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    Bulletin of Botanical Research 2012 Vol.32
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     A new species of Gentiana L.— G.hsinganica from Inner Mongolica,China
    YU Jing-Hua;HE Hao;ZHANG Bao-You;YUAN Shu-Sheng;CHEN Chang-Mei
    Bulletin of Botanical Research    DOI: 10.7525/j.issn.1673-5102.2012.01.001
    Taxonomic Status of Cephalotaxus alpina(Li) L.K.Fu(Cephalotaxaceae) in the Views of Morphological Characteristics of Seeds and Leaves
    LANG Xue-Dong;SU Jian-Rong*;LU Shu-Gang;ZHANG Zhi-Jun;LI Shuai-Feng
    Bulletin of Botanical Research    2012, 32 (1): 4-9.   DOI: 10.7525/j.issn.1673-5102.2012.01.002
    Abstract1624)      PDF(pc) (641KB)(679)       Save
    The significant differences in length, width, thickness, length/thickness ratio, and width/thickness ratio of seeds, as well as in length and width of leaves between Cephalotaxus alpina(Li) L.K.Fu and Cephalotaxus fortunei Hooker were shown in this study. The width range and mean of leaf of C.fortunei Hooker are 3.50-6.20 cm and 5.02 cm, respectively. In contrast, C.alpina(Li) L.K.Fu has only 2.00-3.50 cm and 2.89 cm. The range of seed width/thickness ratio and mean of C.fortunei Hooker are 0.53 and 1.43, respectively, while C.alpina(Li) L.K.Fu is 0.23 and 1.02, respectively. Scatter diagrams of C.alpina(Li) L.K.Fu and C.fortunei Hooker illustrated that the variations in leaves length, seeds length, seeds width, seeds thickness, seeds length/width ratio and seeds length/thickness ratio between these two taxa can not differ C.alpina(Li) L.K.Fu from C.fortunei Hooker, but the variation in leaves width and seeds width/thickness ratio can differ C.alpina(Li) L.K.Fu from C.fortunei Hooker notably. Based on these significant differences, we supported the view that Fu (1984) treated C.fortunei Hooker var. alpina H.L.Li as C.alpina(Li) L.K.Fu, and disagreed with the argument that Silba(2007) thought C.fortunei Hooker var. alpina Li as C.fortunei Hooker subsp. alpina(Li) Silba. On the other hand, there were evidences supporting that C.fortunei Hooker var. globosa S.Y.Hu(1964) should be the same taxon with C.alpina(Li) L.K.Fu. Finally, a new plates 1-6 was provided.
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    Micromorphological Features of Epidermis of 20 Species Ferns from Yunnan
    XU Cheng-Dong;CHEN Yan-Ping;FENG Jian-Meng
    Bulletin of Botanical Research    2012, 32 (1): 10-16.   DOI: 10.7525/j.issn.1673-5102.2012.01.003
    Abstract1668)      PDF(pc) (1255KB)(600)       Save
    Epidermal micromorphology of 20 species ferns leaves from different families and genera in Yunnan were observed by optical microscope. The results showed that: different appendages exist in different leaves surface, some species have no appendage. There are eight stomatal apparatus types: polocytic, axillocytic, aotinocytic, diaisocytic, anomotetracytic, copolocytic, coaxillocytic and anomocytic types. The stomatal apparatus have multi-type phenomena, all the stomata are hypostomatic types, while their distributive modes and sizes also varies. The leaf epidermal cell micromorphology from different genera and families show certain differences, basically have irregular type, multilateral type or regular strip type. The leaf epidermal cell anticlinal walls are mainly corrugated, deep wavy or wavy. The results in this study provide basic references for systematics and evolution of ferns.
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    Morphological Characteristics of Fruits of Koenigia(Polygonaceae) from China
    CHEN Yan-Song;ZHOU Zhong-Ze*
    Bulletin of Botanical Research    2012, 32 (1): 17-21.   DOI: 10.7525/j.issn.1673-5102.2012.01.004
    Abstract1393)      PDF(pc) (1019KB)(499)       Save
    Macro- and micro-morphological characteristics of fruits in eight species of the genus Koenigia L. (Polygonaceae) from China were observed under stereoscope and scanning electron microscope (SEM). Based on the results, the fruits of the studied species are divided into type Ⅰ, Ⅱ and Ⅲ. In type Ⅰ: Achene is ovoid to lati-ovoid, intumescently in the middle to up middle, as a double burning glass. Its surface is smooth or rough, favulariately, or undulately. Four species, K.islandica, K.nummularifolium, K.forrestii and K.fertilis are in this type. In type Ⅱ: Achene is ovoid. Its intumescent in the middle, also have three ridged arrises but not notable. One species, K.delicatula is in this type. In type Ⅲ: Achene is ovoid. Maximum width occurred in middle but non-intumescent. Its have three ridged arrises. Their surface are smooth, rugosely, concave-convexly, or verrucately. This type contained three species, K.nepalensis, K.cyanandra and K.pilosa. According to the morphological characteristics of fruits, K.cyanandra and K.pilosa exist independently; K.islandica might be closely related to K.fertilis.
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    Developmental Anatomy of the Floral Nectaries in Dracocephalum nutans Linn.
    GENG Hua-Mei;FU Qiang;GUO Jun;WANG Hong*
    Bulletin of Botanical Research    2012, 32 (1): 22-27.   DOI: 10.7525/j.issn.1673-5102.2012.01.005
    Abstract1168)      PDF(pc) (1535KB)(424)       Save
    The nectaries in Dracocephalum nutans Linn. were located on receptacles under the ovary. The disk nectar upper split into four pieces a list, base in swollen reflexed round the lap cup-shaped receptacle. The nectary composed of secretory epidermis, nectariferous tissue and vascular bundles, which was typical structural nectary. Histochemistry stain indicated that the starch grain exhibited a regular change, hence the nectary belonged to starch nectary. During the development of nectaries tissue, the vacuole occured regular change and the starch grains and protein decreased and increased regularly. The nectar was secreted out through stomata and cuticle.
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    Effects of Color Plastic Films on Shoot Regeneration of Stems of Phellodendron amurense and on Some Physiological and Biochemical Indexes During Shoot Regeneration
    WANG Hui-Mei;CHEN Ya-Jie;YU Hai-Di;ZU Yuan-Gang
    Bulletin of Botanical Research    2012, 32 (1): 28-31.   DOI: 10.7525/j.issn.1673-5102.2012.01.006
    Abstract1177)      PDF(pc) (817KB)(430)       Save
    The stems of sterile plantlets of Phellodendron amurense were cultured on MS medium supplemented with 1.5 mg·L -1 6-BA and 0.5 mg·L -1 NAA, the effects of different film filters on shoot regeneration from stem explants of P.amurense and some physiological and biochemical indexes changes during shoot regeneration were investigated. The results showed that there was obvious promotion of blue films to shoot regeneration and the highest percentage of shoot regeneration(75.4%) with the maximum number of shoots per callus(14.7) were observed under blue films, followed by fluorescent light and yellow films. However, red and green films were ineffective for shoot regeneration. In this study, it was found that blue films and fluorescent light induced lower chlorophyll a/b ratio and higher level of chlorophyll content than other treatment. Antioxidant enzymes activity and soluble protein were the highest under blue films, followed by fluorescent light, the lowest under green ones.
