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    Bulletin of Botanical Research 2016 Vol.36
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    Two New Species of Ranunculaceae from China
    WANG Wen-Tsai
    Bulletin of Botanical Research    2016, 36 (1): 1-3.   DOI: 10.7525/j.issn.1673-5102.2016.01.001
    Abstract628)      PDF(pc) (501KB)(531)       Save
    Two species of the family Ranunculaceae, Delphinium yongdengense and Thalictrum panzhihuaense, are described as new from Gansu Province and Sichuan Province, and the morphological differences between them and their allies are given respectively.
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    Leaf Epidermal Features of Five Species Plants of Huperzia
    WU Yan-Di1,2;Han Qian-Qi1,2;HE Mei-Na1,2;BAI Zhuang-Dong1,2;GUO Bin1,2;WEI Ya-Hui1,2*
    Bulletin of Botanical Research    2016, 36 (1): 4-9.   DOI: 10.7525/j.issn.1673-5102.2016.01.002
    Abstract635)      PDF(pc) (1258KB)(448)       Save
    We studied the microscopic characters of the leaves of Huperzia serrata(Thunb. ex Murray) Trev, Huperzia crispata(Ching) Ching, Huperzia sutchueniana(Herter) Ching, Huperzia nanlingensis and Huperzia javaensis. with optical microscope and electron microscope. There are no trichome in the leaves of five species. We observed two types of stomatal apparatu(anomotetracytic and anomocytic) in the leaves of the five species. Anomocytic type is a typical stomata of these five species. The stomatal apparatus have multi-type phenomena, most of the stamatal are hypostomatic types, while their distributive modes and size also varies. There are obvious differences of leaf epidermal cell size and shape in five kinds of Huperzia plant, and the anticlinal walls of the epidermal cell are corrugated(undulate and sinuate).
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    Biosystematic Relationships among Taxa below Leymus(Poaceae) from China Based on the External Morphology and Micromorphological Characteristics of Leaf Epidermis
    LI Zhao-Meng;ZHOU Yong-Hui;LIU Yu-Ping;SU Xu*
    Bulletin of Botanical Research    2016, 36 (1): 10-25.   DOI: 10.7525/j.issn.1673-5102.2016.01.003
    Abstract528)      PDF(pc) (3277KB)(276)       Save
    We analyzed the main character differences of seventy-two reprehensive populations from twenty four Leymus species previously reported by scholars by morphological observation and anatomical analysis of leaf epidermises. There are different degrees of difference in fifteen morphological characters including length and width of flag leaves, hairs of leaf blades, straight/curve and length/width of inflorescence, spikelet No. of each inflorescence, floret No. of each spikelet, length of lemma and palea. The tested species can be divided into three sections according to them. The sect. Racemosus species of L.racemosus, L.crassiusculus, L.pluriflorus and L.pseudoracemosus, with sturdy, erect and thick spike, 3-number of spikelets per rachis node, 3-10 florets each spikelet, linear lanceolate glume, etc., is the most primitive one in genus Leymus. Due to the sect. Anisopyrum species including L.pishanica, L.golmudensis and L.ruoqiangensis, having thin, erect and loose spike, only one spikelet per rachis node, usually 2-5 florets each spikelet, linear lanceolate or cone glume, etc., so it is the most advanced of them. Meanwhile, because the external morphological characters of the sect. Leymus species usually lie in the intermediate transition state between the first two groups, naturally its systematics position should also be between the sect. Racemosus and sect. Anisopyrum species. At the same time, based on the evolutionary trends of the external morphological characters of three above sections, we deduced that the sect. Racemosus might produce immediately the sect. Leymus, whereas the sect. Anisopyrum might derive immediately form the sect. Leymus. The leaf epidermis of Leymus from China consists of long cell, stomatal cell, short cell and prickle hair. It belongs to the typical festucoid type. Except that some characters, such as the type of long-cells and thick/thin of cell walls, the volume and type of guard cells from stomatal apparatus, have obvious overlap and show similar, other characters, including the length of long-cells and curve extent of cell walls, distribution pattern of short-cells, shape of subsidiary cells, and type of prickle-hairs, have obvious differences. They can delimitate Leymus species into three sections identical with the morphology. According to the evolutionary trends of leaf epidermal characteristics of three sections and species within each section, their evolutionary positions and relationships were inferred. The results show that the sect. Racemosus is the most primitive one among three sections, the sect. Leymus is slightly more advanced than the former, and the sect. Anisopyrum is the most advanced of them. The sect. Racemosus might produce immediately the sect. Leymus, whereas the sect. Anisopyrum might derive immediately form the sect. Leymus. The biosystematic relationships of taxa below the genus are corroborated by the evolutionary trend of external morphology.
