Table of Content

20 May 2015, Volume 35 Issue 3

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Reduction on Orinus tibeticus Based on the External Morphology and Micromorphological Characteristics of Leaf Epidermis

SU Xu,LIU Yu-Ping,LI Zhao-Meng,ZHOU Yong-Hui,CHEN Ke-Long,CHEN Wen-Li

2015, 35(3):  321-326.  doi:10.7525/j.issn.1673-5102.2015.03.001

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We analyzed the main character differences among nearest related Orinus tibeticus, O.thoroldii and O.anomala in the genus Orinus by morphological observation and anatomical analysis of leaf epidermises. There was very slight disparity between O.tibeticus and O.thoroldii in external morphology, and classifying two species was difficult. The differences between O.tibeticus and O.anomala were distinct, and two taxa were two different species. Anatomically, the most features of O.tibeticus were consistent with or similar to that of O.thoroldii, while for O.anomala, the discontinuity of characters between them was obviously present. Therefore, O.tibeticus and O.thoroldii should belong to the same species, and O.tibeticus can be reduced to synonymy of O.thoroldii.

Cloning and Genetic Transformation of CnCHS Gene from Camellia nitidissima

ZHOU Xing-Wen；LI Ji-Yuan；ZHU Yu-Lin

2015, 35(3):  327-332.  doi:10.7525/j.issn.1673-5102.2015.03.002

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Using total cDNA from the petals of Camellia nitidissima as template, we amplified the full length chalcone synthase(CHS) cDNA of C.nitidissima successfully by the specific primers designed according to the CnCHS sequence in genbank. The amplified fragment of CnCHS was inserted into PMD18-T vector and then transformed into E.coli DH5α. After the compounded plasmid was identified by enzyme digestion and sequencing, it was digested and connected to the expression vector pCAMBIA1300. The compounded plasmid pCAM-CnCHS was identified and the sense expression vector of CnCHS was constructed successfully. Then, the pCAM-CnCHS was transformed into Agrobacterium tumefaciens EHA105 for plant infections, and 18 transgenic tobaccos containing CnCHS were obtained. The transgenic plants were then identified by PCR and Southern blotting, and the transgenic tobaccos which contain single copy of CnCHS gene were obtained with the positive rate of 67%. The genetic transformation system of CnCHS to tobaccos was constructed successfully, which would lay the foundation for further function research and regulation effects on flower color of CnCHS.

Prokaryotic Expression of Caffeine Synthase Gene(TCS1),Its Polyclonal Antibody Preparation and Identification

DENG Wei-Wei；JIN Yang；LI Min；MA Lin-Long；ZHANG Zheng-Zhu

2015, 35(3):  333-339.  doi:10.7525/j.issn.1673-5102.2015.03.003

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Cloing and prokaryotic expressing of TCS1(Caffeine synthase 1) gene, preparing the antibody are the preliminary works for further research of the expression of TCS1 in tea plant. According to the cDNA fulllength sequence of TCS1 from GenBank, we found out the complete ORF(Open Reading Frame). The open reading frame(ORF) of TCS1 gene from the cDNA of leaves of tea plant was amplified. The ORF was then ligated into the expression vector pGEX-4T-2. The recombinant protein was expressed by induction with IPTG. Then, we tested in vitro enzyme activity, and purified the recombinant protein subsequently by affinity chromatography. The TCS1 antibody was further refined by immunizing white rabbits with the purified protein. The antibody titer and specificity of the polyclonal antibody were confirmed by ELISA and Western blot. Induced by optimizing the conditions, the best expression condition of the recombinant protein was at 30℃ for 4 h. After induction of total protein, soluble protein and inclusion body protein showed an obvious foreign proteins stripes. ELISA detection of the antibody recommended dilution ratio of 1∶2 000. We verified the favourable specific of the antibodies by western blot. The prokaryotic expression plasmid pGEX-4T-2-TCS1 was constructed, and the prepared polyclonal antibody had a high titer and good specificity.

Cloning and Identification of a Novel thioredoxin h Gene from Tamarix hisipida

SONG Xin；DONG Jing-Xiang；LI Kai-Long；LI Hong-Jiao；LI Hui-Yu

2015, 35(3):  340-346.  doi:10.7525/j.issn.1673-5102.2015.03.004

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Thioredoxins play an important role in various cellular processes through redox regulation. We isolated a novel Trx cDNA sequence, ThTrx1, from cDNA libraries of T.hisipida to characterize their functions under abiotic stress. By bioinformatic analysis, ThTrx1 is a new member of the thioredoxin h subfamily and belongs to subgroup 1. We used real-time quantitative RT-PCR to examine ThTrx1 gene expression in roots and leaves under different abiotic stresses(NaCl, PEG, CdCl₂, low temperature and ABA). The mRNA levels of ThTrx1 in both leaves and roots were up-regulated under NaCl, PEG, CdCl₂ and ABA treatments. However, the expression of ThTrx1 mRNA decreased during low temperature treatment. We also expressed the ThTrx1 gene in E.coli and examined its effect on bacterial growth and stress tolerance. The E.coli containing recombinant plasmid(pET32a-Trx1) was more resistant to salt, metal ions and drought stress that was the control. Therefore, ThTrx1 is involved in an abiotic stress response and regulated by ABA-dependent signaling pathways.

