Table of Content

20 July 2015, Volume 35 Issue 4

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Two New Species of the Genus Aconitum(Ranunculaceae) from Gansu

WANG Wen-Tsai

2015, 35(4):  481-483.  doi:10.7525/j.issn.1673-5102.2015.04.001

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Two species of the genus Aconitum(Ranunculaceae), Aconitum hezuoense and A.lianhuashanicum, are described as new from Gansu Province. The morphological differences between them and their allies are given respectively.

A Taxonomic Revision of Anomobryum(Bryaceae,Musci) in China

LIU Yong-Ying1,2；WANG Yu-Shui1；LI Lin2；ZHAO Jian-Cheng2*

2015, 35(4):  484-493.  doi:10.7525/j.issn.1673-5102.2015.04.002

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Phylogenetic relationshios between Anomobryum and Bryum, and between Anomobryum and Pohlia were discussed based on the morphological, anatomical and molecular data to determine that Anomobryum was a valid genus. Anomobryum julaceum(Schrad. ex G. Gaertn., B.Mey. & Scherb.) Schimp. and A.nitidum(Mitt.) Jaeg. were available name instead of A.filiforme(Dicks) Husn. and A.gemmigerum Broth., separately. A.concinnatum(Spruce) Lindb. was recognized at the specific level. Newly we updated the checklist of Anomobryum in China, including five species. We provided a key to the species of Anomobryum in China, as well as diagnostic characteristics and habitats for each data. The voucher specimens were deposited in the herbarium of Hebei Normal University(HBNU), and Chinese National Herbarium, Chinese Academy of Sciences(PE).

Intraspecific Variability of Pollen Morphology of A Suspicious Species in the sect. Ranunculoides

LIU Xiao-Mei1；LIU Yun-Yun1；XIE Fei1；HONG Fang-Ling1；LI Zhi-Zhong1；YAN Hao1；SHAO Jian-Wen1,2*

2015, 35(4):  494-498.  doi:10.7525/j.issn.1673-5102.2015.04.003

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We investigated the pollen morphology of 50 individuals(from five populations) of a suspicious species(in sect. Ranunculoides Chen et C.M.Hu), distributing at northwestward in Zhejiang Province. The pollen morphology of this suspicious species were high diversity. Based on the shape and number of aperture and their component, five pollen types could be recognized, including stephanocolpate type, colpi more than pores type, pores more than colpi type, pantoporate type and ringcolpate type. The component of pollen types was comparatively different among studied populations. There might be more than one pollen types within the same individual or the same anther. There might be high variability of pollen morphology at intraspecific level. Pollen morphology characters showed this suspicious species was a distinct and authentic new species, and likely evolved from cross bred progeny of parents with stephanocolpate type and pantoporate type pollens.

Effects of Pb²⁺ on Spore Germination and Gametophyte Development of Three Species of Fern

WANG Sheng；LIU En-De；XIA Yi；DAI Xi-Ling*

2015, 35(4):  499-503.  doi:10.7525/j.issn.1673-5102.2015.04.004

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We studied the effects of heavy metal Pb²⁺ on spore germination of Ceratopteris thalictroides, Alsophila costularis and Neocheiropteris palmatopedata and the gametophyte development of C.thalictroides. With the increasing of Pb²⁺, germination percentage of spores is reduced in C.thalictroides, A.costularis and N.palmatopedata. When the concentration is lower than 10^-4 mol·L^-1, the spore of C.thalictroides will germinate. When the concentration is lower than 10^-5 mol·L^-1, the spore of A.costularis and N.palmatopedata will germinate. Spores of C.thalictroides, A.costularis and N.palmatopedata have a certain tolerance to Pb²⁺. The heavy metal (Pb²⁺) have an effect on gametophyte development of C.thalictroides, which makes gametophyte irregular and produces more male gametophyte. The antheridium is recessed into the edge of prothallium, and the chloroplast is unevenly distributed in the C.thalictroides. The present investigation provided some selective plants which can repair the Pb²⁺ polluted soil.

