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Bulletin of Botanical Research 2013 Vol.33
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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
Bulletin of Botanical Research 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.
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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
Bulletin of Botanical Research 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.
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Leaf Epidermal Cell Micromorphology of Sect.
Conostylae
(Wolf) Yü et Li of
Potentilla
L.
ZHANG Man-Man;NUERMAIMAITI·Mo Ming;WANG Hong*;BUZUORIGULI·Ka Di Er;JIANG Yan-Cheng
Bulletin of Botanical Research 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..
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Resource and Floristic Characteristics of the Key Protection Wild Plants in Shanxi
ZHANG Yin-Bo;ZHANG Xiao-Long;LU Yi-Meng;LI Ming
Bulletin of Botanical Research 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.
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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*
Bulletin of Botanical Research 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.
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Characteristics of Seeds and Hybrid Seeds in Three Sympatric
Cypripedium
Species
WANG Heng-Rong;LI Peng*;LI Jun-Lin;LUO Yi-Bo
Bulletin of Botanical Research 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.
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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
Bulletin of Botanical Research 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 (H
2
O
2
), 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 H
2
O
2
contents, while the treatment of high concentrations of SNP significantly increase the MDA and H
2
O
2
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 H
2
O
2
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 H
2
O
2
contents and membrane lipid peroxidation, and might cause a certain injuries on
T.chinensis
seedlings.
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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*
Bulletin of Botanical Research 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 H
2
O
2
as well as the rate of O
2
-·
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 50mmol·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.
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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*
Bulletin of Botanical Research 2013, 33 (
1
): 51-57. DOI:
10.7525/j.issn.1673-5102.2013.01.009
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1302
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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.
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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
Bulletin of Botanical Research 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.
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Cloning and Expression Analysis of Tea Hydroperoxide Lyase Gene
CsiHPL1
XIN Zhao-Jun;SUN Xiao-Ling;ZHANG Zheng-Qun;CHEN Zong-Mao*
Bulletin of Botanical Research 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 upregulated 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.
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Prokaryotic Expression of TFMV,FVY and LMoV CP Gene,Antiserum Preparation and Virus Detection
WU Qi-Yao;WEI Chuan-Bao*;LI Jin-Hua
Bulletin of Botanical Research 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.
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Impacts of Microenvironment on Pollination Success of an Orchid Species
Phaius delavayi
in Huanglong Valley,Sichuan
HUANG Bao-Qiang;AN De-Jun
Bulletin of Botanical Research 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.
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Allelopathy of Aqueous Extracts from Four Species Plants on
Artemisia wudanica
Wendurihu;WANG Tie-Juan*;HAN Wen-Juan;DAI Ting-Ting
Bulletin of Botanical Research 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
.
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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
Bulletin of Botanical Research 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.
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Determination of Effects of Environmental Factors on Stream Macroalgae in Xin’an Spring by Grey Correlation Analysis
HU Bian-Fang;XIE Shu-Lian*
Bulletin of Botanical Research 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. Fortytwo 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.
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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;
Bulletin of Botanical Research 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), Fouriertransform 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.
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Scutellaria baicalensis
Georgi Cell Apoptosis Regulated by
bax
Gene
SU Hu;XU Mao-Jun;JIANG Hu;LI Yu-Ping
Bulletin of Botanical Research 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.
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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
Bulletin of Botanical Research 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.
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New Research Progress on the Regulation of Stomatal Movement in Plant
QIAN Bao-Yun;LI Xia*
Bulletin of Botanical Research 2013, 33 (
1
): 120-128. DOI:
10.7525/j.issn.1673-5102.2013.01.020
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1545
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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.
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Peristome Morphology Characteristics of 16 Species of Family Pottiaceae and their Systematic Significance
WANG Xiao-Rui;WANG Li-Bao;LI Min;ZHAO Jian-Cheng*
Bulletin of Botanical Research 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.
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Anatomy of An Ephemeroid Plant
Eremurus altaicus
TIAN Li-Li;MA Miao*
Bulletin of Botanical Research 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.
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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
Bulletin of Botanical Research 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
.
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Observation on Megasporogenesis, Microsporogenesis and Development of Female and Male Gametophytes of
Catalpa speciosa
Warder
LI Li-Ping;LIU Hai-Yan;CHEN Fa-Ju*
Bulletin of Botanical Research 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.
