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    05 May 2020, Volume 40 Issue 3
    Research Report
    Contents of Pigments and Anatomical Structure in the Leaves of Forsythia koreana ‘Sun Gold’
    SUN Wang-Wang, MENG Xian-Min, XU Xiu-Yuan, WU Min-Hua, QIN Meng-Yang, LIN Meng-Meng, ZHANG Xin-Yan, YUE Guo-Zhong
    2020, 40(3):  321-329.  doi:10.7525/j.issn.1673-5102.2020.03.001
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    The mature leaves of different canopy layers of Forsythia koreana ‘Sun Gold’ appeared different colors. In order to explore the effect of leaf pigment content and anatomical structure on leaf color, we used deep green leaves of F.koreana as control materials aimed to observe the leaf color of top, middle and bottom canopy, meanwhile measured and analyzed the blade area, chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents. The total chlorophyll contents of top yellow, middle yellow green and deep light green leaves were 0.51%, 4.44% and 66.47% of control respectively, which were extremely significantly lower than control(P<0.01). However, the chlorophyll a/b ratio of yellow green leaves increased significantly(P<0.05), while the total chlorophyll/carotenoid ratio of yellow green leaves decreased extremely significantly(P<0.01). The chloroplast development of the yellow and yellow green leaves was stay in the monolayer stage, the degree of thylakoid differentiation was lower than control, and the grana lamella of the light green leaves was swollen. The anatomical structure of the leaves showed that the organelle was degraded, and the shape of palisade tissue cells was difficult to distinguish. However, the upper epidermal cell of yellow green leaves was convex. F.koreana ‘Sun Gold’ belongs to the mutant of total chlorophyll and reduced chlorophyll b synthesis, which is characterized by serious chlorophyll deficiency and increased relative carotenoid content. The chloroplast development of F.koreana ‘Sun Gold’ was stagnated and the thylakoid structure was abnormal, which was the main factor for the different color of its leaves, which had nothing to do with the microscopic structure of its leaves.
    An Assessment of Phylogenetic Diversity and Dissimilarity of Halophytes Across Different Provincial Region,China
    ZHANG Zhong-Hua, SONG Xiao-Qian, TANG Zhong-Hua, LIANG Zheng-Wei
    2020, 40(3):  330-338.  doi:10.7525/j.issn.1673-5102.2020.03.002
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    Phylogenetic diversity serves regional vegetation history, evolutionary law and the protection of biodiversity. Halophytes are the unique group of regional vegetation evolution and the important material basis for the development of future agricultural germplasm resources. The phylogenetic diversity of halophytes is great significance to reveal regional environmental changes, protection of germplasm resources and regional development,but so far, there is a lack of research in this area. Based on the theory and method of phylogenetic diversity and taking provincial administrative regions as units, we systematically evaluated the phylogenetic diversity and dissimilarity in China. A phylogenetic tree of 484 species, 17 varieties and 8 subspecies of Halophytes in 65 families has been constructed. Only Xinjiang, Ningxia, Gansu and Qinghai,Shaanxi, Inner Mongolia and Beijing have a net relatedness index (NRI) greater than 0. The correlation of phylogenetic diversity with family, genus and species richness was 67.01%, 91.20% and 96.99%, respectively. According to the taxonomic floristicsimilarity and phylogeneticsimilarity of halophytes, Chinese halophytes were divided into four regions. Our results have important guiding significance for provincial halophyte resource assessment, halophyte germplasm collection and halophyte regionalization in China.
    Modeling Association among Multispecies Plant Invasion and Impact Factors in Laohuling Forest,Nanning
    HUANG Xiao-Rong
    2020, 40(3):  339-346.  doi:10.7525/j.issn.1673-5102.2020.03.003
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    A deep understanding of the interactions between multispecies invasions and impact factors facilitates more pertinent control of invasions. Plot survey was carried out in Laohuling Forest, Nanning, and the vegetations were recorded in six districts; and relative cover of all alien plants in a plot, the alien ratio, was used as invasion index. R-lme4 mixed effect model with random district intercept and SPSS GLM-GEN were used to calculate impacts of environmental on invasions, and R-effects package to visualize the complex interaction effects. Multivariate mixed effect model showed that roadside had positive main effect on invasion(P=0.000), canopy cover and dominant plant height both had negative main effect on invasion(P=0.000); native richness' main effect on invasion was not significant, but its interaction with roadside had significant effect on invasion(P=0.007); on roadside, native richness strongly raised resistance to invasion; while within forest, native richness weakly increased resistance to invasion; canopy cover and dominant plant height's interaction effect on invasion was significant(P=0.004), but their filter effects were non-additive. GLM-GEN results indicated that forest age and tending times did not influence invasion; and we did not find significant difference between introduced and native species plantations in impacts on invasions; furthermore, plot locations relative to forest lanes influenced alien invasions, and plots below forest lanes were more vulnerable to invasions. In future works of monitoring and controlling alien plants in forest, emphasis should be focused on stands those below forest lanes.