Sporogenesis and Gametogenesis of Calla Lily(Zantedeschia hybrids)

YANG Liu-Yan1,2；WANG Yu-Bing3；ZHANG Yong-Chun1,2*；SUN Yi1,2

2015, 35(4):  504-508.  doi:10.7525/j.issn.1673-5102.2015.04.005

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We studied the megasporogensis, microsporogensis and the development of female and male gametophytes of Zantedeschia ‘Majestic Red’ through traditional paraffin sections. The ovule was anatropous, bi-integument, and crassinucellar with integument tapeta. The megaspore mother cell meiosis formed a linear megaspore tetrad. The chalazal end megaspore developed into the embryo sac mother cell, which developed to Polygonum type embryo-sac, another 3 megaspores degenerated. Each stamen had 2-3 anthers, with butterfly-shaped pollen sacs, ach side with 2 small sporangial. The wall of anther was completely developed, which consisted of four layers: epidermis, endothecium, middle layers and tapetum. The tapetum was deformation type. The cytokinesis of microspore mother cell during meiosis was simultaneaous type. Microspore tetrads were mainly cross, and the pollen grains had 2-celled when shed.

Characteristics of Leaf Anatomical Structure of Malus sieversii in Different Geographical Distribution and Plant Age

YANG Mei-Ling；YAN Xiu-Na；YU Wei-Wei；LONG Hong；YAN Guo-Rong*

2015, 35(4):  509-514.  doi:10.7525/j.issn.1673-5102.2015.04.006

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We analyzed the characters of leaf anatomical structures of Malus sieversii(wild apple trees) in different populations and different growth ages by paraffin sections. The leaf of M.sieversii is bifacial, composed of upper epidermis, palisade mesophyll, spongy mesophyll and lower epidermis. Thickness of tissues and patterns of cell arrangement is not same in different populations, and those in GongLiu, EMin and TuoLi show thickened palisade mesophyll and higher value of P/S ratio. However, the leaves of wild apple trees in XinYuan correlate with thinner palisade mesophyll and lower P/S. Leaf thickness reduces with the increase of age of tree. The perennial and “The King of Wild Apple” are with less layers of palisade and spongy as well as the thinner leaves. Our results imply that M.sieversii can adapt to different environment and growth status to ensure the efficiency of photosynthesis with the coordination of each part of the anatomical structure. Our data provide evidences for in situ preservation and conservation of M.sieversii.

Sequence Analysis of ITS Regions and ndhA Gene for Determining Phylogenetic Relationship of Diospyros kaki(persimmon) With Other Related Wild Diospyros(Ebenaceae) Species

FU Jian-Min1,2；LIANG Jin-Jun1,2；WUYUN Tana1,2；LIANG Yu-Qin1,2；SUN Peng1,2；LI Fang-Dong1,2*

2015, 35(4):  515-520.  doi:10.7525/j.issn.1673-5102.2015.04.007

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We used eight species(30 genotypes) of Diospyros spp. to study the interspecific relationship and intraspecific variation by ndhA and ITS sequences for calculating substitution model of two sequence and their combination, and then building three phylogenetic trees constructed by ML method based model parameters. The length range of ndhA sequence were 1492 to 1511, 14 informative sites, and the length range of ITS sequence were 660 to 761, 56 informative sites. The best fit models of ITS, ndhA and ndhA+ITS were(TrN+I+G), (F81+I) and (GTR+I+G), respectively. D.kaki was closely related to D.oleifera and D.artrotricha, but distant from D.virginiana and D.cathayensis. The length of D.kaki cultivar was 730, including four variation sites. We analyzed the genetic diversity of selected 21 D.kaki cultivars based on four variation sites. The results suggested that ndhA and ITS sequence variation analysis could explain the relationship between D.kaki and its related species, and the intraspecific variation could be analyzed according to four variation sites.

Carbohydrate,Soluble Protein and Morphometric Changes of Leaves of Populus euphratica Oliv. Individuals under Different Developmental Stages

LI Jia-Hao；FENG Mei；LI Zhi-Jun*

2015, 35(4):  521-527.  doi:10.7525/j.issn.1673-5102.2015.04.008

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By field observation and laboratory analysis, we investigated carbohydrate, soluble protein and morphometric changes of leves of Populus euphratica Oliv. individuals under different developmental stages. With the increase of DBH(age) and canoy levels, leaf index decreased gradually with a very significant negative correlation among leaf index, DBH and tree height. Leaf soluble sugar contents kept steadily in superior level under different developmental stages and canoy levels. But it had no correlation with individual development stages. There were a significant, very significant negative and positive correlations with leaf starch contents, DBH, leaf index, and leaf width, respectively. There were a significant and very significant positive correlation with the leaf soluble protein contents, leaf index, and leaf length, respectively. By comprehensive analysis, leaf change of P.euphratica individual not only existed in different development stages, but also in canoy levelLeaf changes of P.euphratica(heterophylly) is the biological cs. haracteristics under different developmental stages.The leaf indexes deceased with the increase of developmental stages and canoy levels. With the increase of DBH and the decease of leaf index and leaf width, the leaf starch contents decreased gradually. With the decease of leaf index and the increase of leaf length, leaf soluble protein contents decrease gradually. Starch and soluble protein metabolism of P.euphratica leaf had a regulating effect on leaf length and width change.

