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    Bulletin of Botanical Research 2020 Vol.40
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    Two New Species of Aconitum(Ranunculaceae) from Sichuan
    WANG Wen-Tsai
    Bulletin of Botanical Research    2020, 40 (1): 1-4.   DOI: 10.7525/j.issn.1673-5102.2020.01.001
    Abstract605)      PDF(pc) (498KB)(468)       Save
    Two species of the genus Aconitum(Ranunculaceae), A.tenuigaleatum and A.laxifoliatum, are described as new from western Sichuan Province. The diagnostic dlifferences between them and their allices are given respectively.
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    Three new species of Thalictrum(Ranunculaceae) from Sichuan Province
    WANG Wen-Tsai
    Bulletin of Botanical Research    2020, 40 (1): 5-9.   DOI: 10.7525/j.issn.1673-5102.2020.01.002
    Abstract630)      PDF(pc) (770KB)(429)       Save
    Three species of the genus Thalictrum(Ranunculaceae), T.kangdingense, T.tenuicaule and T.jiulongenseare described as new from western Sichuan Province. The diagnostic differences between them and their allices are given respectively.
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    Cyclobalanopsis reclinatocaulis,A New Species of Fagaceae from Fujian Province
    LIN Qin-Wen, ZHONG Yi-Xin, LIN Mu-Mu
    Bulletin of Botanical Research    2020, 40 (1): 10-14.   DOI: 10.7525/j.issn.1673-5102.2020.01.003
    Abstract544)      PDF(pc) (452KB)(435)       Save
    Cyclobalanopsis reclinatocaulis M.M.L, a new species from western Fujian, China, is described and illustrated. Based on morphological characters, it is similar to Cyclobalanopsis grauca(Thunb.) Oerst., but differs in its winter budsreddish-brown; leaf blade subleathery, often elliptic, ovate-elliptic, abaxially often dark gray when dry, margin often shallow serrations; cupule bracts in 8-10 rings, crowded; nut obovoid to obovoid-ellipsoid, apex somewhat depressed, dense pale-pubescent; stylopodium slightly broad 3-4 mm in diam., 7-8-ringed.
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    Notes on the Geographical Distribution of Delphinium mollipilum(Ranunculaceae)
    REN Lu-Ming, CHEN Xue-Lin
    Bulletin of Botanical Research    2020, 40 (1): 15-17.   DOI: 10.7525/j.issn.1673-5102.2020.01.004
    Abstract556)      PDF(pc) (479KB)(407)       Save
    Point out that the inclusion of Gansu Province in the distribution of Delphinium mollipilum W.T.Wang are not correct, a species actually occurring only in Helan Mountain.
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    Geographical Characteristics of Orchidaceae Plants in Qinling Mountains
    GAO Xu-Zheng, KANG Yong-Xiang, ZHANG Li-Li, KANG Yue-Cheng, GUO Ming
    Bulletin of Botanical Research    2020, 40 (1): 18-28.   DOI: 10.7525/j.issn.1673-5102.2020.01.005
    Abstract776)      PDF(pc) (2019KB)(412)       Save
    The Qinling Mountains is one of the most abundant and highly differentiated areas with wild Orchidaceae plants in China. However, the systematic arrangement of Orchidaceae plants in this area has not yet been seen. The aim of this study is to provide certain basic data for the protection of plant biodiversity. On the basis of references and sorted out relevant data, we did supplementary investigation and collected specimen in the key areas of Orchidaceae plants in Qinling Mountains, made a list of Orchidaceae plants in Qinling Mountains, and analyzed their genera and species characteristics, floristic composition and spatial distribution pattern. There were 149 species of Orchidaceae plants belonging to 52 genera in Qinling Mountains, of which 70 species were endemic to China, accounting for 47% of the total species; life forms were terrestrial, epiphytic, semi-epiphytic and saprophytic, mainly terrestrial; Orchidaceae plants in Qinling Mountains were mainly distributed at altitudes of 800-3 400 m; hotspots were Southwest Longnan Mountains, Taibai-Foping Mountains and Funiu Mountains. At the same time, we concluded the floristic characteristics of Orchidaceae plants in Qinling Mountains:(1)rich in species and remarkable temperate traits; (2)ancient origin and coexistence of new and old species; (3)abundant endemic components; (4)accumulation of rare and endangered protected plants; (5)richness varies with habitats.
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    Effects of Shading on Physiological Characteristics and Ultrastructure of Mesophyll Cell of Emmenopterys henryi Leaves
    LI Dong-Lin, JIN Ya-Qin, CUI Meng-Fan, HUANG Lin-Xi, PEI Wen-Hui
    Bulletin of Botanical Research    2020, 40 (1): 29-40.   DOI: 10.7525/j.issn.1673-5102.2020.01.006
    Abstract475)      PDF(pc) (2403KB)(128)       Save
    We studied the effects of light on physiological characteristics and ultrastructure of mesophyll cells of Emmenopterys henryi leaves with four different light treatments:total light(L 0), transmittance 60%(L 1), transmittance 25%(L 2) and transmittance 10%(L 3). The results showed that:Shading affected the leaf water content(LWC) of and leaf morphology. With the increase of shading intensity, LWC increased, single leaf area(SLA) increased, single leaf dry weight(SLW d) and lamina mass per unit area (LMA) decreased, and lamina area per unit mass (LAM) increased. Shading had a significant effect on photosynthesis of seedlings. The diurnal changes of both P n and T r showed obvious bimodal curves for L 0 and L 1, and the "noon break" appeared at 12:00, and the change of both L 2 and L 3 was relatively mild, the phenomena werenot obvious. The variation of G s was basically consistent with that of P n. The order of daily average of both P n and T r was L 1 > L 0 > L 2 > L 3. Therefore, the moderate shading is beneficial to the improvement of photosynthesis efficiency of E.henryi seedlings. Chlorophyll and carotenoid of leaves were increased under shading condition, and the contents of both chlorophyll and carotenoid were increased progressively with the increase of shading intensity. Long-term shading improved the light capture ability of leaves. Under shading condition, the change of MDA was more moderate, the activity of SOD and POD increased first and then decreased, but the MDA content under shading was always lower than that in full light. Soluble sugar showed a similar change. Under full light, the chloroplast and cell wall were distributed parallel to each other, and the thylakoid was evenly arranged, while under shading condition, the number of chloroplasts in cells increased obviously, the percentage of chloroplasts in the whole cell increased significantly, the starch granules increased, the arraying of thylakoids was close and the stacking degree increased, which improved the photosynthetic efficiency of the cells under low light. The seedlings can maintain normal physiological activities under proper shading, but the effective radiation intensity must be above 60% of natural light intensity.
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    Effects of Simulated Nitrogen Deposition on the Growth and the Content of Non-structure Carbohydrate of Sinocalycanthus chinensis Seedlings
    ZHAO Zhe, JIN Ze-Xin
    Bulletin of Botanical Research    2020, 40 (1): 41-49.   DOI: 10.7525/j.issn.1673-5102.2020.01.007
    Abstract574)      PDF(pc) (1655KB)(122)       Save
    With the seedlings of Sinocalycanthus chinensis, four simulated nitrogen deposition levels, including control(CK, 0 g·m -2·a -1), lower nitrogen(N1, 2 g·m -2·a -1), medium nitrogen(N2, 8 g·m -2·a -1), and high nitrogen(N3, 32 g·m -2·a -1). After 1 year, the growth, biomass allocation and the content of the non-structure carbohydrate of S.chinensis seedlings among the different treatments were measured. The results showed that:the plant height and base diameter of the seedling showed a trend of increasing first and then decreasing, with medium nitrogen treatment being the highest. The leaf and shoot biomass of the seedlings showed a trend of gradual increase, while the stem, root, total biomass of the seedlings, and root biomass ratio and root shoot ratio showed a trend of increasing first and then decreasing. The leaf biomass ratio showed a trend of decreasing first and then increasing with the increase of nitrogen concentration. The mean perimeter of the maximum leaf, the mean length and area of the leaf was the highest in the treatment of N2. The width/length ratio of the maximum leaf was the highest in the treatment of N3. The specific leaf area in the treatments of N2 and N3 were significantly lower than that in treatment CK. The contents of starch and non-structural carbohydrates in the leaves were highest in the treatment of N2, while those in the stem were the lowest in the treatment of N3. In conclusion, nitrogen deposition showed negative effects on the growth and physiological traits of S.chinensis seedlings and the most significant effect were found in the treatment of N2.