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    Biosystematic Relationships and Generalized Morphological Characteristics of Two Representative Species from Littledalea(Poaceae)
    ZHOU Yong-Hui1,2;LIU Yu-Ping1,2;LA Ben1,2;LI Zhao-Meng1,2;SU Xu1,2*;MA Yong-Gui3
    Bulletin of Botanical Research    2016, 36 (1): 26-33.   DOI: 10.7525/j.issn.1673-5102.2016.01.004
    Abstract580)      PDF(pc) (1459KB)(303)       Save
    We comprehensively studied the main character differences of 40 reprehensive populations between the nearest related Littledalea racemosa and Littledalea przevalskyi by the morphological observation and anatomical analysis of leaf epidermises. L.racemosa has short rhizomes, panicle and 6-8 florets per spikelet, while L.przevalskyi has no rhizomes, raceme and 3-6 florets per spikelet. They belong to two different species. The former is with more primitive external morphological characteristics, while the latter has more advanced ones. The leaf epidermis consists of long cells, short cells and stomatal cells. There are also differences on the anatomical characteristics of leaf epidermises including length and width of long cells, magnitude and distribution of stomatal apparatus, shape of subsidiary cells, abundance and growing form of short cells. According to the characteristics and evolutionary of trends of leaf epidermises of two species, we inferred their evolutionary positions and relationships. There is obvious discontinuity between them. Especially, L.przevalskyi is more advanced than L.racemosa, so it might derive immediately from the latter. The biosystematic relationships of these two species are corroborated by the evolutionary trend of external morphology.
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    Interspecific Association among Understory Species of the Low-efficiency Plantation in the Jiufeng National Forest Park
    XUE Ou;WEI Tian-Xing*
    Bulletin of Botanical Research    2016, 36 (1): 34-42.   DOI: 10.7525/j.issn.1673-5102.2016.01.005
    Abstract490)      PDF(pc) (1789KB)(335)       Save
    Interspecific relationship can reflect the structure and function of plant community, the type and stage of succession and the degree of community stability. It is of great significance for biodiversity conservation and vegetation restoration. In order to clarify the interspecific relationship of understory species in Jiufeng National Forest Park during restoration period, the experiment was conducted based on the 2×2 table to analyze the interspecific associations among the main species with high frequency by χ 2-test, association coefficient( AC) and Spearman’s rank correlation coefficient test. In shrub layer, 11 species pairs showed significant( P<0.05) or highly significant( P<0.01) association in χ 2-test, while 23 species pairs in Spearman’s rank correlation coefficient test. In herb layer, 15 species pairs showed significant( P<0.05) or highly significant( P<0.01) association in χ 2-test, while 27 species pairs in Spearman’s rank correlation coefficient test. For majority of the species pairs in two layers, their interspecific association did not reach significant level. In shrub layer, there was roughly an equal amount of the species pairs with positive association and those with negative association. In herb layer, the ratio of species pairs having positive association to those having negative association approximately was 1∶2. The species compositions of the two layers were fluctuating. The understory species interactions resulted from many factors including the effects of upper tree, the community succession process, species ecological characteristics and environmental conditions. By principal components analysis, the main species in herb layer and shrub layer were divided into four ecological species groups separately.
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    Taxonomy Significance Based on Pollen Morphology Observation of 19 Species of Manglietia
    SANG Zi-Yang1;ZHANG De-Chun1,2;WANG Yu-Bing2;MA Lü-Yi1*
    Bulletin of Botanical Research    2016, 36 (1): 43-51.   DOI: 10.7525/j.issn.1673-5102.2016.01.006
    Abstract612)      PDF(pc) (2034KB)(298)       Save
    We studied the pollen morphology of 19 species of Manglietia with scanning electron microscope(SEM), and measured the pollen size and densities of cavity and protuberance. The hierachical cluster analysis(HCA) was done based on the metrical data. Under SEM, all the pollens appear as boats, each pollen has a sprout channel whose length almost equal to the longer axis of pollen, the characters of pollen surfaces such as cavity and protuberance are steadily in same species, but there are some visible differences among pollen of different species. The result of HCA shows that the pollen morphology characteristics of the different species distributed in the same geographical region or similar areas have more similarities department, and there is a strong correlation between genetic branches and geographical distribution.
