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    20 March 2013, Volume 33 Issue 2
    Peristome Morphology Characteristics of 16 Species of Family Pottiaceae and their Systematic Significance
    WANG Xiao-Rui;WANG Li-Bao;LI Min;ZHAO Jian-Cheng*
    2013, 33(2):  129-133.  doi:10.7525/j.issn.1673-5102.2013.02.001
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    The peristome morphology of 16 species of the family Pottiaceae was observed by scanning electron microscope (SEM). The results show that the peristomes of these species are similar. They are haplolepideae. Peristome teeth are 1-3 sinistrorse circles or erect, basal membrane is present and densely covered with small papillae. While based on the differences of shape and rotation of peristome teeth and characteristics of basal membrane, three styles of peristome can be distinguished. It shows the hereditary differentiation of genus and species. This paper also discusses the adaptation of peristome morphology to the xerophytic environment.
    Anatomy of An Ephemeroid Plant Eremurus altaicus
    TIAN Li-Li;MA Miao*
    2013, 33(2):  134-138.  doi:10.7525/j.issn.1673-5102.2013.02.002
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    The morphological features and anatomical structures of Eremurus altaicus(Pall.) Stev. were studied by the methods of traditional paraffin wax sections. The results showed: The root system of E.altaicus was composed of storage and absorbing roots with different anotomic structures and physiological functions, which was a result of ecological adaptation of the ephemeroid plant to its habit of fast growth and long-term annual dormancy; It was of lots of xerophyte characteristics, such as significant cortex and pith in root, cuticle on epidermis of leaf and stem, mass fundamental tissue in stem, ring-shaped palisade tissue, and highly-developed water-storage tissue, which suggested that E.altaicus might originate from arid habitat in early time although it is a mesophyte distributed in subalpine meadow; Two kinds of xylems in roots of the plant were found, xylem in storage root was polyarch, but it in absorbing root was diarch or penarch. There was a layer of obvious stripe-like casparian strips along the transverse walls and radial walls on endodermis cells. The diamorphism of xylem and stripe-like casparian strips on endodermis cells suggested that Eremurus might be special taxa among the plant systematic tree.
    Biosystematic Relationships among Three Sects’ Plants in Elymus(Poaceae) Based on the External Morphological Characteristics
    SU Xu;LIU Yu-Ping;YANG Chun-Bin;KE Jun;LA Ben
    2013, 33(2):  139-144.  doi:10.7525/j.issn.1673-5102.2013.02.003
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    The external morphology of the major species of sects. Elymus, Turczaninovia and Macrolepis in Elymus was analyzed, the similarities and the differences of the external morphological characteristics were summarized, and the taxonomic significance of external morphological features among them was discussed. Furthermore, according to the evolutionary trends of external morphological characters including inflorescences, spikelets, florets and paleas of three sects, their evolutionary positions and relationships were analyzed. The results show that the sect. Elymus is most primitive among three sects, the sect. Turczaninovia is advanced than the sect. Elymus, and the sect. Macrolepis is the most advanced sect. The sect. Elymus probably generated the sect. Turczaninovia, whereas the sect. Macrolepis was probably derived from the sect. Turczaninovia. The biosystematic relationships of tested species in Elymus were also discussed. The research provides the theoretic foundation for further studying the systematics and evolution of Elymus.
    Observation on Megasporogenesis, Microsporogenesis and Development of Female and Male Gametophytes of Catalpa speciosa Warder
    LI Li-Ping;LIU Hai-Yan;CHEN Fa-Ju*
    2013, 33(2):  145-148.  doi:10.7525/j.issn.1673-5102.2013.02.004
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    The processes of megasporogenesis, microsporogenesis and the development of female and male gametophytes of Catalpa speciosa Warder were observed by serial paraffin sections. The results are as follows: the ovule has 2 locules and is anatropous, single-integument, and thin-nucellus. Sporogonium developed megaspore mother cell directly. The development of embryo sac is of polygonum type. There are five stamens in C.speciosa Warder, including three degradation and two fertile stamens. Its anthers have 2 mirosporangiates, and the tapetum belongs to glandular type. The tetrads are tetrahedral. The pollen grains are four celled at shedding.