Relationships of 17 Alnus and Identifying Uncertain Species

RAO Long-Bing1；YANG Han-Bo1；GUO Hong-Ying2；DUAN Hong-Ping3；CHEN Yi-Tai1

2015, 35(4):  528-534.  doi:10.7525/j.issn.1673-5102.2015.04.009

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We analyzed the genetic relationships of 17 Alnus, identified the uncertain species by AFLP markers and morphology data, and used UPGMA in clustering analysis. Seven primers were selected to conduct AFLP-PCR, and 369 DNA bands were obtained including 346 bands ofpolymorphism with93.77% of polymorphic bands. All these plants could be divided into four groups by AFLP cluster analysis: the first group was consisted of A.japonica;the second group was consisted of A.viridis, A.cordata, A.glutinosa, uncertain species, A.cremastogyne, A.trabeculosa, A.incana ssp.rugosa, A.hirsuta, A.formosana, A.firma and A.sinuata; the third group was consisted of A.incana, A.rubra and A.tenifolia; the fourth group was consisted of A.nitida and A.nepalensis. All these plants could be divided into three groups by morphology data: A.cordatawas the first group; the second group was consisted of A.japonica, A.formosana, A.nitida, A.hirsuta, A.trabeculosa; the third group was consisted of A.nepalensis, A.incana, uncertain species, A.incana ssp.rugosa, A.rubra, A.glutinosa, A.viridis, A.fima, A.tenifolia, A.sinuata and A.cremastogyne. The uncertain species was A.glutinosaby morphological and AFLP analysis.The results of morphological cluster analysis and AFLP cluster analysis were similar, but also had some difference, that is, Alnus hadvarious genetic background, and there were some differencesbetween the status of molecular and morphological classification between Alnus species.

Expression of A Floral Homeotic Gene PrseSTK from Prunus lannesiana in Single and Double Flower

LIU Zhi-Xiong1,2；LI Feng-Lan2

2015, 35(4):  535-539.  doi:10.7525/j.issn.1673-5102.2015.04.010

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To find out genes involved in the floral organogenesis of Prunus lannesiana, we cloned cDNAs of one MADS-box genes, PrseSTK with GenBank accession No.GU332504 by using homologous cloning. The full length of PrseSTK contained an open reading frame(ORF) of 669 bp and coding for a polypeptide of 222 amino acid residues. By sequence comparison and phylogenetic analysis, PrseSTK was highly homologous to protein STK in Arabidopsis, and belonged to class D. However, by semiquantitative RT-PCR, PrseSTK was highly expressed in stamen and gynoecium of single-flowered P.lannesiana cultivar ‘Makino’, and in sepal, stamen and leaf-like gynoecium of doubleflowered cultivar ‘Albo-rosea’, which was significantly different from the expression pattern of STK in Arabidopsis. The different expression of PrseSTK in double flower showed ectopic ovules formation would result from PrseSTK expression in the sepal.

Variation Analysis of Growth Traits of Transgenic Populus simonii×P.nigra Clones Carrying TaLEA Gene

LIU Dian-Kun1；LIU Meng-Ran1；LI Zhi-Xin1；WANG Guang-Yu2；LI Ying1；ZHENG Mi1；LIU Gui-Feng1；ZHAO Xi-Yang1*

2015, 35(4):  540-546.  doi:10.7525/j.issn.1673-5102.2015.04.011

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We used 11 Populus simonii×P.nigra clones including ten transgenic poplar clones and one control clone in six-year old to study 15 traits including growth traits, shape of stem, leaf traits, and lenticel traits. By ANOVA analysis, there was significant difference in all the traits among 11 clones(P＜0.01). The coefficients of phenotypic variation(PCV) of different traits ranged in 1.92%-39.98%, the coefficients of genetic variation(GCV) were closed to PCV and repeatability(R) ranged in 0.621-0.987, which indicated that the TaLEA gene transfer can influence the plant growth. but the impact were strong controlled by genetic factor. By phenotypic correlation analysis, there were varying degree correlations between growth traits, stem traits, leaf traits and lenticel traits, which indicated that all the traits combined action on plant growth and development. When the selection rate was 10%, XL-1 and XL-9 were selected as the best clones. The average height and diameter at breast height of these two clones were higher than those of CK(average of all the clones) by 16.55%(13.38%) and 15.38%(19.46%), and the genetic gain of these clones were 12.18% and 17.42% in height and diameter at breast height, respectively.