    Effects of Trichoderma on the Growth-related Physiological Indexes of Rocketsalad(Eruca sativa)
    HOU Xue-Yue, DENG Jun-Jie, YAO Zhi-Hong, HUANG Shou-Chen, LI Jia-Zhe, CHA Xin-You, MA De-Zhi, ZHANG Rong-Shu
    2020, 40(3):  347-352.  doi:10.7525/j.issn.1673-5102.2020.03.004
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    Rocketsalad(Eruca sativa) is a kind of new special and aromatic vegetable with great development value in China's vegetable market. In order to develop biocontrol agents of Trichoderma to promote the growth and quality of rocketsalad, the seed soaking and root-watering treatment of rocketsalad with T.harzianum T8 which was isolated and identified in our laboratory were carried out. And the effects of Trichoderma on the growth-related physiological indexes of rocketsalad aged 30 d were analyzed in continuous cultivation of three stubble in a vegetation season. The results showed that the application of Trichoderma can significantly increase the biomass of rocketsalad, improve the photosynthetic characteristics, enhance the resistance-related enzyme activity and proline content, and raise the yield and the nutritional value.
    Ameliorative Effect of Ferulic Acid on Paeonia ostii under Drought Stress
    FANG Zi-Wen, ZHANG Xia-Yan, TAO Jun, ZHAO Da-Qiu
    2020, 40(3):  353-359.  doi:10.7525/j.issn.1673-5102.2020.03.005
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    Paeonia ostii was used to study the ameliorative effect of ferulic acid(FA) on plants under natural drought stress. The results showed that:compared with the control, FA treatment significantly increased leaf water content and antioxidant enzyme activities, reduced the accumulation of reactive oxygen species, proline content and relative electrical conductivity, which reduced the damage of plants under drought stress. In addition, drought stress also caused changes in photosynthetic characteristics and chlorophyll fluorescence parameters, and FA treatment improved the utilization efficiency of light energy in plants and protected of photosynthetic apparatus by enhanced heat dissipation. This study confirmed that FA has some ameliorative effect on P.ostii under drought stress, which would lay a foundation for the cultivation of P.ostii in arid areas.
    Effects of Exogenous Methyl Jasmonate on Photosynthesis and Physiological Characteristics of Scutellaria regeliana under Drought Stress
    WU Jian-Hui, FAN Wei-Fang, NIU Zhe, ZHANG Jing, GAO Peng-Fei, LI Wen
    2020, 40(3):  360-367.  doi:10.7525/j.issn.1673-5102.2020.03.006
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    The effects of exogenous application of different concentrations(0.1, 0.5, 1 and 2 mmol·L-1) of methyl jasmonate on phenotypic characteristics and photosynthesis and physiological characteristics under 15% PEG-6000 stress were studied with the Scutellaria regeliana as experimental materials. The results showed that:exogenous application of methyl jasmonate enhanced the net photosynthetic rate, stomatal conductance and transpiration rate of S.regeliana. It mainly plays a role in promoting these and effectively reduces the damage level of the S.regeliana. The intercellular CO2 concentration was decreased, and the effect of 2 mmol·L-1 methyl jasmonate on the net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 concentration of S.regeliana was the most significant. The free proline content of the treatment group lower than CK. The soluble protein content lower than CK in the first 6 d treatment, and was significantly higher than that of CK after 6 d treatment. Exogenous application of methyl jasmonate can increase the activity of SOD, POD and CAT as a whole. The results of photosynthetic parameters and physiological indexes of S.regeliana showed that exogenous application of methyl jasmonate could effectively alleviate the adverse effects of drought stress on S.regeliana.