Constructingand Function of Plant Expression Vector and RNA Interference Vector of a FAD2 Gene from Camellia oleifera

CHEN Hong-Peng；TAN Xiao-Feng；XIE Yao-Jian；WU Zhi-Hua；ZHANG Dang-Quan；ZENG Yan-Ling

2015, 35(3):  347-354.  doi:10.7525/j.issn.1673-5102.2015.03.005

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As an edible vegetable oil extracted from the seeds of Camellia oleifera, tea oil is rich in oleic acid, linoleic acid, linolenic acid and other unsaturated fatty acidwhich is essentialfor human beings healthy. In the plant biosynthesis pathway of unsaturated fatty acids, monounsaturated fatty acids are catalyzed into poly unsaturated fatty acids step by step by fatty acid desaturase(FAD) family members and FAD2 is mainly in chargeof catalyzing oleic acid(18∶1Δ⁹) and palmitoleic acid(16∶1Δ⁹) into linoleic acid(18∶2Δ^9,11) and hexadecadienoic acid(16∶2Δ^9,11). In order to reveal the function of CoFAD2 gene from C.oleifera, the plant expression vector pBI121-CoFAD2 and RNA interference vector pBI121-CoFAD2 RNAi were constructed on basis of the former research, and pBI121-CoFAD2 and pBI121-CoFAD2 RNAi were transferred into FAD2 mutant and wild type of Arabidopsis plants, respectively. Compared with the wild type, the content of oleic acid(18∶1) and palmitoleic acid(16∶1) was higher, the content oflinoleic acid(18∶2) and hexadecadienoic acid(16∶2) was lower in the FAD2 mutant; moreover,the fatty acid component of the mutant was recovered equal to wild type after transformation by pBI121-CoFAD2.However, the fatty acid component of the wild type plant was changed after transformation by pBI121-CoFAD2 RNAi, oleic acid(18∶1) and palmitoleic acid(16∶1) content increased and the content of linoleic acid(18∶2) and hexadecadienoic acid(16∶2) decreased.Therefore, CoFAD2 gene has function incontrol of catalyzing oleic acid(18∶1Δ⁹) and palmitoleic acid(16∶1Δ⁹) into linoleic acid(18∶2Δ^9,11) and hexadecadienoic acid(16∶2Δ^9,11) and plays an important role in the regulation of tea oil fatty acid composition.

Construction of RNA interference vector carrying two genes of DS and CAS of Betula platyphylla Suk（Birch.）

LIANG Tian；MA Hong-Si；YIN Jing；ZHAN Ya-Guang；ZHANG Meng-Yan

2015, 35(3):  355-362.  doi:10.7525/j.issn.1673-5102.2015.03.006

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Cycloartenol synthase(CAS) and dammarenediol synthase(DS) are the key enzymes of branch pathway of birch in birch triterpenoid synthesis process. Silencing of DS gene and CAS gene help triterpenoid synthesis precursor-2,3-oxidesqualene to synthesize betulin and oleanolic acid. PCR method of homologous sequences and a new vector pRNAi-GG were used to construct RNAi expression vectors of CAS gene and DS gene. The cloning part cDNA sequence of DS and CAS gene, respectively, CAS gene fragment length was 300bp, DS gene fragment length was 509bp, and their similarity reached 100% and 92%, respectively, compared with BPX1 and OSCBPD of Betula platyphylla var. japonica. With the principle of primer design and pRNAi-GG instruction, we connected the fragments on forward and contrary direction to the both sides of PDK intron of pRNAi-GG. The recombinant vectors were transformed into competent E.coli cells Trans1-T1. By PCR and sequence detection, the interference vector of DS gene and interference vector of DS+CAS two genes were constructed successfully. Our study will lay the foundation for the genetic modification of blocking-up triterpenoid branch pathways and promote the precursor to synthetize betulin and oleanolic acid in the triterpenoids metabolism pathway of white birch.

