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    20 January 2013, Volume 33 Issue 1
    Description on the Reproductive Organs of Bambusa multiplex(Lour.) Raeuschel ex J.A.et J.H.Schult. var. incana B.M.Yang
    CHEN Song-He
    2013, 33(1):  1-3.  doi:10.7525/j.issn.1673-5102.2013.01.001
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    There was no description or record for the reproductive organs of Bambusa multiplex(Lour.) Raeuschel ex J. A. et J. H. Schult. var. incana B. M. Yang in the “Chinese Flora”[Tomus 9(1)],《Flora of China》and other related literature. In this paper, the author gathered and manufactured its reproductive organ specimen and described it both in English and in Chinese. Its main features were: the flowering branch 50-160 cm; pseudo spikelets solitary or several clustered at nodes of flowering branches; florets 5-8, rachilla segments flat, 3-4 mm, glabrous; glumes absent; lemma asymmetrical, oblong-lanceolate, glabrous;palea linear; filaments 0.5-1.2 cm; anthers purple; ovary ovoid; stigmas 3 or variable in number, feathery; lodicules 3; stamens 6. Flowering March-April. Mature caryopsis is thin narrow ovale, brown, ca. 0.8-1.0 cm, dia. 2.0-2.5 cm, peel thicker.The specimen was collected from Xiamen Botanical Garden.
    Cephalotaxus talonensis Cheng et Feng ex S.G.Lu et X.D.Lang,a New Name of the Family Cephalotaxaceae and Its Taxonomic Status
    LANG Xue-Dong;SU Jian-Rong*;LU Shu-Gang
    2013, 33(1):  4-6.  doi:10.7525/j.issn.1673-5102.2013.01.002
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    A new name of Cephalotaxaceae, Cephalotaxus talonensis Cheng et Feng ex S. G. Lu et X. D. Lang, is reported to replace the illegitimate name, Cephalotaxus lanceolata K. M. Feng(1975), which is a later homonym of the name Cephalotaxus lanceolata Hort. ex Beiβner (1901). Additionally, based on comparison of leaf morphology and holotypes between Cephalotaxus fortunei Hooker and C.lanceolata K. M. Feng, we support that C.lanceolata K. M. Feng should be treated as rank of species, and disagreed with the argument that Silba (1990) reduced C.lanceolata K. M. Feng to C.fortunei Hooker var. lanceolata (Feng) Silba. In this paper, four new pictures are provided.
    Leaf Epidermal Cell Micromorphology of Sect.Conostylae(Wolf) Yü et Li of Potentilla L.
    2013, 33(1):  7-17.  doi:10.7525/j.issn.1673-5102.2013.01.003
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    The leaf epidermal cell micromorphology of 15 species of Sect. Conostylae(Wolf) Yü et Li of Potentilla L. from Xinjiang were investigated under light microscope (LM) and scanning electron microscope (SEM). The type of epidermal hairs, the size and shape of epidermal cells, the distribution and type of stomatal apparatus were studied. The stomatal size, stomatal density, and stomatal index were analyzed. The results show that there are stomatal apparatus on lower epidermal of all 15 studied species, but no stomatal apparatus on upper epidermal of a few species. The shapes of stomatal apparatus are long oval, oval, wide oval and nearly round. The stomatal apparatus of most species are short paracytic four-cell type, irregular four-cell type, irregular type, around type and radial type. The trichomes are needle-like hairs or ribbon pubescent hairs. The epidermal cells are polygonal to irregular. The characteristics of stomatal apparatus, stomatal index, wax ornamentation and trichomes are all obviously different among different species in genus Potentilla L. The leaf epidermal morphological characteristics could provide evidence to identify subgenus and species of Potentilla L..
