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    Bulletin of Botanical Research 2021 Vol.41
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    Morphological Structure of Vessel Elements of Prunes L. from Northeast Area of China
    Zi-Yu ZOU, Li-Wei GU, Da-Wei ZHANG
    Bulletin of Botanical Research    2021, 41 (1): 4-12.   DOI: 10.7525/j.issn.1673-5102.2021.01.002
    Abstract439)   HTML227)    PDF(pc) (1834KB)(55)       Save

    The scanning electron microscopy(SEM) and resin casting method were used to observe the micromorphological characteristics of vessel molecular types, pits and perforation plates of ten species of Prunus L. in Northeast of China, and the quantitative data of lumen length, width, tail length and end wall angle were measured. The conduit type, tail length and perforation plate type of the same species were stable in different habitats, and did not change with the change of habitats. There were three types of ducts in ten species of Prunus L., including spiral, pitted and reticulate. In addition to P.ussuriensis Kov. et Kost., P.humilis(Bge.) Sok. and P.sibirica(L.)Lam., there are only two types of vessels, including pitted and reticulate. Three types are common in the other seven plants. Helical thickening is common in this genus of conduit molecules. The single perforation plate is common in the plum plants, only in P.maximowiczii(Rupr.) Kom., P.tomentosa(Thunb.) Wall. and P.salicina Lindl.. Only in P.racemosa(Lam.)Gilib. and P.ussuriensis Kov. et Kost. and P.japonica(Thunb.) Lois. and P.sibirica(L.)Lam., the relatively primitive opposite-alternate pitting was observed, and the other six patterns were all alternate pitting. The lumen length, width and end wall area were significantly related to the habitat. The measurement of the tail length shows that the tail length of the vessel elements of the same plant is relatively stable in different habitats, almost unchanged, which can be used in micro plant taxonomy.

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    Pollinia Morphology of 14 Species in Dendrobium(Orchidaceae) and Taxonomic Significance
    Yan-Ping WANG, Lu LI, Chen-Xuan YANG, Yan LUO, Qing-Qing LI
    Bulletin of Botanical Research    2021, 41 (1): 12-25.   DOI: 10.7525/j.issn.1673-5102.2021.01.003
    Abstract633)   HTML42)    PDF(pc) (9877KB)(87)       Save

    The morphology and exine sculpture of pollinia in 14 species of Dendrobium Sw.(Orchidaceae) were investigated using stereoscope and scanning electron microscope in order to provide evidence for classification and systematic evolution of this genus. Under stereomicroscope, the anther is composed of 4 clavated pollinia with a golden, oily, waxy surface and a hard texture in 14 Dendrobium species. According to the general appearance, Dendrobium pollinia observed can be divided into 3 shapes(heart-shaped, nearly heart-shaped and long heart-shaped) and 4 types(extremely small pollinia, small pollinia, large pollinia and oversize pollinia). Based on the curvature of the boundary contour of a pollinium, it can be divided into four types(crescent-like, bow-like, stick-like and rice grain-like). There are 5 kinds of exine sculptures(psilate; psilate with threads; psilate-uneven,coarse reticulate and rugulate) under scanning electron microscope, as well as two types of crystals observed on the surface of pollinia in D.wislsonii Rolfe and D.thyrsiflorum Rchb. The morphology and exine sculpture of pollinia were distinguished in different species of Dendrobium, which is of some significance to its taxonomic clarification.

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    Response of Flowing Time to Temperature Changein Heilongjiang Province
    Xin-Yu CHENG, Zhi-Yuan YAN, Meng-Qiao GUO, Hai-Jun XU, Bin TONG, Hong-Wei NI
    Bulletin of Botanical Research    2021, 41 (1): 26-36.   DOI: 10.7525/j.issn.1673-5102.2021.01.004
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    In order to clarify the responding tendency of florescence to temperature change in Heilongjiang Province, the relationship between plant florescence change and the annual mean temperature change in spring, summer, autumn in the present year and the last year were analyzed according to 188 herbarium plant species(contain infra-species taxa), which were collected from Heilongjiang Province in period of 1951-2007. The results showed that was a significant correlation(P<0.05) between florescence of more than 42% plants and temperature in Heilongjiang Province. In these plants, the relative collection time index of more than 25% species had significant positive correlation with annual mean temperatures of current, last year and season(P<0.05), and most of these were herbs. The relative collection time index of the rest of herbs and woody plants had significant negative correlation with annual mean temperatures of current, last year and season(P<0.05), for example, when the annual average temperature in spring increased to 1℃, the flowering period of woody plants precocious flowered 12.65% on average. Plant florescence was 4.47%, 7.08%, 3.87%, 5.84% and 6.73% ahead of schedule respectively in Heilongjiang Province if the annual average temperature increased to 1℃ in spring, summer, autumn of the current year and last year. The florescence response to temperature changes was relatively obvious in short florescence plants. The florescence response to the change of the annual mean temperature in summer was more sensitive than that in spring, autumn of the current year in Heilongjiang Province.

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    Flower Bud Differentiation and Endogenous Hormone Changes of Rosa ‘Angela’
    Zhi-Yuan LIU, Li ZENG, Xi-Wu DU, PENG-Yong-Zheng, Yi-Wei TAO, Yu-Qing LI, Jun QIN
    Bulletin of Botanical Research    2021, 41 (1): 37-43.   DOI: 10.7525/j.issn.1673-5102.2021.01.005
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    The experiment was conducted to explore the relationship between structure of flower buds and hormones content in Rosa ‘Angela’, and to regulate flowering, to improve landscape quality and breeding work. The flower bud differentiation was divided into five periods within 30 d. The growth cone was conical before the differentiation period. Then the cone became flat, five protrusions appeared around the growth cone at the sepal differentiation period. Multiple petal primordiums and stamen primordiums were respectively appeared around the growth cone during the flower bud differentiation. Several pistil primordiums were appeared at the base of growth cone at pistil differentiation period. The contents of ABA and CTK increased first and then decreased, which contents at sepal primordium differentiation period were significantly higher than other periods. The content of IAA increased gradually trend, and the content of GA decreased gradually. The ratio of IAA/GA and IAA/ABA showed a gradual increasing trend. The ratio of CTK/GA and(ABA+CTK)/GA at sepal primordium differentiation period were significantly higher than that before the differentiation period. The significant increasing of ABA and CTK contents at the sepal primordium differentiation period were related to flower bud induction. Low level of GA and decreasing of IAA were beneficial for flower bud differentiation. The increasing of CTK/GA and(ABA+CTK)/GA ratio at sepal primordium differentiation period were related to the flower bud induction, and high levels of IAA/ABA and IAA/GA ratio may relate to the further development of floral organ primordium.

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    Physiological Responses to Drought Stress of Five Speciesfrom Catalpa Scop
    Xiao-Chi YU, Gui-Juan YANG, Ju-Lan DONG, Jun-Hui WANG, Wen-Jun MA, Peng ZHANG
    Bulletin of Botanical Research    2021, 41 (1): 44-52.   DOI: 10.7525/j.issn.1673-5102.2021.01.006
    Abstract375)   HTML29)    PDF(pc) (1182KB)(96)       Save

    In order to analyze the response to drought stress of the five species of Catalpa Scop., to provide theoretical basis for the selection on drought-resistant breeding and cultivation of drought-resistant tree species of Catalpa Scop., the C.speciosaC.fargesii f.duclouxiiC.fargesiiC.ovata, and C.bungei were selected as materials, and cultivated for 50 days under the water conditions of 700 and 400 mL respectively, and seedling height, diameter, leaf water potential, relative content of chlorophyll, chlorophyll fluorescence parameters, and specific leaf weight were measured respectively. The membership function method was used to comprehensively evaluate drought resistance. The results showed that the C.bungei grew best but the C.ovata was relatively poorest under the two water conditions. Under drought stress, except for the C.speciosa, the leaf water potential(ΨL) of all other species decreased, and the maximum stomatal conductance(Gs) increased respectively. The relative chlorophyll content(SPAD) of different species increased under drought stress. The maximum light energy conversion efficiency(Fv/Fm) of photosystem Ⅱ of each species increased under drought stress, except for C.fargesii f.duclouxii and C.bungei. In addition to C.fargesii, the specific leaf weight(LMA) of other tree species increased under drought stress. The results of comprehensive drought resistance using the membership function method were gray C.fargesii>C.ovata>C.fargesii f.duclouxii>C.bungei>C.speciosa. The results showed that different species of Catalpa Scop. have different drought resistance. Comprehensive analysis showed that the drought resistance of C.bungei and C.fargesii were strong, and that of C.fargesii f.duclouxii and C.ovata were the second, and that of C.speciosa was the worst.