Isolation and Functional Analysis of FLC Gene from Poplar (Populus simonii×Populus nigra)

SUN Qi；QU Chun-Pu；ZHENG Mei-Zhu；LIU Guan-Jun

2015, 35(3):  363-369.  doi:10.7525/j.issn.1673-5102.2015.03.007

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MADS-box family protein is an important class of transcription factors regulating flowering. FLC is a member of MADS-box transcription factors, and the FLC gene is controlled by many pathways, and then to regulate the flowering. We cloned a FLC-homolog gene in male flower buds from Populus simonii×Populus nigra. The open reading frame (ORF) of this gene had 726 bp which encoded a protein containing 241 amino acid. The molecular weight of the predicted protein was 27.559 KD, and the pI was 9.37. By real-time quantitative PCR, vernalization could down-regulate the expression of PnFLC 57.9%-84% in the root, stem and leaves tissues of poplar. By GUS staining, the promoter of the PnFLC is highly expressed in actively dividing tissues. Furthermore, by transforming Arabopsis, PnFLC down-regulated the expression of AtAP1, AtSOC1 and AtFT, while delayed the flowering in Arabidopsis.

Effects of Ammonium and Amio-acid Nitrogen Fertilization on Plant Growth and Carbon Metabolism of Sugar beets(Beta vulgaris L.)

HU Xiao-Hang；ZHOU Jian-Chao；CHEN Li-Xin；WANG Qiu-Hong

2015, 35(3):  370-377.  doi:10.7525/j.issn.1673-5102.2015.03.008

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We studied the effects of different proportions of inorganic nitrogen(ammonium nitrogen) and organic nitrogen(amino acid nitrogen), accounting for 33% and 67% of total nitrogen, respectively, on plant growth and carbon metabolism of sugar beet in pot experiment. We analyzed the leaf chlorophyll content, total nitrogen and dry matter of leaf and root, the sucrose phosphate synthase(SPS), sucrose synthase(SS), invertase in leaf, the soluble sugar, sucrose and reducing sugar of leaf and root, sugar yield and content at different growth stages of sugar beet. Different nitrogen forms and proportion treatments produced various effects on plant growth and carbon metabolisms at different growth stages of sugar beet. Both above-ground and under-ground dry matter accumulations of sugar beet were increased by different ratios of organic nitrogen treatments. Before harvest, organic nitrogen(67% of total nitrogen application) treatment could promote the growth of sugar beet significantly. However, the amount of dry matter in sugar beet was the highest under organic nitrogen(33% of total N application) treatment in harvest. In the whole growth stage, the leaf chlorophyll content of sugar beet was increased under the organic nitrogen(33% of total nitrogen application) and organic nitrogen(67% of total nitrogen application). After sugar increasing period, the nitrogen accumulations in root and leaf of sugar beet under organic nitrogen treatments were higher than those under inorganic nitrogen treatment, which indicated amino acid nitrogen was a good nitrogen source. Various ratios of organic nitrogen treatments could enhance the SPS, SS and invertase activities in sugar beet leaf at different development stages. With the increase of the proportion of organic nitrogen, soluble sugar and sucrose content were increased significantly in root, the reducing sugar content was declined, and such changes played a functional role in the sucrose accumulation. Sugar content was enhanced with the increase of the organic nitrogen proportion in root. By contrast, both root and sugar yield were the highest when organic nitrogen accounted for 33% of total nitrogen.

Carbon Storage of 45-year-old Stands in Different Origins on the West Slope of Zhangguangcai Mountain

SUN Hong-Yang；WANG Qing-Cheng

2015, 35(3):  378-383.  doi:10.7525/j.issn.1673-5102.2015.03.009

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In Wuchang Fenghuangshan Forest Farm on the west slope of Zhangguangcai Mountain, we selected Larch(Larix olgensis L.), Larch and broad-leaved mixed forest(H), and secondary broad-leaved natural forest(C) in different origins(artificial regeneration, artificial-natural regeneration, and natural regeneration) to study the carbon storage of tree layer, shrub and grass layer, the litter layer, soil layer and the total carbon storage of all three types of forest stands, explore the effects of different ways of regeneration on forest carbon storage, and determine the way of forest regeneration with high carbon sequestration capacity. Artificial-natural mixed stand could increase the carbon storage of the shrub and grass layer(P<0.05). L showed the highest carbon storage of undecomposed litter layer, semi-decomposed litter layer and the highest total carbon storage of litter, and its carbon storage in the semi-decomposed litter layer and the total carbon storage of litter were significantly different in the three forest types(P<0.05). The total soil carbon storage and the total carbon storage of the forest ecological system were higher in H than that in the other two forest types, but the difference was not significant(P>0.05). The artificial-natural regeneration was conductive to increasing the carbon sink capacity of vegetation and soil, and Larch artificial regeneration had high carbon storage of litter.