    Resource and Floristic Characteristics of the Key Protection Wild Plants in Shanxi
    ZHANG Yin-Bo;ZHANG Xiao-Long;LU Yi-Meng;LI Ming
    2013, 33(1):  18-23.  doi:10.7525/j.issn.1673-5102.2013.01.004
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    According to List of Wild Plants under State Protection (First Batch) and List of Wild Plants under Provincial Protection in Shanxi (First Batch), the list of key protection wild plants in Shanxi was established. It was recorded that there were 57 protected plants, which belong to 38 families and 45 genera (including 2 species under state protection category Ⅰ, 6 species under state protection category Ⅱ and 49 species under provincial protection). By analysis, the key protection wild plants are not concentrated in Shanxi and scatter in the 61 counties (cumulative statistics). Moreover, it was found that the distribution ranges of the majority of species are small and narrow. The floristic characteristics of the key protected wild plants in Shanxi are complex and diverse, old origin, more relict plants and monotypic genera. Temperate floristic elements are significantly dominant in genera, with obviously tropical nature, it also has the characteristics of transition, and its unique geographical advantage and Conservation value. The aim of the research is to analyze preliminarily resources situation and floristic characteristics of the key protection wild plants in Shanxi, in order to provide scientific and theoretical support for reasonable conservation and effective use of the key protection wild plants in Shanxi.
    Spatial Pattern Analysis of Individuals in Different Age-classes of Pinus bungeana in Wulu Mountain Reserve,Shanxi,China
    YANG Xiao-Feng;MIAO Yan-Ming;ZHANG Qin-Di;ZHANG Ling;BI Run-Cheng*
    2013, 33(1):  24-30.  doi:10.7525/j.issn.1673-5102.2013.01.005
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    Pinus bungeana is an important protective plant. Spatial pattern analysis of individuals in different age-classes of P.bungeana was conducted in the present study. Here we employed the technique of point pattern analysis which can analyze patterns under all scale along a gradient. The results showed: The densities of six age-classes were more different, and age-classes 2~5 had more individuals than the other two age-classes. The individuals of all age-classes were not clumping distributed in space, however their distribution pattern trended to random distribution varied with the change of scale. The co-relationship between age-class 1 and age-class 2 was significantly positive, this significantly positive correlation is beneficial for the development of the group. In the smaller scale, the individuals of different age-classes were a stronger spatial association. When the scale was larger, their spatial association would be weakened greatly. This showed the spatial patterns of different age-classes individuals and the spatial association between the individuals had a close relationship with the spatial scale.
    Characteristics of Seeds and Hybrid Seeds in Three Sympatric Cypripedium Species
    WANG Heng-Rong;LI Peng*;LI Jun-Lin;LUO Yi-Bo
    2013, 33(1):  31-38.  doi:10.7525/j.issn.1673-5102.2013.01.006
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    It is important to study hybrid seeds because it will illuminate hybrid inviability and reproductive isolation mechanism between plant species. Morphological characteristics and viability of seeds derived from self-pollination, cross-pollination and hybridization were analyzed in three sympatric Cypripedium species. Hybrid seeds with high viability were successfully obtained from sympatric plants C.flavum, C.tibeticum and C.calcicola. They were not significantly different in seed shape, surface sculpture compared with seeds from self-pollination and cross-pollination. The width, length, embryo ratio and pigmentation ratio of hybrid seeds were not significantly lower than those of seeds from self-pollination and cross-pollination. The results suggested that three sympatric Cypripedium species could be hybrid compatible and that hybrid inviability could not be the main reason of isolation among them. It was not necessary to produce other isolation mechanisms because pollinators with different sizes functioned as the main isolation mechanism between C.flavum and C.tibeticum or C.calcicola. However, with the same pollinators and hybrid viability, C.tibeticum or C.calcicola would produce other isolation mechanisms or they should be combined into one species.