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    Embryonic Callus Induction and Plant Regeneration of Lilium fargesii
    GAO Jie1;WANG Yuan-Zhong2;HUANG Heng-Yu1*
    Bulletin of Botanical Research    2016, 36 (1): 52-57.   DOI: 10.7525/j.issn.1673-5102.2016.01.007
    Abstract624)      PDF(pc) (1251KB)(424)       Save
    In order to rationally protect and utilize the important wild traditional Chinese medicine resources, we built an efficient and instability regeneration system of Lilium fargesii Franch. by embryonic callus induction. With the scales as explants, by an orthogonal experiment, we studied the effect of different kinds and contents of plant hormone on embryonic callus induction and plantlet regeneration, and established a propagation system by tissue culture. The optimal medium for inducing embryonic callus was MS+6-BA 0.5 mg·L -1+NAA 0.5 mg·L -1+2,4-D 0.1 mg·L -1, the callus induction rate was 89.29%, and the generation coefficient of bulblet was 4.7. The optimal medium for regeneration of embryonic callus and inducing the bulblets was MS+6-BA 1.0 mg·L -1+2,4-D 0.1 mg·L -1, and its regeneration rate was 5.0/35d. The optimal rooting medium was 1/2MS+NAA 0.2 mg·L -1, and the rooting rate was 100%. There were 90% of plantlet surviving after transplanting rooted plantlets into sand when cultured in constant temperature and humidity condition for 35 d. Our study could provide a basis on protecting the improved trait of wild resources of L.fargesii, and lay the foundation for expanding the artificial cultivation and embryoid asexual breeding.
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    Rooting Anatomy of Hardwood Cutting for Albizia julibrissin Duraxx. and Activity Change of Related Enzymes During Rooting Process
    ZHOU Xiang-Ming;LIU Yu-Tang;ZHAO Xian-Zheng;SONG Zhao-Wei;WANG Shu
    Bulletin of Botanical Research    2016, 36 (1): 58-61.   DOI: 10.7525/j.issn.1673-5102.2016.01.008
    Abstract445)      PDF(pc) (943KB)(327)       Save
    With one-year hardwood cutting from Albizia julibrissin Duraxx. trees, we studied the anatomical structure during the formation of adventitious roots by paraffin section, and detected the activities of peroxidase(POD), polyphenol oxidase(PPO) and indoleaceticacid oxidase by colorimetry. The adventitious root of A.julibrissin cuttings was derived from a type of induced root primordium, and the primordium of adventitious root originated from the cambium. The enzyme activities of POD, PPO and IAAO showed the same trends in treatment and control during rooting process, but the activities of three enzymes in treatment were higher than those in control.
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    Variations of Growth and Leaf Traits of Intraspecific Hybridization Clones of Populus tomentosa
    LI Chun-Ming1,2;YAN Dong1;XIA Hui1;LIANG De-Yang1;SI Dong-Jing1;YIN Shao-Peng1;LIU Dian-Kun1;ZHAO Guo-Hui3;ZHAO Xi-Yang1,3*
    Bulletin of Botanical Research    2016, 36 (1): 62-67.   DOI: 10.7525/j.issn.1673-5102.2016.01.009
    Abstract428)      PDF(pc) (1017KB)(273)       Save
    50 intraspecific hybridization Populus tomentosa clones including 49 offspring clones and one male parent clone in one year-old as materials, tree height(H), basal diameter(BD) and 10 leaf traits(leaf length(LL), leaf width(LW), leaf petiole(LP) et al.) were investigated. There were exist highly significant difference( P<0.01) for all the traits among different clone. The average of H, BD, LL, LF and leaf area(LA) were 64.47 cm, 7.53 mm, 9.43 cm, 8.78 cm and 60.05 cm 2, respectively. Coefficients of phenotypic variation(PCV) of all the traits ranged from 2.88%(leaf basal angle, LBA) to 100.36%(leaf sharp angle, LSA), coefficients of genetic variation(GCV) were close to PCV and repeatabilities(R) ranged from 0. 851 to 0. 973. High PCV and R were beneficial for excellent clone selected. There was significant positive correlation among LL, LW, LA, H and BD( P<0.01). Regression analysis results showed that LW, LL and leaf sawtooth number(LSN) were the most important factors for influencing H and BD growth. By the comprehensive evaluation, with a selection rate of 10%, Ph 111, Ph 153, Ph 90, Ph14 and Ph 12 were selected as the excellent clones.Genetic gains from these five clones were 32.20% for tree height and 13.01% for basal diameter, respectively.
