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    05 March 2020, Volume 40 Issue 2
    Research Report
    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
    2020, 40(2):  161-171.  doi:10.7525/j.issn.1673-5102.2020.02.001
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    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.
    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
    2020, 40(2):  172-176.  doi:10.7525/j.issn.1673-5102.2020.02.002
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    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.
    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
    2020, 40(2):  177-188.  doi:10.7525/j.issn.1673-5102.2020.02.003
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    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 CO2 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.
    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
    2020, 40(2):  189-195.  doi:10.7525/j.issn.1673-5102.2020.02.004
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    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.
    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
    2020, 40(2):  196-201.  doi:10.7525/j.issn.1673-5102.2020.02.005
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    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% HgCl2 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%.
    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
    2020, 40(2):  202-208.  doi:10.7525/j.issn.1673-5102.2020.02.006
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    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.
    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
    2020, 40(2):  209-216.  doi:10.7525/j.issn.1673-5102.2020.02.007
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    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(Fv/Fm). Photosynthetic physiology of Pn 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.
    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
    2020, 40(2):  217-223.  doi:10.7525/j.issn.1673-5102.2020.02.008
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    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.
    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
    2020, 40(2):  224-232.  doi:10.7525/j.issn.1673-5102.2020.02.009
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    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.
    PubZIP1 Gene Cloning,Subcellular Localization and Expression Pattern under Drought Stress
    LIU Xiao, YANG Jia, ZHANG Xin, MA Miao-Miao, YANG Jing-Li
    2020, 40(2):  233-242.  doi:10.7525/j.issn.1673-5102.2020.02.010
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    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.
    Cloning and expression analysis of 5 ZFP genes from Poplus trichocarpa
    LI Ya-Bo, Lü Jia-Xin, TAN Bing, GAO Cai-Qiu
    2020, 40(2):  243-250.  doi:10.7525/j.issn.1673-5102.2020.02.011
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    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, PEG6000 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.
    Function of Populus trichocarpa HDA902 Gene in Response to Cold Stress
    GUAN Tao, LIU Chao, LI Kai-Long, XIA De-An, MA Xu-Jun
    2020, 40(2):  251-256.  doi:10.7525/j.issn.1673-5102.2020.02.012
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    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.
    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
    2020, 40(2):  257-265.  doi:10.7525/j.issn.1673-5102.2020.02.013
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    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 T3 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. H2O2 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 H2O2 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.
    Expression Analysis of the FaesAP2B in Mutant Buckwheat with Long Pistil and Long Stamen
    ZHANG Jiao, WANG Xuan, ZHANG Liang-Bo, LIU Zhi-Xiong
    2020, 40(2):  266-273.  doi:10.7525/j.issn.1673-5102.2020.02.014
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    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 FaesAP2B(GenBank accession number is MK290847.1). By sequence analysis, FaesAP2B 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 FaesAP2B 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 FaesAP2B gene expression in root, stem, leaf, perianth, stamen, pistil and juvenile fruit in four-day buckwheat mutant lpls, respectively. The FaesAP2B is expressed in both vegetative tissues and reproductive structures. However, the expression level of FaesAP2B in reproductive organs such as floral organs and fruits is significantly higher than that in vegetative tissues. In addition, the expression level of FaesAP2B in stamens is the highest among seven different organs(LSD, P<0.01). And the expression level of FaesAP2B 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 FaesAP2B may be mainly involved in regulating flower and fruit development in the buckwheat mutant ‘lpls’.
    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
    2020, 40(2):  274-283.  doi:10.7525/j.issn.1673-5102.2020.02.015
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    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 H2O2, NaCl and NaHCO3 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.
    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
    2020, 40(2):  284-292.  doi:10.7525/j.issn.1673-5102.2020.02.016
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    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.
    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
    2020, 40(2):  293-300.  doi:10.7525/j.issn.1673-5102.2020.02.017
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    The BoPDIL1-2 gene was isolated from the stigma of ornamental kale(Brassica oleracea var. acephala) S13-bS13-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.
    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
    2020, 40(2):  301-307.  doi:10.7525/j.issn.1673-5102.2020.02.018
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    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.
    Scientific Notes
    Optimization of Dry Enzymatic Extraction Process and Physicochemical Properties of Pine Seed Oil
    ZU Shu-Chong
    2020, 40(2):  308-313.  doi:10.7525/j.issn.1673-5102.2020.02.019
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    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".
    Preparation Process and Bioavailability Evaluation of β-cyclodextrin Inclusion Korean Pine Seed Oil
    ZU Shu-Chong
    2020, 40(2):  314-320.  doi:10.7525/j.issn.1673-5102.2020.02.020
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    The single-factor method was used to optimize the preparation process of the Korean pine seed oil inclusion compound. Under the optimal conditions, the solidification rate of the Korean pine seed oil inclusion compound was 70.95%, and the oil content of the inclusion compound was 26.88%. The fatty acid composition, particle size, potential, and bioavailability of Korean pine seed oil and Korean pine seed oil inclusion complex were compared. There was no significant difference between the contents of each component of the Korean pine seed oil in the inclusion compound and that of the Korean pine seed oil, among which the content of pinolenic acid was in 14% to 16%. The average particle size of the Korean pine seed oil inclusion complex was 177.3±2.6 nm, and the potential was -33.01±1.4 mV. The blood drug concentration(CMAX), t1/2A, AUC and MRT were 1.27, 1.42, 1.56 and 1.04 times of the Korean pine oil inclusion complex group, respectively. The inclusion complex of Korean pine seed oil and aqueous solution formed an emulsion, the particle size decreased, the water solubility increased, the inclusion complex in vivo t1/2A and the average retention time(MRT) increased, and the absorption of pinolenic acid in vivo increased.