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    Analysis and Evaluation on Genetic Diversity of Scutellaria baicalensis G. by ISSR Markers
    WEN Miao-Miao;LI Gui-Shuang;ZHANG Long-Jin;ZHENG Peng;BAI Cheng-Ke;*
    Bulletin of Botanical Research    2012, 32 (1): 32-37.   DOI: 10.7525/j.issn.1673-5102.2012.01.007
    Abstract1508)      PDF(pc) (1280KB)(525)       Save
    To study the genetic diversity of Scutellaria baicalensis G., 147 individuals from six populations were analyzed by inter-simple sequence repeat (ISSR). Eighteen primers were selected to produce highly reproducible ISSR bands. Among 485 amplified bands, 466 showed polymorphism, the percentage of polymorphic bands reached 96.08%. Nei’s gene diversity index and Shannon information index were 0.244 4 and 0.388 9, respectively. Number of alleles ( Na) and effective number of alleles ( Ne) were 1.993 8 and 1.383 9, respectively. G st was 0.123 3, genetic identity ( I) and genetic distance ( D) were 0.951 5 and 0.050 1, respectively. The results showed that the collected germplasm resources of S.baicalensis had a high genetic diversity in general. There was a certain genetic differentiation and gene flow among different populations and the genetic variation was mainly within populations. Molecular clustering results showed that the same region germplasms were not clustered completely following the collection source. It may be related to folk culture introduction or less than normal for ISSR primers. The genetic diversity of S.baicalensis germplasm resources can provide a basis for the evaluation, preservation and breeding of new varieties in the future.
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    Cloning and Bioinformation Analysis of C3H Gene in Neosinocalamus affinis
    ZHOU Mei-Juan;HU Shang-Lian*;CAO Ying;LU Xue-Qin;REN Peng;WU Xiao-Yu;LI Xiao-Rui
    Bulletin of Botanical Research    2012, 32 (1): 38-46.   DOI: 10.7525/j.issn.1673-5102.2012.01.008
    Abstract1227)      PDF(pc) (1810KB)(494)       Save
    p-Coumarate 3-hydroxylase plays an important role in the biosynthesis pathways of plant lignin. The full-length sequence of C3H in Neosinocalamus affinis( NaC3H) was successfully cloned by RT-PCR, and its bioinformation analysis was carried out. It aims to provide theoretical basis for decreasing the content of lignin in bamboo for making-pulp through engineering technique. The sequence is 1 581 bp in length. The bioinformation analysis showed that the CDS region of the nucleotide sequence was 1 539 bp, encoding 512 predicted amino acids. The protein molecular weight was 58.33 kD and theoretical pI was 9.09. According to the amino acid sequence and structural analysis, it showed that the protein encoding by C3H contained one conserved domain, viz. P450 domain. Phylogenetic analysis showed that NaC3H was most similar to C3H from P.edulis and Oryza sativa. The proteins encoded by C3H genes were hydrophilic ones. Subcellular localizations of these three proteins were in the endoplasmic reticulum (membrane). The secondary structures and tertiary structures were abundant in alpha helixs and random coils, beta turns and extended strains were less. Two relative conserved disordered domains were found in amino acid sequences encoded by C3H genes from N.affinis, P.edulis and O.sativa. The full-length sequence of NaC3H has been submitted to GenBank with the accession number JF693629.
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    Cloning and Analysis of the cat Associated Genes of Hevea brasiliensis and Banana in Hainan
    YU Nai-Tong;SUN Yu-Juan;ZHANG Yu-Liang;LUO Zhi-Wen;LIU Zhi-Xin*
    Bulletin of Botanical Research    2012, 32 (1): 47-53.   DOI: 10.7525/j.issn.1673-5102.2012.01.009
    Abstract1602)      PDF(pc) (1265KB)(523)       Save
    According to the existence of plant cat genes in GenBank, we designed a pair of degenerate primers, and amplified cat genes of banana, Hevea brasiliensis, yellow lantern pepper, pineapple, sugar cane, papaya from Hainan island. Ultimately, we cloned the rubber cat-1 and banana cat-2 genes, while the others not. Sequence analysis showed that open reading frame (ORF) of rubber cat-1 and banana cat-2 genes were both 1 479 bp, and encoded 492 amino acids. GenBank accession number were HQ660587 and HQ660588. Using bioinformatics software, the tertiary structures of rubber’s CAT-1 and banana’s CAT-2 proteins were predicated, which were similar to Exiguobacteriu Oxidotolerans (PDB code:2j2mA0) and Pseudomonas Syringae (PDB code:1m7sA), respectively. Phylogenetic analysis indicated that Hainan H.brasiliensis CAT-1 possessed the highest homology with Ricinus communis CAT-1 (XP_002521709.1), while banana CAT-2 was the highest homology with Elaeis guineensis CAT-2 (ACF06566.1).
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    Relationship Between Contents of Mineral Nutrient Elements and Peroxidase Isoenzyme in Leaf of Wild Cerasus humilis
    MA Jian-Jun;YU Feng-Ming;DU Bin;ZHANG Li-Bin;REN Yan-Jun
    Bulletin of Botanical Research    2012, 32 (1): 54-60.   DOI: 10.7525/j.issn.1673-5102.2012.01.010
    Abstract1320)      PDF(pc) (1085KB)(463)       Save
    In this study, the content variations of peroxidase isoenzyme in Chinese dwarf cherry ( Cerasus humilis) leaves and 6 mineral elements in leaves and fruits as well as their correlations were investigated by the method of polyacrylamide gel electrophoresis (PAGE) using 113 Chinese dwarf cherry which were cultivated at the same conditions in Yanshan Mountain as materials. The results showed that 7 types of POD isoenzyme bands appeared in Chinese dwarf cherry seedling population. Among these bands, two characteristic bands (Rf=0.126, Rf=0.716) appeared in all tested materials. The results of system clustering analysis showed that the relative amounts of the same position isoenzyme bands in different POD isoenzyme band types were different, and the variation was greater. The results of correlation analysis showed that notable or extremely notable correlation existed between the amount of correlative isoenzyme bands in different types and the mineral element contents. The amount of POD isoenzyme band could indicate the genetic difference and diversity of mineral nutrient metabolism of Chinese dwarf cherry leaves in some degree.
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    Cloning and Expression Analysis of HMGR Gene from Taraxacum kok-saghyz
    WANG Qi-Chao;LIU Shi-Zhong;XIAO Xian-Zhou;*
    Bulletin of Botanical Research    2012, 32 (1): 61-68.   DOI: 10.7525/j.issn.1673-5102.2012.01.011
    Abstract1536)      PDF(pc) (1547KB)(632)       Save
    Based on the design of degenerated oligonucleotides according to the conservative regions of nine 3-hydroxy-3-methylglutaryl-CoA reductases from nine plants and the total RNA extracted from Taraxacum kok- saghyz, a HMGR named TKHMGR was first obtained using the techniques of degenerate RT-PCR and RACE. Comparison of the amino acid sequence homology and system evolution analysis show that, TKHMGR belongs to the new members of the HMGR gene families. At the same time, the gene expression in different organizations was analyzed using fluorescence quantitative method. The cloning, bioinformatics analysis and expression analysis of this gene laid a foundation for further research on its function.