    Shoot Regeneration from Transverse Thin Cell Layers of Pseudobulbs and Micropropagation of Bletilla striata
    FAN Li, WANG Ren-Rui, YU Ma, LIU Jin-Huan, HOU Da-Bin, TANG Zhi-Kang
    2020, 40(3):  368-375.  doi:10.7525/j.issn.1673-5102.2020.03.007
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    Pseudobulbs of Bletilla striata were taken as initial explants and cut averagely for three transverse thin cell layers-upper position, middle position and lower position to culture on different media. Different parts of pseudobulbs, plant growth regulators and their concentrations were used to study the effects on shoots regeneration from pseudobulb transverse thin cell layers. The thickness of pseudobulb transverse thin cell layers were also used for explants for comparing the browning rate and the number of shoots. Orthogonal design was used to study the effects of BA and NAA for shoots micropropagation. The micropropagated shoots were cultured on a medium for being strong and rooting prior to transplanting to soil. The results showed that:Position of pseudobulb transverse thin cell layers significantly affected shoot induction. Lower part of pseudobulbs were significant easier to induce shoots than upper part and middle part. TDZ and BA also had obvious effect on shoot induction. NAA had no obvious effect on shoot induction. Lower parts of pseudobulb transverse thin cell layers cultured on basic medium+2.0 mg·L-1 BA+1.0 mg·L-1 TDZ for 4 weeks was the optimal treatment to induce shoots. The induction rate was 93.3% and the number of shoots was 15. Pseudobulb transverse thin cell layers(1.6-2.0 mm in thickness) were scarcely browning. Basic medium+1.5 mg·L-1 BA+0.3 mg·L-1 NAA was the optimal medium for micropropagation. The multiplication factor was 4.3, and the length of shoots was 7.8 cm. Shoots regenerated directly from pseudobulb transverse thin cell layers and micropropagation system of B.striata was established successfully, which laid a foundation for innovation of germplasm resources.
    Evaluating Parents of Pinus koraiensis Seeds Orchard with Growth and Fruiting
    ZHOU Xue-Yan, GAO Hai-Yan, LI Zhao-Min, ZHAO Yin-Kun, GE Li-Li, HOU Qing-Wen, DING Wen-Ya, ZHAO Xi-Yang
    2020, 40(3):  376-385.  doi:10.7525/j.issn.1673-5102.2020.03.008
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    In order to evaluate and select the excellent breeding materials of Pinus koraiensis, the growth traits(height, diameter at breast height) and fruit traits(cone number, fresh cone weight, thousand seed weight and cone seeds weight) of 69 half-sib families in 26 and 28 years old were determined and analyzed. At different determination time, and all traits among different families reached an extremely significant difference level(P<0.01). The heritabilities of each trait was higher(0.87-0.96). The range of phenotypic variation coefficient was 6.30%(thousand seeds weight)-63.00%(cone number). The range of general combining ability of the height, diameter at breast height and cone number were(-1.77-1.83), (-9.31-8.67) and (-15.97-16.03), respectively. The correlation coefficients among growth traits showed high level(0.716-0.998), but most fruit traits and growth traits showed no significant correlation. Used the method of multi-trait comprehensive evaluation, 69 half-sib families were evaluated by growth and fruit traits, respectively. Seven families were selected by growth traits with the selection rate by 10%, the genetic gains of height, diameter at breast height were 15.66% and 35.40%, respectively. Seven families were selected according to fruit traits, the genetic gains of cone number, fresh cone weight and cone seed weight were 87.30%, 1.77% and 3.39%, respectively. The results can provide the basis and materials for the evaluation and selection of elite families.