Function of Non-specific Lipid Transfer Protein Gene PsnsLTP in Polygonum sibiricum Laxm

LI Wei1；QU Chun-Pu1；XU Zhi-Ru1,2；LIU Chun1；LIU Guan-Jun1*

2015, 35(4):  547-553.  doi:10.7525/j.issn.1673-5102.2015.04.012

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We transformed the plant expression vector pROKII-PsnsLTP harboring non-specific lipid transfer protein gene from Polygonum sibiricum Laxm into tobacco by Agrobacterium-mediated genetic method. After transgenic tobacco was identified by kanamycin resistance screening and PCR, we observed phenotypic after induced Colletotrichum nicotianae and Pythium aphanidermatum, respectively, on the leaves of transgenic tobacco and wild type, and measured physiological indicators, the phenotypic in transgenic tobacco, and wild type in the treatments of NaCl, CdCl₂, 7.5% PEG6000 solutions. Compared with wild type tobacco, transgenic tobacco plants had a strong resistance to Colletotrichum nicotianae and Pythium aphanidermatum. SOD and POD activity was significantly enhanced, and MAD content was extremely reduced besides the transgenic tobacco seedlings had better growth traits. Therefore, PsnsLTP improved the ability of transgenic tobacco to resist biological and abiotic stress.

Dynamic Changes of Atmospheric Nitrogen Wet Deposition During Growing Seasons in Maoershan Area in Northeast China

SUN Tao；LI Xing-Huan；LIU Rui-Peng；MAO Zi-Jun；LI Na；HAN Ying-Ying；DING Yi-Yang；DUAN Xi-Hua*

2015, 35(4):  554-558.  doi:10.7525/j.issn.1673-5102.2015.04.013

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We studied the dynamic changes of atmospheric nitrogen deposition concentration and amount in precipitation collected by a standard rain gage, in Maoershan area. The total amount of atmospheric nitrogen deposition from precipitation was 19.16 kg·hm^-2, and the proportions of NH⁺₄-N, NO^-₃-N and DON to total nitrogen deposition in precipitation were 52%, 26% and 22%, respectively. The ratio of NH⁺₄-N to NO^-₃-N was nearly 2.0. On a seasonal basis, the average content of NH⁺₄-N deposition in precipitation was 1.59 mg·L^-1 with the greatest contribution to atmospheric nitrogen wet deposition. There was an obvious seasonal change in the contents of total atmospheric nitrogen wet deposition, being the highest value in May and September and lowest in July. Moreover, the monthly amounts of NH⁺₄-N, NO^-₃-N and total nitrogen input from wet deposition were all positively related with the time of rainfall (P<0.01) with the correlation coefficients of 0.65, 0.63 and 0.76, respectively. However, the monthly amount of DON nitrogen input from wet deposition was not significantly correlated with the time of rainfall (P>0.05) with the correlation coefficient of 0.24. The high annual NH⁺₄/NO^-₃ ratio compared with the more developed region suggested that N deposition in this region was mostly affected by agricultural activities rather than industrial activities.

ISSR Analysis on the Genetic Diversity of Relic Plant Bretschneidera sinensis from Wuyishan Region

LIN Le-Jing1,2；JIN Chu-Qi1；ZHENG Chun-Hao3；ZHOU Ya-Mei1；ZHOU Ying1

2015, 35(4):  559-563.  doi:10.7525/j.issn.1673-5102.2015.04.014

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Bretschneidera sinensis Hemsl., with important research value, is an endangered species endemic in East and South Chin. We used inter-simple sequence repeat(ISSR) markers studied the genetic diversity within and among six populations of B.sinensis. The genetic diversity at species level was high(PPB: 75.70%; H_ES: 0.304 5; H: 0.450 1). However, relatively low genetic diversity existed within populations. Population in Minjiangyuan(MJY) exhibited the greatest level of variability(PPB: 60.13%; H_EP: 0.238 1; H: 0.347 5), whereas the population in Baishanzu(BSZ) showed its own variability at the lowest level(PPB: 44.86%; H_E: 0.173 6; H: 0.255 6). By the genetic analysis of the ISSRs, we revealed a relatively high level of genetic differentiation among populations by Nei’s gene diversity statistics(21.81%), Shannon’s information measure(22.79%) and analysis of molecular variance(AMOVA)(18.62%). The main factors responsible for the relatively high level of differentiation among populations were probably related to the predominant outcrossing reproductive system and recent habitat fragmentation, and a significant correlation was between genetic distance and geographic distance(r=0.626 7, P<0.05). The high diversity would hold promise for conservation and restoration efforts to save the endangered species from extinction.