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    Cloning and Bioinformatics Analysis of Full-length cDNA of Actin Gene( CsActin1) from Tea Plant ( Camellia sinensis(L.) O. Kuntze)
    YANG Ya-Jun;WANG Xin-Chao;MA Chun-Lei
    Bulletin of Botanical Research    2012, 32 (1): 69-76.   DOI: 10.7525/j.issn.1673-5102.2012.01.012
    Abstract1095)      PDF(pc) (1654KB)(686)       Save
    Two suppression subtractive hybridization (SSH) libraries of tea plant( Camellia sinensis(L.) O. Kuntze) were constructed using cDNA from the dormant buds and sprouting buds. A cDNA fragment, which was homologous to the 5′-end of actin gene family, was identified from the sprouting bud SSH library. Using one primer designed on the basis of the fragment, its fulllength cDNA sequence was cloned through 3′-rapid amplification of cDNA ends (3′-RACE). The full length of the actin gene, named CsActin1, was 1 470 bp (GenBank accession No. HQ235647) and contained a 1134 bp open reading frame (ORF) encoding a 377 amino acid residues, a 5′-UTR of 100 bp and a 3′-UTR of 236 bp. The deduced protein molecular weight was 41.70 kD and its theoretical isoelectric point was 5.31. It contained the characteristic actin family signature sequence (YVGDEAQs.KRG and WISKgEYDE) and actin-related proteins signature (LLTEApLNPkaNR). Homologous alignment showed that it shared over 80% nucleotide sequence similarity and over 95% amino acid sequences similarity with actins in other plants. The phylogenetic tree reconstructed on the basis of amino acid sequences suggested that the relationship between C.sinensis and Populus trichocarpa is the most intimate.
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    Phylogeny of Polygonum Sect. Cephalophilon(Polygonaceae) Inferred from the Chloroplast trnL-F, rbcL and Nuclear Ribosomal ITS Sequences
    ZHAO Da-Peng;WANG Kang-Man;HOU Yuan-Tong*
    Bulletin of Botanical Research    2012, 32 (1): 77-83.   DOI: 10.7525/j.issn.1673-5102.2012.01.013
    Abstract1656)      PDF(pc) (1171KB)(484)       Save
    The phylogeny of Polygonum sect. Cephalophilon was analyzed using the chloroplast genes trnL-F, rbcL and nuclear ribosomal ITS sequences. Individual and combined analyses of three datasets were performed with maximum parsimony method and the gene trees were constructed with Rheum officinale as outgroup. The results show that Polygonum sect. Cephalophilon is a more natural group which is the most closely related to Polygonum sect. Persicaria, sect. Echinocaulon and the genus Antenoron. The results support to put Polygonum sect. Cephalophilon into the genus Persicaria as a section and transfer P.filicaule and P.cyanandrum from Polygonum sect. Cephalophilon to the genus Koenigia, do not support to attribute Polygonum sect. Echinocaulon to Polygonum sect. Cephalophilon. There is strong conflict regarding the systematic position of Polygonum delicatulum, which needs further studies.
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    Contamination Control during Tissue Culture of Peony
    WANG Ji-Feng;LI Qing;BAO Xin
    Bulletin of Botanical Research    2012, 32 (1): 84-90.   DOI: 10.7525/j.issn.1673-5102.2012.01.014
    Abstract1304)      PDF(pc) (1457KB)(559)       Save
    To solve the problem of explant contamination during peony tissue culture, the explants of different seasons were studied. The results showed that soaking explants in 0.1%-0.2% HgCl2 for 8 min, etiolation treatment, low temperature and electronic vibration are effective for sterilization; Antibiotics added into the media can inhibit contamination caused by endophytes.
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    Analysis of Spatial Pattern and Spatial Association of Pinus tabulaeformis Populations at Different Developmental Stages
    ZHANG Ling;MIAO Yan-Ming;SUN Yong-Xing;BI Run-Cheng*
    Bulletin of Botanical Research    2012, 32 (1): 91-98.   DOI: 10.7525/j.issn.1673-5102.2012.01.015
    Abstract1408)      PDF(pc) (1523KB)(714)       Save
    In Huoshan Qiliyu woodland, a field survey was made on the typical Pinus tabulaeformis, and the distribution of P.tabulaeformis at different age-classes and its co-relationship were studied. The results show that: (1)The distribution of the age-classes appears kurtosis, i.e. the majority is the middle-aged, 20-30 cm, whereas the numbers of young and old are comparatively less; (2)The spatial distribution of these seven age-classes tends to be in groups, in which ages 4, 5 and 6 are more obvious. Meanwhile, it centralizes in groups in small-scale and tend to be in random distribution in large-scale; (3)The co-relationship between different age-classes is significantly positive correlated, of which ages from 3 to 5 are more significant. This significantly positive correlation is beneficial for the P.tabulaeformis to make full use of the resources in order to guarantee the living and development of the groups.
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    Biomass Allocation Pattern and Water Characteristics of Each Component of Phoebe bournei Container Seedling
    TANG Xiao-Yan;YUAN Wei-Gao*;SHEN Ai-Hua;ZHANG Jun;ZHU Jin-Ru;JIANG Bo
    Bulletin of Botanical Research    2012, 32 (1): 99-104.   DOI: 10.7525/j.issn.1673-5102.2012.01.016
    Abstract1296)      PDF(pc) (1128KB)(602)       Save
    In this research, the moisture content and biomass distribution of 1.5-year old Phoebe bournei container seedling at different height stages were measured and analyzed. The results showed that: (1)With the increase of seedling height, the biomass and its components in P.bournei container seedling increased. The biomass allocated in the order of root biomass>leaf biomass>stem biomass>branch biomass. With the increase of seedling height, the biomass of stem increased, while the biomass of leaf first increased and then decreased, and the biomass of root first decreased and then increased. The biomass as well as its components are significant correlated to basal diameter (D) and height (H) except branch. (2)The moisture contents of stem and leaf reach peak values while the seedling height was between 20 and 25 cm, the moisture content of root reach peak value while it between 35 and 45 cm. (3)Root moisture content as well as leaf, stem and aboveground biomass are significant positive correlated, while a significant negative correlation between leaf moisture content and different organs biomass was identified.