Soil Respiration and Total Belowground Carbon Allocation of Betula costata Secondary Forest and Mixed Broadleaved Korean Pine Forest in Xiaoxing’an Mountains,China

HUANG Wei；MAO Zi-JUN；HAN Ying-Ying；SUN Tao

2015, 35(3):  384-390.  doi:10.7525/j.issn.1673-5102.2015.03.010

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Soil respiration and total belowground carbon allocation are two most important carbon fluxes of forest ecosystems. We measured them in Betula costata secondary forest and mixed broadleaved Korean pine forest. The mean soil respiration of B.costata secondary forest was higher than that of the mixed broadleaved Korean pine forest in the growing season（5.52 and 5.43 μmol·m^-2·s^-1）. The seasonality of soil respiration was driven mainly by soil temperature, which led to 77% and 81% of the variation of soil respiration in B.costata secondary forest and mixed broadleaved Korean pine forest, respectively. Q₁₀ was 2.74 and 2.23, for two-type forest, respectively. The annual flux of soil respiration and total belowground carbon allocation in B.costata secondary forest were both higher than those in mixed broadleaved Korean pine forest.

Effect of Exponential Fertilization on Growth Dynamic,Biomass Allocation and Photosynthesis of Betula platyphylla Bare-rooted Seedlings

LI Wen；ZHANG Cheng；WANG Qing-Cheng；HAO Long-Fei；LIU Shuang

2015, 35(3):  391-396.  doi:10.7525/j.issn.1673-5102.2015.03.011

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We studied the growth(height, ground diameter, biomass) dynamic, biomass allocation, relative growth rate and leaf photosynthesis of Betula platyphylla bare-rooted seedlings under constant fertilization(CF), exponential fertilization(EF) and 1.5 exponential fertilization(EF1.5) regimes in one growing season. Dynamic of seedling height, ground diameter and biomass presented the same tendency, which barely showed difference in the early growing stage between three treatments, while both EF and EF1.5 were significantly higher than CF in the late growing period; however, EF and EF1.5 were not significantly different. At the end of the growing season, EF1.5 seedlings were the tallest, EF and EF1.5 seedlings were 18.2% and 25% taller than CF, respectively(P>0.05). EF seedlings owned the thickest ground diameter, EF and EF1.5 were 11.2% and 5.8% thicker than CF, respectively(P<0.05). The biomass sequence of each plant between three treatments was EF>EF1.5>CF, and EF and EF1.5 were 41.5% and 25.4% higher than CF, respectively(P<0.05). Stem biomass, leaf biomass and root biomass under EF and EF1.5 were both significantly higher than those under CF(P<0.05). Height, ground diameter and biomass RGR under EF and EF1.5 were both significantly higher than those under CF(P<0.05), and biomass RGR was EF>EF1.5(P<0.05). During the fast-growing periods of seedlings, EF treatment promoted seedling leaf photosynthetic characteristics. The seedling leaf area under EF treatment was 16.8% greater than that under CF treatment(P<0.05). Chlorophyll content and photosynthetic rate were increased by 4.5%(P>0.05) and 5.0%(P>0.05). EF and EF1.5 promoted seedling growth and biomass accumulation obviously than CF. EF saved a large amount of fertilizer than EF1.5, and promoted seedling growth similarly. Therefore, EF is the best treatment.

Synergistic Use of ICESat/GLAS and MISR Data for Estimating Forest Aboveground Biomass

WU Di；FAN Wen-Yi

2015, 35(3):  397-405.  doi:10.7525/j.issn.1673-5102.2015.03.012

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Large footprint lidar ICESat/GLAS data contain lots of vertical structure information of ground including forest vertical profile and terrain, and these parameters have high correlation with above ground biomass. Based on waveform data process, we extracted waveform parameters, and used linear stepwise regression model and Erf-BP neural network model to set up the relation between waveform parameters and forest above ground biomass, respectively. We used the Erf-BP neural network model and waveform data of GLAS to inverse the biomass of GLAS footprints. We used a machine learning approach named random forests and the MISR data to build biomass model to extend point to planar and validated the accuracy of this model with the field measured data from plots. The prediction accuracy of Erf-BP neural network model (P=0.965, R_MSE=3.81 t·ha^-1) was better than that of the linear stepwise regression model (P=0.86, R_MSE=4.54 t·ha^-1). The predicting precision of the biomass spatial extensive model was good (P=0.81, R_MSE=2.39 t·ha^-1), the range of inverse biomass in research area was 0-144.4 t·ha^-1, and the mean of the biomass was 59.28  t·ha^-1. The retrieval precision verified by sample data was high (R²=0.72, R_MSE=8.98 t·ha^-1). The estimation was close to real value. Therefore, using few field measurement data to inverse forest ABG in large scale and high precision could provide an effective approach for forestry inventory, ecology research and carbon investigation.