    Flower Characters Variation of Different Individuals of Catalpa bungei
    MA Wen-Jun, YANG Gui-Juan, WANG Jun-Hui, ZHAO Kun
    2020, 40(3):  386-393.  doi:10.7525/j.issn.1673-5102.2020.03.009
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    Catalpa bungei is important hardwood timber and ornamental tree in the central China. It has been cultivated more than 2 600 years. Study on the diversity of flower traits reveals the huge intraspecific phenotype variation of flower in C.bungei, and would provide useful information for identification and protection of new variety and theoretical basis for flower color and excellent ornamental cultivars in Chinese C.bungei breeding project. Superior individual and hybrids F1 were collected in 1985-1990. A total of 27 individuals, and two qualitative and 7 quantitative traits of flower were analyzed using variance analysis and cluster analysis methods. ①The difference of flowering phenology of 27 individuals was up to 5 d. The flower size, inflorescence length, number of flower per plant of the 27 individuals showed significant differences. In addition, different individuals showed a characteristics of explicit flower and implicit flower because of difference of the length of petiole and the length of flowering branch. ②C.bungei is didynamous stamen, which divided into male fertile and sterile. Twelve of the 27 individuals that in survey were male fertile. There was a significant difference in the amount of pollen. ③The flower is trilobation with five lips, which has two-sectional upper lip, three-sectional lower lip, and the length of upper lip flap is larger than the lower lip. The length of flower branch, length of inflorescence, amount of flower of single flower branch, length of upper lip of flower, length of lower lip of flower and corolla diameter of 27 individuals have significant differences. The value changes were 10.7-16.4 cm, 5.6-9.6 cm, 2-13 flower, 3.8-5.2 cm, 3.0-4.3 cm and 3.9-5.4 cm, respectively. The coefficients of variation were 12.8%, 12.5%, 36.7%, 7.5%, 9.1% and 9.2%, respectively. ④The L* value(the value of brightness or black and white), a* value(red and green value), b* value(yellow and blue value), C* value(chroma), and h value(hue) between the 16 individuals were significantly different. The coefficients of variation of the a* value, b* value and chroma C* value were 35.1%, 52.5% and 29.8%, respectively; measuring color with L*, a* and b* value, the 16 individuals were clustered into 3 categories, red, pink and white by cluster analysis. The length of flower branch, length of inflorescence, and size of flower of the 27 individuals of C.bungei have significant differences. The color of the 16 individuals also have obvious variation. According to the L*, a* and b* value by cluster analysis, the 16 individuals were clustered into 3 groups, red, pink and white when the value of Euclidean distance is 15.
    Drought Resistance of Larch NAC Gene by Transient Genetic Transformation
    ZHANG Lei, XIONG Huan-Huan, CAO Qing, ZHAO Jia-Li, ZHANG Han-Guo
    2020, 40(3):  394-400.  doi:10.7525/j.issn.1673-5102.2020.03.010
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    An NAC transcription factor named LoNAC18 was cloned by transcriptome analysis of Larix olgensis. The sequence analysis results showed that the whole gene was 1 101 bp, encoding 366 amino acids, and it was an unstable hydrophilic protein. Compared with the control, gene expression increased of the transient genetic transformation plant under PEG simulated drought stress, while SOD, POD, soluble protein, soluble sugar content was all increased, and the MDA content is lower than the control plants. Therefore, LoNAC18 genes was involved in regulating the L.olgensis response to PEG stress, the results could provide the reference to research on NAC function and mechanism of action of conifer stress response.
    Contents of Active Components of Scutellaria baicalensis Plantlet and Its Biological Activities of Hyperhydricityin Tissue Culture Recvered by AgNO3
    WANG Sheng-Fang, HAN Shu-Lan, DANG Shi-Kun, WANG Hui-Mei
    2020, 40(3):  401-406.  doi:10.7525/j.issn.1673-5102.2020.03.011
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    We studied the reversal effect of AgNO3 on the hyperhydric Scutellaria baicalensis plantlets, and the contents of the active components. AgNO3 had a significant recovery effect on biological activities of the restored hyperhydric S.baicalensis plantlets and the recovery effect was affected by the concentration of AgNO3. When the concentration of AgNO3 was 4 mg·L-1, the recovery rate of hyperhydric plantlets was 88%, and the recovered S.baicalensis plantlets had the best growth state. We studied the contents of active components and biological activities of hyperhydric S.baicalensis plantlets recovered by AgNO3 for one month. The content of baicalin in recovered hyperhydric S.baicalensis plantlets was 68.6 mg·g-1, significantly higher than that in hyperhydric plantlets(38.3 mg·g-1), but lower than that in normal tissue cultured plantlets(108.2 mg·g-1). The results of antioxidant, antimicrobial and anti-tumor experiments showed that the recovered hyperhydric S.baicalensis plantlets were significantly better than the hyperhydric plantlets, but still lower than the normal tissue-cultured plantlets. Therefore, AgNO3 had a good recovery effect on the hyperhydric plantlets of S.baicalensis.