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    Life Table and Dynamic Analysis of Fagus hayatae Population in Micangshan Nature Reserve,Sichuan Province,China
    LI Jin-Xin1;WU Ding-Jun2;ZHANG Shi-Peng2;HE Xian-Xiang2;CHEN Jian2;SHI Qing-Mao2;XU Xiao1*
    Bulletin of Botanical Research    2016, 36 (1): 68-74.   DOI: 10.7525/j.issn.1673-5102.2016.01.010
    Abstract668)      PDF(pc) (1527KB)(590)       Save
    Fagus hayatae, a rare and endangered plant in China(category II), is distributed in China. Understanding the age structure and number of F.hayatae population is useful to reveal population dynamics and life-history traits. We investigated F.hayatae population distributes on the north slope of Laolin Valley in Micangshan Nature Reserve, and obtained a static life table on the basis of age class structure by using the smoothing out technique. Then, we analyzed the population quantitative dynamics of F.hayatae according to the results of survival and spectral analysis. The population was composed of 52% young individuals(including seedlings), 45.5% of mature individuals, and 2.5% of old individuals. Two peaks of higher mortality rate were in 1st and 17th age class periods of the lifespan. In addition to two big drops, the population survival rate decreased gradually from 1st to 24th age class. However, the opposite trend was found in cumulative mortality rate curve. By spectral analysis, the regeneration process of the population showed the marked periodic variations. The amplitude Ak had the largest values at fundamental wave A 1 and harmonic wave A 2 reflecting the lifespan of F.hayatae individuals. The quantitative fluctuation of population exhibited obvious small cycles, such as A 3 and A 7, which closely related with height growth and intense competitions during radial growth phase, respectively. Our results showed that the survival curve of F.hayatae population in Micangshan Nature Reserve was agree with Deevey type Ⅲ, and the population dynamics exhibited a trend that population size decreased sharply at young stage, kept stably at mature stage, and declined at old stage.
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    Transcriptome Analysis for Pinus yunnanensis Based on High Throughput Sequencing
    CAI Nian-Hui1,2,3;DENG Li-Li1,2,3;XU Yu-Lan1,2,3;XU Yang1,2,3;ZHOU Li1,2,3;WANG Da-Wei1,2,3;TIAN Bin1,2,3;HE Cheng-Zhong1,2,3;DUAN An-An1,2,3
    Bulletin of Botanical Research    2016, 36 (1): 75-83.   DOI: 10.7525/j.issn.1673-5102.2016.01.011
    Abstract563)      PDF(pc) (1786KB)(755)       Save
    The transcriptome of Pinus yunnanensis was sequenced by using Illumina Hiseq 2 000. In total 80 000 Unigene with an average length of 890 nt and N50 of 1 881 nt were obtained by de novo assembly. Of the Unigene, 43 434, 46 415 and 29 418 Unigenes had significant similarity with known data bank in NR, NT and Swiss-Prot, respectively. 14 792 Unigenes were annotated in clusters of orthologous groups of proteins(COG) and assigned to 25 clusters. 26 743 Unigenes were annotated in gene ontology(GO) and grouped into biological processes, cellular components and molecular function three functional categories, 55 sub-categories. The biological processes were most commonly existed. A total of 25 873 Unigenes were divided into 128 Kyoto Encyclopedia of Genes and Genomes(KEGG) pathways whose functions focused on metabolism. We found some Unigenes related to lignin biosynthesis. The sequence data for P.yunnanensis wiill be helpful for the gene discovery and utilization, molecular marker development and genetic improvement in the further research.
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    Relationships between the Mortality and Morphoallaxis of Haloxylon ammodendron Seedling
    TIAN Yuan1;TANG Li-Song2,4;QIAO Rui-Ping1,3
    Bulletin of Botanical Research    2016, 36 (1): 84-89.   DOI: 10.7525/j.issn.1673-5102.2016.01.012
    Abstract466)      PDF(pc) (1294KB)(262)       Save
    We studied the growth of Haloxylon ammodendron seedlings to reveal their growth traits and adaptabilities in the southern edge of Gurbantunggut Desert. The the seedlings grew well from early spring when the soil water content was high. With the consumption of soil moisture, the above- and below-ground part of the plant began to fall off, and then both parts recovered growing in August. Seedling mortality showed a downward trend in the whole growth period. H.ammodendron seedling could occupy more resource through rapid growth because of the high mortality rate. The falling of the assimilating branches during the drought period could maintain balance between water supply deficit and water consume of the assimilating organs. This morphological adjustment reduced seedling mortality effectively. Also, the fact that more photosynthate was assigned toward root could ensure seedling survival and recover growth quickly at proper water conditions.