Physical-chemical Properties of Soil,C and N of Three Typical Communities in Tianjin Coastal Wetland

CHEN Guo-Ping；CHENG Shan-Shan；LIU Jing；CONG Ming-Yang；GAO Xin；WANG Hui；SHI Fu-Chen

2015, 35(3):  406-411.  doi:10.7525/j.issn.1673-5102.2015.03.013

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We measured the ecological characteristics including salinity (EC), bulk density, pH, organic carbon density and nitrogen content in different depths of soils from Phragmites australis community, Suaeda glauca community and Aeluropus sinensis community in coastal wetlands. The order of soil salinity in 0-30 cm depth soil was: the highest in the A.sinensis community and the lowest in the P.australis community. For bulk density, the highest value was in the A.sinensis community and the lowest in the P.australis community. The pH was the highest in the S.glauca community and the lowest in the P.australis community. Within these community, organic carbon density in 0-30 cm depth soil was the highest in the P.australis community (7.06 kg·m^-2), followed by A.sinensis community (6.38 kg·m^-2) and the lowest in the S.glauca community (5.70 kg·m^-2). Organic carbon density in the topsoil (0-10 cm) was the highest, and then decreased with the increasing of the soil depth. The highest total nitrogen content in 0-30 cm depth soils was from P.australis community and the lowest was from A.sinensis community. The order of NO^-₃-N content was S.glauca＞P.australis＞A.sinensis community, and the order of NH⁺₄-N was P.australis＞S.glauca＞A.sinensis. There were significant negative correlations between organic carbon density and pH value, and between total nitrogen, pH and bulk density. NH⁺₄-N content had significant positive correlations with both organic carbon density and total nitrogen, and NO^-₃-N content had a significant positive correlation with organic carbon density, but no significant correlation with other factors.

Effect of Fertilization on Woody Characteristics of Betula platyphylla

CUI Zhi-Yuan；LIU Gui-Feng；LI Bo；GUO Hai-Feng；WEI Xing-Biao；WANG Chao

2015, 35(3):  412-417.  doi:10.7525/j.issn.1673-5102.2015.03.014

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To search the effects of different fertilization prescriptions on woody traits of Betula platyphylla, we chose four group B.platyphylla, fertilized by nitrogen, potash, phosphatate and total nutrient elements fertilizer, respectively, for three years’ treatment. We observed the differences between four groups of B.platyphylla in terms of growth characteristics, the content of lignin and cellulose, wood density, microfibril angle, and fiber morphology changes. Two groups fertilized by nitrogen and total nutrient elements fertilizer had significant changes compared to the control group. Diameter at breast height of two groups were 1.6 and 1.9 times more than that of the control group, respectively; crown width were 1.4 times and 1.5 times more than that in the control group, respectively; cellulose contents increased by 8.7% and 14.3% than that in the control group, respectively. In cross-sectional unit area, the number of catheters increased, the diameters of catheters decreased, length and width of fibers decreased significantly. However, lignin contents and aspect ratio of those two groups did not change significantly. In contract, the wood density of the group fertilized by phosphatate fertilizer had increase by 5% than that of the control group, and microfibril angle of the phosphatate fertilized group decreased by 5.4% than that of the control group. The results are that increasing nitrogen fertilizer reasonably can improve the paper performance of birch and the increasing phosphate fertilizer reasonably can improve the mechanical strength of birch.

Development of Pinus koraiensis Tree-ring Chronology and Master Year Analysis in Liangshui National Natural Reserve,China

YANG Qing-Xiao；ZHU Liang-Jun；WANG Xiao-Chun

2015, 35(3):  418-424.  doi:10.7525/j.issn.1673-5102.2015.03.015

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We developed one tree-ring chronology of Korean pine(Pinus koraiensis) in Liangshui National Natural Reserve, Xiaoxing'an Mountain, Northeast China, by using traditional dendrochronology. We identified the relationships between Korean pine chronology and monthly climatic factors(air temperatures and precipitation) by correlation analysis and single year analysis. Korean pine radial growth was mainly limited by both monthly air temperatures and precipitation, and the limited effects of minimum temperatures before growing season(from February to April) and mean temperatures and precipitation during growing season on Korean pine growth were most significant, and Korean pine radial growth was positively correlated with temperatures in the previous June and July and the current February and April. The responses of the chronology to monthly maximum and minimum temperatures showed a similar relationship with monthly mean temperatures, which indicated that Korean pine radial growth were mainly influenced by air temperatures before and during growing seasons. The chronology was negatively correlated with maximum and mean temperature in June of the current year, and positively correlated with precipitations in March and June. The soil moisture in June also had a significant influence on Korean pine growth. High temperature in June accelerates soil evaporation and plant transpiration, resulted in water stress and narrow ring formation of Korean pine in this region. Single year analysis confirmed the results from correlation analysis. Therefore, Korean pine radial growth was synthetically affected by precipitation and temperature during and before growing seasons, and also seriously limited by soil moisture in June.