    Effect of Exogenous Nitric Oxide on Photosynthetic Pigment and Antioxidant Enzymes in Taxus chinensis var. marei Seedlings
    LI Mei-Lan;LI De-Wen*;YU Jing-Hua;ZU Yuan-Gang
    2013, 33(1):  39-44.  doi:10.7525/j.issn.1673-5102.2013.01.007
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    In this experiment, 4-year-old Taxus chinensis var. mairei seedlings were used as test material and treated with different concentrations (0, 0.01, 0.1, 0.5 and 1 mmol·L-1) of sodium nitroprusside (SNP), a donor for exogenous nitric oxide (NO) to study the effect of exogenous nitric oxide on photosynthetic pigment and antioxidant enzymes in T.chinensis seedlings. The contents of photosynthetic pigment, malondialdehyde (MDA) and hydrogen peroxide (H2O2), and the activities of antioxidant enzymes in T.chinensis seedlings were determined. Spraying low concentrations (0.01, 0.1 mmol·L-1) of SNP would significantly increased chlorophyll a/b ratio and the contents of chlorophyll a, chlorophyll b, carotenoids and the total chlorophyll, while the treatment of high concentrations (0.5, 1 mmol·L-1) of SNP would reduce the chlorophyll contents. The activity of catalase(CAT) significantly increased with the increasing of the concentration of exogenous NO donor, while the activity of peroxidase(POD) was first increased, and then decreased. Moreover, at the early stage of treatment, low concentrations of SNP treatment obviously increased the ascorbate peroxide(APX) activity, while high concentrations of SNP treatment significantly decreased the APX activity, and at the later stage of treatment, the activity of APX significantly decreased with the increasing of SNP concentration. Spraying low concentrations of SNP would effectively improve the superoxide dismutase(SOD) activity and increase the soluble protein concentrations, reduced MDA and H2O2 contents, while the treatment of high concentrations of SNP significantly increase the MDA and H2O2 contents. Therefore, treatment of low concentrations of SNP (<0.5 mmol·L-1) would increase chlorophyll contents, the activity of antioxidant enzymes, and reduce the MDA and H2O2 contents in T.chinensis seedlings, while the treatment of high concentrations of SNP (≥0.5 mmol·L-1) would reduce the chlorophyll contents, increase the H2O2 contents and membrane lipid peroxidation, and might cause a certain injuries on T.chinensis seedlings.
    Changes of Physiological Properties in Limonium aureum(Linn.) Hill Seedlings under Salt Stress
    YOU Jia;ZHANG Jin;WANG Wen-Rui;LU Jin;JIA Peng-Xiang;MIAO Jian-Shun;YANG Ying-Li*
    2013, 33(1):  45-50.  doi:10.7525/j.issn.1673-5102.2013.01.008
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    In this study, desert halophyte Limonium aureum(Linn.) Hill was used to investigate the changes of osmolyte content, reactive oxygen species production and antioxidant enzyme activities in the leaves of seedlings under different NaCl concentrations. The levels of proline, soluble sugar and H2O2 as well as the rate of O2 production increased in the seedlings treated with different NaCl concentrations. Compared with the control, the amount of soluble protein decreased in the leaves of seedlings treated with 25 and 50mmol·L-1 NaCl, but enhanced under 100 and 150 mmol·L-1 NaCl treatments. Additionally, all applied NaCl concentrations resulted in the enhancement of superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities. In contrast, the activity of peroxidase increased at first and then decreased, but significant decrease in catalase activity was observed in the leaves of L.aureum treated with NaCl, as compared with the untreated seedlings. These results indicated that the accumulation of osmoregulation substances and the enhancements of SOD and APX activities in response to salt stress elevated the osmotic adjust action and the oxidation resistance ability in the leaves of L.aureum, which might be associated with the elevation of plant tolerance to salt condition.
    Comparative Proteome Analysis of Heterosis in Wheat F1-hybird Bred Through Hybridization of CMS with Restoring Line
    LIU Xiao-Ying;DUAN Shuang;WANG Zhen-Ying;GAO Yue;PENG Yong-Kang*
    2013, 33(1):  51-57.  doi:10.7525/j.issn.1673-5102.2013.01.009
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    In this study, by using the leaves of seedling stage (tillering stage), shooting stage, booting stage and anthers of PMC meiotic stage, microspore 2-3 nuclei stage, pollen grain as experimental materials, proteome approaches were applied to a comparative analysis of heterosis in wheat F1-hybrid which was bred through hybridization of CMS with restoring line. The results indicated that three types of protein expression levels, super-parents type, parent di-type, low-parents type, were observed among F1-hybrid and parents. The frequence of protein expression was showed as parent di-type>low-parents type>super-parents type. A total of 80 protein spots were detected in F1-hybrid and parents, 17 protein spots have been identified by MALDI-YOF MS. These protein spots were involved in diverse biological processes, including DNA and protein synthesis, energy metabolism, stress response, gene transposition and photosynthesis. In the F1-hybrid, at seedling stage, leaf aerial traits such as leaf fresh weight (LFW), leaf dry weight (LDW) and leaf number, show a tendency to its parental lines when compared to parent, these results were consistent to proteome analysis results which showed most of protein spots expression in F1-hybrid exhibiting parent di-type. However, LFW, LDW, and total leaf number exhibited obviously heterosis because of tillering number (TN) increasing in F1-hybrid, it is necessary to further study the relationship between the heterosis and proteome change.