Tree Carbon Storage Increment of Natural Larch Forest in Daxing’an Mountains

PENG Wei；LI Feng-Ri*；JIA Wei-Wei；GAO Hui-Lin

2015, 35(4):  564-571.  doi:10.7525/j.issn.1673-5102.2015.04.015

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We studied the dynamic of annual carbon emissions of mortality trees, the carbon storage of ingrowth trees, the net increment of carbon storage, and the change trend of carbon storage along with site quality and stand density for natural larch(Larix gmelinii) forestwith the data of 602 permanent sample plots(PSPs) from forest resources continues inventory of four periods between 1990-2010 in Daxing’an Mountains. The emissions of carbon for all age groups were 0.133 7-0.484 1 t·hm^-2·a, and it increased with the increasing of age group. The carbon storage of ingrowth trees is larger(0.128 2 t·hm^-2·a) in young stand, it is smaller(0.04 t·hm^-2·a) in near-mature stand and there are not significant for other age group. The net increment of carbon storage is inversely proportional with the age group, and they are 1.374 9, 0.982 1, 0.649 9, 0.538 1 and 0.240 7 t·hm^-2·a for young, middle-aged, near-mature, mature, and over-mature stand, respectively. The emissions of carbon for all age group are positive correlated with stand density under same site quality. The carbon storage of ingrowth trees for young, middle-aged and mature stand decrease with increasing of stand density, and it’s not significant for near-mature stand and over-mature stand. The net increment of carbon storage is the maximum with middle stand density for all age groups except young aged stand. Under the same stand density, the emissions of carbon of all age groups(except near-mature stand and mature stand) are inversely proportional with site quality. For all age groups(except mature stand), the better the site quality is, the smaller the carbon storage for ingrowth trees is. Under the similar stand density, the better the site quality is, the greater the net increment of carbon storage is.

Functional Identification and Promoter Preliminary Analysis of Flavonoid 3′-Hydroxylase Genes in Turnip

XU Zhi-Ru1,2,3；MA Jing1,2,3；CUI Guo-Xin1,2,3；LIU Tong1,2,3；LIU Guan-Jun1,2,3

2015, 35(4):  572-582.  doi:10.7525/j.issn.1673-5102.2015.04.016

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Flavonoid 3′-hydroxylase(F3′H) is a cytochrome P450 monooxygenase, it can convert dihydrokaempferol to dihydroquercetin in the pathway of anthocyanin biosynthesis, and the cyanidin can be formed. The overexpression vectors of BrF3′H1 and BrF3′H2 genes cloned from ‘Tsuda’ turnip and ‘Yurugi Akamaru’ turnip, respectively, were constructed, and then were transformed into tobacco. The flower color of transgenic tobacco plants was darker. The 846 and 851 bp promoter fragments upstream of BrF3′H1 and BrF3′H2 genes were obtained by genome walking method. Many cis-acting elements in BrF3′H1 and BrF3′H2 promoters were identified by bioinformatics analysis, such as TATA box, CAAT box, light responsive elements, MRE, ABRE, ATGCAAAT-motif, ERE, O2-site, RY-element and LTR. The nucleotide sequences of BrF3′H1P and BrF3′H2P had 7 differences. The 35S promoter of pCAMBIA1301 plant expression vector was substituted by BrF3′H1P and BrF3′H2P, respectively, and then the recombinants were introduced into tobacco through the mediation of Agrobacterium tumefaciens. Histochemical staining result of GUS indicated that the expression of GUS reporter gene could be driven by BrF3′H1P and BrF3′H2P sequence. A series of 5′-end deleted fragments of BrF3′H1P and BrF3′H2P were obtained by PCR, which were fused with GUS reporter gene and then were transformed into tobacco. The stain result showed that the deleted fragments of BrF3′H1P and BrF3′H2P had starting activity that could initiate the expression of GUS gene. The functional identification of BrF3′H1 and BrF3′H2 genes and the preliminary analysis of their promoters would lay a foundation for clarifying the mechanism on light inducible expression regulation of F3′H genes in ‘Tsuda’ turnip and ‘Yurugi Akamaru’ turnip.

