1 233 bp promoter contained a variety of cis-acting elements related to plant disease resistance, such as WRKY71OS. Under treatment with pathogens such as Colletotrichum gloeosporioidesMelanconium oblongum, and Xanthomonas campestrisJrNPR1 was significantly induced, and its expression was up-regulated by salicylic acid(SA). The expression level of JrNPR1 under CgTJ+SA, MoZK+SA, XcHB+SA was 1.31~178.89 fold of that under single stress of CgTJ, MoZK, XcHB, respectively. These results suggested that JrNPR1 might be an important disease resistance gene against walnut anthracnose, branch blight and bacterial black spot, and might be involved in the SA signal pathway.

    Cloning and Bioinformatics Analysis of CbuATX1,CbuATX1-like and CbuATX2 Genes from Catalpa bungei
    Jiaming Zhao, Erqin Fan, Yi Liu, Zhi Wang, Junhui Wang, Guanzheng Qu
    2022, 42(1):  47-61.  doi:10.7525/j.issn.1673-5102.2022.01.006
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    Flowering is a complex life process that is regulated by both genetic factors and environmental factors in higher plants. It is a symbol of plants genetic and reproductive ability after it enters into reproductive growth. The genes of CbuATX1CbuATX1-like and CbuATX2 were cloned respectively by PCR with cDNA as a template from Catalpa bungei flower bud, and the related bioinformatics software was used to predict the protein structure and subcellular location of these three genes respectively. The three genes promoter sequence were analyzed in cis-acting elements, and the expression levels of the three genes in different stages of C. bungei flower bud development were detected by qRT-PCR. The results showed that the CDS of CbuATX1 was 726 bp, encoding 241 amino acids with transmembrane transport structure. The protein was belonged to the HMA protein family and closely related to Sesamum indicum and Beta vulgaris. The CDS of CbuATX1-like was 801 bp. encoding 266 amino acids with transmembrane transport structure. The protein was belonged to the HMA protein family, which was closely related to Daucus carota and Olea europaea. The CDS of CbuATX2 was 1 554 bp, encoding 517 amino acids with no transmembrane transport structure. The protein was belonged to the PWWP protein family and closely related to Erythranthe guttatus. The promoters of three genes contained several basic elements of eukaryotic promoters respectively, such as CAAT box and TATA box. In addition, they also contained light response factors, low temperature response factors and biological activity elements. The results of qRT-PCR showed that the expression levels of the three genes were significantly different in different developmental stages. It was hoped to further elaborate the flowering characteristics of C. bungei, and provided theoretical support for the flowering mechanism and directional genetic improvement of C. bungei.

    Regulation of Soil Salinity by Vegetation Coverage in Urban Greenbelt Saline-alkali Land
    Xiaoli Peng, Wangze Wu, Juan Shen, Hongbin Cheng, Jiangtao He
    2022, 42(1):  62-70.  doi:10.7525/j.issn.1673-5102.2022.01.007
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    Soil salinization was a global problem, which not only seriously affected agricultural production, and also greatly threatened landscape and urban planning projects. In present study, different landscape combinations and fertility measures were used to investigate the adjustable mechanism of the vegetation coverage to the urban green belt saline-alkali soil in the Lanzhou New District. The results showed that “Aesculus chinensis Bunge + Catalpa bungei C. + Rosa rugosa Thunb.” landscape combination Ⅱ, treated with organic fertilizer and microbial inoculants,the average vegetation coverage reached to 70.06% during a growth season. And the topsoil desalination rate could reach to 53.51%, while the "bare land" was only 20.62% and 5.46% respectively. The pH value and electrical conductivity of the landscape combination Ⅱ decreased by 12.54% and 63.83%, respectively, while the “bare land” only decreased by 3.14% and 12.28%. Correlation analysis(P<0.05) shown that vegetation coverage was highly negatively correlated with soil salinity(r= -0.949), but highly positively correlated with soil organic matter(r=0.887) and with total nitrogen(r=0.853) respectively. These results indicated that can effectively improve the salt content of saline-alkali soil in the Lanzhou New District, by increasing the vegetation coverage of the green belt and improving soil fertility.