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    Effects of 24-Epibrassinolide on the Chlorophyll Fluorescence Transient in Leaves of Pepper under Drought Stress
    Wen-Hai HU, Xiao-Hong YAN, Xiao-Hong LI, Zao-Gui CAO
    Bulletin of Botanical Research    2021, 41 (1): 53-59.   DOI: 10.7525/j.issn.1673-5102.2021.01.007
    Abstract295)   HTML30)    PDF(pc) (1028KB)(55)       Save

    In order to investigate the role of exogenous 24-epibrassinolide(EBR) in improving drought tolerance in pepper (Capsicum annuum L. cv.Chaola 9), the chlorophyll fluorescence transient curve(OJIP) was examined in plants under drought stress(15% PEG6000). Drought reduced the photochemical efficiency and photosynthetic performance of pepper leaves, and induced photoinhibition. Drought not only disrupted the oxygen-evolving complex(OEC), but also impaired the PSⅡ reaction and acceptor side of PSⅡ, which inhibited the photosynthetic electron transportation. Drought decreased the number of active PSⅡ reaction centers per unit area. Drought also decreased the specific energy fluxes per unit area for absorption(ABS/CS), trapping(TRo/CS), electron transport(ETo/RC), but increased dissipation energy fluxes per unit area(DIo/CS). These results indicated that drought initiated defense mechanism by decreasing the absorption and transportation of light energy by reversible deactiveation of PSⅡ, and reducing the accumulation of excess excitation energy by promoting heat dissipation. EBR treatment improved the electron transport of PSⅡ acceptor side, alleviated the drought-induced decrease of active reaction center per unit area, optimized the process of photosynthetic electron transport, maintained high heat dissipation capacity in pepper leaves under drought, and alleviated drought-induced photoinhibition. In conclusion, EBR could optimize photochemical efficiency and photosynthetic performance, and protect the photosynthetic electron transport chain and PSⅡ reaction center in pepper leaves under drought stress.

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    Morphological Variations of Different Populations from Psammochloa villosa,a Peculiar Sandy Plant
    Ting LÜ, Tao LIU, Rui-Fang LIANG, Xu SU, Yu-Ping LIU
    Bulletin of Botanical Research    2021, 41 (1): 60-66.   DOI: 10.7525/j.issn.1673-5102.2021.01.008
    Abstract418)   HTML34)    PDF(pc) (1073KB)(65)       Save

    An experiment was conducted to analyze the morphological variation in terms of 12 phenotypic traits,including height of plant,length of spike,length of flag leaf,width of flag leaf,length of spikelet et al,using 20 natural populations of Psammochloa villosa. F value of 12 trait populations was 1.832?8.958,which reached a significant or extremely significant level,the phenotypic traits of different populations of P.villosa were widely varied,and the degree of variation was different. By principal component analysis,the first four principal components represented 82.277% of the morphological diversity. The PCA results indicated that the length of flag leaf,width of flag leaf,length of glume and length of spikelet were the main factors leading to morphological variations of different populations. When the genetic distance is 20.5,20 natural populations of P.villosa could be divided into 2 groups by UPGMA cluster analysis,and the phenotypic characters were not clustered according to the geographical distribution of the populations.

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    Effects of Exogenous MeJA on Antioxidant System and Stress Genes of Pinellia ternata under High Temperature Stress
    Yun-Hao ZHU, Meng-Jia ZHANG, Cheng-Ming DONG
    Bulletin of Botanical Research    2021, 41 (1): 67-73.   DOI: 10.7525/j.issn.1673-5102.2021.01.009
    Abstract310)   HTML30)    PDF(pc) (1304KB)(50)       Save

    We studied the effects of exogenous MeJA on the activities of superoxide dismutase(SOD), peroxidase(POD) and ascorbic acid peroxidase(APX) of Pinellia ternata under high temperature stress, effects on proline(Pro), malondialdehyde(MDA), soluble sugar content and active ingredient content, and the effect on the expression of stress-related genes such as small molecule heat shock protein(sHSP). The P.ternata plants with consistent growth status were subjected to stress treatment at 40℃, the experimental group was sprayed with 50 μmol/L exogenous MeJA solution, and the blank group was sprayed with the same amount of water. Samples were taken at 2, 6, 12, 24 and 48 h of treatment, respectively, and the indexes of the samples were determined. The results showed that spraying exogenous MeJA at a certain concentration at 40℃ could improve SOD, POD and APX activities of P.ternata leaves, reduce MDA content and increase proline and soluble sugar content. There was no obvious effect on the organic acid content of P.ternate. The expression levels of 2 cytoplasmic small molecule heat shock proteins and GRRBP proteins increased significantly after MeJA treatment. Therefore, spraying exogenous MeJA can partially enhance the activity of antioxidant enzymes in P. ternate, protect the cell membrane of P. ternate, and enhance the osmotic adjustment ability of cells. Moreover, exogenous MeJA may increase the expression of P. ternate high temperature stress response genes.

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    Effects of Sucrose on Growth and Physiological Characteristics in vitro Plantlets of 84K Poplar
    Sen SHI, Na MIAO, Yu-Tong SHI, Hui-Mei WANG
    Bulletin of Botanical Research    2021, 41 (1): 74-78.   DOI: 10.7525/j.issn.1673-5102.2021.01.010
    Abstract360)   HTML28)    PDF(pc) (1360KB)(65)       Save

    Carbon source has an important effect on the growth and quality in vitro plantlets. The effects of different sucrose concentrations(10,20,30 and 40 g·L-1) on the growth and physiological characteristics in vitro plantlets of 84K poplar were studied. The results showed that with the 30 g·L-1 sucrose was the most favorable for the growth in vitro plantlets of 84K poplar with the culture period increased,and the dry weight was 1.5 times that of 10 g·L-1 sucrose concentration treatment. Secondly,in the treatment of higher sucrose concentration,with the culture period increased,the in vitro plantlets of 84K poplar had higher chlorophyll content,Rubisco enzyme activity and net photosynthetic rate,of which 30 g·L-1 sucrose treatment in vitro plantlets of 84K poplar had the highest net photosynthetic rate,which was 3.617 μmol?m-2?s-1. The in vitro plantlets of 84K poplar treated with high concentration of sucrose had higher soluble sugar content,but no significant change in starch content. To sum up,sucrose concentration has an important influence on the growth and physiological characteristics in vitro plantlets of 84K poplar. Higher sucrose concentration was conducive to promoting the growth in vitro plantlets of 84K poplar.

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    Tissue Culture via Seed Embryo for Michelia macclurei
    Ying LIU
    Bulletin of Botanical Research    2021, 41 (1): 79-88.   DOI: 10.7525/j.issn.1673-5102.2021.01.011
    Abstract327)   HTML27)    PDF(pc) (1605KB)(61)       Save

    In order to explore tissue culture and micro-propagation technical system of Michelia macclurei, the seeds from its elite families were used to carry out experimental studies at aspects of explants selection and disinfection, basic medium, PVP concentration, type and concentration of rooting promoter. The results showed that:①The embryos were well sterilized with 0.1% mercury solution for 6-9 min, and the aseptic germination rate could reach 60%-65%; ②LY medium was the best basic medium, and suitable for the proliferation and growth of most embryo lines; ③Application of PVP with concentration of 4-8 g·L-1 could significantly reduce browning degree of medium and promote growth of the shoots; ④There were significant differences in rooting ability and rooting rate among tested embryo lines, the optimal concentration of both rooting promoters was 8.5 mg·L-1, and the rooting induction effect of ABT2 root powder was significantly better than that of IBA(P<0.01); ⑤The survival rate reached above 90% for rooting plantlets in 30 d, and more than 80% for plantlets with no root in 60 d after transplanted. This study provides a theoretical and technical basis for study on tissue culture and micro-propagation of M.macclurei clones and their industrial production.