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    Establishing Tissue Culture and Regeneration System of Scutellaria regeliana
    ZHANG Jing, NIU Zhe, FAN Wei-Fang, GAO Peng-Fei, WU Jian-Hui
    Bulletin of Botanical Research    2020, 40 (1): 50-57.   DOI: 10.7525/j.issn.1673-5102.2020.01.008
    Abstract479)      PDF(pc) (1494KB)(68)       Save
    We studied the effects of different disinfectant treatments and different plant growth hormone ratios on axillary bud induction, callus induction, cluster bud differentiation, proliferation, rooting and transplantation of Scutellaria regeliana stem segments with the stem segments of S.regeliana as explants. The best disinfection method was 0.1% HgCl 2 for 5 min, with the lowest pollution rate of 8.25%; the best medium for inducing axillary buds was MS+1 mg·L -1 6-BA+1 mg·L -1 NAA, with the induction rate of 73.66%; the best medium for inducing callus was MS+1 mg·L -1 6-BA+1 mg·L -1 2-4D, with the induction rate of 91.33%; and the best medium for inducing callus differentiation was MS+1 mg·L -1 6-BA+0.5 mg·L -1 NAA, with the differentiation rate of 44.71%. The optimum medium for bud multiplication was MS+1 mg·L -1 6-BA+0.5 mg·L -1 NAA, and the multiple of bud multiplication was 5.85; the optimum medium for rooting was 1/2MS+0.2 mg·L -1 IBA, and the rooting rate could reach 74.07%; the volume ratio of vermiculite:perlite:pearlite:garden soil was 1:1:3, the highest survival rate of transplantation was 79.24%, and the plant grew vigorously. In this study, the regeneration system of S.regeliana was established to provide theoretical support in the development and application of S.regeliana wild resources on the basis of proper protection.
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    Cytosolic Ca 2+ Involved in Seed Germination of Naked Oat Enhanced by Exogenous H 2S under Saline-Alkali Stress
    LIU Jian-Xin, OU Xiao-Bin, WANG Jin-Cheng
    Bulletin of Botanical Research    2020, 40 (1): 58-65.   DOI: 10.7525/j.issn.1673-5102.2020.01.009
    Abstract499)      PDF(pc) (1455KB)(81)       Save
    Hydrogen sulfide(H 2S) is a new gas signal molecule and calcium(Ca 2+) is an important second messenger. Both play important roles in plant adversity response, respectively. The experiment was conducted to clarify the role of cytosolic Ca 2+ in germination of crop seed promoted by exogenous H 2S under saline-alkali stress. Naked oat( Avena nude) was selected as experimental material which was cultured in petri dishes in incubator. Mixed saline-alkaline solution(NaCl:Na 2SO 4:Na 2CO 3:NaHCO 3 with the mole ratio of 12:8:1:9) was used to simulate the saline-alkaline stress of naked oat growing areas in Gansu Province, extracellular Ca 2+ chelator ethylene glycol-bis-(2-aminoethylether)-N, N, N', N'-tetraacetic acid(EGTA), plasma membrane Ca 2+ channel blocker lanthanum chloride(LaCl 3), tonoplast Ca 2+ release inhibitor ruthenium red(RR), and endoplasmic reticulum calcium pump blocker thapsigargin(Thaps) were combined with sodium hydrosulfide(NaHS, a donor of H 2S), respectively, and distilled water treatments were set as control. Seven germination indexes including germination potential, germination rate, germination index, vigor index, average germination speeds, radicle length, and plantule length of naked oat under control, saline-alkaline stress and co-treatments were determined. The membership function analysis was performed to comprehensively evaluate the effect of cytosolic Ca 2+ on seed germination of naked oat enhanced by H 2S under saline-alkali stress. The germination potential, germination rate, germination index, vigor index, average germination rate, radicle length and plantule length of naked oat seeds decreased significantly with the increase of saline-alkaline concentration. Compared with the control, the 15-75 mmol·L -1 salt-alkaline stresses resulted in a significant decrease in the membership function comprehensive evaluation value D, D value decreased by 73.1% under 30 mmol·L -1 saline-alkali stress. The 100-1 000 μmol·L -1 NaHS increased the D value to varying degrees, and 100 μmol·L -1 NaHS had the most significant mitigation effect on the decrease of D value under 30 mmol·L -1 salt-alkaline stress. This mitigation effect of NaHS was weakened by adding EGTA, LaCl 3 and RR, respectively; however, Thaps had no significant effect on the alleviating effect of NaHS to the decrease of D value under 30 mmol·L -1 salt-alkaline stress. These results show that Ca 2+ involved signal transduction in promoting of naked oat seed germination induced by H 2S under saline-alkali stress, and cytosolic Ca 2+ mainly comes from the influx of extracellular Ca 2+ and the release of Ca 2+ in vacuoles.
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    Optimization of SCoT-PCR System and Screening of Effective Primers for Castanopsis wenchangensis,an Extremely Small Population Endemic to Hainan Province
    YANG Li-Rong, SUN Xiu-Xiu, CHEN Jia-Li, YUN Yong, CHEN Xuan, ZHENG Dao-Jun
    Bulletin of Botanical Research    2020, 40 (1): 66-72.   DOI: 10.7525/j.issn.1673-5102.2020.01.010
    Abstract529)      PDF(pc) (1308KB)(62)       Save
    The effects of DNA, primers and dNTPs, Taq DNA polymerase on the SCoT-PCR amplification results of Castanopsis wenchangensis were analyzed by single factor test and orthogonal test, and the SCoT-PCR system of C.wenchangensis was optimized. The polymorphism primer combination was screened based on the established reaction system, which provided the conditions for the study of SCoT molecular markers in the study of C.wenchangensis germplasm resources. The results showed that the influence of each factor on SCoT-PCR amplification was DNA > dNTPs > primer > Taq enzyme, and the optimum reaction system was as follows:when the total system was 20 μL, DNA content was 2.5 ng, primer concentration was 0.8 μmol·L -1, dNTPs concentration was 0.2 mmol·L -1, Taq content was 1 U, and the amplified products were clear and stable. In addition, 15 primers with good polymorphism and suitable for C.wenchangensis were screened out from 80 SCoT primers by this system, which provided theoretical basis and technical support for the related research of C.wenchangensis and other plants of Fagaceae by SCoT molecular marker technique in the future.
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    Optimization Peony Seed Oil Extraction Process at Suitable Temperature and Physicochemical Property Analysis
    LI Wen-Gang, SUN Xiao-Li, ZU Yuan-Gang, ZHAO Xiu-Hua
    Bulletin of Botanical Research    2020, 40 (1): 73-78.   DOI: 10.7525/j.issn.1673-5102.2020.01.011
    Abstract534)      PDF(pc) (1132KB)(71)       Save
    In order to improve the yield and the quality of peony seed oil extraction process, and provide technical and theoretical basis for further exploring the feasibility of peony seed oil cold pressing, we applied box-behnken experimental design to optimize the peony seed oil extraction process at suitable temperature. The fitting degree of the model was high and the experimental error was small. Finally, the optimums of peony seed oil in extraction process at suitable temperature were the pressure 4.6 MPa, feed rate 1 600 g·min -1, temperature 73℃, and moisture content 4.6%. The maximum oil yield and residual oil rate were 23% and 7.02%, respectively. The unsaturated fatty acid content was 90.36% at the optimal preparation conditions, and the content of linolenic acid, linoleic acid, oleic acid, palmitic acid and stearic acid was 40.71%, 27.25%, 22.40%, 7.23% and 1.60%, respectively. The acid value and peroxide value of peony seed oil were 2.61 mg·g -1 and 1.56 mmol·kg -1, respectively. All the test results are higher than the requirements of the peony seed oil quality standard "Standard of the People's Republic of China for Food Industry LS/T 3242-2014".