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Megasporegenesis and Female Gametophyte Development in
Berberis pruinosa
Franch.
YANG Gui-Ying;MA Shao-Bin*
Bulletin of Botanical Research 2013, 33 (
2
): 149-153. DOI:
10.7525/j.issn.1673-5102.2013.02.005
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1388
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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.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 154-158. DOI:
10.7525/j.issn.1673-5102.2013.02.006
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1231
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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.
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Effect of Grazing on Plant Functional Group Characteristics in
stipa
Steppe between China and Mongolia
BAO Xiu-Xia;YI Jin;LIAN Yong;XIAO Xia;BAO Xiu-Ping
Bulletin of Botanical Research 2013, 33 (
2
): 159-165. DOI:
10.7525/j.issn.1673-5102.2013.02.007
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1299
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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.
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Provenance Analysis of Mongolian Oak at Seedling Stage
QU Hong-Jun;MENG Qing-Bin;ZHANG Zhong-Lin;YANG Chuan-Ping;ZHANG Jie*
Bulletin of Botanical Research 2013, 33 (
2
): 166-173. DOI:
10.7525/j.issn.1673-5102.2013.02.008
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1321
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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.
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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*
Bulletin of Botanical Research 2013, 33 (
2
): 174-180. DOI:
10.7525/j.issn.1673-5102.2013.02.009
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1176
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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/kinetochorebinding 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
AtZW
10 gene was amplified by PCR using
Arabidopsis
cDNA as a template, and the binary vectors consisting of
AtZW
10 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.
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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;*
Bulletin of Botanical Research 2013, 33 (
2
): 181-185. DOI:
10.7525/j.issn.1673-5102.2013.02.010
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1178
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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,
F
v
/F
m
,
Φ
PSⅡ
,
F
v
′/
F
m
′ and
qP
, and the obvious increase in chl a/chl b,
F
o
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.
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Effect of Enhanced UV-B Radiation on
Arabidopsis
Mesophyll Cell Protein
WEI Xiao-Li;ZHENG Na;LI Xiao-Yang;HAN Rong*
Bulletin of Botanical Research 2013, 33 (
2
): 186-190. DOI:
10.7525/j.issn.1673-5102.2013.02.011
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1182
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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, B
2
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 (B
2
), 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.
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Transformation and Expression of 1Dx5 without Selectable Markers in Common Wheat Xindong No.26
WANG Yong;QIN Jian-Bing;FAN Ling;ZHU Chang-Qing*
Bulletin of Botanical Research 2013, 33 (
2
): 191-196. DOI:
10.7525/j.issn.1673-5102.2013.02.012
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1408
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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.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 197-201. DOI:
10.7525/j.issn.1673-5102.2013.02.013
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1441
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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.
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Effects of Simulated Acid Rain on Physiological and Ecological Characteristics of
Sinocalycanthus chinensis
PENG Li-Qiong;JIN Ze-Xin*;WANG Qiang;
Bulletin of Botanical Research 2013, 33 (
2
): 202-207. DOI:
10.7525/j.issn.1673-5102.2013.02.014
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1069
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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.
F
v
/
F
m
and
F
v
/
F
o
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
.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 208-213. DOI:
10.7525/j.issn.1673-5102.2013.02.015
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(
1016
)
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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.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 214-219. DOI:
10.7525/j.issn.1673-5102.2013.02.016
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1315
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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
.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 220-224. DOI:
10.7525/j.issn.1673-5102.2013.02.017
Abstract
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1363
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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 (H
2
O
2
) 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.
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Vegetation Net Primary Productivity in Qinling Mountains and Its Response to Climate Change
YUAN Bo;BAI Hong-Ying*;ZHANG Jie;MA Xin-Ping
Bulletin of Botanical Research 2013, 33 (
2
): 225-231. DOI:
10.7525/j.issn.1673-5102.2013.02.018
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1495
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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.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 232-237. DOI:
10.7525/j.issn.1673-5102.2013.02.019
Abstract
(
1065
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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.
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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
Bulletin of Botanical Research 2013, 33 (
2
): 238-242. DOI:
10.7525/j.issn.1673-5102.2013.02.020
Abstract
(
1101
)
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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, Mg
2+
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
Mg
2+
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
.
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