Ecoregion based Height-diameter Models for Larix gmelinii Rupr. in Daxing’an Mountains

DAI Zhong-Di；JIANG Li-Chun

2015, 35(4):  583-589.  doi:10.7525/j.issn.1673-5102.2015.04.017

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We used 12 height curve models using height-diameter data from felled trees for Larix gmelinii Rupr. from three ecoregions in Daxing’an Mountains Region to evaluate the coefficient determination, bias, and root mean square error for model accuracy and precision. The functions for Korf, Tang, Hossfeld were better for height curve from Region 1 and Region 3. Richard, Weibull, Mitscherlich were better for height curve from Region 2. We used full model and reduced model test method for regional comparison. The height curves were significantly different among different ecoregions. Height curve in Region 2 had significant difference with Regions 1 and 3, and height curve in Region 1 had small difference with Region 3. Incorrectly applying height curve in different ecoregions would result in larger prediction error.

Soil Carbon-budget Mechanism in Different Origin Stands on the West Slope of Zhangguangcai Mountain

SUN Hong-Yang1,2；WANG Qing-Cheng1*

2015, 35(4):  590-596.  doi:10.7525/j.issn.1673-5102.2015.04.018

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In Wuchang Fenghuangshan Forest Farm on the west slope of Zhangguangcai Mountain, we studied the seasonal dynamics of soil carbon input and output of three different forests of secondary broad-leaved natural forest(C), Larch and broad-leaved mixed forest(H), and Larch(Larix olgensis)(L) in different origins(artificial regeneration, artificial-natural regeneration, and natural regeneration) to explore the effect of different forest origins on soil carbon-budget. The soil organic carbon content of H was significantly higher than that of L(P＜0.05) with C in the middle, which showed no significant difference from H or L(P＞0.05). The sequence of litter-full leaf biomass of three different origin stands was C>H>L, with C significantly higher than L(P＜0.05). The sequence of litter-full decomposition rate of three different origin stands was C>H>L. C presented the highest soil respiration rate between three different origin stands in different seasons, and L presented the lowest. The June-October average of soil microbial biomass carbon(MBC) and soil readily oxidized organic carbon(ROC) both showed H>C>L, whereas water-soluble organic carbon(WSOC) showed the tendency of C>H>L. Litter decomposition weight loss rate was significantly correlative(P<0.05) with soil active carbon in three different origin stands, and highly negatively correlative between soil respiration rate, ROC and WSOC(P<0.01). Therefore, in the soil carbon-budget processes the artificial-natural regeneration was conductive to the turnover and storage of soil carbon.

Carbon Storage Maturity of Larch Plantation in Heilongjiang Province

JIANG Jia-Mei；JIA Wei-Wei*；LIU Qiang

2015, 35(4):  597-603.  doi:10.7525/j.issn.1673-5102.2015.04.019

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With the data of 42 sample trees of carbon storage and biomass investigation data of fixed sample plots of larch plantation from 2006 to 2007, we studied the carbon storage compatibility model of larch was builded and the regular of carbon storage in different site conditions. The model built by carbon storage directly had the higher precision. The larch plantation atⅠ, Ⅱ, Ⅲ level site conditions had a carbon storage maturity age of 37.8, 39.2 and 42.5 years. Research suggests that the carbon storage of same stand age rises with the increascent of site condition. The carbon storage of same site condition increased with the increase of the age. The average carbon accumulation and successive year carbon accumulation increased first, and then decreased. The increment of carbon would change after some years, and it would decrease with the increase of the age.

Comparison of Soil Organic Carbon Stock of Betula costata Secondary Forest and Mixed Broadleaved Korean Pine Original Forest in Xiaoxing’an Mountains.