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    Diameter Structural Distribution of Coniferous Broad-leaved Mixed Forest in Taxus cuspidate Habitat in Changbai Mountain
    Xiao-Hong FAN, Li-Tao WANG, Juan-Yan AN, Dong MENG, Qing YANG, Chun-Jian ZHAO, Yu-Chun YANG, Yu-Jie FU
    Bulletin of Botanical Research    2021, 41 (1): 89-97.   DOI: 10.7525/j.issn.1673-5102.2021.01.012
    Abstract471)   HTML27)    PDF(pc) (2268KB)(53)       Save

    This research aimed at the management of Taxus cuspidate stand by selecting suitable models for imitating diameter structural distribution of coniferous broad-leaved mixed forest in T.cuspidate stand, Changbai Mountain Nature Reserve. On the basis of 15 coniferous broad-leaved mixed forest in T.cuspidate stands in Longhuanggou Forest Farm, Changbai Mountain Nature Reserve, negative exponential model, three-parameter Weibull model and χ2 test were used to analyze the diameter structural distribution of 15 plots. The density of 15 plots was 400-981 trees·hm-2, and the mean DBH was 10.5-19.9 cm. The skewness of all plots was positive, which showed the diameter distribution function curves was partial to left. The three-parameter Weibull distribution model was better for the diameter distribution of the 9-11 plots, while negative exponential model was suitable for other plots. The irregular diameter distribution can be fitted better by three-parameter Weibull model, while the inverse J-shaped curves can be fitted well by negative exponential model. It showed that the diameter structure of coniferous broad-leaved mixed forest of Taxus cuspidate stand in the study area were not reasonable, and the management of T.cuspidate stand should be strengthened.

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    Cloning and Expression Analysis of E2 Ubiquitin-conjugating Enzyme Gene TkUBC2 in Taraxacum kok- saghyz Rodin
    Xiao-Xiao WANG, Bi QIN, Yu-Shuang YANG, Qiu-Hai NIE, Ji-Chuan ZHANG, Shi-Zhong LIU
    Bulletin of Botanical Research    2021, 41 (1): 98-106.   DOI: 10.7525/j.issn.1673-5102.2021.01.013
    Abstract274)   HTML26)    PDF(pc) (3854KB)(80)       Save

    In order to analyze the function of E2 ubiquitin-conjugating enzyme(UBC) in stress response and signal transduction, an UBC gene was cloned from Taraxacum kok-saghyz Rodin(TKS) line 1151 and named as TkUBC2. The ORF of this gene was 459 bp and encoded 152 aa. Sequence alignment analysis showed that UBC2 shared high level homology among different species, and TkUBC2 had more than 99% similarity with Lactuca sativa LsUBC2 and Helianthus annuus HaUBC2. qRT-PCR was used to analyze the expression pattern of TkUBC2 in TKS. The results indicated that TkUBC2 was ubiquitously expressed in different tissues. Moreover, TkUBC2 expression was down-regulated by treatments of PEG6000 simulated drought stress, mannitol mediated osmotic stress and plant hormone i.e. methyl jasmonate(MeJA), abscisic acid(ABA) and ethylene(ET) respectively, but its expression was up-regulated by the treatments of NaCl salt stress and UV radiation respectively. The results of this study suggested that TkUBC2 was involved in response to various stress conditions, hormone signaling and DNA damage repair process, which lay a good foundation for further revealing the function of TkUBC2 in TKS.

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    Genome-wide Identification and Expression Analysis GATA Family of Populus trichocarpa
    Meng-Xuan REN, Yang ZHANG, Shuang WANG, Rui-Qi WANG, Cong LIU, Zhi-Gang WEI
    Bulletin of Botanical Research    2021, 41 (1): 107-118.   DOI: 10.7525/j.issn.1673-5102.2021.01.014
    Abstract451)   HTML30)    PDF(pc) (5569KB)(168)       Save

    The GATA transcription factor gene family played an important role in plant growth and development, cell differentiation, and response to environmental changes. However, no genome-wide analysis of this gene family has been reported in woody plants so far. We systematically analyzed the number, gene structure, chromosome positioning, phylogenetic evolution, physicochemical characteristics and conserved motifs of the GATA family members from the genome level respectively. The results showed that the GATA family contains 39 genes distributed on 15 chromosomes, and 6 genes distributed on chromosome 5, and only 1 gene distributed on chromosomes 9, 13 or 19 respectively. The rest chromosomes had no gene distributed on. The genes structure in this family and the basic characteristics encoded protein were somewhat heterogeneous, and could be divided into 4 subfamilies. By the qRT-PCR, there were significant differences in the expression levels of the genes of the GATA family at different developmental stages, and salt stress significantly affected the expression characteristics of the genes. The above results indicated that the structure and function of the genes were significantly differentiated after the replication of GATA family genes, and some of these genes might play an important role in the secondary growth of Populus trichocarpa and the response to salt stress. This study laid the foundation for a comprehensive analysis of the biological functions of the members of the GATA family of P.trichocarpa in their growth and development and response to salt stress.

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    To Explore the Transcription Factor in Response to Low Temperature Stress in Nicotiana alata by Transcriptome Sequencing
    Lin LU, Shang-Yu YANG, Wei-Dong LIU, Li-Ming LU
    Bulletin of Botanical Research    2021, 41 (1): 119-129.   DOI: 10.7525/j.issn.1673-5102.2021.01.015
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    In order to identify the transcription factors involved in the low temperature stress of Nicotiana alata seedlings, 4℃ low temperature was carried out, and samples of the seedlings were collected in 12-hour treatment, then the total RNA was extracted, and high-throughput sequencing technology was used to sequence the transcriptome. For differentially expressed genes, GO and KEGG analysis were applied, the transcription factors involved were classified, and the results of transcriptome sequencing were verified by qRT-PCR. There were 8 388 genes(P<0.01) whose expression changed more than twice after low temperature treatment, of which 4 229 genes were up-regulated and 4 159 genes were down regulated. The functions of these differentially expressed genes were classified into three categories: biological process, cell components and molecular functions, 69 items in go entries, and were significantly enriched in 40 KEGG metabolic pathways. At the same time, under low temperature stress, the expression of 118 transcription factors in N.alata changed significantly, among which 82 genes were up-regulated and 36 genes were down regulated. These transcription factors belonged to 28 families, of which 19 genes to NAC family, 16 genes to ERF family, 15 genes to MYB family and 15 genes to WRKY family. The results provided a reference for the study of the molecular mechanism of low temperature response in N.alata.

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    An Optimized HPLC Procedure for Analyzing Three Endogenous Hormones in Tulip Bulbs
    Yang-Jing ZHAO, Nan TANG, Dao-Cheng TANG, Jing ZHANG
    Bulletin of Botanical Research    2021, 41 (1): 130-137.   DOI: 10.7525/j.issn.1673-5102.2021.01.016
    Abstract315)   HTML27)    PDF(pc) (1165KB)(48)       Save

    Agilent HPLC was used to establish a method for simultaneous determination of endogenous hormones GA3, IAA and ABA in tulip bulbs, and an extraction method of isopropanol was established. The external standard method was used for determination, C18 reverse phase column, mobile phase A(methanol)∶B(phosphate buffer pH=3.5)=45∶55; flow rate 1 mL·min-1; detection wavelength 0-3.2 min 265nm, 3.0-4.5 min 212 nm, 4.5-6.5 min 218 nm; 6.5-13.0 min 265 nm; column temperature 20℃ for HPLC determination. Isopropyl alcohol extractant and dichloromethane low temperature shaker to purify the endogenous hormones in the bulb. The whole process was simple to operate, which took only 2.0-2.5 h to complete the hormone purification. The linear correlation of the detection methods was above 0.995. Detection limits of GA3, IAA and ABA were 200 ng·mL-1, 5 ng·mL-1 and 20 ng·mL-1, respectively. Recovery rate of Purification method was 84.812%-95.679%. The relative standard deviation was 6.432%-2.831%. A stable baseline and high accuracy, and a satisfactory peak shape, simple operation, less loss of hormones can be obtained in each link by extracting the endogenous hormone.