    Plant Leaf Traits in Minqin Oasis-desert Transition Zone
    Fei Wang, Shujiang Guo, Baoli Fan, Fugui Han, Fanglin Wang, Weixing Zhang, Yunian Zhang
    2022, 42(1):  71-80.  doi:10.7525/j.issn.1673-5102.2022.01.008
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    To explore the adaptability of plants to the environment and provide a theoretical basis for the restoration and reconstruction of vegetation in Minqin oasis-desert transition zone, 20 kinds of main plant were used, 6 functional characters of leaves were determined, variation characteristics and correlation of leaf traits were analyzed, and the difference of leaf traits of different functional groups were compared respectively. The results showed that: ①There were interspecific differences and different degrees of variation in leaf traits, the variation range from 14.11% to 47.63%, the variation coefficient of ChlC was the largest, while that of LCC was the smallest. ②LDMC with ChlC or LCC showed extremely significant positive correlation(P<0.01), ChlC with LCC showed extremely significant positive correlation(P<0.01), while the correlation between other leaf traits was not significant. ③There were significant differences in leaf traits among different functional groups. LDMC, ChlC and LCC in legumes were significantly higher than those in chenopodiaceae plants(P<0.01). The SLA of herb leaves was significantly higher than that of shrub plants(P<0.01). SLA and LNC of annual plants were significantly higher than those of perennial plants(P<0.05). The LDMC of perennial leaves was significantly higher than annual leaves(P<0.01). The content of C in leaves of perennial plants was significantly higher than that of annual plants(P<0.05). LDMC and ChlC of C3 plants were significantly higher than that of C4 plants(P<0.05). The δ13C of C4 plants was significantly higher than that of C3 plants(P<0.01). LCC content in leaves of C3 plants was significantly higher than that of C4 plants(P<0.05).

    Allelopathy and Its Mechanism of Three Natural Diterpenes
    Zhaowei Huan, Qiaoming Ou, Lan Ding
    2022, 42(1):  81-92.  doi:10.7525/j.issn.1673-5102.2022.01.009
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    To evaluate the allelopathic potential of three diterpenes, including wangzaozin A, leukamenin E and weisiensin B, Lolium perenne L. and Lacutuca sativa L. were taken as materials and the effects of three ent-kaurene diterpenes on the seed germination, growth, chlorophyll, membrane damage of tested plants were studied respectively. It was found that three diterpenes with different concentrations had a slightly decrease on germination of L. perenne L. and L. sativa L. seeds, but the germination time was delayed obviously; three diterpenes at higher concentrations had significant negative effects on the root length, seedling length, fresh weight and dry weight of plants respectively, and it showed clearly concentration-dependent trend; meanwhile, the chlorophyll a and chlorophyll b contents were significantly reduced but the malondialdehye(MDA) content and relative electricity conductivity were increased, however, the relative water content showed downward trends. The results showed that three diterpenoids inhibited the growth of tested plants by affecting its photosynthetic capacity, causing oxidative damage to the cell membrane and reducing the water retention of the leaves.

    Genetic Transformation of BpERF98 Gene and Abiotic Stress Response of Transgenic Plant in Betula platyphylla
    Qi Li, Siyu Yan, Su Chen
    2022, 42(1):  93-103.  doi:10.7525/j.issn.1673-5102.2022.01.010
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    Abiotic stresses seriously affected plant growth and development, and plants had evolved sophisticated internal molecular regulatory mechanisms to deal with, in which transcription factors played a vital role. In this study, BpERF98 gene was cloned from the leaves of Betula platyphylla and overexpressed using Agrobacterium-mediated. The physiological indicators of the transgenic lines and wild-type under low temperature, freezing damage and salt stress were measured respectively, the results showed that the malondialdehyde content and relative conductivity of the transgenic lines under abiotic stress were lower, while the SOD and POD enzyme activities were higher than those of the wild-type. The results showed that overexpression of BpERF98 gene could improve the tolerance of white birch to abiotic stress, and provided an important theoretical basis for studying the molecular mechanism under abiotic stress and molecular breeding for tolerance of white birch.

    Tissue-specific Expression and Analysis of Exogenous Hormone Response of BpPIN5 Gene Promoter in Betula platyphylla
    Yunli Yang, Chang Qu, Yang Wang, Guifeng Liu, Jing Jiang
    2022, 42(1):  104-111.  doi:10.7525/j.issn.1673-5102.2022.01.011
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    Being the polar carrier of IAA, PIN family proteins played a key role in plant embryonic development, organ development and tropic growth, especially in the formation of plant phyllodes, veins and vascular tissue differentiation.In order to identify the response characteristics BpPIN5 gene to exogenous hormones, the upstream 1447 bp sequence of BpPIN5 was cloned from the genome DNA of Betula platyphylla. The cis-acting elements in the sequence were predicted by PLACE online software. The results showed that the promoter sequence of BpPIN5 contained different types of auxin responsive elements such as auxin, gibberellin, Salicylic acid, abscisic acid, methyl jasmonate, ethylene respectively. Transgenic vector pro-BpPIN5::GUS to B. platyphylla was constructed. GUS histochemical staining analysis reflicted that BpPIN5 promoter had transcriptional activity in the tip of leaf crack, venule and root of B. platyphylla respectively. After transgenic Betula platyphylla was treated with IAA, GA, MeJA, SA and ABA respectively, the results showed that BpPIN5 promoter responded to the five hormones above at the crack edge of the first leaf, petiole and root tissue of the second leaf, and the response changes were basically the same. The results would help to clarify the function of BpPIN5 in B. platyphylla.