    Megasporegenesis and Female Gametophyte Development in Berberis pruinosa Franch.
    YANG Gui-Ying;MA Shao-Bin*
    2013, 33(2):  149-153.  doi:10.7525/j.issn.1673-5102.2013.02.005
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    The megasporogensis and female gametophyte formation in Berberis pruinosa were first studied through paraffin sectioning. The main results can be concluded as follows: solitary pistil is monocarpellary with 2 ovules, anatropous,marginal placenta, crassinucellate. The zig-zag micropyle is formed by both the inner and the outer integuments. Single archesporial cell is under nucellar epidermis. The tetrad of megaspores is linear shaped, with the first cell from the chalazal end being functional. The development of embryo sac belongs to the Polygonum type. In mature embryo sac, two polar nucleus fuse into the secondary nucleus before fertilization, three antipodal cells are ephemeral. Inside the triangular egg apparatus, two synergids with fifiform apparatus are developed.
    Fruit Types of Angiosperm and Their 4 Life Forms in Tibet and Its Southeastern Region
    YU Shun-Li;FANG Wei-Wei;ZEREN Wangmu*;Ni Zhen;ZHANG Xiao-Feng
    2013, 33(2):  154-158.  doi:10.7525/j.issn.1673-5102.2013.02.006
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    This article calculated the percentages of different fruit types of total angiosperms (5810 species, 28 varieties) and their life forms (trees, shrubs, herbs and lianas) distributed in Tibet and southeastern Tibet. The results showed that in Tibet capsule is the dominant fruit type(37.74%), followed by achene, nut and berry. The percentage of dry fruits is far bigger than that of fleshy fruits in Tibet. There are great variations in 4 life forms in fruit type spectrum. Drupe is dominant in tree, berry in liana, achene in shrub, and capsule in herb. Tree possesses the greatest percentage of fleshy fruit in 4 life forms, followed by liana, shrub and herb. In southeastern Tibet, capsule is also the dominant fruit type (37.54%), followed by achene, berry, drupe, nut and caryopsis. The larger differences occur among the percentages of various fruit types.Drupe is dominant in tree, berry in liana, capsule in shrub and in herb. In the 4 life forms tree possesses the largest percentage of fleshy fruits followed by liana, shrub and herb. The percentage of fleshy fruits is higher in southeastern Tibet than in the whole Tibet, and on the contrary, dry fruits is not. Such fruit characteristics are adapted to their environment and are results of long-term evolution. This study can help us in understanding a long term adaptation and evolution of fruits to their ecological environment.
    Effect of Grazing on Plant Functional Group Characteristics in stipaSteppe between China and Mongolia
    BAO Xiu-Xia;YI Jin;LIAN Yong;XIAO Xia;BAO Xiu-Ping
    2013, 33(2):  159-165.  doi:10.7525/j.issn.1673-5102.2013.02.007
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    In our study, the plant functional characteristics in Chinese and Mongolian typical steppe under different grazing methods were investigated. The results showed that the proportion of perennial under settled grazing (SG) was significant lower than the others, the ratio is 57.576%, while the proportion of xerophytes was significantly higher than the others, the ratio is 69.697%. The diversity of functional groups showed that Shannon-wiener and Simpson index at four season nomadic grazing (FSNG) were higher than the others, while Pielou was contrary to diversity index. Correlative analysis of functional groups diversity showed that there was a significantly positive correlation between Shannon-wiener and Simpson index, while negative correlation between Pielou and diversity index. Niche breaths between perennial and annual plants, xerophytes and intermediate xerads were higher, it affected niche overlap. It showed irrational grazing led to degradation of vegetation, which was clearly found to be of lesser economic and ecological value.