DING Yi-Yang1；MAO Zi-Jun1*；ZHANG Ling2；DING Li3

2015, 35(4):  604-611.  doi:10.7525/j.issn.1673-5102.2015.04.020

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Soil organic carbon content plays an important role in the global ecosystem carbon stock revolution. By space replacing time series method, we chose two types of forests of mixed broadleaved Korean pine original forest and Betula costata secondary forest to measure the soil organic carbon(SOC) stock, soil nitrogen(TN) content, microbial biomass carbon(MBC) and other soil chemistry and physical properties. The SOC, TN, MBC contents are gradually decreased with the increase of the depth of soil layer, and finally to a stable value. On the contrary, the soil bulk density increases with the increase of depth of the soil layer. There is no significant difference in 0-10 cm soil layer’s SOC,TN content and 10-20 cm soil layer at original forest, but there are significant relations in e two soil layers at secondary forest. The soil organic carbon densities(SOCD) of original forest and secondary forest are 21.46 and 21.3 kg·m^-2, respectively. There is no significant difference between two forest types. The mean soil organic carbon contents of original forest and secondary forest are 35.79 and 28.6 g·kg^-1, respectively. Soil total nitrogen contents of original forest and secondary forest are 2.86 and 1.83 g·kg^-1, respectively. The liner correlation of microbial biomass carbon and soil organic carbon in secondary forest is higher than original forest. The soil fertility of original forest is more than secondary forest. It is better to plant mixed coniferous with pine species in the secondary forest, and have appropriate thinning in original forest to balance the underground carbon system.

qRT-PCR Analysis of Gene Expression of AG and SOC1 During Flower Development of Fraxinus mandshurica Rupr.

REN Xiao-Long1；ZHAN Ya-Guang1,2；LIANG Xue1；QI Feng-Hui1,2*

2015, 35(4):  612-617.  doi:10.7525/j.issn.1673-5102.2015.04.021

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We analyzed the temporal expressions of four housekeep genes, 18S rRNA, GAPDH, β-actin and TU, in ash(Fraxinus mandshurica Rupr.) flowers with real-time quantitative PCR. With NormFinder software, TU could be selected as inner reference genes to study the expression of two genes, AG and SOC1, related to the flowering of ash. The expressions of both AG and SOC1 were different between male and female. The expression of AG started with a peak in female during the early period of flowering, followed by a decrease till mature embryo sac stage. In male, the expression of AG reached the peak(1.53) at meiosis stage. The expression of SOC1 in female was lower than that in male. Our results would provide evidence to study the expressions of flowering-related genes during the period of flower development in ash.

Remote Sensing Evaluation and Analysis of Hippophae rhamnoide Plantation Biomass

WANG Hai-Bo；XIN Ying；ZHAO Yu-Sen*

2015, 35(4):  618-622.  doi:10.7525/j.issn.1673-5102.2015.04.022

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Weestimated Hippophae rhamnoide plantation biomass using satellite images of Landsat TM(2011)based on RS and GIS. In one-dimensional linear regression analysis models,the NDVI-based and RVI-based simple linear model were the optimal model for estimating biomasswith the coefficient correlation(R²) of 0.908 6 and 0.868 5, respectively. The multiple regression model established by four kinds of VI could be used for estimating forest biomass with the coefficient of determination(R²) of 0.909. Themultivariable linear regression models were better than one-dimensional linear regression analysis models, and the predicted biomass based on regression analysis models was higher than the theoretical biomass.

Effects of Sugar content on Cell Growth,Accumulation of Rosemary Acid, and Antioxidative Enzyme Activity in Suspension Cells of Rosemary

QU Dan；WANG Hui-Mei*；REN Jie

2015, 35(4):  623-627.  doi:10.7525/j.issn.1673-5102.2015.04.023

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We investigated the effects of different concentrations of sucrose, maltose and glucose on cell growth, rosemary acid accumulation in rosemary suspension cells. And we also studied the different concentration of sucrose on antioxidant enzyme activity in rosemary suspension cells. The optimum sugar treatment for cell biomass accumulation was 30 g·L^-1 sucrose, 70 g·L^-1 maltose and 40 g·L^-1 glucose in rosemary suspension cells. Moreover, the growth rates of suspension culture cells were 74.08% and 72.33% under processing of 30 g·L^-1 sucrose and 70 g·L^-1 maltose, respectively，but was nearly three times more than that of 40 g·L^-1 glucose. The content of rosemary acid in 30 g·L^-1 sucrose suspension culture cells was 228 times more than that in 70 g·L^-1 maltose, but slightly lower than that in 40 g·L^-1 glucose. Under different sucrose treatment, rosemary acid content showed an increase trend with the increase of sucrose content, which meant that a high concentration of sucrose was conducive to rosemary acid accumulation. H₂O₂ and MDA content obviously increased under the treatment of higher sucrose concentration, while antioxidant enzymes such as catalase(CAT), guaiacol peroxidase(G-POD) and SOD also increased. The osmotic stress under high concentration of sucrose was bad for the growth of rosemary suspension culture cells, but was conducive to the accumulation of secondary metabolites. Considering the growth rate and rosemary acid content, the medium supplemented with 30 g·L^-1 sucrose was most suitable for rosemary suspension cultured cells.