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    Growth Responses of Pinus sylvestris var. mongolica Seedlings under Simulated Nitrogen Deposition and Different Inoculation of Ectomycorrhizal Fungi Treatments
    Xu-Fu WANG, Long-Fei HAO, Jia-Xin HAO, Wen-Ying HAO, Hui-Ga BAO, Shu-Lan BAI
    Bulletin of Botanical Research    2021, 41 (1): 138-144.   DOI: 10.7525/j.issn.1673-5102.2021.01.017
    Abstract203)   HTML27)    PDF(pc) (887KB)(30)       Save

    The growth responses of Pinus sylvestris var. mongolica seedlings in pot culture conditions were discussed under different inoculation treatments and simulated nitrogen deposition. The different inoculation treatments were performed after 2-month seedling, which included a strain inoculation of Suillus bovinus(SB) and 8 strain mixed inoculation(HJ), including Suillus luteusS.grevilleaTricholoma fulvumB.grisellusS.bovinusLeucocortinarius bulbiger Rhizopogon luteolus and Pisolithus tinctorius. The simulated nitrogen deposition treatments were performed for six months after mycorrhizal colonization, which included on nitrogen(CKN), 1-time nitrogen concentration(CN) and 2-time nitrogen concentration(2CN) treatments. The seedling growth index, root morphology and mycorrhizal colonization rate were measured after the 5-month interaction treatments. The results showed that: ①With the increase of nitrogen concentration, the mycorrhizal infection rate showed a downward trend under SB and HJ treatments. The mycorrhizal infection rate of SB inoculation was affected significantly by nitrogen deposition treatments. ②The seedlings aboveground and underground biomass were accumulated significantly after inoculation treatments, which were the best effect under mixed inoculation treatment. The aboveground biomass in the same inoculation treatments were inhibited by the increase of nitrogen concentration. The seedlings ground diameter and above-ground biomass were affected significantly by interaction of inoculation and nitrogen concentration, but the seedlings height and underground biomass had no effects. ③Compared with CK and SB treatments, the seedlings average root diameter, total root length and total surface area of CKN and CN treatments were promoted significantly under HJ treatment. With the increase of nitrogen content, the seedlings average root diameter, total root length and total surface area were promoted significantly under CK and SB treatments, but the morphological indexes were inhibited under HJ treatment. The seedling root morphology were affected significantly by the inoculation, nitrogen concentration, and the interaction treatments.

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    Effects of Nitrogen Deposition and Ectomycorrhizal Fungi on Root Architecture and Rhizosphere Soil Enzyme Activities of Larix olgensis Seedlings
    Ting-Yan LIU, Long-Fei HAO, Xu-Fu WANG, Hai-Xia YAN, Shu-Lan BAI
    Bulletin of Botanical Research    2021, 41 (1): 145-151.   DOI: 10.7525/j.issn.1673-5102.2021.01.018
    Abstract354)   HTML25)    PDF(pc) (939KB)(38)       Save

    In order to explore the responses of root morphology and rhizosphere soil enzyme activities of one-year-old Larix olgensis seedlings to different simulated nitrogen deposition and ectomycorrhizal fungi, the morphology of roots with diameter of 0-0.5 mm(including the total root length, surface area, volume and tips) and soil enzyme activities(including β-1,4-glucosidase(BG), L-leucine aminopeptidase(LAP), β-1,4-N-acetylglucosaminidase(NAG), acid phosphatase(ACP) and alkaline phosphatase(ALP)) were measured respectively. The different inoculated seedlings of L.olgensis were selected, including the not noculated(-M) and inoculated with 8 ectomycorrhizal fungi(+M) respectively. Four nitrogen deposition treatments were set, including no nitrogen(0N, 0 kg·N·hm-1·yr-1), low nitrogen(LN, 15 kg·N·hm-1·yr-1), middle nitrogen(MN, 30 kg·N·hm-1·yr-1) and high nitrogen(HN, 60 kg·N·hm-1·yr-1). The results indicated that: ①The nitrogen deposition concentration had a negative effect on the total root length, surface area, volume, tips with a diameter 0-0.5 mm of L.olgensis seedlings under +M and -M treatments. The root morphology indexes of -M was higher than that of +M treatment under 0N, LN, and MN treatments, respectively. ②With the increasing of nitrogen deposition, the activities of BG, LAP, ACP, ALP in the rhizosphere soils showed a tendency of first rising and then decreasing under the two inoculation treatments, but the activities of NAG showed a decreasing tendency. ③The morphology of roots with diameter of 0-0.5 mm was significantly negatively correlated with the activities of BG in the rhizosphere soils(P<0.05), under the +M and -M treatments. There was significantly positive correlation between the activities of NAG and root morphology(P<0.05), except the number of the tips. The effect of nitrogen deposition on root morphology of L.olgensis seedlings was weakened by ectomycorrhizal fungi inoculation treatment. However, the activation of soil enzyme activity by +M was higher than that of -M treatment under low nitrogen deposition.

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    Preparation and Slow Controlled Release of Porous Starch Solidifying Rhododendron tomentosum Oil
    Qian ZHANG, Xiu-Hua ZHAO, He-Di KANG, Xiao-Xue ZHANG, Jian-Hang YANG
    Bulletin of Botanical Research    2021, 41 (1): 152-160.   DOI: 10.7525/j.issn.1673-5102.2021.01.019
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    Rhododendron tomentosum was abundant plant resources in the northeast of China. The plant was a distinctive aroma and high oil content in leaves. For the disadvantages of high volatility, poorly stable, short fragrance and inconvenient carry of R.tomentosum oil, it limited the development and utilization of essential oil related products. We explored the preparation and slow controlled release of porous starch solidifying R.tomentosum oil. The solidified essential oil powder was prepared by grinding and adsorption. DSC and TG were used to investigate the thermal stability of the solidified essential oil powder and the effect of the slow controlled release fragrance at different temperatures. The results showed that the maximum adsorption capacity of porous starch was 1.42 g?g-1. When the mass ratio of porous starch to essential oil was 1∶0.75 (g·g-1), the solidified essential oil powder smelt well and presented yellowish powder. The thermal stability of the solidified essential oil was 3.57 times higher than the retention rate of essential oil. At the same temperature, the slow controlled release fragrance effect of solidified essential oil powder was more significant than that of uncured essential oil powder at the same time. The preparation process of the porous starch solidified essential oil was simple, and the effect of sustained release of the sample was obvious with the application and promotion values.

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    A New Species and a New Variety of Delphinium(Ranunculaceae) from China
    Wen-Tsai WANG
    Bulletin of Botanical Research    2021, 41 (2): 161-163.   DOI: 10.7525/j.issn.1673-5102.2021.02.001
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    A species, belonging to Sect.Delphinastrum DC. subsect. Grandiflora W.T.Wang, and a variety of D. iliense Huth were described as new.

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    Oreorchis bashanensis Wang Yong, a New Species of Orchidaceae from Shaanxi, China
    Yong WANG
    Bulletin of Botanical Research    2021, 41 (2): 164-167.   DOI: 10.7525/j.issn.1673-5102.2021.02.002
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    A new species of Orchidaceae, Oreorchis bashanensis Wang Yong from Dabashan mountain area in Shaanxi was reported. The species was closely related to O.patens. Comparing with O.patens, the new species was decedious. The leaves fell during its flowering period. During its nutritional growth period, there were 2-3 leaves on each individual. It had larger pseudobulb and leaf blade. The lateral petals were erect. The claw of lip petal was about half the length of the whole lip petal. The orange callus protruded between lateral lobes on lip. However, O.patens was evergreen, and it had 1-2 leaves on each individual. The claw of lip petal was about 1/4 the length of the whole. The lateral petals were curved. The longitudinal lamellae was white or pale yellow. The type specimens were deposited in Herbarium of Shaanxi University of Technology(HZTC).

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    Phenological Observation of the Extremely Small Population Plant Styrax zhejiangensis
    Guo-Lin FU, Ting-Ting LI, Ye-Hua ZHANG, Xiang-Ying WEN, Xue-Chu MA, Chu-Ping WU
    Bulletin of Botanical Research    2021, 41 (2): 168-173.   DOI: 10.7525/j.issn.1673-5102.2021.02.003
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    Styrax zhejiangensis was a species with extremely small population, and endemic in Taohuawu, Longjiang, Jiande Forest Farm, Zhejiang Province. To clarify the reason this species endangered, phenology of S.zhejiangensis was observed, including its flowering and seed setting pattern, as well as the biological characteristics of seeds. The results showed that flowers and leaves shared the same proliferation period. The characteristics of flower organs and pollen were similar to those plants of the same genus, and the seed setting rate was 21%-32%. Seeds had high morphological variations, and their insufficient plumpness and poor water permeability caused by dense and hard seed coat may affect seed germination. Abundant rainfall, and temperature differences between day and night, were two major drivers that would aggravate the falling of flowers and fruits. More attention should be paid to their seeds, flowers and fruits if we aim to conserve this endangered species.