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    Decreasing the Hyperhydricity and Rooting of Scutellariae baicalensis Georgi
    HAN Shu-Lan;WANG Hui-Mei*;ZHANG Dan
    Bulletin of Botanical Research    2016, 36 (1): 90-96.   DOI: 10.7525/j.issn.1673-5102.2016.01.013
    Abstract424)      PDF(pc) (1423KB)(280)       Save
    For reducing hyperhydric rate and improving rooting percentage of in vitro regenerated shoots from Scutellariae baicalensis Georgi, with the regenerated shoots induced from aseptic seedling stems, we studied the effect of N 6-Benzyladenine(6-BA), sucrose, agar and paclobotrazol(PP333) on hyperhydricity of regenerated shoots in S.baicalensis and the effects of IBA on rooting of regenerated shoots. The hyperhydric rate significantly decreased with lower 6-BA concentration on the medium. The lowest hyperhydric rate and the highest multiplication rate were on the medium containing 0.2 mg·L -1 6-BA. With the sucrose concentration increasing, the hyperhydric rate of S.baicalensis adventitious shoots was reduced, while their multiplication coefficient was also reduced. The best multiplication coefficient in vitro shoots without hyperhydricity were found on the medium containing 25 g·L -1 sucrose. On the medium containing 7.5 g·L -1 agar, the hyperhydric rate of S.baicalensis adventitious shoots was the lowest with higher proliferation coefficient. The medium containing PP333 positively affected the hyperhydric occurrence of regenerated shoots. With the increase of PP333 concentration on the medium, the hyperhydric rate was decreased gradually and in vitro shoots were stronger and healthier. The optimal rooting was got from the medium containing 0.1 mg·L -1 IBA, with the rooting rate of 100% and seven roots per shoot. The survival rate of rooting plantlets was over 95% after transferred to soil.
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    Relationship between H 2S Signal and WRKY in ABA Regulating Root Growth and Stomatal Movement
    SUN Li-Min;PEI Yan-Xi;LIU Zhi-Qiang*
    Bulletin of Botanical Research    2016, 36 (1): 97-104.   DOI: 10.7525/j.issn.1673-5102.2016.01.014
    Abstract387)      PDF(pc) (1654KB)(518)       Save
    Gaseous signal molecule hydrogen sulfide(H 2S) and plant hormone abscisic acid(ABA) modulate plant seed germination, root growth, stomatal movement, stress responses and other physiological processes, but the understanding of the relation between H 2S and ABA signaling is very limited. The transcription factors of WRKY18, WRKY40 and WRKY60 involve in ABA signaling in Arabidopsis. We used the Arabidopsis thaliana wild type(WT), lcd(L-cysteine desulfhydrase enzyme encoding gene konckdown mutant) and wrky18wrky40wrky60 mutant(WRKY18, WRKY40 and WRKY60 encoding gene knockout triple mutants) to study the interactions of H 2S, ABA and WRKY transcription factors in regulating seedling root growth and stomatal movement. The expression level of LCD and the content of endogenous H 2S in WT were increased by ABA treatment. The expression level of WRKY40 was decreased and WRKY60 was increased in WT by exogenous H 2S treatment. The expression level of WRKY18 and WRKY40 were increased, and WRKY60 was decreased in the lcd mutant. Compared with WT, the effect of ABA on seedling root growth and stomatal movement in lcd was weaker, the promoting effect of H 2S on root growth was more stronger in wrky18wrky40wrky60, but there were no obvious difference between WT and wrky18wrky40wrky60 on H 2S promoting the stomatal closure.
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    AtJ3 is Involved in ABA Response through Interaction with the PKS5 Kinase in Arabidopsis
    ZHAO Fei-Yi;JIAO Cheng-Jin;CHEN Quan;JIA Zhen;WANG Tai-Shu;ZHOU Hui
    Bulletin of Botanical Research    2016, 36 (1): 105-115.   DOI: 10.7525/j.issn.1673-5102.2016.01.015
    Abstract437)      PDF(pc) (2224KB)(428)       Save
    The phytohormone abscisic acid(ABA) has a wide range of important roles in plant growth and development as well as abiotic stress response. Previous studies identified the enormous components involved in ABA responses in plants. We identified a chaperon AtJ3(Arabidopsis thaliana DnaJ homolog 3; heat shock protein 40-like) using yeast two-hybrid assays in which PKS5 was used as bait. The AtJ3 T-DNA mutants atj3-1 and atj3-2 displayed ABA phenotypes. Seed germinations of atj3-1 and atj3-2 decreased, and the seedlings of them showed stunted growth and leaf chlorotic symptoms under ABA. Double mutants of atj3-1 pks5-1, atj3-1 pks5-3 and atj3-1 pks5-4 had similar ABA phenotypes as mutants of AtJ3 or PKS5. Moreover, the assays of subcellular location and transgenic plants showed that AtJ3 has overlapping expression pattern with PKS5. By co-immunoprecipitation and in vitro phosphorylation, AtJ3 physically interacts with PKS5 and represses the PKS5 kinase activity. Therefore, AtJ3 interacts with PKS5 in response to ABA by repressing the PKS5 kinase activity.