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    Competition Relationship of Populations of Natural Secondary Acer mono Forest
    YIN Dong-Sheng;GE Wen-Zhi;ZHANG Feng-Hai;SHEN Hai-Long
    Bulletin of Botanical Research    2012, 32 (1): 105-109.   DOI: 10.7525/j.issn.1673-5102.2012.01.017
    Abstract1386)      PDF(pc) (1106KB)(565)       Save
    The intraspecific and interspecific competition intensities in the secondary Acer mono forest were quantitatively analyzed by Hegyi single-tree competition index model. The results showed that the intraspecific competition intensity in A.mono was bigger, accounting for 26 percents of total competition, and the competition intensity gradually increased with the increasing tree diameter. However, the competition intensity decreased after diameter at breast height DBH was greater than 30 cm. The interspecific competition (89.061) in A.mono was more intensive than the intraspecific competition (31.487). The competition intensity of interspecific competition with A.mono followed the order of A.mono>Tilia mandshurica>Quecus mongolica>Phellodendron amurense>Juglans mandshurica>Tilia amurensis. The competition intensity of A.mono declined with the increasing DBH, and the stress was more intense for trees less than 15 cm in DBH. The relationship between competition intensity and DBH of target trees followed the exponential function. The tending management should be actualized to increase the timber utilization quotiety when DBH of A.mono individual near 15 cm in natural conditions.
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    Characteristics of Fruit Development and Flower and Fruit Drops of Artificial Population of Xanthoceras sorbifolia
    CHAI Chun-Shan;LU Juan;CAI Guo-Jun;WANG Zi-Ting
    Bulletin of Botanical Research    2012, 32 (1): 110-114.   DOI: 10.7525/j.issn.1673-5102.2012.01.018
    Abstract1164)      PDF(pc) (1037KB)(602)       Save
    The characteristics of fruit development and flower and fruit drops of Xanthoceras sorbifolia in Dingxi district of Gansu were studied. The results showed that: (1) X.sorbifolia fruit need about 2.5 months from developing to ripening, and its main development and growth stage was in the 40 days after flowering. Its fruit development feature was as follows: fruit growth was the fastest in 10 days after blossom period and its growth increment rate reached 193%, in the next month the fruit still keep faster growth and had a growth increment rate of 120%, but it grew slowly in the last month and the increment rate was only 7%, and during the whole growth process, the width of fruit grew always faster than the length of fruit; (2) X.sorbifolia flower and fruit drops occurred in the early stage of its fruit developing and growing process, and it had a higher drop rate of flower and fruit of 94.65% (flower drop rate 5.74%, fruit drop rate 88.91%), but a very low fruit-bearing rate of 5.35%. Its feature of flower and fruit drops was as follows: the fruit drop was an ongoing process and there was always fruit drop from fruit expanding to ripening after blossom stage. However, there were some peak days in fruit dropping, namely in the 10 days after blossom stage the fruit drop was very serious and the fruit drop rate nearly reached 40%. According to this study, it was regarded that, for the consideration of preserving flowers and fruits, the late flowering stage was the key period for X.sorbifolia to enhance the water and fertilizer management. However, considering the facts that X.sorbifolia profuse flowers and the concentration of fertile flower could lead to heavy drops of flower and fruit because of its high consumption nutrients of tree and top inflorescence, it should thin flower and fruit or let it drop naturally in late flowering stage and young fruit stage, but strengthening water and fertilizer management is needed in the middle and late periods of fruit development to promote its fruit-bearing rate.
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    Establishment of SSR Reaction System and Fingerprinting for Some Aechmea Species
    ZHANG Fei;WANG Wei-Yong;ZHANG Zhi;TIAN Dan-Qing;LIU Jian-Xin;LIU Xiao-Jing
    Bulletin of Botanical Research    2012, 32 (1): 115-119.   DOI: 10.7525/j.issn.1673-5102.2012.01.019
    Abstract1263)      PDF(pc) (979KB)(576)       Save
    The SSR reaction system was established by orthogonal design and the SSR fingerprints were constructed for some ornamental bromeliads in Aechmea in this study. The results showed that the optimal SSR reaction system of Aechmea was a total of 10 μL containing 1×PCR buffer, Mg 2+ 2.0 mmol·L -1, dNTPs 200 μmol·L -1, primers 2.5 μmol·L -1, template DNA 90 ng and Taq DNA polymerase 1.5 U. Three informative and polymorphic SSR primers, namely M1, M3 and M4, were screened out from 6 SSR primers based on the established SSR reaction system. Three molecular fingerprints were preliminarily constructed for 15 samples of Aechmea species by these three SSR primers, respectively. The establishment of SSR fingerprint proved the SSR reaction system was stable and reliable, and could be further used in germplasm identification of Aechmea.
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    Effects of TDZ on Polyamines Concentration Changes during the Construction of Floral-buds in Derooted Seedlings of Dendrobium officinate Kimura et Migo in vitro
    HUANG Zuo-Xi;CHEN Yi;XIAO Xiao-Jun;LI Ting-Ting;PENG Wen-Lu;TANG Xu
    Bulletin of Botanical Research    2012, 32 (1): 120-123.   DOI: 10.7525/j.issn.1673-5102.2012.01.020
    Abstract1387)      PDF(pc) (830KB)(404)       Save
    The effects of tridiazuron (TDZ) on endogenous polyamines concentration changes in derooted seedlings of Dendrobium officinate Kimura et Migo in vitro and the construction of floral-buds were researched in this experiment, in which MS basal medium supplemented with TDZ was used. It was found that the formation rate of floral-bud in derooted seedlings cultured on MS mixed with 0.1 mg·L -1 TDZ accounted for 21.1%, whereas the control was 0. During the construction of floral-bud, the endogenous concentrations of Spd, Spm and the ratios of Spd to Put, Spm to Put in top shoots of derooted seedlings cultured on MS mixed with 0.1 mg·L -1 TDZ exceeded those in the control, and remarkably exceeded that of control when the floral-bud emerged. In addition, the levels of Put and DAP in top shoots of derooted seedlings were apparently lower than that in the control. It was seen that exogenous TDZ could promote the construction of floral-bud by means of the changes of endogenous polyamines concentration and its ratio in D.officinate Kimura et Migo.
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    Effects of Copper Stress on Seedlings Growth and Active Ingredients of Salvia miltiorrhiza
    WANG Chuan;LI Kun-Wei;WEI Yu-Kun;CUI Lang-Jun*;LI Fa-Rong
    Bulletin of Botanical Research    2012, 32 (1): 124-128.   DOI: 10.7525/j.issn.1673-5102.2012.01.021
    Abstract1497)      PDF(pc) (1061KB)(522)       Save
    The effects of copper stress on seedlings growth and active ingredients of Salvia miltiorrhiza were investigated. The results showed that copper stress inhibited seedlings growth, increased Cu 2+ accumulation in both soil and seedlings. Moreover, a reduction in photosynthetic pigments and an increase in the levels of TBARS content of stressed seedlings showed that oxidative stress and lipid peroxidation were provoked. The effects of the treatment on active ingredients accumulations in above-ground parts and roots were different. In above-ground parts of copper stressed seedlings, the accumulations of caffeic acid, tanshinol and protocatechuic acid increased, while the contents of rosmarinic acid, protocatechuic aldehyde and salvianolic acid B decreased. However, the accumulations of both six phenolic acids and four tanshinone ingredients all decreased in stressed roots. All these results showed that copper stress had a toxic effect on both S.miltiorrhiza growth and 10 mainly active ingredients accumulations.