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    Identification of Key Amino Acid Sites in AtGDPD-Like3 with Role in Arabidopsis Root Hairs
    WANG Shuang, CHENG Yu-Xiang, XIA De-An
    Bulletin of Botanical Research    2020, 40 (1): 79-84.   DOI: 10.7525/j.issn.1673-5102.2020.01.012
    Abstract341)      PDF(pc) (1157KB)(98)       Save
    AtGDPD-Like3 encode glycerophosphodiester phosphodiesterase-like(GDPD) proteins and shv3, its family gene AtGDPD-Like3 mutant, has serious root hair defects. To identify key amino acid sites in AtGDPD-Like3, we constructed S 538A, V 556A and D 628A single site mutation of AtGDPD-Like3, which were expressed in atgdpdl3 mutants, respectively. The results showed that AtGDPD-Like3 with V 556A or D 628A mutation could completely restore root hairs growth defects of atgdpdl3, whereas, AtGDPD-Like3 with S 538A mutation partially restored root hairs growth defects. These indicate that Ser 538 is the key amino acid site in AtGDPD-Like3, which affects function of its protein. Meanwhile, it implies other key amino acid sites in AtGDPD-Like3. The results of this study would lay a foundation for further exploring the functional mechanism of AtGDPD-Like3 protein.
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    Effects of Extracellular adenosine-5'-triphosphate on Cellular damage, H 2O 2 Content,and Activities of H 2O 2 Detoxifying Enzymes under Lead Stress
    CAO Jia-Xin, DA Meng-Ting, PANG Hai-Long, JIA Ling-Yun, FENG Han-Qing
    Bulletin of Botanical Research    2020, 40 (1): 85-92.   DOI: 10.7525/j.issn.1673-5102.2020.01.013
    Abstract546)      PDF(pc) (1402KB)(165)       Save
    The extracellular adenosine-5'-triphosphate is an important signaling molecule for plant cells. By using the tobacco( Nicotiana tabacum L.cv.Bright Yellow-2) cell suspension cultures, we studied the effects of extracellular adenosine-5'-triphosphate on the cellular damage, H 2O content, and the activities of H 2O detoxifying enzymes underlead stress. With the increase of concentrations of Pb(NO 3) 2(from 30-400 μmol·L -1), the levels of the extracellular adenosine-5'-triphosphate were decreased, whilethe levels of the intracellular adenosine-5'-triphosphate and cellular damage were increased. The content of H 2O 2 and the activity of the CAT(catalaseenzyme) were increased and reached their maximum at 200 μmol·L -1Pb(NO 3) 2, while the activityof the POD(peroxide enzyme) was decreased, with the increase of concentrations of Pb(NO 3) 2. Compared with the cells under lead(200 μmol·L -1) stress, the lead-stressed cells treated with exogenous adenosine-5'-triphosphate(30 μmol·L -1) had lower levels of cellular damage, H 2O 2 content, andthe activity of CAT, but had higher level of the activity of POD. Therefore, the cellular damage, H 2O content, and the activities of H 2O detoxifying enzymesunderlead stress could beregulated by extracellular adenosine-5'-triphosphate.
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    Transcriptome of Single and Double Tepals of Clematis patens
    XIONG Yang-Yang, LUO Lin-Li, ZHAO Shi-Hao, ZHOU An-Long, WANG Jin
    Bulletin of Botanical Research    2020, 40 (1): 93-105.   DOI: 10.7525/j.issn.1673-5102.2020.01.014
    Abstract527)      PDF(pc) (2185KB)(115)       Save
    With the typical Clematis patens ‘Vyvyan Pennell’ which can open either single petal or double petals as the research object, we chose high throughput sequencing to splice and annotate the sequencing data of three different petal types(single petal, semi-double petal and double petal) on the same plant at the same period of C.patens ‘Vyvyan Pennell’, then selected differentially expressed genes to real-time fluorescence quantitative PCR. There were 13.8 GB raw data. A total of 3 075 differentially expressed genes(DEG) were obtained from the three transcriptome libraries by paired comparison, including 649 upregulated DEG and 605 upregulated DEG in the comparison between single and semi-double petals samples(A vs B). The comparison of semi-double flowers with double flowers(B vs C) included 1 046 up-regulated DEG and 721 down-regulated DEG. There were 1 129 upregulation DEG and 859 upregulation DEG in the comparison between single and double flowering(A vs C). There were 134 differentially expressed genes coexisting under three different perianth. By gene functional annotation, 26 genes that might be related to the double petal trait were screened out from the total DEG for cluster analysis, and 10 target genes were randomly selected for fluorescence quantitative PCR verification. PCR results showed that the expression levels of these genes were significantly different in the three floral envelope types of the same plant at the same time of C.patens ‘Vyvyan Pennell’. Finally, the key genes associated with double tepals Clematis conversion were screened out, including MADS-BOX genes PMADS1, AP3, FRUITFULL and FLC. Auxin reactive protein IAA9, auxin input vector, abscisic acid 8' hydroxylase, indoleacetic acid-induced protein ARG7, etc. This study provides basic data and theoretical basis for exploring the molecular mechanism of Clematis double flower.
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    Structural and Functional Analysis of R2R3 MYB Transcription Factor HbMYB88
    FAN Song-Le, WANG Ji-Kun, XIE Gui-Shui, WANG Meng, WANG Li-Feng
    Bulletin of Botanical Research    2020, 40 (1): 106-116.   DOI: 10.7525/j.issn.1673-5102.2020.01.015
    Abstract1142)      PDF(pc) (1870KB)(199)       Save
    The R2R3-MYB transcription factors are involved in biological processes, such as plant growth and development, hormone signaling transduction, and stress response, etc. To explore the structure and functions of the MYB family members in the rubber tree, we cloned the full-length cDNA of HbMYB88 from the leave of rubber tree Reyan 73397(RY73397). It is 1 848 bp in length, containing a 1 440 bp ORF which encodes 479 amino acid residues. The deduced amino acid sequence of HbMYB88 contains two SANT conserved domains, has HTH tertiary structure, share highly similarity to Arabidopsis AtMYB88, AtMYB124. AtMYB88 and AtMYB124, not divided into subfamilies, are related to drought stress response, etc. By qRT-PCR analysis, HbMYB88 was mainly expressed in the stem and flower of rubber tree, whilst its expressions in root, leaf, bark and latex were extremely low. The expression level of HbMYB88 in tissue culture seedling leave was significantly upregulated under the treatment of hydrogen peroxide(H 2O 2), and abscisic acid(ABA). These indicated that HbMYB88 has a relationship with stress response such as drought in rubber trees, which providing a foundation for further study of its structure and function.
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    Temporal and Spatial Distribution of Florescence and Capsule Development of Introduced Eucalyptus cloeziana in Sichuan Province
    HUANG Zhen, YANG Yong-Zhi, CHEN Zhi, LI Jia-Man, WANG Li-Hua, GUO Hong-Ying
    Bulletin of Botanical Research    2020, 40 (1): 117-124.   DOI: 10.7525/j.issn.1673-5102.2020.01.016
    Abstract411)      PDF(pc) (1572KB)(117)       Save
    In order to explore the regularity of florescence and capsule development of Eucalyptus cloeziana, the flowering stages, were studied the blossom buds and capsules development of 7-year trees from 5 provenances which were introduced in Yibin of Sichuan. The results showed that:(1)Most of the blossom buds distributed in the lower canopy, few blossom buds germinated directly from the main stem, the buds on the same branch randomly opened. (2)The time gaps of the initial flowering stage from the same provenance was no more than 5 d, the time gaps of the determined stage from the same provenance was no more than 10 d, flowering stage last for 41-62 d. The florescence of 5 E.cloeziana provenances were from mid to late December to early May of the next year, so there was asynchronous flowering period from different provenances. (3)There was a significant difference in the flowering rate of provenances, No.72 had the highest rate of flowering, No.74 had not starting reproductive growth yet. (4)The external characteristics of capsule development can be divided into four stages:early development stage, mericarps formation period, mericarps opening period and seed dispersal period.