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    Distribution and Molecular Marker of SSR in EST Resource of Casuarinaceae
    HU Pan1;ZHONG Chong-Lu1;ZHANG Yong1;JIANG Qing-Bin1;CHEN Yu1;CHEN Zhen1;HAN Qiang1;Khongsak Pinyopusarerk2;Didier Bogusz3
    Bulletin of Botanical Research    2016, 36 (1): 116-122.   DOI: 10.7525/j.issn.1673-5102.2016.01.016
    Abstract630)      PDF(pc) (1422KB)(355)       Save
    There are 34 752 ESTs of Casuarinaceae in the database of NCBI, resulting in 12 062 non-redundant sequences(Unigene) with total length of 7 278.578 kb. 367 SSR were discovered to distribution on 353 Unigenes, the frequency and repeat motif types number of these EST-SSRs was 2.93% and 39, and the mean distance was 19.83 kb. Dinucleotide and trinucleotide repeats were the dominant types among the EST-SSRs, accounting for 57.77% and 34.06%, respectively. AG/CT(93.87%) and AAG/CTT(44.09%) were the most abundant motifs for dinucleotide and trinucleotide. There were 97 pairs of primers designed for the obtained EST-SSR, and 32 of all could be effectively amplified. By Blast analysis, 77.3% ESTs sequences containing SSR locus were homologous with functional sequences in non-redundant protein sequences databases. The homology with functions from Vitis vinifera had the greatest proportion(10.4%). By GO functional classification, 47.3% EST sequences containing SSR locus had at least one GO ontology, the sequences relevant to cellular components were most, and the functional items relevant to cytoplasm and nucleus were highest.
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    Culture System on Loose, Homogeneous, Embryogenic Callus of Populus× canadensis Moench ‘Tower’
    LIU Yan-Jun1;ZHANG Chao2;YANG Jing-Hui1*;LI Bing2;LIU Ting1;QIN Yang-Jun1
    Bulletin of Botanical Research    2016, 36 (1): 123-128.   DOI: 10.7525/j.issn.1673-5102.2016.01.017
    Abstract461)      PDF(pc) (1273KB)(227)       Save
    To get loose, homogeneous, embryonic callus(LHEC) from Populus× canadensis Moench ‘Tower’, an efficient system was set up from the loose callus induction(LC) to successive transfer culture and embryonic callus induction with leaves of seedlings in tube and different kinds and concentration of hormones(BA,KT,2,4-D), sucrose and inorganic salt. The loose, homogeneous callus(LC) was induced completely(induction rate of 100%) with leaves and on the culture medium of low sugar and low inorganic salt(1/4MS+2 mg·L -1 2,4-D+1 mg·L -1 BA+10 g·L -1 sugar). The globule embryonic callus(formerly embryoid) was induced after LC was transferred for 2-3 times on successive culture medium(MS+2.0 mg·L -1 2,4-D+2.0 mg·L -1 BA+Vc 100 mg·L -1+30 g·L -1), then transferred onto the medium of LHEC(MS+0.5 mg·L -1 2,4-D+3 mg·L -1 BA+Vc 100 mg·L -1+40 g·L -1 sucrose), and cultivated for 4-5 weeks(one time transfer every seven days). The cells of LC was kept in the characteristics of loose without roots, browning and with fast growth under the appropriate concentration of 2,4-D and BA. LC grew better and LHEC was produced more in medium containing 30 g·L -1 sucrose. Vitamin C suppressed browning of LC in subculture. BA was conducive to the formation of LC, but KT was beneficial to the formation of compact callus, while the callus growth was faster by treatment of BA than by KT. When the concentration of 2,4-D was 0.5-1 mg·L -1, the number of LHEC was increased with the increase of BA concentration. The number of LHEC reached to the maximum of 100%, when BA was 3 mg·L -1. The main factors were discussed in callus embryogenic.
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    Reactive Oxygen Metabolism of the TheIF1A Transgenic Tobacco under Methyl Viologen Stress
    ZHAO Yu-Lin1;YANG Gui-Yan1,2;YU Li-Li1;GUO Yu-Cong1;ZHAO Zhen1;GAO Cai-Qiu1*
    Bulletin of Botanical Research    2016, 36 (1): 129-133.   DOI: 10.7525/j.issn.1673-5102.2016.01.018
    Abstract521)      PDF(pc) (1107KB)(302)       Save
    Eukaryotic translation initiation factor eIF1 involved abiotic stress regulation in plant. Previous studies showed that overexpressed TheIF1A gene from Tamarix hispida improved drought and salt resistance of genetically modified plants. We further studied the function of TheIF1A response to methyl viologen(MV). The results of histochemical stainning showed that ROS were less accumulated in leaves, guard cells and root tips of the TheIF1A transgenic lines than WT tobacco under MV stress. The H 2O 2 content in transgenic lines were also less than WT under MV stress. Accordingly, the CAT and POD activity in transgenic lines were obviously higher than WT under MV stress. Therefore, TheIF1A could improve plant oxidative stress resistance by regulating the protecting enzymes activity to increase the ROS scavenging ability.