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    Responses of Water Absorption and Germination of Angelica sinensis Seeds to Winged Perianth Removal
    Yong-Wei MI, Cheng-Wen GONG, Guo-Xiang WANG, Yong-Hong QI
    Bulletin of Botanical Research    2021, 41 (2): 174-179.   DOI: 10.7525/j.issn.1673-5102.2021.02.004
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    The water absorption and germination process for intact and de-winged seeds(winged perianth removed) of Angelica sinensis were determined to provide a theoretical basis for its artificial propagation. With intact and de-winged seeds of A. sinensis, and intact seeds being used as a control group(CK), the 1000-grain weight, water content, volume, volume weight and water absorption ratio of seeds were measured, in the meanwhile, the germination test was proceeded under natural room temperature and in laboratory. Compare with CK, 1000-grain weight and volume of de-winged seeds decreased by 46.87% and 90.78%(P<0.01), respectively. The volume weight and water content of de-winged seeds increased by 475.92%(P<0.01) and 20.15%(P<0.05), respectively. The time of water absorption to reach stationary phase of de-winged seeds was 2.48 h, and was shorter than that of CK by 3.38 h, because the winged perianths significantly inhibited the water absorption capacity of seeds, and the soaking process fitted the logistic equation. Compared with CK, the germination potential, germination rate and germination index of de-winged seeds increased by 256.70%, 12.28% and 27.77%(P<0.01), respectively. The volume and weight of A. sinensis seeds were decreased, the water absorption capacity and germination quality of seeds were improve by removing winged perianths. Therefore, the de-winged seeds of A.sinensis should be applied in agricultural production.

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    Effects of Micro-habitats on Water Retention and Loss of a Drought-tolerant Saxicolous Moss Grimmia pilifera
    Dong QIU, Jing ZHANG, Nan WU, Ye TAO
    Bulletin of Botanical Research    2021, 41 (2): 180-190.   DOI: 10.7525/j.issn.1673-5102.2021.02.005
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    The Grimmia pilifera is a typical drought-tolerant saxicolous moss species, and the water is the key factor influencing its survival and reproduction. To explore the effects of micro-habitats to water physiology of G.pilifera, three typical micro-habitats(habitat S-1 is the bare rock shaded by bamboo individuals; habitat E is the exposed rock; habitat S-2 is the bare rock shaded by shrub Ficus pumila) were chosen in the Dalongshan National Forest Park in Anhui Province, then the differences in individual plant size, saturated water content and water content indices of G.pilifera during dehydration in different micro-habitats, and finally the impact of micro-habitats on moss water stress tolerance and trade-off characteristics were comprehensively assessed. It was found that the plant size and saturated water content indices varied significantly among three micro-habitats; there into, the moss plant size in habitat S-1 was low but the internal and external water contents were relatively high; the plant sizes and external water contents in habitat E and habitat S-2 were similar but the external water content was much more higher of the former. The external water contents of moss plants in the two shaded habitats S-1 and S-2 were seven times the internal water contents, and their ratios were notably higher than that in habitat E. The water content indices of G.pilifera in habitat E the same time during dehydration were higher than that in the two shaded habitats, and the time differences reaching the same water content gradually expanded as the dehydration proceeded; thus it won more time for moss plants in habitat E to carry out effective photosynthesis(here the relative water content is not less than 35%) and physiological and structural adjustment during the later dehydration. In total, G.pilifera plants in exposed rocks represented much stronger water stress tolerance than that in shaded rocks, but the later was able to increase the external water to make up the deficiency of rapid water loss. This might be the trade-off strategy of water absorption and retention for G.pilifera lived in different micro-habitats.

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    Spatial Distribution Characteristics of All Genera in Rhamnaceae of China
    Ya-Li LI, Li-Chuan QIU, Jian WANG
    Bulletin of Botanical Research    2021, 41 (2): 191-198.   DOI: 10.7525/j.issn.1673-5102.2021.02.006
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    Spatial distribution characteristics of all genera in Rhamnaceae of China are based on the distribution data of species of Rhamnaceae in China and county of China. Spatial distribution data of genera were made in ArcGIS software. Area, perimeter and number of every patch were gotten from the maps. Simpson’s evenness indexes, shape indexes and largest patch indexes were calculated. Spatial distribution characteristics of all genera in Rhamnaceae can be analyzed. The experiment was conducted to determine the minimum range of spatial distribution, quantitative research, plant protection, utilization, resource development and restoration. Results showed that: ①Alphitonia Scutia Chaydaia Berchemiella Colubrina Gouania and Ventilago are stenochoric genera. Paliurus Rhamnella Sageretia Hovenia Berchemia Ziziphus and Rhamnus are eurychoric genera. Both the stenochoric genera and eurychoric genera account for 50% of the total genera of Rhamnaceae in China. The total area of stenochoric genera is less than 30×104 km2Berchemiella mainly distribute in the central subtropical zone of China. Other stenochoric genera mainly distribute near the tropic of cancer and south of it. The total area of eurychoric genera is greater than 40×104 km2.. They are distributed in most parts of our country, and scattered in the northwest.②Rhamnus has the largest distribution range and total area in Rhamnaceae of China, which almost covers the distribution range of all other genera. It can reflect the spatial distribution and characteristics of Rhamnaceae in China. ③Alphitonia is the only genus of continuous and the narrowest distribution of Rhamnaceae in China, and other genera are disjunction. Chaydaia Gouania Berchemia and Rhamnus only have the main distribution area. ScutiaSageretia and Hovenia have main and secondary area. Berchemiella ColubrinaVentilago Paliurus Rhamnella and Ziziphus are all scattered area.

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    Observation and Comparison of Root Primary Xylem Archy in Different Chlorophytum Varieties
    Xue-Peng FU, Le WANG, Jia-Le WEN, Kai-Fang YANG, Xiao-Jie YANG
    Bulletin of Botanical Research    2021, 41 (2): 199-204.   DOI: 10.7525/j.issn.1673-5102.2021.02.007
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    Three Chlorophytumcomosum varieties were used to investigate the relationship between the root primary xylem archy and C.comosum varieties, diameter of the roots and living conditions by using sections cut free-hand and stained with Safranin. The root primary xylem archy in the roots of different varieties were different. The root primary xylem archy in the roots of C.comosum in soil culture were also different from those under hydroponic condition, and the number of xylem bundles in the roots of C.comosum under hydroponic conditions was much less than that in soil culture. Under hydroponic conditions, there was a positive correlation between the number of xylem bundles in the roots of C.comosum and the diameter of the roots, however, the root xylem archy in C.comosum ‘Yinxin’ and ‘Yinbian’ has no correlation with the roots diameter of C.comosum.

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    Photosynthetic and Stomatal Characteristics of Pinus koraiensis and P.sibirica under Low Temperature Stress
    Fang WANG, Zhi-Min LU, Jun WANG, Shi-Kai ZHANG, Yu-Xi LI, Shao-Chen LI, Jian-Qiu ZHANG, Yu-Chun YANG
    Bulletin of Botanical Research    2021, 41 (2): 205-212.   DOI: 10.7525/j.issn.1673-5102.2021.02.008
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    Both Pinus koraiensis and P.sibirica with strong cold resistance are famous forest species in the cold temperate zone. Compared with P.koraiensis P.sibirica has stronger cold resistance. In order to explore the physiological response and cold resistance mechanism of the two species under low temperature stress, 5-year-old P.koraiensis and P.sibirica seedlings were used to be conducted by low temperature treatment. By setting three stress temperatures(0℃, -20℃ and -40℃) and three stress times(6, 24 and 48 h), taking 20℃ as control, which were used to study the photosynthetic characteristics and stomatal characteristics of P.koraiensis and P. sibirica under low temperature stress. By t test and variance analysis, there were significant differences (P<0.05) between P.koraiensis and P.sibirica for each photosynthetic index and stomatal density, both low temperature and low temperature stress time had an extremely significant effect on each photosynthetic index of P.koraiensis and P.sibiricaP<0.01), and low temperature also had an extremely significant effect on stomatal opening size and stomatal area of P.koraiensis and P.sibiricaP<0.01). The net photosynthetic rate, stomatal conductance and transpiration rate in P.koraiensis were significantly higher than those in P.sibirica at 20℃(without stress) and 0℃(low temperature stress). However, under -20℃ for 6 h, each photosynthetic index in P.sibirica was higher than that in P.koraiensis. With the decrease of temperature and the extension of stress time, each photosynthetic index of the two species showed a decreasing trend. The stomatal density in P.koraiensis was significantly higher than that in P.sibirica. Before stress(20℃), the stomas of P.koraiensis and P.sibirica were all oval. The stomatal opening size and stomatal area of the two species decreased significantly with the decrease of temperature.