    Effect of Exogenous Nitric Oxide on Photosynthetic Pigments and Chlorophyll Fluorescence under Enhanced UV-B Radiation in Larix gmelinii
    MENG Qing-Huan;ZU Yuan-Gang;GUO Xiao-Rui;DUAN Xi-Hua;*
    2013, 33(2):  181-185.  doi:10.7525/j.issn.1673-5102.2013.02.010
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    Under enhanced UV-B radiation, 3-year-old seedlings of Larix gmelinii were chose as target to study the impact of exogenous nitric oxide on photosynthetic pigments (Chl a, Chl b and Car) and chlorophyll fluorescence. The results of variance analysis showed 0.5 mmol·L-1 sodium nitroprusside (SNP), a NO donor, have significant impact on nursery stocks of L.gmelinii under enhanced UV-B radiation stress. 0.5 mmol·L-1 SNP can significantly inhibited the apparent decline in photosynthetic pigments, Fv/Fm, ΦPSⅡ,Fv′/Fm′ and qP, and the obvious increase in chl a/chl b, Fo and NPQ after supplement UV-B radiation. All these results indicated that exogenous nitric oxide could reduce the physical damage of photosynthetic reaction centers in seedlings of L.gmelinii under UV-B radiation stress, and enhancement of environmental adaptability of the seedling to supplementary UV-B radiation stress.
    Provenance Analysis of Mongolian Oak at Seedling Stage
    QU Hong-Jun;MENG Qing-Bin;ZHANG Zhong-Lin;YANG Chuan-Ping;ZHANG Jie*
    2013, 33(2):  166-173.  doi:10.7525/j.issn.1673-5102.2013.02.008
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    In this study, the heights and diameters of 25 provenances of Quercus mongolica from Dailing sample plots were investigated and analyzed. The conclusions were as follows: (1) There were significantly positive correlations in height and diameter between 1-year and 6-year Q.mongolica seedlings, respectively. The growth characteristics from 25 provenances of Q.mongolica were significantly different, indictated that it is important to carry out provenance selection for Q.mongolica. Therefore, it was initially stated that Weihe and Jian were excellent seed resources in Dailing sample plots for forestry production. (2) The main trend of geographical differentiation was controlled by both latitude and longitude, while latitude influenced more. (3) There were certain correlations among growth characteristics and geographical climatic factors of Q.mongolica provenances. (4) 25 provenances were divided into 3 provenance areas. The first area was mainly located in Longgang hill and the south of the Changbai hill. The second one is mostly in Changbai hill and Laoyeling. The majority of the third one was located in Xiaoxing’anling and Zhangguangcailing.
    Preliminary Study on the Subcellular Localization of Arabidopsis thaliana AtZW10
    LU Yu-Jian;LIU Heng;LI Yu-Xi;LIANG Da-Wei;ZHAO Peng;JIA Peng-Fei;NIU Song-Zhao*
    2013, 33(2):  174-180.  doi:10.7525/j.issn.1673-5102.2013.02.009
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    The bioinformatics analysis of AtZW10 protein was performed by using different approaches. The results showed that the Arabidopsis thaliana AtZW10 protein is much conserved during the evolution of high eukaryotes. In AtZW0 protein, there is a similar conserved region to the centromere/kinetochorebinding protein ZW10. Moreover, the high similarity was found between AtZW0 and the Drosophila DmZW10 protein. AtZW10 was considered to be a hydrophilic protein by hydrophobicity analysis, and no obvious transmembrane domains. Prediction results also indicated that AtZW10 is likely to be located in the nucleus and cytoplasm. In order to further determine the subcellular localization of AtZW10, the full-length of AtZW10 gene was amplified by PCR using Arabidopsis cDNA as a template, and the binary vectors consisting of AtZW10 fused with the yellow (green) fluorescent protein (YFP/GFP) coding sequence were further constructed. The recombinant vectors were transformed into wild-type Arabidopsis protoplasts of mesophyll cells and Nicotiana benthamiana epidermal cells, respectively. Subsequently, the analyses on the distribution of the fusion proteins suggest that AtZW10 is an nucleoplasm positioning protein, and is mainly distributed in the nucleus, consistent with the conclusions of bioinformatics analysis. The molecular features and the subcellular localization of AtZW10 were analyzed in the present study, which might provide a basis for studying the function of AtZW10 gene in the future.