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    Clone and Characterization of A Glutathione-s-transferase Gene in Muscari armeniacum
    YANG Hui-Ping;LIU Ya-Li*;LOU Qian;LIU Ni-Ni
    Bulletin of Botanical Research    2016, 36 (1): 134-140.   DOI: 10.7525/j.issn.1673-5102.2016.01.019
    Abstract443)      PDF(pc) (1840KB)(187)       Save
    Based on the latex the transcriptome database, the full length cDNA of glutathione-S-transferase(GST) from Muscari armeniacum was cloned by reverse-PCR and PCR, designated as MaGST. The full length cDNA of MaGST was 711 bp, and the ORF(Open Reading Frame) length was 666 bp, encoding a protein polypeptide of 221 amino acids with a predicted molecular weight of 54.1 kD and pI of 5.13. By phylogenetic tree analysis, the putative MaGST protein displayed identities to the GSTs of Allium cepa and Triticum aestivum of 76.72% and 62.50%, respectively, and contained the Tau GST-specific N-terminal domain(G site) and the C-terminal domain(H site), belonging to the family of GST Tau. By realtime PCR, the expression pattern of MaGST in different organs were similar, belonging to constitutive expression. The expression of MaGST was regulated by salicylic acid, but not by the NaCl.
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    Optimization of Microwave-assisted Extraction of Syringin and Oleuropein from Syringa reticulata
    WANG Hua1,2;ZHOU Li-Ping1,2;LI Meng-Sha1;ZHU Liang-Yu1,2;MU Li-Qiang2;NI Hong-Wei1*
    Bulletin of Botanical Research    2016, 36 (1): 141-145.   DOI: 10.7525/j.issn.1673-5102.2016.01.020
    Abstract495)      PDF(pc) (1158KB)(339)       Save
    We used microwave-assisted extraction to extract syringin and oleuropein from Syringa reticulate, and optimized the extraction process by the Box-Behnken response surface method. The order of the effect of factors on the yield of syringin and oleuropein was the concentration of ethanol>the microwave time>microwave power. The optimum conditions for the extraction were as follows: soaking time of 1 h, ethanol concentration of 45%, ratio of solid to liquid of 1∶20 g·mL -1, microwave output power of 600 W, and extraction time of 5 min. With the optimum extraction conditions, the yield of syringin and oleuropein from Syringa reticulate was 5.28%±0.102%. The microwave-assisted extraction was efficient and rapid for extraction syringin and oleuropein from Syringa reticulate, and was benefit for the comprehensive utilization of Syringa reticulate.
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    Economic Dynamics Performance Evaluation of Lignin Diesel
    LI Wen-Gang;SUN Xiao-Li;GE Yun-Long;ZHAO Xiu-Hua;ZU Yuan-Gang*
    Bulletin of Botanical Research    2016, 36 (1): 146-150.   DOI: 10.7525/j.issn.1673-5102.2016.01.021
    Abstract491)      PDF(pc) (1022KB)(372)       Save
    We studied the economic dynamics performance of the lignin diesel conforming to the national standard<GB 252-2000 light diesel oil>. From the engine full load speed characteristics contrast experiment, the measured power and torque of lignin diesel and No.0 diesel oil were equivalent, in accordance with the amount of pure diesel in lignin diesel, and the oil-saving rate of lignin diesel was 10%-15% less than that of No.0 diesel fuel on average. From the engine load characteristics contrast experiment, under the same torque and in accordance with the amount of pure diesel in lignin diesel, oil-saving rate of lignin diesel was 10%-15% less than that of No.0 diesel fuel on average. By the engine free acceleration condition emissions test, there was no statistical differences about exhaust emission by lignin diesel and No.0 diesel.
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    Hydroxymethylation of Lignin from Ethanol Pulping
    LU Qi1,2;MENG Yong-Bin1,2;XU Lei1,2;ZHANG Zi-Dong1,2;LIU Ying1,2;MENG Qing-Huan1,2;ZU Yuan-Gang1,2*
    Bulletin of Botanical Research    2016, 36 (1): 151-155.   DOI: 10.7525/j.issn.1673-5102.2016.01.022
    Abstract534)      PDF(pc) (981KB)(203)       Save
    We extracted the lignin by ethanol method, with the residues of Acanthopanax root and stem as raw materials. By single factor and orthogonal experiments, the optimum of ethanol lignin hydroxymethylation alkali dose of 2.0%, reaction temperature of 80℃, reaction time of 3.5 h, and mass ratio of kraft lignin to formaldehyde of 3∶1. The modified lignin reached 11.56% hydroxymethyl groups and 0.257% free formaldehyde.