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    Variation of Flower Phenotypic Traits on 195 Syringa oblata SeedSeedings Single Trees
    Rui-Qing HE, Yun-Yao YANG, Zeng-Hui HU, Jing WU, Ping-Sheng LENG
    Bulletin of Botanical Research    2021, 41 (2): 213-220.   DOI: 10.7525/j.issn.1673-5102.2021.02.009
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    To accurately grasp the variation degree of floral traits and the phenotypic diversity level of Syringa oblata, so as to provide a theoretical basis for the breeding of S.oblata excellent varieties. The 195 seed seedlings of S.oblata trees around 8 years old were used to analyze the genetic variation, trait correlation and clustering relationship of 5 quantitative traits and four quality traits. Five quantitative traits were extremely significant differences among the 195 trees(P<0.01), and accorded with the normal distribution. The genetic variation coefficients were from 19.72%-30.22%, with an average of 24.97%, and the inflorescence length was the largest. According to the ISCC-NBS method, the group colors were divided into six color groups, namely white, pale pink, purple, purplish red, purplish pink and violet groups. Purple group(54.35%) and purplish red group(32.31%) accounted for the largest proportion; the diversity index of four quality traits ranged from 0.36 to 1.09, with an average of 0.86, of which the corolla lobes(state) were the largest, indicating that the quality traits were abundant within 195 trees. By correlation analysis, there were significant positive correlations between the five quantitative traits, and the correlation coefficient between inflorescence length and inflorescence width was the largest(0.767). By cluster analysis, the 195 samples of S.oblata can be divided into three categories based on euclidean distance. The first type was characterized by corolla tube length; the second group was characterized by long inflorescence; the last type was characterized by corolla lobes. The phenotypic variation of floral traits in the tested genetic resources of S.oblata was richer.

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    Establishing an Efficient Regeneration Protocol in vitro for Rapid Micro-propagation of Two New Varieties of Ilex verticillata L.
    Rui LIU, Jia-Yi ZHANG, Chao-Qi GE, Lu-Lu XIE, Ying CHEN, Can ZHANG, Xi CHEN
    Bulletin of Botanical Research    2021, 41 (2): 221-231.   DOI: 10.7525/j.issn.1673-5102.2021.02.010
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    An efficient and rapid micro-propagation system using tissue culture for two new varieties “Red Sprite” and “Winter Gold” of Ilex verticillata L. was established with the young shoots as explants, and an optimum method produced large numbers of genetically uniform seedlings and maintained its excellent character for I.verticillata L was offered. The optimal medium for axillary bud induction was MS+1.0 mg·L-1 6-BA+0.2 mg·L-1 NAA, and the induction rate of both varieties were 100% respectively. The preferred medium for cluster bud proliferation was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.5 mg·L-1 CPPU. The highest proliferation coefficient was 5.7 for “Red Sprite”, and 4.5 for “Winter Gold” for 25 d culturing respectively. The optimal seedling medium for “Red Sprite” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+1.0 mg·L-1 TDZ+1.0 mg·L-1 GA, and that of “Winter Gold” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.05 mg·L-1 TDZ+1.0 mg·L-1 GA, the seedling height was up to 6.80 and 4.38 cm respectively. The optimal rooting induction medium was 1/2MS+0.4 mg·L-1 NAA+0.4 mg·L-1 IBA+1.0 mg?L-1 activated carbon, the rooting rate of “Red Sprite” and “Winter Gold” were 95.0%, or 97.0% respectively. The best transplanting substrates were peat∶perlite=3∶2, with a survival rate of 95.7% for “Red Sprite” and 96.8% for “Winter Gold” respectively.

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    Dynamic Changes of Endogenous Hormones in Rootstocks and Scions within Nurse Seedling Graft in Camellia oleifera Under Wound
    Wei LONG, Xiao-Hua YAO, Le-Yan LÜ
    Bulletin of Botanical Research    2021, 41 (2): 232-242.   DOI: 10.7525/j.issn.1673-5102.2021.02.011
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    In order to analyze the physiological response to the surface healing of rootstocks and scions, and to provide the theoretical support for the growth mechanism, the dynamic changes of endogenous hormones in post-traumatic stress response in Camellia oleifera A. under wound were studied. The six-year-old Changlin 18 and Changlin 53 trees were used, and the contents of IAA, ABA, TZR, Zeatin, SA, JA were assayed using HPLC-MS at different wound stages, and the changes of endogenous hormone in different period and relationship between varieties were analyzed. The results showed that the content of IAA, SA and JA decreased with the increasing time of wound in scions of Changlin 18, and the content of TZR and Zeatin increased first and decreased later which they were respectively reached the highest in S10 and S0. The content of ABA rose continually and reached the peak in S10. In scions of Changlin 53, the contents of IAA and JA would decrease after peak; the TZR, Zeatin, SA were gradually increased after minimum. The content of ABA was gradually decreased in S0 after maximum. The contents of endogenous hormones were higher in Changlin18 than in Changlin53 in stem of rootstock between varieties. The endogenous hormones were decreased in Z0 and increased in Z10 in varieties except the contents of ABA and SA in Changlin53 increased first and then decreased. In stem and root of rootstocks, the contents of endogenous hormones were higher in Changlin 18 than in Changlin 53 except JA; The contents of IAA and ABA in stem of rootstocks were higher than roots which other endogenous hormones in root of rootstocks were higher than in stem. There was difference in the ratio of hormone in varieties and part of rootstocks. The ratios of IAA/ABA, IAA/TZR, IAA/Zeatin, IAA/JA, ABA/TZR, ABA/Zeatin, ABA/JA in scions and rootstocks were higher in Changlin 53 than in Changlin 18. The ratio of SA/IAA in scions was higher in Changlin 18 than Changlin 53, and it was higher in Changlin 18 than Changlin 53 in ratios of SA/JA, SA/ABA of scions while it was higher in Changlin 18 in stem of rootstocks. The ratio of TZR/SA, TZR/JA in scions and stem of rootstocks were higher in Changlin 18 than Changlin 53. There was the significantly positive correlation in IAA and JA after wound, and the correlation of IAA and SA, SA and JA had difference between varieties, the significantly positive correlation in Changlin 18 and the significantly or significantly negative correlation in Changlin 53. There was the significantly positive correlation among TZR, Zeatin, SA and the significantly negative correlation among TZR, Zeatin, SA. In stem of rootstocks, there was very significantly positive correlation among hormones in Changlin 18, and there was the significantly positive correlation among TZR, IAA and JA, Zeatin and IAA and JA while there was the significantly negative correlation in TZR and SA, Zeatin and SA. There was significantly negative correlation in SA and JA. In stem and root of rootstocks, there was the difference in correlation of SA and other hormones among varieties, and other hormones in Changlin 18 and Changlin 53 had the same significantly positive or negative correlation. To sum up, it was easy to callus formation in surface of wound in Changlin 18, but it was difficult in Changlin 53 because there was much more high-level hormone of stress-resistant so that it would be possible that affects healing after grafting wound 10 min. The differences in ratio and contents of hormones between rootstocks and scions may be affected by the morphological reconstruction of grafted unions and the growth of scions in later stage.

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    Cloning and Sequence Analysis of Squalene Synthase( SQS) Gene from Dioscorea bulbifera L. Microtuber
    Wei-Zhong KE, Wen-Juan ZHONG, Kai-Ying LIU, Xiang-Yuan LI, Ming-Hua YIN
    Bulletin of Botanical Research    2021, 41 (2): 243-250.   DOI: 10.7525/j.issn.1673-5102.2021.02.012
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    The core fragment of SQS gene was screened from the transcriptome database of Dioscorea bulbifera L. microtuber, the conservative fragment of SQS gene was obtained by RT-PCR, and the 3' and 5' terminal sequences of SQS gene were obtained by race technique, and the sequence was analyzed by bioinformatics. The SQS gene of D.bulbifera L. microtuber was 1 548 bp long and 415 bp amino acid sequence, with a theoretical molecular weight of 46 786.38 D and an isoelectric point of 5.97. SQS protein is a hydrophobic protein without signal peptide. It contains the functional domain necessary for SQS belonging to Isoprenoid_Biosym_C1 superfamily. The homology of SQS protein between D.bulbifera and other plants is high, and the amino acid similarity of SQS protein between D.bulbifera and D.zingiberensis is 96.4%. The full-length sequence of SQS gene was obtained for the first time from D.bulbifera L. microtuber, which has the typical characteristics of SQS homologous gene. It provides the basis for further study on SQS gene structure, gene expression and gene mutation of D.bulbifera microtuber, and provides data support for the correlation analysis of positive selection site and function of SQS, a key enzyme of triterpene synthesis pathway in Dioscorea.