Branch Dynamics for the Korean Pine Plantation Based on Linear Mixed Mode

JIA Wei-Wei；SUN Shou-Qiang；LI Feng-Ri；GAO Hui-Lin

2015, 35(3):  425-430.  doi:10.7525/j.issn.1673-5102.2015.03.016

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With the branch analysis data of 4 987 branches for 79 Korean pine plantations in Mengjiagang Forest Farm, we established the branch diameter and branch length of the linear mixed model and the branch diameter and branch length growth of the nonlinear mixed model. We used SAS9.22 to solve parameters of the model and used AIC, BIC and LRT to give significance test for the converged model. For branch diameter and branch length linear mixed model, all the parameters were significant for t test of the branch diameter and branch length and the standard error of the parameters was low, and it performed well as for the stability of the models. For branch diameter growth nonlinear mixed model, the combination for b₁ and b₃ for the model performed well based on the low value for AIC and BIC, when the sample plots effect was not considered. When the sample plots effect was considered, the combination for b₁ and b₃ for the model performed well compared to the other combination form. For branch length growth nonlinear mixed model, the parameter combination for b₁ and b₃ of the model performed well based on the AIC and BIC when the sample plot effect was not considered, and if considered, the combination for b₃ and b₄ performed best for the model.

Extraction and Antioxidant Activity of Tannic in Juglans mandshurica Maxim.

ZAN Zhi-Hui；GAO Yan-Mei；SUN Mo-Long

2015, 35(3):  431-435.  doi:10.7525/j.issn.1673-5102.2015.03.017

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Juglans mandshurica Maxim. tannins was extracted by ultrasonic method. Tannins content was determined by UV spectrophotometry. The effects of the volume fraction of acetone, ultrasonic temperature, ultrasonic time, and ratio of material to liquid on tannins were evaluated by orthogonal test method in the ultrasonic experiment. The optimal extraction conditions for the tannins in green peel of J.mandshurica were the volume fraction of acetone of 40%, ultrasonic temperature of 40℃, extraction time of 30 min, and solid-liquid ratio of 1∶15. The optimal extraction conditions for the tannins in leaves of J.mandshurica were the volume the fraction of acetone of 50%, ultrasonic temperature of 60℃, extraction time of 40 min, and solid-liquid ratio of 1∶20. The tannins in both green peel and leaves had excellent scavenging capacity for hydroxyl radical, nitrite ions and superoxide anion. The ability of scavenging three kinds of free radicals of the tannins in leaves was stronger than that of the green peel. J.mandshurica tannins had strong antioxidation, and the tannins was with potential pharmaceutical value as a good natural antioxidant.

Structure Characteristics of Dead Tree in Dominant Species Populations in An Evergreen Broad-leaved Forest of Baishanzu

WU You-Gui；XU Da-Ming；LAN Rong-Guang；WU Tian-Ming；WU Yi-Shong；LUO Zheng-Rong

2015, 35(3):  436-442.  doi:10.7525/j.issn.1673-5102.2015.03.018

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An experiment was conducted to study the dead tree structure characteristics of dominant species populations in an evergreen broad-leaved forest with the dead trees of ten years and living trees form ten dominant species in a 5 hm² permanent plot in north slope of Baishanzu Mountain. In the age class of dead trees with inverted-J distribution, they were Cleyera pachyphylla, Sycopsis sinensis, Ilex editicostata, Cyclobalanopsis stewardiana and C.multinervis, Lithocarpus brevicaudatus and Fagus lucida, Schima superba. In the age class of dead trees with peak type distribution(with the peak in the second age class), they were Rhododendron latoucheae and Sorbus folgneri. The dead trees mainly concentrated in the top three age classes. The death ratio of every dominant species in the first age class was biggest, especially while the death ratio of Rhododendron latoucheae and Lithocarpus brevicaudatus in the first age class were relatively larger, being 47.61% and 50.00%, respectively. The death ratio of dominant species decreased with the increasing of age class, and it changed after the fourth age class. The average death ratio of Sorbus folgneri was biggest(30.42%), and that of Fagus lucida was smallest(13.70%). The dead trees were denser in the habitats where the living trees were denser. The density of dead trees of Sycopsis sinensis was higher in gully, and that of Cyclobalanopsis multinervis had little difference among four kinds of habitats. The dead trees of the other eight dominant species populations were denser in ridge and sparser in gully. The death ratios of Sorbus folgneri in steep slope(42.86%) and Ilex editicostata in gully(29.78%) were higher, while that of Fagus lucida in ridge(12.40%) was lowest.