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    Seed Vigor of Different Half-sib Families in Betula schmidtii
    LIN Lin, GAO Yu-Ting, CHENG Fu-Shan, XIN Ben-Hua, WANG Gui-Chun, XIA Fu-Cai, MU Huai-Zhi
    Bulletin of Botanical Research    2020, 40 (1): 125-132.   DOI: 10.7525/j.issn.1673-5102.2020.01.017
    Abstract454)      PDF(pc) (1267KB)(57)       Save
    Seed vigor can be applied to predict the growth and adaptability of sapling, which is usually taken as an important evaluation index for seed trees. Betula schmidtii as one of the hardest tree species in northeast forest area, the seed vigor has not been reported. The seed vigor of different families was compared, and the seed trees were preliminarily evaluated, which could provide reference for selecting B.schmidtii superior family. Nested ANOVA test, heritability estimation, multiple comparison, correlation analysis on thousand seed wright, germination rate, germination energy, germination index, vigor index, emergence rate and robust rate were conducted by taking 30 half-sib families from Jiaohe, Ji'an and Hunchun in B.schmidtii, and principal component analysis was used to evaluate comprehensively the tested families. There was significant difference in robust rate and extremely significant difference in vigor index among seed sources, and there were significant or extremely significant family effects for all studied traits. Except emergence rate and robust rate, the family heritability in each of the other traits was higher than 0.68. The positive correlation in all studied traits was significant or extremely significant in different families. Based on the results of principal component analysis, JH6, JA1, JA7, JH10, JH3, HC1 and HC10 were identified as the superior families. On average, the seed wright, germination rate, germination energy, germination index, vigor index, emergence rate and robust rate of superior families were 3.87%, 30.29%, 30.22%, 31.99%, 58.22%, 11.88% and 30.60% higher than those of the 30 families, respectively. We recommend the seed trees of these superior families as preferred seed trees in the future.
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    Screening of Excellent Ectomycorrhizal Fungi-tree for Drought Resistant with Pinus sylvestris var. mongolica
    ZHAO Min, HAO Wen-Ying, NING Xin-Zhe, HAO Long-Fei, YAN Hai-Xia, MU Ya-Nan, BAI Shu-Lan
    Bulletin of Botanical Research    2020, 40 (1): 133-140.   DOI: 10.7525/j.issn.1673-5102.2020.01.018
    Abstract425)      PDF(pc) (1512KB)(71)       Save
    In order to screening the excellent ectomycorrhizal fungi-tree for drought resistant with Pinus sylvestris var. mongolica in Honghuaerji. Five dominant ectomycorrhizal fungi strains in P.sylvestris var. mongolica forest were used as inoculants to artificially inoculate five-month-old seedlings of P.sylvestris var. mongolica. The mycorrhizal infection and morphology were observed after eight months, and the mycorrhizal seedlings growth and physiological and biochemical indexes were detected under non-drought stress and drought stress conditions. The results show that five native strains could successfully infect P.sylvestris var. mongolica and form typical ectomycorrhiza. All growth indexes of mycorrhizal seedlings were significantly higher than CK under drought stress condition( P<0.05). It was the highest colonization rate, height, aboveground biomass, underground biomass, and root-shoot rate that seedlings were inoculated with Suillus bovinus. Ectomycorrhizal symbionts can improve drought tolerance of P.sylvestris var. mongolica by increasing SOD and POD activities and reducing MDA content. The wilting time of all inoculated seedlings was delayed compared with the control under drought stress. The longest delay was the inoculation of S.bovinus, which was 96.3 h later than the control. In addition, inoculation treatment could significantly prolong the critical lethal time of the host, especially the inoculation of S.bovinus could prolong 113.8 h. Thererfore, S.bovinus and P.sylvestris var. mongolica were an ideal combination of drought-resistant ectomycorrhizal fungi-tree.
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    Overexpression of K Transporter Gene trkH in Enhancing K Nutrition in Maize
    DING Bao-Juan, AN Li-Jia, SU Qiao
    Bulletin of Botanical Research    2020, 40 (1): 141-147.   DOI: 10.7525/j.issn.1673-5102.2020.01.019
    Abstract492)      PDF(pc) (1263KB)(49)       Save
    Potassium is a necessary nutrient for plant growth and development, which is involved in various physiological and biochemical processes. Potassium deficiency seriously affects the quality and yield of maize. To improve K + nutrition of maize by genetic modification is one of the effective approaches to solve this problem. The bacterial K + transporter gene trkH was cloned from marine microbial metagenomic DNA, and its function was verified in yeast and tobacco. To further analyze the function of trkH gene in maize, the trkH gene was transferred into Hi-Ⅱ variety by Agrobacterium-mediated transformation method, and 21 independent baster resistant seedlings were obtained. By PCR, the trkH gene was successfully introduced into maize genome. By Bar strip test, the Bar gene was expressed at the protein level. Partial T 0 transgenic plants were crossed with the maize backbone inbred line PH6WC and eight hybrid progeny were obtained. By semi-quantitative RT-PCR, trkH gene was expressed at the transcriptional level. Herbicide was sprayed at the three-leaf and one-heart stage, and L3, L5 and L7 lines with a ratio close to 1:1 were selected for PCR detection. Chi-square was performed with PCR data and the results indicate that the ratio is in accordance with Mendelian segregation law. K + depletion experiment showed that overexpression of trkH gene improved K + absorption of L3, L5 and L7 transgenic seedlings, which would lay the foundation of breeding new maize variety with high efficient potassium nutrition.
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    Optimization of Ultrasound Assisted Acid Hydrolysis Process for Extracting Vitexin from Crataegus pinnatifida Bge. var. major N.E.Br. Leaf
    XING Wen-Miao, CHEN Min, FENG Chang-Yin, MENG Xian-Ming, TIAN Meng-Fei, MU Fan-Song, LUO Meng
    Bulletin of Botanical Research    2020, 40 (1): 148-152.   DOI: 10.7525/j.issn.1673-5102.2020.01.020
    Abstract484)      PDF(pc) (875KB)(90)       Save
    Ultrasound-assisted extraction of vitexin from Crataegus pinnatifida Bge. var. major N.E.Br. leaf was optimized by orthogonal design. Through the single factor experiments of six factors the HCl content, extraction time, solid-liquid ratio, ultrasonic power, ultrasonic temperature and ethanol concentration, the regularity trend between them and the vitexin extraction yield was studied. The optimization experiment with orthogonal design was conduted to study the effects of HCl content, extraction time, solid-liquid ratio and ultrasonic power on the extraction yield of C.pinnatifida Bge. var. major N.E.Br. and the optimal extraction process were determined. When the HCl content was 2 mol·L -1, the extraction time was 40 min, the solid-liquid ratio was 1:20, the ultrasonic power was 500 W, the ultrasonic temperature was 50℃, and the ethanol concentration was 50%, the best result was obtained and the yield was 2.603 mg·g -1. The HCl content was significant, and the order of influence of each factor was:HCl concentration > ultrasonic power > extraction time > solid-liquid ratio.
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    Rapid Propagation of Virus-free “Dan Mei 1” Strawberry Using Tissue Culture Technology
    WANG Xin, GAI Qing-Yan, JIAO Jiao, FU Yu-Jie, LIU Jing, WANG Zi-Ying
    Bulletin of Botanical Research    2020, 40 (1): 153-160.   DOI: 10.7525/j.issn.1673-5102.2020.01.021
    Abstract682)      PDF(pc) (1589KB)(62)       Save
    The superior "Dan Mei 1" strawberry variety that is suitable for growing in Northeast China were selected as the research object, and the tissue culture technology for detoxification and rapid propagation of strawberry was carried out systematically. The effects of different plant hormones on adventitious bud induction, proliferation and rootage were investigated using the stem tip of "Dan Mei 1" strawberry as explants. Also, the effects of transplanting medium on the domestication survival of strawberry plantlets were investigated. The results showed that MS with 1 mg·L -1 6-BA+0.1 mg·L -1 2,4-D was suitable for adventitious bud induction. The best medium for adventitious bud proliferation was MS with 1 mg·L -1 6-BA+0.3 mg·L -1 NAA. And, MS with 0.4 mg·L -1 IBA was the best medium for adventitious bud rootage. Perlite:vermiculite:nursery soil with the ratio of 1:2:1 was the best transplanting medium for the domestication of strawberry plantlets. Overall, this study provided a theoretical foundation for the large-scale propagation of the virus-free "Dan Mei 1" strawberry variety.