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    Biosystematic Relationships among Sect.Elymus,Sect.Turczaninovia and Sect.Macrolepis in Elymus(Poaceae) Based on the Micromorphological Characteristics of Leaf Epidermis
    SU Xu;LIU Yu-Ping;CHEN Ke-Long
    Bulletin of Botanical Research    2012, 32 (2): 129-136.   DOI: 10.7525/j.issn.1673-5102.2012.02.001
    Abstract1503)      PDF(pc) (1484KB)(582)       Save
    The leaf epidermises of the major species of sect.Elymus, sect.Turczaninovia and sect.Macrolepis in Elymus were examined, their similarities and differences on the leaf epidermis were summarized, and the taxonomic significance of leaf epidermal features among them was discussed as well. Furthermore, according to the evolutionary trends of leaf epidermal characteristics of three sects, their evolutionary positions and relationships were inferred. The results show that the sect. Elymus is the most primitive one among these three sects, the sect.Turczaninovia is slightly more advanced than the former, and the sect.Macrolepis is the most advanced of them; the sect.Elymus might produce immediately the sect.Turczaninovia, whereas the sect.Macrolepis might derive immediately form the sect.Turczaninovia. The biosystmatic relationships of these three sects are corroborated by the evolutionary trend of external morphology.
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    Pollinarium Morphology of 12 Species of Cynanchum(Asclepiadaceae)
    LI Yu-Ling;QIN Xin-Sheng*;FENG Yi-Qing
    Bulletin of Botanical Research    2012, 32 (2): 137-142.   DOI: 10.7525/j.issn.1673-5102.2012.02.002
    Abstract1108)      PDF(pc) (1031KB)(521)       Save
    The pollinarium morphology of 12 species of Cynanchum Linn. were observed under light microscope (LM). Three types of pollinia (spheroidal, prolate and perprolate) and six types of retinaculum (oblong, square, near square, regular triangle and near triangle) were observed. This is important for the classification of Cynanchum otophyllum and supports the treatment of Cynanchum Sect. Vincetoxicum in Flora of China(Vol. 63). As the pollinarium in Cynanchum is saggy while that of Tylophora R. Br. is horizontal, it supports that Tylophora should be a unique group.
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    Leaf Anatomical Structure and Its Ecological Adaptability of Five Species of Rhizophoraceae Plants
    WU Tian;YE Chang-Hui;HAN Wei-Dong*
    Bulletin of Botanical Research    2012, 32 (2): 143-146.   DOI: 10.7525/j.issn.1673-5102.2012.02.003
    Abstract1469)      PDF(pc) (945KB)(743)       Save
    Using the paraffin-section method, the leaf anatomical structures of five species of Rhizophoraceae plants ( Rhizophora apiculata Bl., R.stylosa Griff., Kandelia candel(Linn.) Druce, Bruguiera gymnorrhiza(Linn.) Savigny and B.sexangula(Lour.) Poir) were observed in this study. The results showed that thick cuticles and lower epidermis were commonly existed in the leaves of all these five species of mangrove plants, while the epidermis cells contained tannin. This study also concluded that there was no stomata apparatus on the upper epidermis, and the layers of palisade tissues were also observed. Besides, the xylems in the leaves were identified to be well developed. All these results indicated that the leaves of mangrove plants exhibited strong drought-resistant capacity, and the leaves of homogeneous plants displayed similar characteristics in their anatomical structures.
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    Clonal Reproductive Property and Community Characteristic of Invasive Species Coreopsis lanceolata
    ZENG Jian-Jun;XIAO Yi-An;ZHOU Xiao-Jun
    Bulletin of Botanical Research    2012, 32 (2): 147-150.   DOI: 10.7525/j.issn.1673-5102.2012.02.004
    Abstract1392)      PDF(pc) (928KB)(619)       Save
    The clonal reproductive property and community characteristic of invasive species Coreopsis lanceolata in different habitats in Lu Shan, Jiangxi province were studied. The results showed that there was no significant difference in clonal architecture under habitats with different resources. Whether in rock or deserted farmland, the clonal architecture was mainly phalanx which developed from the base of plant, it seemed that there was no correlation between the guerrilla clonal architecture and the level of environment resources. Those guerrilla clonal growth traits made C.lanceolata had collectivize growth strategy, which improved its resistance to barren and drought environment. Thus, naturalized C.lanceolata could dominate various indigenous plants in Lushan, the companion species of C.lanceolata populations were weeds with wider adaptability.
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    Comparison Study on Soil Nutrient and Photosynthetic Characteristics of Ferula ferulaeoides at Different Growth Periods
    LI Xiao-Bo;ZHUANG Li*;WANG Zhong-Ke;XU Zhi-Quan
    Bulletin of Botanical Research    2012, 32 (2): 151-158.   DOI: 10.7525/j.issn.1673-5102.2012.02.005
    Abstract1119)      PDF(pc) (1635KB)(307)       Save
    In this paper, the dynamic changes of soil water content, pH, organic substance and soil nutrient, as well as the diurnal variations of net photosynthetic rate, transpiration rate and water use efficiency of Ferula ferulaeoides at different growth periods were studied. We found that the soil water content, organic substance and soil nutrient showed downward tendency with the growth of F.ferulaeoides while the pH was reverse, but changed slightly. In different soil depths, the soil water content and pH gradually increased with the soil depth, while organic substance and soil nutrient gradually reduced. The net photosynthetic rates at three growth periods varied in a double-peak curve and presented obvious phenomenon of “midday rest” at 14:00, with the daily mean values of leaf expansion period>maturation period>fruit period; The transpiration rates appeared to vary in a single-peak curve and had the highest value at 14:00, with the daily mean values of leaf expansion period>fruit period>maturation period; The water use efficiency showed a increased tendency at growth period and had the lowest value at 14:00, with the diurnal mean values of maturation period>fruit period>leaf expansion period. The correlation analysis revealed that stomatal conductance, photosynthetic active radiation had positive correlation with photosynthetic rate; Light intensity had evident positive correlation with transpiration rate.
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    Effect of Strong Light Stress on the Growth,Biomass and Chlorophyll Fluorescence Parameters in Seedlings of Endangered Plant Camellia nitidissima
    CHAI Sheng-Feng;WEI Xiao*;SHI Yan-Cai;WANG Man-Lian;ZOU Rong;TANG Hui
    Bulletin of Botanical Research    2012, 32 (2): 159-164.   DOI: 10.7525/j.issn.1673-5102.2012.02.006
    Abstract1366)      PDF(pc) (1322KB)(606)       Save
    Camellia nitidissima is shadetolerant and has a limited photosynthetic potential of acclimating high light. The effects of different shading levels (8%, 25%, 50% and 100% of full sunlight) on the growth, biomass, chlorophyll fluorescence parameters of C.nitidissima seedlings were examined in order to know the growth and physiological responses of C.nitidissima to strong light. The growth of C.nitidissima seedlings were inhibited under different strong light stress. As the strong light stress was enhanced, the color of C.nitidissima seedlings leaves turned into light green and yellow green from dark green, leaf burning was getting worse, time of shooting was delayed and the shoots grew poor, the death rate increased. The biomass of root, stem, leaf and total plant significantly decreased with the strengthened stress. The strong light stress had the most influence on leaf biomass, followed by root biomass, and then the stem biomass. With the increasing strong light stress, the contents of total chlorophyll (Chl), chlorophyll a (Chla) and chlorophyll b (Chlb) in leaves of C.nitidissima seedlings significantly decreased, while Chla/Chlb and Car/Chl increased. The chlorophyll fluorescence parameters F o, F m, F v, F v/ F o, F v/ F m all decreased under strong light stress, which indicated that with the increasing of strong light stress, the reaction center of PSⅡ was destroyed, photosynthetic electron transform capability was weakened and photosynthesis was restrained.