The Relationship between Crown Size and Pine Nut Yield for Korean Pine Plantation

ZHANG Qiang；DONG Li-Hu；LI Feng-Ri

2015, 35(3):  443-449.  doi:10.7525/j.issn.1673-5102.2015.03.019

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With the data of the branch analysis of 70 sample trees from 12 sample plots and stand pine nut yields(2008-2010) from 21 permanent sample plots for Korean pine(Pinus koraiensis) plantation in Mengjiagang Forest Farm, Jiamusi, Heilongjiang Province, we used allometric growth equation to develop the predicting models of crown volume(C_V) and crown surface area(C_A), and analyzed the correlation between pine nut yield of a bad harvest year, an average harvest year, a good harvest year and crown size. The correlations between crown volume and crown surface area with crown length, diameter at breast height, crown width and tree height were all significant, the maximum correlation coefficient was with diameter at breast height, and the minimum correlation coefficient was with tree height. The determination coefficients(R²) of the models were all above 0.55, and most of the prediction accuracies were above 85%. We used the models for crown volume and crown surface area to analyze their influence on the pine nut yield, and the result indicated that the pine nut yield had the linear correlation with crown volume and crown surface area for Korean pine plantation, in which the relationship between pine nut yield in a good harvest year with crown volume and crown surface area was the most obvious(r=0.726 7-0.733 8), followed by in an average harvest year(r=0.688 8-0.667 5), and the worst was in a bad harvest year(r<0.5). Generally, the models of crown volume and crown surface area with univariate and multivariate variables for Korean pine plantation standing tree are suitable for estimating the crown size of Korean pine, and providing further study of pine nut yield and sustainable management of Korean pine plantation.

Genetic Variation within Ten Accessions of Dipsacus in China by ISSR Analysis

LI Xiao-Ling；YANG Jin；ZHANG Xiong；WANG Xue-Song

2015, 35(3):  450-456.  doi:10.7525/j.issn.1673-5102.2015.03.020

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Dipsacus L., a member of Dipsacaceae, is an important plant for the traditional herbal medicine in China. But there is still inaccurate identification in the botanical origin and genetic relatedness of Dipsacus plant. We used ten highly polymorphic ISSR(Inter-simple sequence repeat) markers to assess the genetic relatedness of ten accessions of Dipsacus, and used ten highly polymorphic ISSR primers from 100 primers screened to conduct PCR amplification with the genomic DNA isolated from ten accessions of Dipsacus. A total of 947 DNA bands were produced, of which 828 were polymorphic(87.2%) among ten accessions, and Dipsacus plant had abundant genetic variation. The Nei’s genetic coefficient ranged in 0.557 0.806 with an average of 0.661. By UPGMA cluster, ten Dipsacus accessions were grouped into two main groups(Group 1 and Group 2 ) with a similarity level of 0.582 and then into five subgroups at the similarity level of 0.702. Group 1 contained DL(L), DL(W), DL(H), DL(WS), ES, RB, C(J) and C(D) and was further divided into three subgroups(Subgroup 1, Subgroup 2 and Subgroup 3) and Group 2 consisted of DT and SZ, which was distance from any other accessions and constituted a separate minor subgroup(Subgroup 4 and Subgroup 5), respectively. Subgroup 1 contained DL(L), DL(W), DL(H) and DL(WS), Subgroup 2 with ES and RB and Subgroup 3 with C(J) and C(D), which indicated that the genetic relationship among them was very close and the gene flow among them was frequent. The results of principle coordinate analysis(PCA) were consistent with those of the UPGMA analysis. The distance between ES and RB was extremely narrow(0.210), which showed that RB and ES had a close relationship between them. The geographical distributions of them had overlapped intensively. The molecular cluster result corresponded to the morphology analysis. The results also showed that ES may be a variant of RB.

A Simple and Efficient Method of DNA Extraction from Different Plant Samples

DING Hao；ZHENG Tang-Chun；LIANG De-Yang；QU Guan-Zheng

2015, 35(3):  457-461.  doi:10.7525/j.issn.1673-5102.2015.03.021

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It is difficult to extract high quality DNA from plants for molecular work, since there are a great deal of secondary metabolities. We provided a simple improved CTAB protocol for extracting plant DNA. RNA was digested with RNase A after the step of incubation at 65℃. Then, DNA was extracted, precipitated and washed. The purity and quality of total DNA was identified by agarose gel electrophoresis and nucleic acid detection instrument. PCR amplification of the extracted DNAs as templates could present clear bands of the target gene. Compared with the previous methods, it was simple and high throughput to be used for extraction of purified DNA from different plant samples, which would set a technical foundation of subsequent analysis of molecular biology.