    Resistance Analysis of Transgenic PnDof30 Arabidopsis under Abiotic Stress
    ZHANG Yu-Qing, LIU Ye, QU Chun-Pu, LIU Guan-Jun, YANG Tian-Tian, YANG Cheng-Jun
    2020, 40(3):  407-415.  doi:10.7525/j.issn.1673-5102.2020.03.012
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    Dof(DNA-binding with one finger) transcription factor is a unique transcription factor in plants,it is a family of zinc finger proteins with many members,the length of amino acids is generally between 200-400, and it contains very conservative N-terminal and relatively changeable C-terminal. Previous studies have shown that Dof transcription factor family plays an important role in regulating carbon and nitrogen metabolism, improving nitrogen uptake and utilization, and enhancing abiotic stress tolerance. To explore the resistance of PnDof30 gene in Populus simonii×P.nigra, this study used transgenic PnDof30 Arabidopsis thaliana as a research object to compare the physiological and biochemical indices of over-expressed PnDof30 Arabidopsis line L2 and wild type A.thaliana under drought, salt and osmotic stress. It was found that the seed germination rate, root length and fresh weight of L2 were higher than those of WT; the contents of SOD, POD and proline were higher than those of WT, while the contents of chlorophyll and MDA were decreased; the PnDof30 gene in L2 was significantly increased after stress. These results indicate that PnDof30 gene has the ability to resist drought, salt and osmotic stress. The results of this experiment will be of great significance for understanding the stress resistance of Dof transcription factors.
    Communities and Protection Strategy of Endangered Species Wild Taxus cuspidata
    DONG Ming-Zhu, WANG Li-Tao, Lü Mu-Jie, MENG Dong, YANG Chun-Yu, ZHAO Chun-Jian, FU Yu-Jie, YANG Qing
    2020, 40(3):  416-423.  doi:10.7525/j.issn.1673-5102.2020.03.013
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    With the wild Taxus cuspidata communities in Changbai Mountain as the research object, we set up the four standard areas(50 m×50 m), then analyzed the tree characteristics(height, DBH, crown width, etc.), site conditions, stand status by forest mensuration and population statistics, and studied the species diversity characteristics of T.cuspidata community in the distribution area. The four communities contained 32 woody species belonging to 17 families and 24 genera, 53 herbaceous species belonging to 23 families and 41 genera. And the changes of Simpson index and Shannon index are consistent, as for Pielou index in four communities were herb layer > shrub layer > arbor layer, and another sample areas were shrub layer > arbor layer > herb layer. The highest Margalef index in four community was herb layer, and in the rest of the plots was arbor layer. Besides, the important value of T.cuspidata was lower in each community. T.cuspidata were mainly composed of mixed forests of Pinus koraiensis Sieb. et Zucc., Picea jezoensis Carr. var. komarovii and Tilia amurensis Rupr.. Thus, the effective measures and countermeasures for the conservation, breeding and utilization of wild T.cuspidata resources were provided.
    Zinc Finger Protein OsZFP6 Expression Features and Functions in Saline-alkali Stress Response
    LIU Jia-Li, HE Ming-Liang, LIU Chen-Xi, LIAO Xu, LI Xiu-Feng, GUAN Qing-Jie
    2020, 40(3):  424-432.  doi:10.7525/j.issn.1673-5102.2020.03.014
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    C2HC type zinc finger protein was screened, encoded 304 amino acids and named OsZFP6. The expression level of OsZFP6 was increased by abiotic stress, including alkali(NaHCO3) and H2O2 treatment in two-week old rice leaves but decreased in roots. Thus, it was speculated that OsZFP6 was involved in the regulation corresponding reaction pathways of resistance. We fused a green fluorescent protein(GFP) with it and found that OsZFP6 was located in the nucleus. Moreover, rice overexpressed OsZFP6 were more tolerant to NaHCO3 treatments. The overexpressed plants had strong tolerance, and were higher than WT in plant height and fresh weight, but the contents of MDA and H2O2 in vivo were lower than non-transgenic plants, which indicated that OsZFP6 could enhance the resistance of plants by scavenging peroxide in response to NaHCO3 stress. Preliminarily, it was speculated that OsZFP6 gene participated in the regulation of resistance and it may play a vital role in transcription regulation.