    Effect of Enhanced UV-B Radiation on Arabidopsis Mesophyll Cell Protein
    WEI Xiao-Li;ZHENG Na;LI Xiao-Yang;HAN Rong*
    2013, 33(2):  186-190.  doi:10.7525/j.issn.1673-5102.2013.02.011
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    Four-week-old wild-type Arabidopsis seedlings (Columbia-0) was treated using different doses of UV-B radiation, the protein in the mesophyll cells was extracted using acetone precipitation and TCA-acetone, then Arabidopsis mesophyllcellular protein content and composition of the different intensity of UV-B radiation response were analyzed. The results showed that comparing two methods, the protein content of the TCA-acetone extract is relatively higher, which was more suitable for the analysis of enhanced UV-B radiation on Arabidopsis mesophyll cell protein; The changes of protein contents by two extraction methods showed the same trend, along with the increasing of UV-B radiation dose, the protein content increased first and then reduced, B2 group reached a maximum. In addition, the number of protein bands and expression has taken place significant change, the most obvious changes were also in the middle dose treatment group (B2), both new bands and disappearance bands. This may be due to the Arabidopsis thaliana can activate some of its own resistance gene expressions and induce resistance protein by low doses of UV-B radiation, and thus resist the damage of UV-B; however, when subjected to high doses of UV-B radiation, damage their own protein synthesis pathway, and affect protein synthesis.
    Transformation and Expression of 1Dx5 without Selectable Markers in Common Wheat Xindong No.26
    WANG Yong;QIN Jian-Bing;FAN Ling;ZHU Chang-Qing*
    2013, 33(2):  191-196.  doi:10.7525/j.issn.1673-5102.2013.02.012
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    To improve the bread-making quality of wheat, the Xinjiang winter common wheat Xindong No.26 were transformed with high-molecular-weight glutenin subunits (HWM-GS) gene 1Dx5 that without selectable markers by particle bombardment. Transformed plants were screened by PCR, three transgenic plants were detected among 1 000 transformed plants, yielding a transformation efficiency of 0.3%. The compositions of HWM-GS were analyzed by SDS-. The HMW-GS gene 1Dx5 was expressed in some seeds of transgenic plants. The high molecular weight glutenin subunit gene 1Dx5 was transformed into wheat XinDong No.26 and was expressed in some transgenic progenies. Our studies lay the foundations for the improvement of bread-making quality of wheat.
    Establishment and Optimization of Genetic Transformation and Introduction of TaNHX2 Gene by Agrobacterium tumefaciens in Albizia julibrissin
    ZHOU Xiang-Ming;;HAO Zhi-Yu;XIA Shi-Yun;WANG Shu;SONG Jian;CHEN Shou-Yi;LIU Zhong-Qi
    2013, 33(2):  197-201.  doi:10.7525/j.issn.1673-5102.2013.02.013
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    Some factors influencing the transformation efficiency, such as the concentration and infection duration of Agrobacterium tumefaciens, pre-culture and co-culture duration, were optimized to establish a stable Agrobacterium mediated gene transformation protocol of Albizia julibrissin Duraxx. The TaNHX2 gene was transformed into A.julibrissin. Mediated by A.tumefaciens, lots of regenerated plants resistant to Kanamycin (Kan) were obtained. PCR analysis and real time PCR detection of some transgenic plants showed that the TaNHX2 gene was evidently integrated into the genome of transgenic plants and could be transcribed properly.
    Effects of Simulated Acid Rain on Physiological and Ecological Characteristics of Sinocalycanthus chinensis
    PENG Li-Qiong;JIN Ze-Xin*;WANG Qiang;
    2013, 33(2):  202-207.  doi:10.7525/j.issn.1673-5102.2013.02.014
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    The physiological and ecological characteristics of the endangered Sinocalycanthus chinensis seedlings with the treatments of simulated acid rain of pH2.5, pH3.5, pH4.5 and pH5.6 (control) were determined and analyzed. The results showed that, in all treatments, the day variation of net photosynthetic rate of S.chinensis was a curve with one peak in July. The mean daily net photosynthetic rate reduced with the increasing of the acidity with significant differences among different treatments. The maximum net photosynthetic rate, light saturation point and the relative chlorophyll content decreased gradually with the increasing of the acidity, while the dark respiration rate and the light compensation point increased gradually with the increasing of the acidity. The apparent quantum yields in the treatments of pH2.5 and pH3.5 were lower that the control, while that in the treatment of pH4.5 was higher than the control. Fv/Fm and Fv/Fo reduced with the increasing of the acidity. The activity of peroxidase and superoxide dismutase, the content of malonydialdehyde and the membrane permeability significantly increased with the increasing of the acidity. The results suggested that simulated acid rain could reduce the photosynthetic capacity and had detrimental effect on the physiological traits of S.chinensis.