Removal Ability of Floating Macrophytes on N and P from Eutrophicated Water

PAN Bao-Yuan；YANG Guo-Ting；MU Li-Qiang；MA Yun；LI Jing

2015, 35(3):  462-466.  doi:10.7525/j.issn.1673-5102.2015.03.022

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We studied the effects of floating macrophytes on eutrophication water under different nutrition conditions. Eichhornia crassipes, Pistia stratiotes and Dracaena sanderiana can eliminate total nitrogen(TN) and total phosphorus(TP) in water. After growing 30 days, TN and TP removal rates were 50%-78.46% and 68.16%-89.56% by E.crassipes, respectively. TN and TP removal rate were 57.58%-76.87% and 72.28%-76.47% by P.stratiotes, respectively. TN and TP removal rate were 0.60%-10.80% and 2.48%-10.04% by D.sanderiana, respectively. TN and TP removal rate were 31.17% and 20.48% by E.crassipes and P.stratiotes from treated water in the outdoor experiment. Thus, E.crassipes and P.stratiotes have good removal effect on nitrogen and phosphorus of eutrophication water.

Tara Oil Extraction by Supercritical Carbon Dioxide and Its Quality

LI Zhan-Jun；ZHANG Lin；ZU Yuan-Gang；YANG Feng-Jian

2015, 35(3):  467-470.  doi:10.7525/j.issn.1673-5102.2015.03.023

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We used three groups of parallel single factor experiment to determine Tara seed oil extraction process conditions to improve Taraoil yield. We focused on supercritical CO₂ fluid extraction technology to extract Tara oil, optimized the Tara oil’s extraction process and methods, and analyzed the Tara oil quality. With oil extraction yield as index, on the basis of a single factor preliminary experiments, we studied extraction time, extraction temperature, extraction pressure, and their interactions on Tara seed oil extraction yield systematically. The optimum extraction conditions were extraction time of 120 min, extraction temperature of 45℃, and extraction pressure of 35 MPa. By single factor experiment for three repeated experiments, under optimum conditions the Tara oil actual extraction yield was 21.07%, which was consistent with the single factor experiment results. The extracted oil showed transparent and yellowish, belonging to drying oil with high unsaturated fatty acid content, with high quality and valuableuse.

Preparation, Characterization and Function Evaluation of Ultrafine Resveratrol Powder

WANG Kun-Lun；ZHAO Xiu-Hua；ZU Yuan-Gang；LI Jia-Lei

2015, 35(3):  471-475.  doi:10.7525/j.issn.1673-5102.2015.03.024

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By supercritical SAS, we made the resveratrol anti solvent recrystallization to prepare ultrafine resveratrol powder under supercritical condition. The conditions were under 50℃, 20 MPa, resveratrol ethanol solution concentration of 9 mg·mL^-1, and sample solution flow rate of 1 mL·min^-1. We analyzed the particles by Dynamic Light Scattering(DLS) equipment and Scanning Electron Microscopy(SEM). The particle size of the particles was greatly reduced and the average particle size was 0.68±0.03 μm. In addition, characterization of ultrafine Res and raw-resveratrol were tested by Fourier transform infrared spectroscopy(FT-IR), X-ray diffraction(XRD), High performance liquid chromatography(HPLC) and Liquid chromatography-mass spectrometry(LC-MS). SAS process didn't change the chemical structure of Res. The chemical structure of resveratrol was unaffected by the SAS process. The degree of crystallinity of the u-Res particles greatly reduced. We used the purity of the u-Res particles increased from 98.5% to 99.2% during the SAS process. Using drosophila anti-aging experiments to detect the oxidation resistance of resveratrol. The antioxidant capacity of resveratrol powder were higher than that in the second-stage and blank experiment under the concentration of resveratrol solution, 2.5, 5 and 10 mg·mL^-1. The resveratrol had the oxidation resistance, and ultrafine resveratrol powder was better than raw-resveratrol powder.

Enzymatic Hydrolysis Technology from Tala Fiber Residues

LU Qi；GAO Yue；XIANG Feng-Ying；NIE Si-Ming；ZHANG Ying；ZU Yuan-Gang

2015, 35(3):  476-480.  doi:10.7525/j.issn.1673-5102.2015.03.025

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Tannin from Tara pods is hydrolyzed for preparation of gallic acid, whose residues is rich in cellulose. We discussed the effects of various factors on the yield of reducing sugar from Tara fiber residues. By the single factor experiment, we optimized four factors including the ratio of solid-liquid, enzymatic temperature, pH and enzymatic time by orthogonal experiments L₁₆(4)⁵. The optimum conditions were under the ratio of solid-liquid of 1∶6 g·mL^-1, enzymatic temperature of 55℃, pH of 6 and enzymatic time of 48 h, and the yield of reducing sugar from Tara fiber residues was 43.95%.