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    Pollinia Development in a New Species of Pennilabium yunnanense(Orchidaceae)
    LI Lu, ZHANG Jin, LI Chu-Ran, WANG Yan-Ping, TAN Qing-Qin, LUO Yan
    Bulletin of Botanical Research    2020, 40 (2): 161-171.   DOI: 10.7525/j.issn.1673-5102.2020.02.001
    Abstract899)      PDF(pc) (2228KB)(1391)       Save
    Pennilabium J. J.Sm. is a newly recorded genus in China, including two species founded in Yunnan and Hainan. One of them is a new species of P.yunnanense. In this paper, floral morphology and pollina development in P.yunnanense were investigated. The column of a mature flower is short without column foot. Pollinarium unit is composed of two subglobose pollinia, stipe and viscidium. In the early stage, anther primordium is differentiated into a pair of lateral juxtaposed thecae. At the stage of microsporocyte, an incomplete sterile septum is formed adjacent to the inner anther wall between two thecae. During the microsporogenesis, the sterile septum becomes gradually degraded, resulting two pollinia with pore toward the inner anther wall disorganized. The hypodermal sporogenous cells produce 4-layered anther wall by periclinal divisions, including epidermis, endtothecium, middle layer, and a tapetum, which was of monocotyledonous type. Tapetum is uninuclear and secretory. Epidermis is not persistent, which becomes degraded as well as middle layer and tapetum. Fibrous thickenings occur on the entdothecium. Simultaneous cytokinesis results in tetrahedral and isobilateral microspore tetrads. Microspores undergo a mitosis leading to 2-celled pollen tetrads. Pollen tetrads arranged tightly and formed a subglobose pollinium in each theca. The embryological data on pollinia development of P.yunnanense could provide new insight for a better understanding of biodiversity and conservation in Orchidaceae.
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    A New Understanding of the Spines in Genus Berberis: Morphology and Occurrence of Spines in Berberis thunbergii var. atropurpurea
    MIAO Qing, ZHAI Qiang, QU Bo, SHI Mian-Mian, YAN Bi-Yu, SHAO Mei-Ni
    Bulletin of Botanical Research    2020, 40 (2): 172-176.   DOI: 10.7525/j.issn.1673-5102.2020.02.002
    Abstract1283)      PDF(pc) (1018KB)(1229)       Save
    In view of the different opinions on the origin of the spines of Berberis, we studied the morphology and occurrence of the spines of Berberis by means of entity anatomy and paraffin section. The spines of Berberis descended from leaves, origins from leaf primordial, are leaf spines, not stem spines. It was clear that the spines of Berberis belong to one of the leaf metamorphosis types.
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    Photosynthetic and Stomatal Morphological Variation of Poplar Clones in Seedling Stage under Drought Stress
    QIAO Bin-Jie, WANG De-Qiu, GAO Hai-Yan, LI Zhao-Min, GE Li-Li, DING Wen-Ya, ZHAO Xi-Yang
    Bulletin of Botanical Research    2020, 40 (2): 177-188.   DOI: 10.7525/j.issn.1673-5102.2020.02.003
    Abstract827)      PDF(pc) (1918KB)(1279)       Save
    The experiment was conducted to study the changes of leaf stomatal morphological and photosynthetic characteristics of Populus spp. under different drought stress. Four poplar clones were taken as materials and investigated their photosynthetic and leaf stomatal morphological characteristics under drought stress. By ANOVA analysis, there were significant differences( P<0.01) for all traits among different source of variation in different treatment except stomatal apparatus length. With the reduction of soil relative water content, net photosynthetic rate, transpiration rate and stomatal conductance decreased, however, the intercellular CO 2 concentration increased at first and then decreased. These results suggested that that drought-induced photosynthetic inhibition was due to stomatal limitation when the relative water content of soil was above 40%; However, the main limiting factor of photosynthetic inhibition of plants under severe drought stress was due to non-stomatal limitation. Drought decreased all traits of stomatal morphological characteristics. Using the method of subordinate function analysis, the drought-tolerance of Populus psedosimonii×P.nigra ‘baicheng-1’ was better than other clones.
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    Effect of Light Quality on Seed Germination and Seeding Growth of Emmenopterys henryi
    XIAO Zhi-Peng, YIN Chong-Min, GUO Lian-Jin, WU Yuan-Rong, HU Jin-Ping, LIU Yan-Yan, ZHONG You-Chun, XUE Ping-Ping
    Bulletin of Botanical Research    2020, 40 (2): 189-195.   DOI: 10.7525/j.issn.1673-5102.2020.02.004
    Abstract908)      PDF(pc) (1195KB)(1250)       Save
    We studied the effect of light quality on seed germination and seedling growth of the endangered plant Emmenopterys henryi by setting eight light quality treatments including the 940 nm(far red), 850 nm(far red), 730 nm(far red), 630 nm(red), 610 nm(orange), 590 nm(yellow), 525 nm(green), 460 nm(blue) and a nature light as CK, and setting six light quality treatment include 730, 630, 610, 590, 525 and 460 nm, respectively. The results showed that no seed germinate under 940 and 850 nm, and only 1.33% seeds germinate under 730 nm. The germination percent under 525 nm was significantly higher than that under other treatments include natural light control, and there was no significant difference between the final germination percent under natural light and that under 610 and 590 nm. The final germination rate under 460 and 610 nm was significantly lower than that of other treatments. At 120 d, the dry weight of the seedlings was 590 nm > 630 nm > 610 nm > 730 nm > 525 nm > 460 nm. At 120 d, the dry weight of the seedlings under 590 nm was significantly higher than that under other light treatments. The relative growth rate under 630 nm was significantly higher than that of other treatments between 30-90 d, and that under 590 nm was significantly higher than that of other treatments between 90-120 d. It was significantly higher than other treatments at 460 nm between 120-150 and 150-180 d. Root mass ratio was between 0.17-0.25 on the 30th day of treatment, and with no significant difference between treatments. Root mass ratio under 460 nm was significantly higher than that of other treatments on the 30th day of treatment. At 30 d, stem mass ratio under 730 nm was significantly higher than that of other treatments. Leaf mass ratio in each treatment was between 0.53-0.68 on the 30th day of treatment, and there was no significant difference between the treatments.
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    Tissue Culture Technology of Stem Segment of Cinnamomum bodinieri var. citralifera
    XIAO Zu-Fei, WANG Ling-Ling, CAO Lu-Yao, LIAO Wen-Xuan, JIN Zhi-Nong, LI Feng, Lü Xiong-Wei, ZHANG Bei-Hong, ZHAO Jiao
    Bulletin of Botanical Research    2020, 40 (2): 196-201.   DOI: 10.7525/j.issn.1673-5102.2020.02.005
    Abstract973)      PDF(pc) (1173KB)(1179)       Save
    Cinnamomum bodinieri var. citralifera is an important tree species for landscaping, timber and oil. We studied the effects of collection season, disinfection time and hormones on tissue culture of annual branches of C.bodinieri var. citralifera. The axillary bud germination rate of stem segments from semi-lignified stem segments of C.bodinieri var. citralifera was highest with 0.1% HgCl 2 disinfected for 5 min. in the Mid-May, and the browning rate and pollution rate were low. The suitable medium for stem segment germination was MS+1.0 mg·L -1 6-BA+0.05 mg·L -1 IBA, with germination ratio 60%. The suitable medium for multiplication was MS+1.0 mg·L -1 6-BA+0.2 mg·L -1 IBA, with multiplication coefficient of 4.33, plant height of 3.67 cm and ground diameter of 1.04 mm. Adventitious roots appeared at 11 d after culture of the base of tissue culture seedlings. The suitable medium for rooting was MS+1.5 mg·L -1 IBA. The rooting rate was 75.00%, the length of roots was 5.06 cm, the number of roots was 3.50, and the diameter of roots was 0.82 mm. Tissue culture seedlings with roots more than 3 had a survival rate of more than 80%.