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    Comparison of Photosynthesis Characteristics in Five Provenances of Chinese Pine( Pinus tabulaeformis Carr.)
    DI Xiao-Yan;CHI Xi-Mei;CHEN Jian-Wen;WANG Meng-Ben*
    Bulletin of Botanical Research    2012, 32 (2): 165-170.   DOI: 10.7525/j.issn.1673-5102.2012.02.007
    Abstract1235)      PDF(pc) (1179KB)(567)       Save
    In order to reveal the photosynthetic characteristic variations of different provenances and provide the theoretical basis for the work of provenance selection, the seedlings of Chinese pine ( Pinus tabulaeformis Carr.) from five provenances, including provenances Luonan (LN) and Huangling (HL) from Shaanxi, Lvliang Mountain (LL) and Lingkong Mountain (LK) from Shanxi and Qian Mountain (QM) from Liaoning were planted in Loess Plateau. The photosynthetic parameters and their correlations to environmental factors were studied with Li-6400 portable photosynthesis system under natural condition in order to reveal the mechanism of adaptation of different provenances. The results indicted that the diurnal dynamics of net photosynthetic rate ( P n), transpiration rate ( T r) and stomatal conductance( G s) for five provenances were double-peak curves with obviously midday depression in P n. The water use efficiency ( WUE) of porvenance QM had only one peak, while those of provenances LN, LL, LK and HL had two peaks. The first peaks for all provenances occurred at 8:00 or 10:00, and the second peaks of provenances LN, LL, LK and HL occurred between 12:00 and 16:00. Based on the tendency of changes in P n, G s, intercellular CO 2 concentration ( C i) and stomatal limitation ( L s), we speculate that the midday depressions of provenance HL was not controlled by stomatal factors, while those of the other four provenances were. The daily mean values of P n, T r and WUE for provenance LK was the highest and those for provenance HL was lower than the other provenances. The correlation analyses showed that the main factors affecting P n were air temperature ( T a) and vapour pressure deficit (VPD). The main factor affecting T r was relative humidity (RH) for provenances LN, LL and LK, while for provenances HL and QM, the main factors were T a and VPD.
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    Effects of Drought Stress on Ultrastructure and Stress Physiology of Nostoc flagelliforme
    ZHOU You-Wen;ZHANG Ya-Ping;YANG Jun;XU Qing;LEI Xiao-Ting;LIANG Wen-Yu*
    Bulletin of Botanical Research    2012, 32 (2): 171-176.   DOI: 10.7525/j.issn.1673-5102.2012.02.008
    Abstract1274)      PDF(pc) (1311KB)(454)       Save
    Nostoc flagelliforme is a terrestrial nitrogen-fixing cyanobacteria, which has strong xeric ecological adaptability. The ultrastructure and physiological resistance of N.flagelliforme under drought stress were studied. The results showed that with increasing drought stress, the cell size and wall thickness were not significantly changed, the sheath tended to be shrunk, the arrangement of thylakoids tended to be disordered, and the polyhedron tended to be blurred and even disappeared, the amount of glycogen granules were decreased, but the amount of structural granules did not change significantly. SOD and CAT activities were firstly increased and then decreased, SOD activity reached the peak when the content of water reached 120%, but CAT activity reached the peak when the content of water reached 445%. With the increasing of drought stress, MDA content and oxygen free radical level were increased, H 2O 2 content firstly increased and then declined, it reached the peak when water content reach 120%. There were obvious differences between N.flagelliforme which dried storage for 1 year and resuscitative N.flagelliforme, the SOD and CAT activities, MDA and H 2O 2 contents were all lower than those of resuscitative N.flagelliforme, but oxygen free radical level reached the highest value. The results laid the foundation for the further studies on tolerance mechanism in N.flagelliforme.
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    Effects of Light and Nitrogen Supply on the Carbon Balance of Seedling of Platycarya strobilacea
    WANG Chuan-Hua;WANG Yuan;LI Jun-Qing*
    Bulletin of Botanical Research    2012, 32 (2): 177-182.   DOI: 10.7525/j.issn.1673-5102.2012.02.009
    Abstract1456)      PDF(pc) (1121KB)(368)       Save
    Platycarya strobilacea is widely distributed in the sub-tropical area of China. To study the potential effect of atmosphere nitrogen deposition on its seedling regeneration, the seedling leaf functional traits, biomass allocation, gas exchange and seedling carbon balance to low light (3.0%, 6.0%, 12.0% and 25.0% PAR of open sky light) and nitrogen (15.0 and 3.0 mmol·L -1) treatments were determined in a greenhouse experiment. The results showed that light deeply affected leaf functional traits and biomass allocation. In contrast, nitrogen supply had no influence on them, though the interactions of low light and nitrogen supply could significantly affect RMR. We also found that the maximum photosynthesis rate ( A max) and apparent photon efficiency ( Ф) decreased with increasing nitrogen supply under low light (3.0% and 6.0% of full sun light), vice versa in a relatively higher light (12.0% and 25.0% of full sun light). Hence, the interactions of light and nitrogen deeply influenced leaves gas exchange of seedlings of P.strobilacea. The results simulated with a whole plant carbon balance model indicated that light compensation point of P.strobilacea seedlings ( LCP -wholeplant) increased with increasing nitrogen under 3.0%, 6.0% PAR of open sky light, while it decreased with increasing nitrogen supply under 12.0% and 25.0% conditions. Therefore, we suggested that atmosphere nitrogen deposition would deteriorate P.strobilacea regeneration under lower light environment conditions, but increase its survival under disturbance and gap conditions.
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    Effects of Ethephon on the Expression of Genes Related to Anthocyanin Biosynthesis in Grapevine
    YU Miao;ZHAO Quan;WANG Jun;*
    Bulletin of Botanical Research    2012, 32 (2): 183-190.   DOI: 10.7525/j.issn.1673-5102.2012.02.010
    Abstract1185)      PDF(pc) (1291KB)(444)       Save
    The present study investigated the effects of different concentrations of ethephon on the changes of anthocyanin contents and the transcription of anthocyanin biosynthesis enzyme genes during the fruit coloring period in Jingyou grapevine by means of real-time PCR. The results demonstrated that the anthocyanin biosynthesis pathway genes, including CHSs, CHIs, F3Hs, F3H, F35H, DFR, LDOX, UFGT, OMT, GST and transcriptional factors ( MybA1, MybA1- 2) were up-regulated expressed at veraison. The transcription of CHS3, CHI2 and F3H2 in gene families of CHSs, CHIs and F3Hs is closely related to the anthocyanin synthesis. Besides, UFGT, GST and MybA1 were transcribed in great quantity with anthocyanin synthesizing. The transcription of the anthocyanin biosynthesis pathway genes was enhanced by ethephon treatments, which accelerated transcribing earlier and enhanced the quantity of transcription, especially GST, UFGT and MybA1. Correlation analysis of the principal related genes and anthocyanins demonstrated that the genetic transcriptional level of some anthocyanin biosynthesis genes ( CHS3, CHI2, F3H2, F35H, UFGT and GST) and transcription factor ( MybA1) were positively correlated to the content of anthocyanin. The genetic transcriptional level of CHS1, CHS2, CHI1, F3H1, DFR, F3H, LDOX and OMT was not correlated to the content of anthocyanins. The results of this experiment will provide a theoretical basis for further clarifying the mechanism of anthocyanin biosynthesis and application.