    Transcription Factor Gene ERF11 Response to Osmotic Stress in Populus simonii×P.nigra Poplar
    LIU Yue, ZHAO Kai, Lü Guan-Bin, JIANG Ting-Bo, ZHOU Bo-Ru
    2020, 40(3):  433-440.  doi:10.7525/j.issn.1673-5102.2020.03.015
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    The ERF11 transcription factor gene(Potri.011G05700.1) cDNA is encoded with 243 amino acids in Populus simonii×P.nigra. ERF11 gene protein is an unstable pro-water protein, has no signal peptide, has no transmembrane ability, and contains AP2/ERF family conservative structure. The systematic evolution tree shows that PsnERF11 protein is close to Populus trichocarpa, Euphratica, Ricinus communis, Hevea brasiliensis, Jatropha curcas, Manihotesculenta, indicating that the genetic relationship is relatively close. The results of subcellular positioning show that ERF11 protein is located in the nucleus. ERF11 gene expression is tissue-specific and induced by coercion. Its relative expression level in the root was significantly higher than in the stem and leaf, and the gene was expressed higher under the pressure of salt and glycol. It is shown that ERF11 transcription factor genes may be associated with plant response to high salt and drought stress.
    Expression and Function of BoNR8 lncRNA in Cabbage under Cold Stress
    YANG He, ZHANG Nan, LIU Zi-Guang, LIN Jian-Hui, SUN Shi-Chen, LIU Sheng-Yi, CEN Xi, WU Juan
    2020, 40(3):  441-446.  doi:10.7525/j.issn.1673-5102.2020.03.016
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    Long non-coding RNA(lncRNA) with longer than 200 bp in length do not contain open reading frames, and they generally do not have a protein-coding function. They are ubiquitous in the body and play a variety of biological roles. There are few studies on lncRNA specifically expressed in the germination stage of plant seeds. BoNR8 lncRNA is a long non-coding RNA(about 272 bp) transcribed by RNA polymerase Ⅲ in cabbage. Previous studies have found that it is specifically expressed in the seed germination stage, and the overexpression of BoNR8 lncRNA in Arabidopsis inhibits seed germination under normal conditions and reduces the sensitivity of germinated seeds to ABA. In this study, we analyzed the sequence of BoNR8 lncRNA and found that there was a cold stress motif(ATATAAATAAAT) in the transcribed region, and this motif exists in the largest stem loop of the secondary structure of BoNR8 lncRNA. Biochemical experiments further showed that BoNR8 lncRNA responded to low temperature environment, and its overexpression inhibited seed germination of Arabidopsis thaliana under low temperature stress. The discovery of low-temperature related functions of BoNR8 lncRNA provides new materials for low temperature resistance research of plant and enriches the study of low temperature resistance mechanism of plant.
    Codon Usage Bias of Chalcone Synthase Gene CHS in Pinus koraiensis
    LI Xiang, FAN Zuo-Yi, WANG Jing-Yuan, WANG Qi, LI Xi-Peng, WANG De-Qiu, KONG Ling-Yuan, CAO Sen-Lin, MENG Qing-Gang, ZHAO Xi-Yang
    2020, 40(3):  447-457.  doi:10.7525/j.issn.1673-5102.2020.03.017
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    Chalcone synthase(CHS) is widely found in plants and is a key enzyme in anthocyanin synthesis pathway. It can catalyze the binding of malonyl coenzyme A and coumaryl coenzyme A to generate flavonoids. CodonW and EMBOSS were used to analyze the codon usage bias of CHS gene in Pinus koraiensis. The CHS gene of P.koraiensis were compared with those of the other 24 plants and model plant genome, which laid a foundation for the selection of appropriate expression system. The effective codon number(ENC) and GC content of CHS gene coding region were 48.92 and 0.351, respectively. The content of C+G was higher than that of A+T, and the codon preferred to end with A/T. The G+C content of most plants CHS genes is higher than that of A+T, and the codon preferred to end with C/G. By cluster analysis, the codon usage bias of P.koraiensis was similar to P.massoniana and P.densiflora pdchsx. In addition, it was found that Arabidopsis thaliana and Escherichia coli might be ideal receptor systems for CHS genetic transformation and heterogeneous expression.