Enzyme Assisted Ethanol Extraction of Total Flavonoids from Peony Episperm

MENG Qing-Huan1；ZU Yuan-Gang1；WANG Hua2；WANG Hong-Zheng1；JIANG Tao3；WU Wei-Wei1；ZHONG Chen1；DUAN Xi-Hua1*

2015, 35(4):  628-631.  doi:10.7525/j.issn.1673-5102.2015.04.024

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We optimized enzyme assisted ethanol extraction of total flavonoids from peony episperm. We studied the effects of factors including enzymolysis temperature, enzyme content, enzymolysis pH, enzymolysis time on the extraction rates of total flavonoids by singlefactor and orthogonal test. The optimal extraction conditions were after enzyming pretreatment under the pectinase content of 0.05 mg·mL^-1, enzymolysis temperature of 45℃, enzymolysis pH of 4.5, enzymolysis time of 180 min, total flavonoids yield was up to 74.839 mg·g^-1.Compared with the traditional ethanol extraction, the total flavonoids yield was improved significantly.

Optimization of Ultrasonic-assisted Extraction of Total Flavonoids from Crataegus pinnatifida Leaves and Its Antioxidant Activities

LUO Meng1,2；SONG Zhuo-Yue1,2；HU Jiao-Yang1,2；MU Fan-Song1,2*；QIAO Qi1,2；ZU Yuan-Gang1,2

2015, 35(4):  632-637.  doi:10.7525/j.issn.1673-5102.2015.04.025

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We used ultrasonic-assisted extraction(UAE) to extract the total flavonoids from Crataegus pinnatifida leaves, and used Box-Behnken design(BBD) combined with response surface methodology(RSM) to optimize extraction conditions. We determined the effects of four variables in terms of ratio of liquid/solid, temperature, ethanol concentration and time on the total flavonoids contents(TFC). The optimized extraction parameters obtained the highest total flavonoids yield were under the ratio of liquid/solid of 15, 40℃, and 40% of ethanol for 32 min. Under these conditions, the yield was 15.50 mg·g^-1. By in vitro antioxidant assays, the extract exhibited a strong DPPH radical-scavenging in ability with IC₅₀ value of 0.69 mg·mL^-1. Compared with conventional maceration extraction(ME) and heat-refluxing extraction(HE) methods, UAE method showed remarkable advantages in terms of better antioxidant activities. Overall, the present results indicated that UAE was a suitable method for the extraction of total flavonoids from C.pinnatifida leaves and its extracts were potential antioxidant ingredients.

Distribution of Resveratrol in Different Parts of Several Plants

WANG Kun-Lun1；ZHAO Xiu-Hua1；ZU Yuan-Gang1*；LI Jia-Lei2

2015, 35(4):  638-640.  doi:10.7525/j.issn.1673-5102.2015.04.026

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By HPLC resveratrol in grape purple peony, peanuts and giant peak distribution of each part the development and utilization of these plants can provide valuable data. The content of purple peony seed and pod was far higher than that in the peanut and the giant peak grapes. Peanut resveratrol content was the lowest. Grape purple peony, peanuts and giant peak resveratrol distribution of each part were as follows: purple peony leaf, purple peony collateral, purple peony stems and purple peony root did not contain resveratrol, and the purple peony seeds and purple peony pod contained resveratrol, the purple peony seed resveratrol content was 0.87‰, and purple peony pod resveratrol content was 0.26‰. Peanut leaf, peanuts, peanut embryo and endosperm did not contain resveratrol, peanut collateral, peanut stem, peanut shell, peanut coat, peanut lateral root and peanut root contains resveratrol contains resveratrol, and the contents were 3, 11, 12, 15, 5 and 10 mg·kg^-1 in lateral branch, peanut stalk, peanut shells, peanut coat, lateral root and peanut taproot, respectively. The grape flesh, grape skins and grape seed, grape pedicle, resveratrol were in four parts, including the resveratrol content in grape flesh 0.017‰, grape skin resveratrol content of 0.028‰, grape seed extract resveratrol content of 0.005‰, grape pedicle and its adjacent branch resveratrol content was 0.12‰. Therefore, peanut coat, peanut shell, grape skin and grape seed, grape pedicle, peony seed, and peony pod have the medical value and economic value.