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    05 January 2020, Volume 40 Issue 1
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    Two New Species of Aconitum(Ranunculaceae) from Sichuan
    WANG Wen-Tsai
    2020, 40(1):  1-4.  doi:10.7525/j.issn.1673-5102.2020.01.001
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    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.
    Three new species of Thalictrum(Ranunculaceae) from Sichuan Province
    WANG Wen-Tsai
    2020, 40(1):  5-9.  doi:10.7525/j.issn.1673-5102.2020.01.002
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    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.
    Cyclobalanopsis reclinatocaulis,A New Species of Fagaceae from Fujian Province
    LIN Qin-Wen, ZHONG Yi-Xin, LIN Mu-Mu
    2020, 40(1):  10-14.  doi:10.7525/j.issn.1673-5102.2020.01.003
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    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.
    Notes on the Geographical Distribution of Delphinium mollipilum(Ranunculaceae)
    REN Lu-Ming, CHEN Xue-Lin
    2020, 40(1):  15-17.  doi:10.7525/j.issn.1673-5102.2020.01.004
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    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.
    Geographical Characteristics of Orchidaceae Plants in Qinling Mountains
    GAO Xu-Zheng, KANG Yong-Xiang, ZHANG Li-Li, KANG Yue-Cheng, GUO Ming
    2020, 40(1):  18-28.  doi:10.7525/j.issn.1673-5102.2020.01.005
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    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.
    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
    2020, 40(1):  29-40.  doi:10.7525/j.issn.1673-5102.2020.01.006
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    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(L0), transmittance 60%(L1), transmittance 25%(L2) and transmittance 10%(L3). 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(SLWd) 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 Pn and Tr showed obvious bimodal curves for L0 and L1, and the "noon break" appeared at 12:00, and the change of both L2 and L3 was relatively mild, the phenomena werenot obvious. The variation of Gs was basically consistent with that of Pn. The order of daily average of both Pn and Tr was L1 > L0 > L2 > L3. 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.
    Effects of Simulated Nitrogen Deposition on the Growth and the Content of Non-structure Carbohydrate of Sinocalycanthus chinensis Seedlings
    ZHAO Zhe, JIN Ze-Xin
    2020, 40(1):  41-49.  doi:10.7525/j.issn.1673-5102.2020.01.007
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    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.
    Establishing Tissue Culture and Regeneration System of Scutellaria regeliana
    ZHANG Jing, NIU Zhe, FAN Wei-Fang, GAO Peng-Fei, WU Jian-Hui
    2020, 40(1):  50-57.  doi:10.7525/j.issn.1673-5102.2020.01.008
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    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% HgCl2 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.
    Cytosolic Ca2+ Involved in Seed Germination of Naked Oat Enhanced by Exogenous H2S under Saline-Alkali Stress
    LIU Jian-Xin, OU Xiao-Bin, WANG Jin-Cheng
    2020, 40(1):  58-65.  doi:10.7525/j.issn.1673-5102.2020.01.009
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    Hydrogen sulfide(H2S) is a new gas signal molecule and calcium(Ca2+) is an important second messenger. Both play important roles in plant adversity response, respectively. The experiment was conducted to clarify the role of cytosolic Ca2+ in germination of crop seed promoted by exogenous H2S 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:Na2SO4:Na2CO3:NaHCO3 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 Ca2+ chelator ethylene glycol-bis-(2-aminoethylether)-N, N, N', N'-tetraacetic acid(EGTA), plasma membrane Ca2+ channel blocker lanthanum chloride(LaCl3), tonoplast Ca2+ release inhibitor ruthenium red(RR), and endoplasmic reticulum calcium pump blocker thapsigargin(Thaps) were combined with sodium hydrosulfide(NaHS, a donor of H2S), 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 Ca2+ on seed germination of naked oat enhanced by H2S 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, LaCl3 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 Ca2+ involved signal transduction in promoting of naked oat seed germination induced by H2S under saline-alkali stress, and cytosolic Ca2+ mainly comes from the influx of extracellular Ca2+ and the release of Ca2+ in vacuoles.
    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
    2020, 40(1):  66-72.  doi:10.7525/j.issn.1673-5102.2020.01.010
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    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.
    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
    2020, 40(1):  73-78.  doi:10.7525/j.issn.1673-5102.2020.01.011
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    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".
    Identification of Key Amino Acid Sites in AtGDPD-Like3 with Role in Arabidopsis Root Hairs
    WANG Shuang, CHENG Yu-Xiang, XIA De-An
    2020, 40(1):  79-84.  doi:10.7525/j.issn.1673-5102.2020.01.012
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    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 S538A, V556A and D628A single site mutation of AtGDPD-Like3, which were expressed in atgdpdl3 mutants, respectively. The results showed that AtGDPD-Like3 with V556A or D628A mutation could completely restore root hairs growth defects of atgdpdl3, whereas, AtGDPD-Like3 with S538A mutation partially restored root hairs growth defects. These indicate that Ser538 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.
    Effects of Extracellular adenosine-5'-triphosphate on Cellular damage, H2O2 Content,and Activities of H2O2 Detoxifying Enzymes under Lead Stress
    CAO Jia-Xin, DA Meng-Ting, PANG Hai-Long, JIA Ling-Yun, FENG Han-Qing
    2020, 40(1):  85-92.  doi:10.7525/j.issn.1673-5102.2020.01.013
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    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, H2O content, and the activities of H2O detoxifying enzymes underlead stress. With the increase of concentrations of Pb(NO3)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 H2O2 and the activity of the CAT(catalaseenzyme) were increased and reached their maximum at 200 μmol·L-1Pb(NO3)2, while the activityof the POD(peroxide enzyme) was decreased, with the increase of concentrations of Pb(NO3)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, H2O2 content, andthe activity of CAT, but had higher level of the activity of POD. Therefore, the cellular damage, H2O content, and the activities of H2O detoxifying enzymesunderlead stress could beregulated by extracellular adenosine-5'-triphosphate.
    Transcriptome of Single and Double Tepals of Clematis patens
    XIONG Yang-Yang, LUO Lin-Li, ZHAO Shi-Hao, ZHOU An-Long, WANG Jin
    2020, 40(1):  93-105.  doi:10.7525/j.issn.1673-5102.2020.01.014
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    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.
    Structural and Functional Analysis of R2R3 MYB Transcription Factor HbMYB88
    FAN Song-Le, WANG Ji-Kun, XIE Gui-Shui, WANG Meng, WANG Li-Feng
    2020, 40(1):  106-116.  doi:10.7525/j.issn.1673-5102.2020.01.015
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    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(H2O2), 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.
    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
    2020, 40(1):  117-124.  doi:10.7525/j.issn.1673-5102.2020.01.016
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    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.
    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
    2020, 40(1):  125-132.  doi:10.7525/j.issn.1673-5102.2020.01.017
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    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.
    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
    2020, 40(1):  133-140.  doi:10.7525/j.issn.1673-5102.2020.01.018
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    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.
    Overexpression of K Transporter Gene trkH in Enhancing K Nutrition in Maize
    DING Bao-Juan, AN Li-Jia, SU Qiao
    2020, 40(1):  141-147.  doi:10.7525/j.issn.1673-5102.2020.01.019
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    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 T0 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.
    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
    2020, 40(1):  148-152.  doi:10.7525/j.issn.1673-5102.2020.01.020
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    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.
    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
    2020, 40(1):  153-160.  doi:10.7525/j.issn.1673-5102.2020.01.021
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    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.