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    Effect of Three Growth Regulators on Rooting of Cuting of Crateva unilocularis
    ZHENG Xin-Hua, DONG Qiong, DUAN Hua-Chao, CHA Xiao-Fei, ZHOU Chun, ZHOU Zhi-Chao
    Bulletin of Botanical Research    2020, 40 (2): 202-208.   DOI: 10.7525/j.issn.1673-5102.2020.02.006
    Abstract738)      PDF(pc) (1281KB)(139)       Save
    Different kinds and mass concentrations of plant growth regulators were used to deal with the cuttings of Crateva unilocularis, in order to know the cutting mechanism and to screen out the growth regulator and concentration ratio for the propagation of the C.unilocularis, so as to provide a real and feasible scientific basis for the expansion propagation and production of C.unilocularis. By taking C.unilocularis annual branches as materials, orthogonal test was conducted with three kinds of plant growth regulator(IAA, NAA, ABT-1) in three kinds of different mass concentration, and water treatment(CK) was kept as contrast, cutting three months after the shoot growth and root traits(longest rooting rate, root volume, root length, average root length, etc.) were measured and comprehensively analyzed. The test screening of C.unilocularis cuttings, the optimal scheme for A1B2C2, namely ABT1 mass concentration of 50 mg·L -1, mass concentration of 200 mg·L -1 NAA, IAA concentration of 200 mg·L -1, the rooting rate(83.33%), average root length(24.03 mm), the longest root length(46.13 mm), the average root number(24), root effect index(19.06) for CK 2.27 times, 2.97 times and 3.03 times, 4.50 times and 12.79 times, The root system was well developed. The rooting ability of C.unilocularis can be significantly increased by mixing different plant growth regulators at the appropriate concentration, thus accelerating the growth and increasing the rooting rate of cuttings.
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    Evaluation of Photosynthetic Characteristics and Growth Traits in Teak( Tectona grandis L.f) Clones
    LIU Bing-Yu, HUANG Gui-Hua, LIANG Kun-Nan, WANG Xi-Yang, CHEN Tian-Yu, ZHOU Zai-Zhi, YANG Guang
    Bulletin of Botanical Research    2020, 40 (2): 209-216.   DOI: 10.7525/j.issn.1673-5102.2020.02.007
    Abstract605)      PDF(pc) (1327KB)(116)       Save
    This study was to analyze the genetic variation of photosynthetic physiology and growth trait among teak clones, and to select superior teak clones. The gas exchange, chlorophyll fluorescence parameters and tree height of 19 teak clones and one CK which selected from different provenances were measured based on a field testing forest, the photosynthetic characteristics and their relationships were analyzed. The gas exchange, chlorophyll fluorescence and growth traits had very significant difference among teak clones and provenances. There were abundant genetic variation in measured traits in teak clones except maximum photochemical ef?ciency of PSⅡ( F v/ F m). Photosynthetic physiology of P n had no significantly correlation with height growth at early stage. The clones 71-7 and FS3 were selected with relatively higher photosynthesis rate and growth. Emphasis should be put on the resources introduced from India in further breeding research that aims at high photosynthetic efficiency in teak. Photosynthetic physiology and field growth should be evaluated to selected superior teak clones.
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    Variation Analysis of Growth Traits and Coning Quantity of Pinus sylvestris var. mongolica Clones
    LI Jia-Qi, HAN Xi-Dong, MA Ying-Hui, LI Yue-Ji, WANG Li-Xiang, HAN Xi-Tian, LIU Zhi, LI Hai-Min, ZHAO Xi-Yang
    Bulletin of Botanical Research    2020, 40 (2): 217-223.   DOI: 10.7525/j.issn.1673-5102.2020.02.008
    Abstract544)      PDF(pc) (1115KB)(78)       Save
    In order to obtain high yield and excellent quality resources of Pinus sylvestris var. mongolica, 304 P.sylvestris var. mongolica clones from the forest tree seed orchard in Baicheng City of Jilin Province were taken as materials, growth traits(tree height, basal diameter, diameter at breast height, diameter at 3 m height, canopy, branch angle and lateral branch thickness) and coning quantity characteristics(coning quantity of 2015, 2016 and 2017) of different clones were investigated. By variance analysis, all the traits were significant different( P<0.01) except for canopy. The coefficients of phenotypic variation of all the traits ranged from 3.79%-65.22%. Repeatability range from 0.24-0.70; By correlation analysis, the existed significantly positive correlation among all the growth traits(0.181-0.896) except for lateral branch thickness with diameter at 3 m height(0.082). Most of growth traits with the different age of coning quantity was not significantly correlation By growth traits, with the selected rate by 5%, 15 clones were selected as excellent clones, the genetic gains of height, basal diameter, diameter at 3 m height, branch angle and lateral branch thickness of the selected clones were 5.47%, 4.48%, 15.18%, 11.78%, 2.38% and 6.66%, respectively. When evaluated clones by coning quantity characteristics, with the selected rate by 5%, 15 clones were selected as excellent clones, the genetic gains of coning quantity of 2015, 2016 and 2017 of the selected clones were 2.89%, 46.32% and 13.88%, respectively. This study provides materials for the selection of excellent clones of P.sylvestris var. mongolica seed orchard, and also provides a basis for the breeding of P.sylvestris var. mongolica for west of Baicheng, Jilin Province.
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    Carbon, Nitrogen and Phosphorus Contents and Their Ecological Stoichiometry Characteristics in Leaves of Casuarina equisetifolia and Ipomoea pes-caprae in the Coastal Zone of Hainan Island
    ZHANG Shu-Qi, XU Quan, YAO Hai-Rong, YANG Qiu, LIU Wen-Jie, WANG Meng
    Bulletin of Botanical Research    2020, 40 (2): 224-232.   DOI: 10.7525/j.issn.1673-5102.2020.02.009
    Abstract753)      PDF(pc) (1503KB)(134)       Save
    The experiment was conducted to study the stoichiometry characteristics of the plant leaves and their influencing factors for improving the ecological environment of the coastal zone. The Casuarina equisetifolia and the Ipomoea pes-caprae on the nearby beaches of 12 cities(counties) along the coast of Hainan Island were selected as research objects. The stoichiometry characteristics and differences of C, N and P in the leaves of two plants were analyzed to explore the different environmental factors by measuring the contents of carbon(C), nitrogen(N) and phosphorus(P) in leaves of C.equisetifolia and I.pes-caprae. And the effects of C, N, P content, C:N, C:P and N:P on the leaves of the two plants were conducted. These could find the main limiting factors affecting the growth of coastal zone plants. The results showed that the average contents of C, N and P in leaves of C.equisetifolia in Hainan Island were 399.06±20.29, 12.55±1.03, 12.55±1.03 g·kg -1, respectively. The ratios of C:N, C:P and N:P were 32.02±2.8, 420.65±121.27 and 13.10±3.47, respectively. The average contents of C, N and P in leaves of I.pes-caprae were 364.31±30.20, 12.84±1.96 and 2.06±0.64 g·kg -1, respectively. The ratios of C:N, C:P and N:P were 29.13±4.95, 198.74±79.41 and 6.92±2.69, respectively. The N content in leaves of C.equisetifolia was significantly correlated with annual mean temperature(AMT) and annual mean precipitation(AMP). The P content in leaves of C.equisetifolia was significantly positive correlated with AMP. The C:P and N:P in leaves of C.equisetifolia were significantly negatively correlated with AMP. There was a significantly positive correlation between C content in leaves of I.pes-caprae and AMT. The C:N of I.pes-caprae leaves showed a significantly negative correlation with AMP. The N content in leaves of C.equisetifolia was significantly negatively correlated with the SOC of the soil at the depth of 10-20 cm. C:N was significantly correlated with the SOC of the soil at the depth of 10-20 cm. C:P was significantly correlated with C:N of the soil at the depth of 0-10 cm. The C content in leaves of I.pes-caprae was significantly negatively correlated with the SOC of the soil at the depth of 10-20 cm. The P content were significantly correlated with the TN content, N:P of the soil at the depth of 0-10 cm, and the SOC of the soil at the depth of 10-20 cm. C:N was significantly correlated with C:N of the soil at the depth of 0-10 cm. C:P was significantly negatively correlated with TN content of the soil at the depth of 0-10 cm. C:P was significantly correlated with C:N of the soil at the depth of 0-10 cm. N:P was significantly negatively correlated with TN content of the soil at the depth of 0-10 cm. The results show that the carbon and nitrogen contents of the leaves are low in the coastal zone of Hainan Island, and N may be the main factor affecting plant growth in this area. At the same time, vegetation growth is affected by the AMT and AMP. Vegetation growth was less affected by soil nutrient content. The environment factors have differently effects on different plants.