    Vegetation Net Primary Productivity in Qinling Mountains and Its Response to Climate Change
    YUAN Bo;BAI Hong-Ying*;ZHANG Jie;MA Xin-Ping
    2013, 33(2):  225-231.  doi:10.7525/j.issn.1673-5102.2013.02.018
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    Based on the NDVI data and meteorological data from 1999 to 2009, the vegetation net primary productivity (NPP) in Qinling mountains was estimated using CASA model , and the characteristics of spatial and temporal variation of NPP and the response to climate change in Qinling mountains were analyzed. The results showed that: 11-year average annual NPP was 542.24 gC·m-2·a-1; NPP in Qinling mountains was remarkably increased (P<0.01) during the studied period, and the maximum of NPP (718.77 gC·m-2·a-1) was in 2008, the minimum (471.78 gC·m-2·a-1) was in 2001; the size of contribution rate of four seasons for annual NPP followed by summer (49.90%)>spring (26.16%)>autumn (18.87%)>winter (5.07%); there was a significant correlation of monthly NPP with temperature and precipitation, and the correlation with temperature was higher than that with precipitation, the influence of temperature on NPP was larger than precipitation on NPP in the month level; the correlation of NPP with temperature and precipitation was mainly characterized by positive value in spatial distribution during growing season.
    SNP Analysis of 4CL Gene in Larix olgensis Henry Populations
    XU Yue-Li;ZHANG Han-Guo*;SHI Tian-Yuan;YAO Yu;ZHANG Lei;WANG Yan-Hong;LIU Ling
    2013, 33(2):  208-213.  doi:10.7525/j.issn.1673-5102.2013.02.015
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    With the off-site preservation forest 21-year old Larix olgensis in Qinshan forestry centre in Linkou, in Heilongjiang province as the materials, the resources of Baidaoshan were analyzed using SNP molecular markers and he software DNAMAN. There is genetic diversity in L.olgensis. SNP marker has proved the provenance of L.olgensis. Rupr has a wealth of genetic diversity from the molecular level, and the marker indicates that the genetic variation mainly exists within populations, smaller variation exists among population SNP Primers detected of 178 SNPs. It has sequenced a total of 240 EST sequencing, and the successfully are 193 EST sequencing, in which the successful percentage is 80.42%. Using the software DNAMAN analysis the sequencing result found that there are 178 SNPs, 126 belong to transition type, accounting for 70.79% of the total variance; with 52 belong to transversion type, accounting for 29.21% of the total variance, the mutant type roughly in line, transition∶transversion=7∶3. In the transition type, A/G and C/T transition account for 37.08% and 33.71%, respectively; some types of transversion G/C, A/C, A/T, and G/T respectively in 7.87%, 8.99%, 7.87% and 4.99%. But within each family, transition and transversion are quite different, there is no law. The largest proportion of conversion and transversion is No.554 family, a ratio of 7∶1, the smallest pedigree is 855 pedigree with ratio 3∶4.
    Primary Study on the Endangered Mechanism of the Rare Plant Keteleeria evelyniana var. pendula
    MOU Feng-Juan;DAI Xing-Fen;MA Shi-Zhu;DEND Chang-Xian
    2013, 33(2):  214-219.  doi:10.7525/j.issn.1673-5102.2013.02.016
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    The geographical distribution and natural habitats, as well as the population size and the age structure of the population of Keteleeria evelyniana var. pendula in Huaning County, Yunnan Province were studied in this paper, and the biological characteristics, such as the morphological features, the development rule of the growth and its spreading and germination of seeds were well knew. It was concluded that K.evelyniana var. pendula was mainly distributed in the mountain areas near villages, such as Pingdi and Mada in Huaning County; the existing groups of this variety were the small populations, the clustering populations were characterized by the illogical age structure, and the populations were in a relatively unsteady stage. Although the characteristics of the seed wing were good for spreading over long distances, the seed yield was extremely low and the transformation of seeds to seedling is difficult. Therefore, the sexual reproduction was suffocated and the life history tended to break, which is an important one of the internal reasons that results in its being endangered condition at last. Furthermore, the survival of this species is severely threatened by the natural habitats being seriously damaged and the adults being excessively excavated, which is the main external reasons that cause the dramatical decreasing of the individuals in the wild population. The effective measures of protection and utilization were proposed, based on the endangered mechanism of K.evelyniana var. pendula.