    Cloning of Camellia nitidissima Flavonol Synthase cDNA and Construction of Sense,RNA Interference Expression Vectors
    ZHOU Xing-Wen;LI Ji-Yuan*;Yin Heng-Fu;FAN Zheng-Qi;Li Xin-Lei
    2013, 33(1):  58-65.  doi:10.7525/j.issn.1673-5102.2013.01.010
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    A full-length cDNA sequence of flavonol synthase (FLS) gene was obtained from petals of Camellia nitidissima using the methods of Reverse Transcription PCR and RACE, and named CnFLS(GenBank accession No.JF343560.1). The sense expression vector pCAM-CnFLS was constructed successfully while the gene FLS amplified by the primers P3/P4 and inserted into PMD18-T vector was connected to expression vector pCAMBIA1300. The RNA interference (RNAi) expression vector was established by many times enzyme digestion and connection on the base of intermediate vector pUCCRNAi, expression vector pCAMBIA1300 and the combined vector PMD18-T with a conserved segment of 250 bp which was amplified by primers G1 and G2. The successful construction of sense and RNAi expression vectors provides a foundation for further studying on CnFLS gene functions and its effects to flower color.
    Cloning and Expression Analysis of Tea Hydroperoxide Lyase Gene CsiHPL1
    XIN Zhao-Jun;SUN Xiao-Ling;ZHANG Zheng-Qun;CHEN Zong-Mao*
    2013, 33(1):  66-72.  doi:10.7525/j.issn.1673-5102.2013.01.011
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    Primers were designed according to the cDNA sequence of tea CsiHPL1. RT-PCR method was used to clone CsiHPL1 from Longjing 43′. The expression of CsiHPL1 under biotic and abiotic stress and subcellular localization were analyzed. The results showed CsiHPL1 contains an open reading frame of 1 476 bp which encodes a protein of 491 amino acids. This gene was predicted as a 13-HPL gene. Tea geometrid feeding, wounding and JA treatment upregulated the expression levels of CsiHPL1. The total sequence of this gene was fused with GFP to construct a binary vector for tobacco transient transformation. Under confocal laser-scanning microscopy, green fluorescent signals were localized in chloroplasts in transgenic tobacco plants, suggesting that the gene encodes a protein targeting to chloroplast.
    Prokaryotic Expression of TFMV,FVY and LMoV CP Gene,Antiserum Preparation and Virus Detection
    WU Qi-Yao;WEI Chuan-Bao*;LI Jin-Hua
    2013, 33(1):  73-79.  doi:10.7525/j.issn.1673-5102.2013.01.012
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    According to Genbank, specific primers were designed to amplify the CP genes of TFMV, FVY and LMoV infecting Fritillaria thunbergii Miq. and the CP sequences were analyzed. Then the CP genes were inserted into pSBET vector and expressed in Escherichia coli BL21(DE3)plys E strain. The object proteins were purified by 12% SDS-PAGE firstly and subsequently 5%-20% gradient SDS-PAGE. The antiserum against the CPs was raised in mouse. The specificity and serological relationship were confirmed by Western blot analysis and the ability to combine with nature virus particles was confirmed by ELISA analysis. Western blot, indirect ELISA and Dot-ELISA techniques were used for the detection of viruses infecting F.thunbergii. The results indicated that the antiserum is specific to its CP and no acrossing-reaction with others. It could combine with nature virus particles. Indirect ELISA and Dot-ELISA techniques are suitable for detecting three kinds of viruses.