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    Expression Profile Analysis ofXylem Development Regulated by BpNAC012 Gene from Betula platyphylla
    Yi-Ping GUO, Jia-Xin LIU, Ying YU, Chao WANG, Chuan-Ping YANG
    Bulletin of Botanical Research    2021, 41 (2): 251-261.   DOI: 10.7525/j.issn.1673-5102.2021.02.013
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    NAC transcription factors have been considered to be the switches that regulate the formation of plant secondary cell walls. Our previous work indicated that the gene BpNAC012 was involved in the secondary cell wall biosynthesis of birch. In order to identify the downstream target genes regulated by BpNAC012, transcriptome sequencing of stems from BpNAC012 over-expression and RNAi plants was conducted, and the differentially expressed genes were identified compare to the wild type birch. The results showed that, in contrast to the wild type plants, 627 genes were up-regulated and 229 genes were down-regulated in the over-expression transgenic line, and 299 up-regulated genes and 207 down-regulated genes were identified in the RNAi lines. The expression changes of downstream genes in the overexpression transgenic lines were much greater than that in the RNAi plants. Inhibition of BpNAC012 expression significantly affected the expression of genes related to protein modification and transport. The differentially expressed genes were enriched in pathways such as receptor signaling pathways, nutritional metabolism, amino acid synthesis, and phenylpropane biosynthesis. In addition, BpNAC012 can regulate the expression of genes related to cellulose, lignin bio-synthesis and xylem development, as well as a variety of transcription factors. This study laid the foundation for further analysis of the molecular regulatory mechanism of BpNAC012 in secondary cell wall bio-synthesis of birch.

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    Effects of Different Canopy Density on the Chlorophyll Fluorescence and Water Storage Capacity of Sphagnum palustre in the Edge of the Tuntianjing Sinkhole in Bijie City
    Kuang-Zheng SHI, Zhao-Hui ZHANG, Chun-Mei HE, Zhi-Hui WANG
    Bulletin of Botanical Research    2021, 41 (2): 262-269.   DOI: 10.7525/j.issn.1673-5102.2021.02.014
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    In order to enrich the understanding of the growth environment of Sphagnum, explore its possible impact on the formation of erosional karst tiankeng, and find more suitable growth conditions for it, we used gradient method, with S.palustre of different canopy density environments in the edge of the Tuntianjing sinkhole in Bijie City, to definite the chlorophyll fluorescence parameters and water storage capacity. Results showed: ①The initial fluorescence(Fo) and maximum fluorescence(Fm) of S.palustre increased significantly with increasing canopy density(Fo:224.67-264; Fm:716-785.33). The maximum photochemical efficiency(Fv/Fm) of PSⅡ decreased firstly and then became constant; photochemical quenching coefficient(qP) and photosynthetic electron transport rate(ETR) increased significantly(qP:0.17 -0.46; ETR:5.72-9.68) with increasing canopy density; non-photochemical quenching coefficient(NPQ) decreased significantly from 1.57-0.8. ②With canopy density increased, the rate of water absorption and water storage capacity of S.palustre were significantly increased, the natural water absorption and the saturated water absorption increased from 14.36-26.37 and from 21.7-27.01, respectively; the natural water storage increased from 3 032.97-12 816.01, and the saturated water storage increased from 4 576.34-13 175.97. ③Although S.palustre has a good sunproof, they are more suitable for living in the low light conditions. As canopy density increased, it was beneficial to accumulate chlorophyll, promote photosynthesis and increase the accumulation of biomass, leading to a stronger water storage capacity. Competition can inhibit the growth of S.palustre, share soil moisture, and a certain moisture was very important for the growth of S.palustre. ④The good water-holding properties and water absorption of S.palustre have an important role in slowing surface runoff that slowing down in geological processes. The difference in water holding capacity of S.palustre under different canopy densities can affect the distribution of water flow to a certain extent, thus affecting the formation of erosion-type sinkholes.

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    Respond of Soil Nutrient to Plowing and Reseeding of Achnatherum splendens in the Qinghai Lake Region
    Tao LIU, Di ZHU, Ting LÜ, Rui-Fang LIANG, Feng LIU, Zi-Lan MA, Yi-Kang LI, Yu-Zhi MA, Xu SU
    Bulletin of Botanical Research    2021, 41 (2): 270-280.   DOI: 10.7525/j.issn.1673-5102.2021.02.015
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    In order to explore the vertical variation character of soil nutrients caused by plowing and reseeding treatments, the grassland of Achnatherum splendens was used to analyze the effects of two restoration with plowing in 1958 or reseeding in 1992 nutritional distribution characteristics of 0?10, 10?20, 20?30, 30?40 and 40?60 cm soil in the zone of the Qinghai Lake of the Qinghai-Tibet Plateau. The results showed that the soil organic carbon and the content of total K, the plot under plowing and reseeding disturbance, were significantly higher than natural A.splendens grassland. Both disturbances were beneficial to the shallow layers of soil to enrich available phosphorus and organic carbon nutrients, not to the restoration of total N(P<0.05). The contents of available K, total N and organic carbon decreased as the depth of the soil increases, but the soil bulk density, soil pH, the content of total P and K were not significantly different among each layers of plowing soil. Compared with the natural group, the content of available N was significantly decreased at each layers of reseeding soil(P<0.01), but both disturbances caused significantly higher total K content, and significantly lower total N content in the soil 40?60 cm(P<0.05). Compared with the natural group, there were significantly positive correlation between the contents of total N and K(P<0.05), not significant correlation between the contents of available P and N after reseeding, both disturbances aroused the negative correlation between the soil nutrients and the soil bulk density. The pH values of the soil decreased significantly after plowing and reseeding, and plowing treatment could concentrate the soil available nutrients in the surface layer. The reseeding treatment accelerated the turnover of soil nutrients, and the increase of output promoted the restoration of aboveground vegetation, Beside the total potassium content. Both disturbances caused the recovery of total nutrients in different soil layers will be an extremely slow process.

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    Leaf Phenotype Variation and Heterosis in F 1 Progeny of Cross Between Castanea mollissima and C.henryi
    Ping-Sheng ZHANG, Xi-Bing JIANG, Bang-Chu GONG, Yang XU, Jun-sheng LAI, Cong-Lian WU
    Bulletin of Botanical Research    2021, 41 (2): 281-294.   DOI: 10.7525/j.issn.1673-5102.2021.02.016
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    In order to explore the genetic variation and heterosis of leaf traits in cross F1 progeny of Castanea, 14 traits of leaf phenotype and photosynthetic physiology of F1 progeny(including 235 individuals) of 9 combinations (including interspecific and intraspecific cross between C.mollissima and C.henryi) and their parents were studied by using the statistical methods of Nested anova, Pearson correlation analysis and principal component analysis. The results showed: ①The average coefficient of variation of 14 traits ranged from 4.40%-27.76%, indicating that there were abundant genetic diversity in leaf phenotype and photosynthetic physiological traits in F1 hybrid progeny of Castanea. ②The average of phenotypic differentiation coefficient among combination were 32.32%, which showed that the variation within combinations was the main source of variation. ③The mid-parent heterosis rate of 14 traits ranged from -27.16%-90.53%, and the heterosis of different combinations was different. ④The Pearson correlation analysis showed that there were 46 and 13 pairs of correlations reached extremely significant levels(P<0.01) and significant levels(P<0.05), respectively, and 5 leaf phenotype traits, such as leaf length and leaf width, and 4 photosynthetic physiological indexes, such as chlorophyll a and chlorophyll b, had a great influence on each other. ⑤2 crossing combinations ‘Kuili’×YLZ No.15 and ‘Kuili’×YLZ No.1 were finally selected by principal component analysis, and their offspring had the characteristics of large leaf area, high dry matter and photosynthetic pigment content, which can provide the main materials for further screening of excellent offspring.