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    PubZIP1 Gene Cloning,Subcellular Localization and Expression Pattern under Drought Stress
    LIU Xiao, YANG Jia, ZHANG Xin, MA Miao-Miao, YANG Jing-Li
    Bulletin of Botanical Research    2020, 40 (2): 233-242.   DOI: 10.7525/j.issn.1673-5102.2020.02.010
    Abstract524)      PDF(pc) (2075KB)(119)       Save
    The PubZIP1 gene was cloned from Populus ussuriensis Kom. The results of gene sequencing showed that the PubZIP1 gene has 1 083 bp coding sequences which encodes a polypeptide of 360 amino acids. By the conserved domains analysis, there are conserved BZIP and DOG1 in PubZIP1 protein. And its secondary structure consists of the 62.50% α-helix, 29.72% random coil, 5.56% extended strand and β-sheet(2.22%). Subcellular localization analysis of the PubZIP1 gene indicates it localized in the nucleus. The result of qRT-PCR analysis showed that the expression of PubZIP1 was decreased after PEG6000 treatment, while it was increased in stem and leaves, especially for leaves. It was predicted that this gene maybe mainly express and function in leaves.
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    Cloning and expression analysis of 5 ZFP genes from Poplus trichocarpa
    LI Ya-Bo, Lü Jia-Xin, TAN Bing, GAO Cai-Qiu
    Bulletin of Botanical Research    2020, 40 (2): 243-250.   DOI: 10.7525/j.issn.1673-5102.2020.02.011
    Abstract680)      PDF(pc) (1349KB)(86)       Save
    ZFPs are a class of transcription factors in plants that have a loop domain. In this study, 5 ZFP genes(named PtrZFP1-5) were identified from Populus trichocarpa, and their sequence characteristics and expression patterns were analyzed to understand the stress response function. The bioinformatics analysis of PtrZFP1-5 gene was carried out, and the expressions of 5 PtrZFP genes in root, stem and leaf of Populus under NaCl, PEG 6000 and ABA treatment were further analyzed by qRT-PCR. The number of amino acid residues encoded by PtrZFP1-5 gene was 258-338 aa, the molecular weight of the encoded protein was 27.7-37.3 kDa,and the theoretical isoelectric point was 4.87-8.61. 5 PtrZFP genes were unevenly distributed on the three chromosomes of Populus genome. The results of qRT-PCR showed that the expression patterns of 5 PtrZFP genes in roots, stems and leaves of Populus were significantly different after treatment with 0.2M NaCl, 15%(w/v) PEG6000or 100μM ABA treatment. The expressions of PtrZFP1 were all up-regulated in three kinds of stresses. The expressions of PtrZFP2 were significantly inhibited in leaves after salt, osmotic and ABA treatment. The expression changed of PtrZFP3 in roots was most obvious under drought stress. In leaves and stems, the expression level did not change significantly at most of the time points of most stress. The PtrZFP4 gene also responded to drought stress in roots and stems. The expression of PtrZFP5 gene in leaves was significantly reduced after exposure to salt and ABA stress. PtrZFP1-5 genes can respond to at least one stress treatment in one organ, but the type and mechanism of the involved stress response may be different.
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    Function of Populus trichocarpa HDA902 Gene in Response to Cold Stress
    GUAN Tao, LIU Chao, LI Kai-Long, XIA De-An, MA Xu-Jun
    Bulletin of Botanical Research    2020, 40 (2): 251-256.   DOI: 10.7525/j.issn.1673-5102.2020.02.012
    Abstract481)      PDF(pc) (1143KB)(86)       Save
    Histone deacetylases plays an important role in plant abiotic stress responses. Histone deacetylase gene HDA902 was cloned from Populus trichocarpa by RT-PCR. The HDA902 gene was transformed into tobacco by Agrobacterium tumefaciens-mediated method. The tolerance of transgenic plants to cold stress was analyzed. The results showed that the expression of HDA902 in tobacco significantly improved the tolerance of transgenic lines to low temperature. The results of NBT and DAB staining showed that transgenic tobacco produced less reactive oxygen species than the wild type after cold treatment. The proline content in the leaves of transgenic plants were much higher than that of wild-type plants, whereas the MDA contents in the leaves of transgenic plants were much lower at low temperature. These results indicated that HDA902 was involved in the response to cold stress, and its overexpression enhanced the tolerance of transgenic plants to cold stress.
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    Study on Salt Tolerance of AtUNE12 Gene in Arabidopsis thaliana
    LI Zi-Yi, HE Zi-Hang, LU Hui-Jun, WANG Yu-Cheng, JI Xiao-Yu
    Bulletin of Botanical Research    2020, 40 (2): 257-265.   DOI: 10.7525/j.issn.1673-5102.2020.02.013
    Abstract674)      PDF(pc) (1597KB)(123)       Save
    Members of the bHLH transcription factor family plays an important role in plant growth, metabolism and abiotic stress response. The salt tolerance of AtUNE12 gene that was part of stress-related bHLH transcription factor family from Arabidopsis thaliana was analyzed. Firstly, the plant overexpression vector of AtUNE12(pROKII- AtUNE12) was constructed, the transgenic T 3 generation A.thaliana was obtained by Agrobacterium-mediated dip then verified by qRT-PCR. Under salt stress, the growth, root length and fresh weight of overexpression and wild-type A.thaliana were analyzed. The MDA content, electrolyte leakage and water loss rate of overexpression and wild-type A.thaliana were compared. H 2O 2 content determination and SOD, POD activity was used to study overexpression and wild-type A.thaliana plants. The results showed that overexpression of AtUNE12 gene could significantly decrease MDA content, electrolyte leakage and the water loss rate, improved POD and SOD activity, and decreased H 2O 2 content in A.thaliana plants. The results indicated that AtUNE12 gene protected the integrity of cell membrane structure, and then enhanced the ROS scavenging ability of A.thaliana plants, thereby improving the salt tolerance of A.thaliana.
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    Expression Analysis of the FaesAP2 B in Mutant Buckwheat with Long Pistil and Long Stamen
    ZHANG Jiao, WANG Xuan, ZHANG Liang-Bo, LIU Zhi-Xiong
    Bulletin of Botanical Research    2020, 40 (2): 266-273.   DOI: 10.7525/j.issn.1673-5102.2020.02.014
    Abstract517)      PDF(pc) (1601KB)(72)       Save
    In order to uncover the molecular mechanism of flower and grain development of buckwheat( Fagopyrum esculentum Moench.) with long pistil and long stamen mutant ‘ lpls’ of a 1 788 bp long AP2 homologous cDNA sequence was cloned from F.esculentum Moench. and designated as FaesAP2 B(GenBank accession number is MK290847.1). By sequence analysis, FaesAP2 B gene contains a full Open Reading Frame with 1 380 bp long, and encodes an AP2/ERF family transcription factor consisting of 459 amino acid residues. The FaesAP2 B transcription factor contains two highly conserved AP2 domains, with a nuclear localization signal region consisting of ten amino acid residues lying before the first AP2 domain. qPCR was performed to detect FaesAP2 B gene expression in root, stem, leaf, perianth, stamen, pistil and juvenile fruit in four-day buckwheat mutant lpls, respectively. The FaesAP2 B is expressed in both vegetative tissues and reproductive structures. However, the expression level of FaesAP2 B in reproductive organs such as floral organs and fruits is significantly higher than that in vegetative tissues. In addition, the expression level of FaesAP2 B in stamens is the highest among seven different organs( LSD, P<0.01). And the expression level of FaesAP2 B in the perianth, pistil and four-day fruit was significantly higher than that in the vegetative organs such as roots, stems and leaves( LSD, P<0.01), but there was no significant difference in the expression of the gene between roots, stems and leaves. Our data suggested that FaesAP2 B may be mainly involved in regulating flower and fruit development in the buckwheat mutant ‘ lpls’.