    Rice Disease Resistance Induced by New Fungal Activator Protein and Its Physiological Mechanism
    YUAN Xiao-Han;GU Cheng-Bo*;QIU De-Wen;FU Li-Nan;LI Wang;WANG Qiu-Xue;GUO Dong-Jie;CAI Man
    2013, 33(2):  220-224.  doi:10.7525/j.issn.1673-5102.2013.02.017
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    In order to clarify new fungal activator protein induced resistance in rice and its physiological mechanism, changes of peroxidase (POD), polyphenol oxidase (POD), superoxide dismutase (SOD) and catalase (CAT) activities and hydrogen peroxide (H2O2) content in rice seedlings were determined at various time points after activator protein treatment. The results showed that activator protein could induce resistance in rice against blast fungus and bacterial blight. The resistance induced by activate protein at the concentration of 1-6 μg·mL-1 against blast fungus and bacterial blight was 45.2%-71.4% and 47.6%-66.3%, respectively, and the best results were achieved at the concentration of 6 μg·mL-1 activate protein. Compared with control group, the activities of POD, PPO and SOD and hydrogen peroxide content in rice seedlings were increased with different levels, but CAT activities inhibited 3-15 d after the rice seedlings treated with the concentration of 2 μg·mL-1 activator protein. These findings showed that activator protein-induced resistance in rice was closely related to active oxygen metabolism.
    ISSR Identification of Clone Plants of Solanum tuberosum Hybrids F1
    GAN Lin;YU Xiao-Xia;JU Tian-Hua;YU Zhuo*;MENG Mei-Lian;ZHANG Zi-Qiang;LI Chang-Qing
    2013, 33(2):  243-247.  doi:10.7525/j.issn.1673-5102.2013.02.021
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    In order to provide reliable materials for potato varieties breeding, the study was to identify the authenticity of clones of hybrids F1 of two potato combinations, ‘J07-4’בLongshu No.6’ and ‘J07-6’בLongshu No.6’, using ISSR molecular markers. The results were that five primers were selected from 152 ISSR primers, primers AF18550, AW75511 and AW20617 were suitable to identify 5 clones of ‘J07-4’בLongshu No.6’ hybrids F1, and AW20607 and AF18549 were suitable to identify 7 clones of ‘J07-6’בLongshu No.6’ hybrids F1. A total of 12 clones were identified as true hybrids based on the paternal-specific bands in progeny’s bands. ISSR fingerprints of different clones of hybrids F1 were established. It showed that it was feasible to use ISSR molecular markers to identify clones of potato hybrids F1.
    Establishment of Molecular ID for Grape Germplasm Based on SSR Markers
    DU Jing-Jing;LIU Guo-Yin;WEI Jun-Ya;LIU De-Bing*;YANG Xiao-Zhen
    2013, 33(2):  232-237.  doi:10.7525/j.issn.1673-5102.2013.02.019
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    Using a total of 80 grape germplasms selected from national grape repository of Zhengzhou as the material, the methods of establishing the grape germplasms molecular ID were studied. Germplasms were distinguished by the selected SSR markers, and then coded the size of augmentation strip molecular based on the different germplasms. The results showed that after screening of the 62 primers, 169 alleles were detected using 28 selected SSR markers located on each chromosome of grape, with a mean value of 6.0 alleles/locus. After alleles assignment, there were only 9 pairs of primers selected out for the construction of germplasms molecular ID with an average value of 8.9 germplasms/primers. The purpose of distinguishing the variety of grape was achieved by a minimum of primers. The results suggested SSR marker techniques can be used for establishing molecular identity of grape germplasm.