    Cloning and Expression Analysis of Transcription Factor LobHLH34 from Larix olgensis
    Yuning Yang, Hao Dong, Shiwei Dong, Nairui Wang, Yue Song, Hanguo Zhang, Shujuan Li
    2022, 42(1):  112-120.  doi:10.7525/j.issn.1673-5102.2022.01.012
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    In order to understand the function of transcription factor LobHLH34 and explore its expression pattern in different tissues and under different stress conditions, the full-length sequence of LobHLH34 gene was obtained from transcriptome data of root, stem and leaf of Larix olgensis, by using the primers designed. The complete open reading frame(ORF) of LobHLH34 gene was 696 bp and encoded 231 amino acids. Subcellular expression vector was constructed and transformed into protoplasts of Populus trichocarpa. The result showed that the LobHLH34 gene was located in the cell nucleus under the laser confocal microscope. The results of phylogenetic tree analysis showed that larch had the closest genetic relationship with Picea asperata and Selaginella tamariscina. By qRT-PCR, the tissue-specific expression of LobHLH34 gene and its response to abiotic stress were analyzed respectively. The results showed that LobHLH34 gene was expressed in the roots, stems and leave, and the lowest expression level was found in the stem and the highest relative expression in the leaf respectively. The expression level of LobHLH34 was different in different organs under NaCl, PEG and ABA stress. It was speculated that LobHLH34 gene could be involved in the process of plant growth, development and respond to stress, and it had special expressional pattern in different organs.

    Effects of Three Kinds of Agrobacterium and Different Transformation Conditions on the Transient Expression of GFP in Nicotiana benthamiana
    Yuejing Zhang, Ying Li, Juanjuan Wang, Hailong Pang, Lingyun Jia, Hanqing Feng
    2022, 42(1):  121-129.  doi:10.7525/j.issn.1673-5102.2022.01.013
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    By using Nicotiana benthamiana as plant material, the influence of the different Agrobacterium strains(LBA4404, EHA105, GV3101), bacterial concentration and infection time on GFP(as the reporter gene) the fluorescence transient expression after transformation were analyzed respectively. The results showed that the expression of GFP with different Agrobacterium strains were various in the optimal concentration and time for transient: the highest transient expression efficiency with LBA4404 strain was achieved when the OD600 of bacterial suspension was 0.8; while the highest transient expression efficiency with EHA105 and GV3101 strains was achieved when the OD600 of the bacterial suspension reached 0.6. The level of transient expression by strain LBA4404 was the highest at 2 days later Agrobacterium infiltration while the level of transient expression by strains EHA105 and GV3101 was the highest at 4 days later Agrobacterium infiltration. The comparative analysis among these different strains showed that the transient expression efficiency by using LBA4404 strain was the highest. The above results indicated that Agrobacterium strains and transformation conditions such as bacterial concentration and infection time were important factors that affected the transient expression efficiency.

    Variation Analysis of Growth Traits of Larix olgensis Parental Clones in Seed Orchards
    Dandan Zhang, Xiang Li, Biying Wang, Xihe Wang, Quan Sun, Yunyang Wu, Pingyang Li, Deyao Li, Yulei Li, Xiyang Zhao
    2022, 42(1):  130-137.  doi:10.7525/j.issn.1673-5102.2022.01.014
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    The growth of parental clones was neglected in seed orchards though many Larix olgensis seed orchards had been built for several years in Northeast China. In this research, 49 L. olgensis parental clones were used as materials, many growth traits were investigated and analyzed respectively. ANOVA results showed that there was significant difference among 49 clones in each growth traits. The phenotypic variation and genetic variation coefficients of different traits ranged from 7.71%-31.93% and 2.05%-12.55%, respectively. The repeatability of different traits ranged from 0.44-0.76, high repeatability and variation coefficients were beneficial for selection of elite clones. The correlation of most growth traits including tree height was extremely significant showed that each growth traits affected each other. The results of principal component analysis showed that the contribution rate of the first principal component was 61.44%, which mainly represented biomass traits. Comprehensive evaluation of L. olgensis parental clones were conducted based on the results of principal component analysis, clone L76, L5, L7, L12 and L4 were selected as elite clones, the genetic gain of different growth traits ranged from 3.55%-15.50%, the selected clones could provide materials for tree breeding of L. olgensis.