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    Cloning of LpPEX7 Gene from Lilium pumilum and Its Expression Characteristics under Salt Stress
    HE Hao, ZHU Guo-Qing, CHEN Shi-Ya, XU Chang, JIN Shu-Mei
    Bulletin of Botanical Research    2020, 40 (2): 274-283.   DOI: 10.7525/j.issn.1673-5102.2020.02.015
    Abstract551)      PDF(pc) (1986KB)(73)       Save
    The peroxidasome biosynthetic gene( LpPEX7) of Lilium pumilum DC was cloned, and its open reading frame length is 957 bp, encoding 318 amino acids. The deduced amino acid sequence of LpPEX7 contains six WD40 conserved domains, through sequence alignment analysis of homologous proteins and evolutionary tree analysis, it was found that LpPEX7 had higher homology with some PEX7 from other plants. The expression of LpPEX7 was higher in seeds, leaves and bulbs, but lower in roots and flowers of L.pumilum. The expression of LpPEX7 changed under H 2O 2, NaCl and NaHCO 3 stress conditions. Subsequently, the seeds germination of LpPEX7 over-expression Arabidopsis thaliana lines as earlier than that of wild seeds under the treatment of salinity and oxidative stress. These results show that the LpPEX7 gene has a certain response relationship with salinity and oxidative adversity. It provides a very important candidate gene for the study of salt-tolerant alkaline molecular mechanism of Lily.
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    Sequencing Analysis of Transcriptome of Male Floral Bud at Two Development Stages in Eucommia ulmoides
    ZHU Li-Li, DU Qing-Xin, HE Feng, QING Jun, DU Hong-Yan
    Bulletin of Botanical Research    2020, 40 (2): 284-292.   DOI: 10.7525/j.issn.1673-5102.2020.02.016
    Abstract584)      PDF(pc) (1557KB)(133)       Save
    The male floral bud Eucommia ulmoides is rich in healthily kinds of nutrition compositions and active matter, which with a high nutritional and medicinal value. The transcriptome of male floral bud during two differentiation stages was sequenced, aimed to comprehend the expression of genes involved in the stamen development of E.ulcommia. lllumina high-throughput sequencing technology was applied to detect the transcriptome sequencing data of the male flora bud of ‘Huazhong No.11’ during bract differentiation and stamen differentiation stages. Obtained clean data was analyzed including gene function annotation and differentially expressed genes(DEGs) screening was analyzed by bioinformatics methods. A total of 40.48 Gb clean data were obtained, approximately 90.00% of the clean reads were mapped to the E.ulmoides reference genome. It was found that 315 genes were significantly up-regulated and 269 genes were significantly down-regulated in the male flower buds at stamen differentiation stage. GO and KEGG analyses of DEGs showed the DEGs were enriched in development process, photoperiodism, hormone biosynthetic and signal transduction process, carbon metabolism and other processes and pathways associated with flower induction. The photoperiodic was perhaps the most important pathway for the floral induction of E.ulcommia, and the carbohydrates, plant hormone and other metabolic substances was required in the process of male floral bud differentiation. Importantly, the members of MADs-box including FLC, SOC1, AGL3 and AGL8 regulated the stamen formation of E.ulcommia. It provides a significant reference for studying the genes involved in the floral organs and molecular breeding of E.ulcommia.
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    Gene Cloning and Expression Analysis of PDIL1-2 of Ornamental Kale( Brassica oleracea var. acephala)
    LI Yang, SHI Ya-Kun, GAO Shi-Ke, Li Xiao-Yu, LAN Xing-Guo
    Bulletin of Botanical Research    2020, 40 (2): 293-300.   DOI: 10.7525/j.issn.1673-5102.2020.02.017
    Abstract651)      PDF(pc) (1487KB)(69)       Save
    The BoPDIL1-2 gene was isolated from the stigma of ornamental kale( Brassica oleracea var. acephala) S 13-b S 13-b homozygotes by RT-PCR and RACE. The BoPDIL1-2 coding region sequence was inserted into the prokaryotic expression vector pET-14b and transformed into the E.coli cell for purification.We prepared the polyclonal antibodies against recombinant BoPDIL1-2 through immune mouse. The expression level of BoPDIL1-2 in the various tissue anddifferent developmental stigmas were detected by Western blot. The deduced amino acid sequence of BoPDIL1-2 shared 97.3% and 85.5% identity with Brassica napus BnPDIL1-2 and Arabidopsis thaliana AtPDIL1-2, respectively. SDS-PAGE results showed that the recombinant BoPDIL1-2 fusion protein about 58 kDa was induced by IPTG.Western blot results revealed BoPDIL1-2 wasspecifically expressed inthe stigma,the expression of BoPDIL1-2 was lower in the early stage of stigma, while it was higher in the stigma at theflowering stage.
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    Preliminary Selection of Potential DNA Barcodes for Rubus
    WU Wei-Feng, SHEN Xi-Long, CHEN Zhe, YANG Ding-Yuan, LI Yong-Xia, WANG Yao, ZHANG Qun-Ying
    Bulletin of Botanical Research    2020, 40 (2): 301-307.   DOI: 10.7525/j.issn.1673-5102.2020.02.018
    Abstract690)      PDF(pc) (1186KB)(142)       Save
    In order to establish a molecular identification technique of DNA barcoding for Rubus, and screening universal barcode sequences suitable for Rubus. With GenBank data we analyzed the genetic variation, barcoding gap and NJ consensus tree of six DNA barcoding sequences that contain ITS, ITS2, matK, rbcL, trnH-psbA and trnL-trnF. The intraspecific variation and interspecies variation of trnH-psbA, matK, rbcL and rtnL-trnF were different, and the variation resolutions were 97.32%, 83.33%, 79.07% and 64.95%, respectively. And trnH-psbA, matK, rbcL and rtnL-trnF had obvious barcoding gap; NJ consistent tree showed that matK had the highest proportion of solopathogenicity(67%), and then trnH-psbA(64%), rtnL-trnF(43%), rbcL(30%). The intraspecific variation and interspecies of matK and trnH-psbA sequences for Rubus are different, which can distinguish different species well with great identification potential. It is recommended to use matK and trnH-psbA as the core barcode sequences identified for Rubus, and rtnL-trnF and rbcL are used as auxiliary barcode sequences.
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    Optimization of Dry Enzymatic Extraction Process and Physicochemical Properties of Pine Seed Oil
    ZU Shu-Chong
    Bulletin of Botanical Research    2020, 40 (2): 308-313.   DOI: 10.7525/j.issn.1673-5102.2020.02.019
    Abstract613)      PDF(pc) (1078KB)(124)       Save
    Used a single factor experiment to optimize a dry enzyme extraction process of pine seed oil. The best preparation process for dry enzymatic extraction of pine seed oil was:0.2% of amylase, the ratio of material to liquid of 6:1, the hydrolysis temperature of 55℃, and the hydrolysis time of 8 h, and the maximum oil yield 90.2% with the residual oil rate of 6.2%. The content of unsaturated fatty acids in pine seed oil extracted after testing for the best preparation conditions was 90.09%, of which oleic acid content was 26.84%, mediating acid content was 2.41%, linoleic acid content was 46.25%, and pinolacol acid was 14.59%. The content of palmitic acid in saturated fatty acids was 6.5%, and the stearic acid was 3.41%. The pine oil had an acid value of 2.61 mg·g -1, a peroxide value of 1.56 mmol·kg -1, and a malondialdehyde content of 0.41 mg·kg -1. All the test results are higher than the requirements of the quality standards in pine seed oil in the "Food Industry Standard of the People's Republic of China LS/T 3242-2014".
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