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    Tree-ring Climatology of Larix gmelinii in Maoershan Region,Northeastern China
    ZHENG Guang-Yu;WANG Wen-Jie*;WANG Xiao-Chun;YU Jing-Hua;LIU Dan;QIU Ling;ZU Yuan-Gang
    Bulletin of Botanical Research    2012, 32 (2): 191-197.   DOI: 10.7525/j.issn.1673-5102.2012.02.011
    Abstract1176)      PDF(pc) (1198KB)(483)       Save
    Through collecting the meteorological data, sampling and standardizing the data of tree rings of larch ( Larix gmelinii), the climatic changes in Maoershan region were analyzed and their influences on tree ring features were studied in this paper. In the past 50 years, significant increases in temperature were found ( p<0.05), i.e. about 0.4℃ per decade in average annual temperature, 0.3℃ per decade in average annual maximum temperature as well as 0.5℃ per decade in annual average minimum temperature were observed. However, no significant change in annual precipitation was found ( p>0.05). Average monthly temperature (average temperature, average maximum temperature, average minimum temperature) also showed increasing tendencies and the most significant month was in February of the winter, about 0.9-1℃ per decade. Much smaller temperature increases, ca. 0.2-0.7℃ per decade was found in June to August of the summer). No significant change in monthly precipitation was found in most months ( p>0.05). During the coming climate warming, widths of early and whole tree rings ( p<0.05) might be decreased with the temperature increment in summer (June to July), while widths of early and the growth of tree rings would increase with the temperature increases in spring (May). The width of late might be increased with the temperature increases in autumn (September). These might cause the non-obvious change in ring width with annual temperature changes ( p>0.05). Although non-significant changes in precipitations, the significant correlations between annual precipitation and variable tree rings indices indicated the growths of early wood and whole tree rings might be positivity affected by the amount of annual precipitations. Considering the warming trend of the climate change is significant in northeast China in future but the rainfall does not. The growth of tree rings caused by spring and autumn temperatures could be offseted by too high temperatures in summer which inhibited the tree ring growth. Therefore, the influence on the radial growth of the Larix might be little.
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    Molecular Cloning and Function Analysis of HMGR Gene from Tilia miqueliana Maxim
    ZHENG Zhu-Jun;CAO Xiao-Ying;LIU Qun;LI Chang-Gen;LU Fang;JIANG Ji-Hong*
    Bulletin of Botanical Research    2012, 32 (2): 198-203.   DOI: 10.7525/j.issn.1673-5102.2012.02.012
    Abstract1346)      PDF(pc) (1327KB)(509)       Save
    The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) is the key enzyme in the pathway for isoprenoid biosynthesis in plants. In this paper, a new HMGR gene, TmiHMGR, was first isolated from the leaves of Tilia miqueliana Maxim by rapid amplification of cDNA end technique. The full length cDNA of TmiHMGR was 2 160 bp, with a 1 758 bp open reading frame encoding a peptide of 585 amino acid residues. The deduced protein TmiHMGR has a calculated molecular weight of 62.9 kD and a pI value of 6.11. Phylogenetic tree analysis indicated that TmiHMGR had the closest relationship with HMGR from Malusx domestica. Tissue expression analysis revealed that TmiHMGR was strongly expressed in stems, while weakly in roots and leaves. The functional color complementation assay indicated that TmiHMGR could accelerate the biosynthesis of carotenoids in the Escherichia coli transformation, suggesting that TmiHMGR plays an influential role in isoprenoid biosynthesis.
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    Antibody Preparation and Protein Expression of NdhK in Nostoc flagelliforme
    GUO Zheng-Hong;WEI Lan-Zhen;WANG Quan-Xi*
    Bulletin of Botanical Research    2012, 32 (2): 204-207.   DOI: 10.7525/j.issn.1673-5102.2012.02.013
    Abstract4496)      PDF(pc) (814KB)(500)       Save
    Nostoc flagelliforme, a terrestrial cyanobacterium, is distributed in arid and semi-arid steppes of the west and northwest of China. Cyanobacterial NADPH dehydrogenase (NDH-1) is an important photosynthetic membrane protein complex, and is essential to CO 2 uptake, cyclic electron transport around photosystem Ⅰ and cellular respiration. However, little is know about the function roles of NdhK subunit in cyanobacteria. In this study, the ndhK gene was PCR amplified from the N.flagelliforme, and the expression plasmid pET32a- ndhK was generated and transformed into BL21(DE3)pLysS, and the expression of NdhK protein was induced by IPTG. After purification, the fusion protein pET-NdhK was used to immunize Japanese white rabbit to obtain the polyclonal antibody. The titer of the polyclonal antibody was detected and analyzed by Western blotting, the result showed that the polyclonal antibody possessed a high specificity. Therefore, the antibody of NdhK obtained in this study will further help us to reveal the functional roles of cyanobacterial NdhK subunit.
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    Genetic Diversity and Relationship Among 7 Species of Genus Epimedium in Sichuan Revealed by ISSR Analysis
    CHEN Yan-Ying;LI Yun-Xiang*;QUAN Qiu-Mei;LIAO Xing-Li;CHENG Xian-Ge
    Bulletin of Botanical Research    2012, 32 (2): 208-212.   DOI: 10.7525/j.issn.1673-5102.2012.02.014
    Abstract1312)      PDF(pc) (1007KB)(528)       Save
    In order to investigate the genetic diversity and relationship of genus Epimedium in Sichuan province, 7 species including 12 populations were used to study the genetic variation by the ISSR marker technique. The result showed that 194 DNA fragments among 12 populations were amplified using 23 reliable ISSR primers, among which 169 DNA bands were polymorphic (the percentage of polymorphic bands reached 87.11%). The genetic distance and similarity among 12 populations were 0.298 0-0.620 7 and 0.653 2-0.748 9, indicating variation of genetic diversity existed among different populations. The observed effective number of alleles ( Ne), Nei’s gene diversity index ( He) and Shannon’s information index ( I) were 1.534 2, 0.314 4 and 0.469 7, respectively, showing the geographical distribution was very obvious. The result of cluster analysis using the unweighted pairgroup method with arithmetic average (UPGMA) and Principal Component analysis (PCA) based on ISSR data both showed that these tested resources were divided into two main groups. In addition, the ISSR marker technique is suitable for the analysis of the genetic diversity and relationship of genus Epimedium in Sichuan.