    The Stoichiometric Characteristics of Desert Ephemeral Plants in Different Growth Periods and Its Association with Soil Factors
    Mingming Wang, Weiwei Zhuang
    2022, 42(1):  138-150.  doi:10.7525/j.issn.1673-5102.2022.01.015
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    To explore the stoichiometric characteristics of desert ephemeral plants, and to better understand the relationship between habitat soil factors and plant survival strategies, 4 widely distributed plant species (Erodium oxyrrhynchum, Nonea caspica, Hyalea pulchella, Lactuca undulata) were collected, and soil physical and chemical properties were measured at different depths (0-5, 5-10, 10-15 cm), and the dynamic change and coupling relationship between stoichiometric characteristics of 4 plants and soil factors were compared and analyzed through in-situ multi-period sampling in the field. The results showed that: ①The carbon(C), nitrogen(N), and phosphorus(P) contents of the 4 ephemeral plants were generally low, and there were differences in stoichiometric characteristics among species, but the changes were roughly similar in different growth periods. The content of N and P gradually decreased from the seedling stage to the fruit setting stage, while the C content tended to be stable for a long time. During the whole growth period, the variation range of P and C∶P were larger, the variation range of C and N∶P were smaller. The differences in the stoichiometric characteristics of the 4 plants in different growth periods were significantly related to the growth period and plant species. ②The content of soil organic carbon(SOC), total nitrogen(TN) and total phosphorus (TP) in the 0-5 cm soil layer were the highest respectively, and gradually decreased with the depth of the soil layer. With the growth of plants, the content of SOC and TN in the 0-5 cm soil showed an obvious increasing trend, while the content of TP showed a parabolic change trend. During different growth periods of plants, the stability of soil TP content was the strongest, and the variability of SOC and TN was strong respectively. The lower N content and TN∶TP ratio showed that the soil in the area belonged to the type of N deficiency. The differences in the stoichiometric characteristics of the 3 soil layers in different growth periods were significantly affected by the soil layers and growth periods. ③There was no consistent law in the correlation of stoichiometric characteristics between plants and soils at each level. Plant stoichiometric indicators were only strongly correlated with SOC and SOC∶TP in the 0-10 cm soil, and most of the stoichiometric characteristics did not show correlation, indicating that the stoichiometric characteristics of plants were not directly determined by the characteristics of soil nutrient content. These results indicated that the stoichiometric characteristics of plants were not directly determined by soil nutrient characteristics, and the obvious interspecific differences indicated the importance of plant genetic characteristics in the coupling relationship of soil-plant stoichiometric characteristics.

    Effects of Climate Change on the Suitable Area and Niche of Cunninghamia lanceolata
    Xinggang Tang, Yingdan Yuan, Jinchi Zhang
    2022, 42(1):  151-160.  doi:10.7525/j.issn.1673-5102.2022.01.016
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    Cunninghamia lanceolata is the main afforestation tree species in China, which has important functions of medicine, economy and ecological service. With the trend of continued global warming, climate has become an important factor restricting the survival and development of species, among which spatial distribution and niche changes are crucial to the interpretation of ecological processes. To analyze the response of C. lanceolata to future climate warming, 211 C. lanceolata distribution points and 20 environmental variables were taken, MaxEnt model and ecospat R software package were used as methods. The results showed that the current potential suitable area of C. lanceolata was 2.196 7 million km2, about 22.88% of the total land area, mainly distributed in the east of 800 mm equivalent precipitation line. Mean annual temperature, diurnal mean temperature difference and driest monthly rainfall were the main environmental factors affecting the distribution of C. lanceolata. The suitable area of C. lanceolata would migrate along the latitude and longitude directions, and the area of highly and moderately suitable habitats continue to decrease. Principal component analysis(PCA) showed that the climatic niches of C. lanceolata shifted and expanded under different representative concentration paths in different periods, and the centers of the climatic niches would move to the average annual temperature and the warmest seasonal rainfall. The niche overlap index would show a trend of continuous decline, and the niche overlap rate of RCP8.5 show the most obvious decline. The results showed that global warming would change the spatial distribution of species, and affect the existing ecosystem to varying degrees. The research on the relationship between C. lanceolata and climate change expanded people’s understanding of climate change and the ecological characteristics of plant species, and provided a theoretical basis for the protection and utilization of C. lanceolata and even arbors species.