    Impacts of Microenvironment on Pollination Success of an Orchid Species Phaius delavayi in Huanglong Valley,Sichuan
    HUANG Bao-Qiang;AN De-Jun
    2013, 33(1):  80-85.  doi:10.7525/j.issn.1673-5102.2013.01.013
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    There are thousands of Phaius delavayi individuals, which mainly distributed in two different habitats, .i e. travertine areas and the remained forests, in Huanglong Valley, Sichuan, and the individual pollination successes were different between these two habitats. However, little is know about the causes and mechanisms leading to the differences. The differences of the microenvironmental conditions, morphological traits, male and female reproductive success of P.delavayi in two habitats were measured. The results showed that the microenvironmental conditions were found to be quite different between these two habitats, tree cover, soil depth, soil moisture, soil organic matter, total nitrogen, and relative air humidity were higher in forest areas, on the contrary, shrub cover, herb cover, moss cover, air temperature, relative light intensity, soil temperature, total calcium, and pH were higher in travertine areas. The results also showed that floral traits and reproductive success of P.delavayi had significant differences between these two habitats. The individuals in travertine areas had larger length and width of the largest leaf, more numbers of flowers within inflorescence, larger length of spur, and longer life span of both single flower and inflorescence than those in forest areas. However, both male and female reproductive successes were lower in travertine areas than those of that in forest areas. We supposed that the differences of temperature and light between two habitats may play important roles in leading to the differences of pollinia removal and nature fruit set of P.delavayi in the studied area.
    Allelopathy of Aqueous Extracts from Four Species Plants on Artemisia wudanica
    Wendurihu;WANG Tie-Juan*;HAN Wen-Juan;DAI Ting-Ting
    2013, 33(1):  86-90.  doi:10.7525/j.issn.1673-5102.2013.01.014
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    Sand-fixing pioneer plant Artemisia wudanica that distributed in the western part of Horqin sandy land showed a declining trend in recent years. Three main aerial seeding plants A.sphaerocephala, Caragana korshinskii, Hedysarum leave and A.wudanica itself were selected to study allelopathy. The results showed that: the aqueous extracts of three aerial seeding plants and A.wudanica itself all had different allelopathy on the growth of A.wudanica, and allelochemicals in different parts (root, leaf, peel or bracts) had different functions. Furthermore, the germination rates of A.wudanica seeds in the root aqueous extracts of all donor plants were significantly declined except H.leave. A.sphaerocephala bracts and H.laeve peel aqueous extracts also had the same result, while the leaf extract of A.wudanica had promoting effect. The aqueous extracts from different parts of four species had obvious inhibitory effects on A.wudanica seed germination speed and root growth. By synthetical effect, four species all had inhibitory effects, the order of inhibitory effects from strong to weak was as follows: A.sphaerocephala>C.korshinskii>A.wudanica>H.laeve.
    Comparison of Three Protein Extraction Methods for Tung trees (Vernicia fordii) Seed
    ZHAN Zhi-Yong;WANG Yang-Dong*;CHEN Yi-Cun;HAN Xiao-Jiao;CUI Qin-Qin
    2013, 33(1):  91-97.  doi:10.7525/j.issn.1673-5102.2013.01.015
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    An effective protein extraction method is crucial to separate the proteins by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Tung tree (Vernicia fordii) seed, which is used for producing the tung oil, contains large amounts of compounds which can interfere with the extraction of proteins, and can even affect the resolution of 2D-PAGE images. In the present study, we developed a suitable protein extraction method from three tested methods, including trichloroacetic acid (TCA)-acetone (Method A), phenol (Method B), TCA-acetone combined with phenol (Method C). The results showed that the protein concentration of samples by Method C was high and it achieved 8.1 μg·μL-1. Method C has a strong capability to extract the high-molecular-weight and the low-molecular-weight proteins from tung tree seeds. In addition, the 2D-PAGE image resolution of Method C was also better than the other two methods. Thus, Method C was used to extract proteins, and it would lay the foundation for proteomic analysis on the tung tree seeds.