    Establishment and Optimization of ISSR-PCR Reaction System for Brassica campestris L. var. rosularis
    SONG Jiang-Hua;ZHAO Ying;WANG Cheng-Gang*;ZHANG Ling-Feng;ZHANG Hui
    2013, 33(2):  238-242.  doi:10.7525/j.issn.1673-5102.2013.02.020
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    For optimizing ISSR-PCR reaction system of Brassica campestris L. var. rosularis, single factor gradient and orthogonal design experiments were conducted. The main factors affecting ISSR-PCR amplification i.e. suitable concentration of primer, dNTP, Mg2+ and Taq DNA polymerase were studied. Furthermore, the annealing temperature and cycling numbers were optimized on the base of the above tests. An ideally ISSR-PCR reaction system was established, namely 20 μL reaction system containing template DNA 30 ng, 0.50 μmol·L-1 primer, 0.25 mmol·L-1 dNTP, 1 mmol·L-1 Mg2+ and Taq DNA polymerase 1.0 U. The optimal PCR amplification program was:3 min at 94℃ for predenaturation, followed by 35 cycles of 30 sec at 94℃ for denaturation, 1 min at 50℃ for annealing, 90 sec at 72℃ for extension, finally extension at 72℃ for 7 min and holding the samples at 4℃. This optimized ISSR-PCR reaction system would provide the basis for the analysis of germplasm classification and identification in B.campestris.
    A Direct Experimental Proof for Long-term Persistent Spore Bank in Hani Peatland of the Changbai Mountains
    FENG Lu;TANG Yuan;BU Zhao-Jun*;ZHAO Gao-Lin;
    2013, 33(2):  248-251.  doi:10.7525/j.issn.1673-5102.2013.02.022
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    Sexual propagule bank is of great importance for persistence of plant populations. To this day, no direct experiment proof of long-term persistent spore bank of bryophytes in peatlands was reported. In Hani Peatland of the Changbai Mountains, a 50 cm deep peat core was drilled and burial time of spores in peat layers was estimated by larch dating method. After the spores of Sphagnum capillifolium being extracted from the peat layer by layer, the spores were cultivated to investigate the effect of burial time on spore germination rate. The results showed that, with the increase of burial time, the germination rate of Sphagnum spores decreased exponentially. The results indicated that there was a long-term persistent spore bank in peatlands since spores of S.capillifolium still have the potential to germinate after 112 years of burial. According to our calculation, the maximum longevity in spores of S.capillifolium could be up to 396.4 years.
    Analysis of the Contents of Total and Three Major Flavonoids in Different Tissues of Mosla chinensis
    JIANG Hua-Jun;LIN Yong;CHANG Qiu;WANG Ying-Zi;HUANG Zhi-Qiang;LIU Zhong-Hua*
    2013, 33(2):  252-256.  doi:10.7525/j.issn.1673-5102.2013.02.023
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    The content of active constituents in Chinese herbal medicine is a key indicator for judging their qualities. In this study, the methods were established for analyzing total and three major flavonoids in Mosla chinensis, respectively, and the differences of the flavonoid contents in different tissues were also compared, which will favour the exploitation of the flavonoids in M.chinensis. The results showed that the optimized spectrophotometry for the determination of total flavonoid content was specific and stable, and the HPLC method for the analysis of luteolin, apigenin and negletein presented a good linear relationship (r>0.999) in the range of the determination and the recovery ranges were 98.77%~102.13% with less than 5% RSDs. The study also found that the content of total flavonoids in different parts of M.chinensis from high to low was bud>tip>stem>root, and the total flavonoid content in M.chinensis from Jiangxi was higher than that from Zhejiang. These prompt that genunie medicinal materials should be selected for the industrial production of the flavonoids from M.chinensis as more as possible, and the root and the lower end of stem may be considered to discard, so as to improve the extraction efficiency and increase economic benefits. In brief, the developed methods are accurate and steady for detecting total and three major flavonoids in M.chinensis, and we also confirmed total flavonoids were mainly enriched in and near the bud of M.chinensis, which would provide a theoretical basis and scientific guidance for further exploiting the resources in M.chinensis.