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    Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa
    Shu-Ping PENG, Cheng-Ming DONG, Yun-Hao ZHU
    Bulletin of Botanical Research    2021, 41 (2): 294-301.   DOI: 10.7525/j.issn.1673-5102.2021.02.017
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    Allene oxide synthase(AOS) and 12-oxophytodienoate reductase(OPR) were the key genes of jasmonic acid biosynthesis in the plants. The genes of AOS and OPR responding to endophytic fungus infection were screened from the interaction transcriptome of Rehmannia glutinosa with endophytic fungus GG22. Specific primers were designed to the open reading frame of RgAOS and RgOPR. Bioinformatics analysis of the sequences and theirs encoded product were performed, and real-time fluorescent quantitative PCR was used to analyze the expression patterns of RgAOS and RgOPR in different tissues infection by endophytic fungus GG22. The open reading frame of RgAOS was 1 626 bp, encoding 541 amino acids with a molecular weight of 60.2 kD. The open reading frame of RgOPR was 1 197 bp, encoding 398 amino acids with a molecular weight of 44.07 kD. QPCR analysis showed that the expression of RgAOS was the highest in roots, the lowest in flowers, and the expression of RgOPR was higher in flowers and leaves, respectively. Endophytic fungus GG22 induced the expression of RgAOS and RgOPR in R.glutinosa. The RgAOS and RgOPR were successfully cloned from R.glutinosa, laying the foundation for further study on the biological activity of Jasmonic acid substances in R.glutinosa and claiming the molecular mechanism of endophytic fungi in the regulation of secondary metabolites of R.glutinosa.

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    Effect of Exogenous ATP on Cold Tolerance of Brassica campestris Seedlings
    Xin-Dan HU, Yao LI, Xiao-Hua ZHANG, Juan-Hong LIANG, Teng-Guo ZHANG
    Bulletin of Botanical Research    2021, 41 (2): 302-311.   DOI: 10.7525/j.issn.1673-5102.2021.02.018
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    The effects of exogenous ATP on the cold tolerance of rape seedlings were studied with “Longyou 7”Brassica campestris L. as experimental material. Compared with low temperature stress alone, MDA content and content of rapeseed seedlings were significantly decreased after exogenous ATP pretreatment and low temperature stress, while chlorophyll content and antioxidant enzyme activities(SOD, POD, CAT, APX) were observed. The expressions of RBOHDRBOHFCPK4 and CPK5 were increased. Compared with exogenous ATP+ low temperature, MDA content was significantly increased under EGTA+exogenous ATP+ low temperature treatment. Total chlorophyll content, T-AOC enzyme activity, RBOHD and RBOHF gene expression were all increased. Significantly decreased, MDA content was increased significantly under DMTU+exogenous ATP+low temperature treatment, total chlorophyll content, Ca2+-ATPase activity, CPK4CPK5 gene expression were decreased, indicating that exogenous ATP affected cold tolerance of rape seedlings by Ca2+ and H2O2-dependent mechanisms.

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    Establishment and Optimization of SSR-PCR Reaction System for Casuarina
    Zhen LI, Chong-Lu ZHONG, Yong ZHANG, Yong-Cheng WEI, Jing-Xiang MENG
    Bulletin of Botanical Research    2021, 41 (2): 312-320.   DOI: 10.7525/j.issn.1673-5102.2021.02.019
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    In order to obtain the optimal reaction medium of SSR-PCR from casuarina, 24 somaclonal clones from 4 species including Casuarina equisetifoliaC.junghuhnianaC.glauca and C.cunninghamiana, were selected as experimental materials. L16(45) orthogonal design was used to determine the optimum concentrations of four factors(primer, Mg2+, dNTP, Taq polymerase) within four concentration levels in the SSR-PCR reaction system of Casuarina respectively, and visual analysis and analysis of variance were used as evaluation. The results showed as follows: The three factors(primer, Mg2+, dNTP) had significant differences on the SSR-PCR reaction systems(P<0.01) respectively, and the order of significance was primer>Mg2+>dNTP, while Taq polymerase had no significant difference. The two optimal SSR-PCR reaction systems(10 μL) of Casuarina were determined, including system 6, which consisted of 1×PCR buffer, 2ng template DNA, 0.5 μmol·L-1 primer, 1.5 mmol·L-1 Mg2+, 0.1mmol·L-1 dNTP, 0.5 U Taq polymerase, and system 15, which contained 1×PCR buffer, 2 ng template DNA, 0.5 μmol·L-1 primer, 1.75 mmol·L-1 Mg2+, 0.2 mmol·L-1 dNTP, 1.25 U Taq polymerase. The system 6 was better than system 15. 60℃ was determined as the optimal annealing temperature of the fluorescent primers M26 and M36.

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    Plant Regeneration System from Cotyledon of Camellia japonica (Naidong)
    Hong-Mei TIAN, Xiao GUO, Kui-Ling WANG, Ying-Kun SUN
    Bulletin of Botanical Research    2021, 41 (3): 321-328.   DOI: 10.7525/j.issn.1673-5102.2021.03.001
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    As the floral emblem of Qingdao,Camellia japonica(Naidong) is a precious ornamental species during the winter of the north, but the quantity of C. japonica(Naidong) cultivars cannot meet requirements for application in landscape at present, so it is necessary to enlarge propagation by the means of tissue culture. In order to establish the regeneration system of C. japonica(Naidong), different factors affected on callus induction and plantlet regeneration were studied from mature cotyledons of C. japonica (Naidong). The optimum timing of sterilization in C. japonica(Naidong) seeds was soaked for 8 min in the 2% NaClo, callus induction rate was 100%. The optimal hormone combination of cotyledon callus subculture was 0.5 mg·L-1 NAA+1.5 mg·L-1 6-BA, the multiplication rate was 4.06. After 2-time subcultured, the callus was transferred to MS+1.0mg·L-1 NAA+10.0 mg·L-1 6-BA for differentiation of adventitious buds, the rate of adventitious bud differentiation was 10.7%. The shoot base was immersed in 500 mg·L-1 IBA solution for 70 min, then transferred to 1/2MS, the rate of rooting could reach 40%. Regeneration system from mature cotyledons of C.japonica(Naidong) by inducing callus, adventitious shoots and roots was established, which would provide technical support for the vegetative production and innovation of its germplasm resources.

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    Sporogenesis and Gametogenesis on Endangered Dayaoshania cotinifolia
    Hui-Ai SHI, Fa-Ju CHEN, Hong-Wei LIANG, Yu-Bing WANG
    Bulletin of Botanical Research    2021, 41 (3): 329-335.   DOI: 10.7525/j.issn.1673-5102.2021.03.002
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    To explore the possible endangered embryology causes of Dayaoshania cotinifolia , the paraffin section method to observe mega-microsporogenesis, development of female and male gametophyte, and embryogenesis was employed. The main results as follows: The ovules were anatropous, single integument, thin nucellus, and with an integument tapetum. The megaspore mother cell had undergone meiotic divisions and developed into a linear tetrad. The chalazal megaspore of the tetrad was functional one and developed into a Polygonum type embryo-sac, another 3 megaspores degenerated. The anther was tetraposiform, the anther wall was composed with an epidermis, endothecium, two-layered middle layer, and a glandular tapetum. In the meiosis of microsporocyte, cytokinesis was a modified simultaneous type. The tetrads were tetrahedral and mature pollen grain was two-celled. The development of endosperm was cellular type and exhausted during embryo development. The process of the mega-microsporogenesis and development of female and male gametophyte of D.cotinifolia were normal, which was not the cause of its endangerment. The embryo developed to the globular embryonic stage at ripe fruit, the morphological after-ripening process of the embryo could be completed after dormancy. The undeveloped embryo might be one of the reasons for the difficulty of natural regenerates naturally.

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    Stability Analysis and Genetic Variation in Seedling Growth of 51 Larix olgensis Clones from Different Sites
    Fang WANG, Lu-Ping JIANG, Qin-Hui ZHANG, Zhi-Min LU, Yu-Chun YANG, Jian-Qiu ZHANG, Xi-Yang ZHAO
    Bulletin of Botanical Research    2021, 41 (3): 336-343.   DOI: 10.7525/j.issn.1673-5102.2021.03.003
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    Larix olgensis is one of the most important fast-growing timber species in northeast China. In this study, 51 L. olgensis clones from different stites were used as materials, the height and ground diameter of the 2-year-old seedlings were investigated and analyzed on the genetic stability and variation respectively. The results showed that the differences of seedling height and ground diameter for L. olgensis were extremely significant(P<0.01) within sites, clones or sites×clones respectively. The phenotypic variation coefficient of seedling height and ground diameter at different sites were from 23.89%-36.82%; and the repeatability of the ground diameter was 0.172 at Siping, but the repeatability of the rest sites were from 0.573-0.891 respectively. Genetic stability analysis results showed that 28 clones including LO 13 had the higher genetic stability respectively. The results of correlation analysis showed that there was an extremely significant positive correlation between seedling height and ground diameter. 10 clones with good comprehensive performance were selected from different site using the method of comprehensive evaluation of multiple traits with a selection rate of 20%; Both clones LO 10 and LO 26 were selected as elite clones showing the excellent performance and the stronger genetic stability in the different cite respectively.

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