    Determination of Effects of Environmental Factors on Stream Macroalgae in Xin’an Spring by Grey Correlation Analysis
    HU Bian-Fang;XIE Shu-Lian*
    2013, 33(1):  98-103.  doi:10.7525/j.issn.1673-5102.2013.01.016
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    The effects of environmental factors on stream macroalgae were investigated. Data were collected from July 2009 to April 2010 from Xin’an Spring, Shanxi Province, north China. The environmental variables consisted of water temperature, pH, specific conductance, dissolved oxygen, surface current velocity, maximum depth and width. Fortytwo species of stream macroalgae were identified in this spring. The correlation between these seven environmental factors and the number of stream macroalgal species were determined using grey correlation analysis. The results showed that water temperature and current velocity were the key factors affecting the number of stream macroalgae. This result was consistent with previous studies. Therefore, it indicated that the grey correlation analysis can be used in the study of algal ecological environment.
    Preparation and Characterization of Micronized Vitexin by Antisolvent Recrystallization Method
    LI Wang;;WANG Qiu-Xue;;GU Cheng-Bo;*;ZU Yuan-Gang;;ZHAO Xiu-Hua;;FU LI-Nan;;GUO Dong-Jie;
    2013, 33(1):  104-108.  doi:10.7525/j.issn.1673-5102.2013.01.017
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    To improve the dissolution rate of vitexin, the micronization of vitexin in an antisolvent recrystallization process using N-methylpyrrolidone (NMP) as solvent and water as antisolvent was studied in this paper. The effects of operating parameters including concentrations of vitexin solution, system temperatures, stirring rates and kinds of surfactants (PVP, Tween80, SDS) on the particle size of micronized vitexin were investigated. Particles of micronized vitexin with a mean particle size of 291.1 nm were obtained under the optimized process conditions (vitexin solution concentration 20 mg·mL-1, volume ratio of antisolvent to solvent 10∶1, stirring rate 1 500 r·min-1 and concentration of PVP 8 mg·mL-1). The processed vitexin particles and the unprocessed one were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), Fouriertransform infrared spectroscopy (FT-IR). The results showed that antisolvent recrystallization process did not induce degradation of vitexin and the micronized vitexin had lower crystallinity. The chemical properties of the processed vitexin did not change when compared with the original drug, and there was a significant increase of the dissolution rate of the processed vitexin comparing with the unprocessed vitexin.
    Scutellaria baicalensis Georgi Cell Apoptosis Regulated by bax Gene
    SU Hu;XU Mao-Jun;JIANG Hu;LI Yu-Ping
    2013, 33(1):  109-113.  doi:10.7525/j.issn.1673-5102.2013.01.018
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    Bax gene as a member of the bcl-2 genes family plays the role of the promotion of apoptosis. In this experiment, Scutellaria baicalensis Georgi cell was transformed with bax gene, the transformed cell treated with 25 and 40 μmol·L-1 estradiol show the features of apoptosis. The browning occurred obviously after 5 days treated with 40 μmol·L-1 estradiol, the features of nucleus marginalization, chromatin pyknosis and the formation of nuclear apoptotic bodies were observed.
    Progress on the Structure and Physiological Functions of Apoplastic Barriers in Root
    YANG Chao-Dong;ZHANG Xia;LIU Guo-Feng;ZHANG Jun-Wei;BAO Man-Zhu;ZHOU Zhi-Xiang
    2013, 33(1):  114-119.  doi:10.7525/j.issn.1673-5102.2013.01.019
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    The progress on the structure and physiological functions of apoplastic barriers in root is presented for recently ten years. The apoplastic barriers consist of the cell wall with Casparian bands, suberin lamellae or lignin which showed as protective tissue, that tight block water, ion and oxygen between plant and environment. The molecular information of Casparian bands indicated the endodermis similar to tight junctions of epithelial barrier. The apoplastic barriers are the key structure for plant adapted to adverse environment that guarantee various physiological and biochemical reactions processing under steady inner surroundings. The apoplastic barriers reacted well to drought, flooding, ion deficiency, and disease stresses, and broadly applied to explore plant resource to adapt and restore the extreme environment.
    New Research Progress on the Regulation of Stomatal Movement in Plant
    QIAN Bao-Yun;LI Xia*
    2013, 33(1):  120-128.  doi:10.7525/j.issn.1673-5102.2013.01.020
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    The stomatal development and regulation mechanism of stomatal movement were reviewed in the present paper. Moreover, a brief analysis on the relationship between the stomatal regulation mechanism of pepc transgenic rice and high photosythetic efficiency was made. Finally, the research of the regulation mechanism of plant stomata movement was prospected.