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    Optimization of SSR Reaction System in Juglans nigra
    ZHAO Peng;;WOESTE E. Keith;CHENG Fei;ZHANG Shuo-Xin;*
    Bulletin of Botanical Research    2012, 32 (2): 213-221.   DOI: 10.7525/j.issn.1673-5102.2012.02.015
    Abstract1215)      PDF(pc) (1586KB)(548)       Save
    The optimization of the SSR-PCR (simple sequence repeat-polymerase chain reaction) reaction system is an important basic protocol when SSRs are used for pedigree construction, genotyping or population genetics research in black walnut ( Juglans nigra L.). We systematically tested the concentrations of Taq DNA polymerase, BSA, dNTPs, Mg 2+, primers, and template DNA concentration in the PCR reaction to determine the optimal reaction system. The results indicated that the optimal SSR-PCR reaction conditions for black walnut ( J.nigra L.) included a total volume of 10 μL containing 1 μL of 10 ng·μL -1 DNA, 1 μL of 10 X Taq DNA polymerase reaction buffer (1.5 mmol·L -1 Mg 2+), 1.25 μL of 200 mmol·L -1 dNTPs (0.3 mmol·L -1), 1 μL of 1 mg·mL -1 BSA (Bovine Serum Albumin, 0.1 mg·mL -1), 1.0 μL of 10 μmol·L -1 each primer (when using one pair primer in each PCR reaction) or 0.5 μL of 10 μmol·L -1 each primer (when using three pairs primer in each PCR reaction), 0.5 units Taq polymerase, and 4.5 μL sterilized distilled water. The thermal cycling conditions were as follows: denaturation 3 min at 94℃; 32 cycles of 15 s at 93℃, 1 min at the annealing temperature for the primer, 30 s at 72℃; and a final extension of 10 min at 72℃ at the end of the amplification. The results showed that this SSR-PCR protocol resulted in clear, reproducible results suitable for the analysis of population genetics, genotyping, and for molecular ecology.
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    Construction of SSH cDNA Library and the Corresponding EST Analysis in Petals and Labellum of Cymbidium lianpan
    WU Yun-Yun;TIAN Lu;ZHU Yong-Ping;HE Feng-Mei*
    Bulletin of Botanical Research    2012, 32 (2): 222-226.   DOI: 10.7525/j.issn.1673-5102.2012.02.016
    Abstract1413)      PDF(pc) (1051KB)(315)       Save
    In order to explore the molecular mechanism of orchid labellum formation, the suppression subtractive hybridization (SSH) cDNA library was constructed with petals and labellum of Cymbidium lianpan in this study. One hundred and forty-seven randomly selected clones of positive library were sequenced, assembled and spliced, 37 unique sequences were found, containing 25 single sequences and 12 spliced sequences. After homology comparison, these ESTs were classified as protein folding, binding, unknown function, stress response, photosynthesis, transcription regulation, signal transduction, biosynthesis, metabolism, MADS and other 10 categories. Through the analysis of phylogenetic tree about MADS-box genes, we can initially speculate that MADS-Contig1 and MADS-Contig2 belong to A class SQUA-like subfamily genes.
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    Analysis on the Factors Influencing Transformation of Cassava ( Manihot esculenta Crantz) Mediated by Agrobacterium tumefaciens
    YAO Qing-Rong;GUO Yun-Ling;KONG Hua;HE Li-Ka;GUO An-Ping*
    Bulletin of Botanical Research    2012, 32 (2): 227-231.   DOI: 10.7525/j.issn.1673-5102.2012.02.017
    Abstract990)      PDF(pc) (1139KB)(394)       Save
    Agrobacteriurn tumefaciens-mediated transformation system was used to study the factors influencing transformation in cassava. Histochemical GUS assay showed that the transient expression rate was significantly improved by using the following transformation conditions: GV3101 as A.tumefaciens strains, infection time 30-45 min, co-culture time 3-4 d, engineering bacterium concentration (OD 600) 0.45 and application of 200 μmol·L -1 AS. However, pre-culture showed negative influence on the transient expression rate. Eight transgenic cassava plants from 453 explants were obtained in this study and the integration of glgC336 transgene was confirmed by PCR and Southern blot analysis. The transformation frequency was 1.77%.
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    Chemical Compound Characteristic Analysis of Wood forming Tissue from Reaction Wood of Pinus koraiensis
    ZHU Li;SHI Jiang-Tao*
    Bulletin of Botanical Research    2012, 32 (2): 232-236.   DOI: 10.7525/j.issn.1673-5102.2012.02.018
    Abstract1429)      PDF(pc) (902KB)(433)       Save
    The acetyl bromide lignin content of wood-forming tissue from Pinus koraiensis reaction wood was detected, and the FTIR and X-ray diffraction patterns were analyzed. The results showed that the lignin content of wood-forming tissue was lower than the mature wood, and the compression wood was higher than normal. The infrared spectrum results indicated that the characteristic absorption peak sites of hydroxyl and absorption band in 1 034-1 510 cm -1 were obviously different between immature and mature woods. The relative absorption intensity of chemical functional group in immature wood was lower than mature one. The site and shape of characteristic absorption peak showed no significant difference among compression, opposite and normal woods. However, the relative absorption intensity of functional group in compression wood was higher than normal one. The diffracted intensity of wood-forming tissue was significantly lower than mature wood by X-ray diffracted pattern. Meanwhile, the diffracted intensity of compression wood was lower than opposite and normal ones. Moreover, the relative crystallinity of wood-forming tissue was significantly lower than mature xylem and it was lower in compression wood than opposite and normal ones. Hence, the chemical properties of wood-forming tissue are dynamic changing in wood formation and differences are existed between reaction and normal woods.
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    Analysis on the Aroma Components of Different Flower Parts in Magnolia officinalis Wild and Cultivated Species
    WANG Jie;YANG Zhi-Ling*;YANG Xu
    Bulletin of Botanical Research    2012, 32 (2): 237-242.   DOI: 10.7525/j.issn.1673-5102.2012.02.019
    Abstract1324)      PDF(pc) (1218KB)(360)       Save
    The aroma components in flower bud samples of Magnolia officinalis wild and cultivated species whose bracts had just opened were studied by solid phase micro-extraction and GC/MS technology. The aroma components and their relative contents in different flower parts were compared and analyzed. The results showed that there were 39 components in M.officinalis wild species, 26 in pistils and stamens, 22 in petals. There were 75 components in M.officinalis cultivated species, 49 in pistils and stamens, 54 in petals. Terpenes were the major compounds in the flower bud of M.officinalis, camphene, (Z)-3, 7-dimethyl-1,3,6-octatriene, caryophyllene and 3,7-dimethyl-1,6-octadien-3-ol were identified in both wild and cultivated species and their relative contents were high. Both the aromatic compounds and their relative contents in M.officinalis wild and cultivated species were distinctly different, the aromatic compounds and their relative contents were also significantly different in pistils and stamens, petals of M.officinalis wild or cultivated species.
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