    Transcriptome Analysis for Medicinal Plant Anisodus tanguticus
    ZHANG Yu, XIA Ming-Ze, ZHANG Fa-Qi
    2020, 40(3):  458-467.  doi:10.7525/j.issn.1673-5102.2020.03.018
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    The transcriptome sequencing analysis of Anisodus tanguticus was carried out by high throughput sequencing technology to understand resource plant A.tanguticus deeply. The 71 463 Unigenes were obtained after processing. By comparing with several databases,we classified and analyzed the genes, and finally succeeded in annotating 47 624 genes. The 13 110 genes with 26 subclasses were annotated after comparing with KOG protein library.Compared with NR library, 39 621unigenes were annotated.The transcripts were compared with Swissprot and TrEMBL to obtain GO functional annotations. The 29 309 unigenes obtained from the annotations could be divided into three categories:molecular function, biological process, and cellular components, with 62 subcategories.Referring to the KEGG database, 3 679 genes were annotated. The metabolic pathways involved can be classified into four categories:metabolic related pathways, genetic information processing,cellular processes, and environmental information processing. Among them,metabolic related pathways are the most, accounting for about half of all metabolic pathways.Statistical results of metabolic pathways and related unigenes of the active ingredients of A.tanguticus showed that the number of metabolic pathways related to alkaloids was the most, and the number of unigenes corresponding to terpenoids and phenylpropanoids was the most.In addition, 31 382 SNP loci and 6 SSR repeat types were detected. Among them, single base repeat types accounted for the highest proportion, with 56.52 SSR repeats per million bases, accounting for 45.30%.These results enrich the transcriptome information of A.tanguticus and lay a foundation for the study of the molecular biology, which contribute to the further development and utilization of A.tanguticus.
    Research Method
    An Efficient Method for Separation and Purification of Betulin from Birch Bark
    LI Ying, LI Lin-Xi, ZHANG Yu-Qi, WANG Yu, ZHAN Ya-Guang, YIN Jing
    2020, 40(3):  468-475.  doi:10.7525/j.issn.1673-5102.2020.03.019
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    The experiment was conducted to obtain a simple and efficient method for extracting and purifying betulin from the bark of birch by comparing different methods. By comparing three extraction schemes of betulin by different extraction time, ultrasonic oscillation method and rotary evaporator recovery. And methanol-chloroform recrystallization method, absolute alcohol recrystallization method and macroporous resin chromatography were further used to purify the initial extract of betulin. The best extraction and purification scheme of betulin was obtained. The birch bark samples were soaked in 95% ethanol solution with a liquid-solid ratio of 50:1(mL:g) for 120 h. The filtrate was filtered by filter paper. The filtrate was heated and refluxed at 50℃ for 5 h. The white solid powder was obtained by vacuum distillation. Then 70 mL ethyl acetate was used as solvent, and the solid was heated and refluxed for 90 min, then filtered. The filtrate is concentrated to dry at 50℃ in a crucible. Then recrystallize with anhydrous ethanol as solvent. Anhydrous ethanol:solid=30 mL:1 g, recrystallize at -20℃. The recrystallization yield can reach 38.00%, and the highest purity can reach 99.80%. The optimized extraction and purification method of betulin is simple, high yield and safe reagent. It is an effective method for large-scale extraction of betulin.
    Scientific Notes
    Root CO2 Efflux Variations of Betula platyphylla Among Sites and Root Diameter Classes in 12 Different Provenances
    SUN Shuo, WANG Xiu-Wei, DU Meng-Tian, LI Jing-Hang, WANG Bo-Yi, LIU Gui-Feng
    2020, 40(3):  476-480.  doi:10.7525/j.issn.1673-5102.2020.03.020
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    The 13 years old Betula platyphylla from 12 provenances in Caohekou and Maoershan were used as experimental materials. In the growing season, three diameter classes roots <0.5 mm, 2.0-5.0 mm and >5.0 mm were taken, respectively. The CO2 efflux rate of root system was measured by infrared gas analysis in vitro, in order to study the effect of root diameter, experiment site and provenance on CO2 efflux of B.platyphylla root. The root diameter and environment had significant(P<0.01) effects on root CO2 efflux rate, while provenance had no significant(P>0.05) effects on root CO2 efflux rate. Root CO2 efflux rate decreased significantly(P<0.01) with the increase of diameter class. All diameter class root CO2 efflux rate in Caohekou was greater than that in Maoershan(P<0.01). The root CO2 efflux rates of B.platyphylla in Caohekou and Maoershan with diameters <0.5 mm were 104.21 and 81.07 mol·g-1·s-1, the root CO2 efflux rates with diameters 2.0-5.0 mm were 41.08 and 30.91 mol·g-1·s-1, and the root CO2 efflux rates with diameters >5.0 mm were 24.87 and 11.37 mol·g-1·s-1, respectively. There were significant(P<0.05) differences in tree height and DBH among 12 provenances of B.platyphylla, and both tree height and DBH were negatively correlated with root CO2 efflux. The results of relevant studies will provide an important basis for more accurate assessment of CO2 efflux fluxes from tree roots.