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    Ethanol Production Technology by Fermentation form Fiber Residues
    MENG Yong-Bin1,2;XU Lei1,2;ZHANG Zhi-Dong1,2;LU Qi1,2;LIU Ying1,2;NIE Si-Ming1,2;ZU Yuan-Gang1,2*
    Bulletin of Botanical Research    2016, 36 (1): 156-160.   DOI: 10.7525/j.issn.1673-5102.2016.01.023
    Abstract331)      PDF(pc) (1039KB)(341)       Save
    We studied the effects of various factors on the yield of ethanol from enzymatic hydrolysate of Tara fiber residues by the single factor experiment, and then optimized ethanol production technology by orthogonal experiments. The optimum conditions were the ratio of sodium alginate-yeast of 1∶2 g·g -1, substrate sugar concentration of 30%, the amount of yeast of 1.5 g, and fermentation time of 120 h, with the yield of ethanol of 83.68%.
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    Metapilea,a New Genus of Urticaceae from Guangxi
    WANG Wen-Tsai
    Bulletin of Botanical Research    2016, 36 (2): 164-166.   DOI: 10.7525/j.issn.1673-5102.2016.02.002
    Abstract629)      PDF(pc) (458KB)(270)       Save
    A genus belonging to the tribe Lecantheae Wedd. of the family Urticaceae, Metapilea, is described as new from Guangxi Autonomous Region. This genus is closely related to Pilea Lindl., and from the latter differs in its piatillate capitate inflorescences with oblong discoid receptacles and with numerous bracts and pistillate flowers borne on the adaxial surface of receptacle. Besides, it is also related to Lecanthus Wedd., and from the latter differ in its sessile pistillate capitula with conduplicate oblong receptacles and lacking involucres and in its neither cucullate nor corniculate pistillate tepals. Without doubt, these two advanced genera, Metapilea and Lecanthus, have both originated from the primitive genus Pilea.
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    Spore Propagation of Four Ferns
    ZHU Xiao-Feng1,2;SHEN Hui1*;JIN Dong-Mei1;ZHAO Guo-Hua1,3; LIU Bao-Dong3;LING Tie-Jun2;YAN Yue-Hong1
    Bulletin of Botanical Research    2016, 36 (2): 167-176.   DOI: 10.7525/j.issn.1673-5102.2016.02.003
    Abstract971)      PDF(pc) (2334KB)(328)       Save
    The development of fern resources is attracting more attention in recent years, however, current spore reproductive technology is immature, and has become a bottleneck. We cultured the spores of four typical ferns, including Blechnum orientale, Matteuccia struthiopteris, Ceratopteris thalictroides and Adiantum malesianum, observed spore propagation characteristics and time of the gametophyte development at each stage. By compiling the reported data, we analyzed the culture conditions, such as light and temperature, in four key development stages, including spore germination, and the formation of prothalli, sexual organs, and juvenile sporophyte. Longer illumination time may promote spore germination. Higher light intensity may prolong the formation of prothallus. Growth rate and temperature had no correlation during gametophyte formation in 20-25℃. Our research is useful for establishing the system of fern propagation and may promote the exploration of fern resources.
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    Optimization of Leaf Regeneration and Genetic Transformation System of Populus tomentosa
    LI Chun-Li;WANG Xiao-Jing;DING Qiang-Qiang;WU Han*
    Bulletin of Botanical Research    DOI: 10.7525/j.issn.1673-5102.2016.02.004
    Main Anthocyanin Profiles in Petals of Freesia hybrida
    XU Yi-Qian1;YUAN Yuan1;TAO Xiu-Hua2;YANG Juan3;SHI Yi-Min1;TANG Dong-Qin1*
    Bulletin of Botanical Research    2016, 36 (2): 184-189.   DOI: 10.7525/j.issn.1673-5102.2016.02.005
    Abstract565)      PDF(pc) (1290KB)(230)       Save
    We analyzed the anthocyanin profiles in 6 cultivars of freesia( Freesia hybrida). The petal color was observed according to the Royal Horticultural Society Color Card(RHSCC). Petal pigment type was identified by characteristic color reaction. UPLC-PAD and UPLC-Q-TOF-MS were used for qualitative and quantitative analysis of anthocyanins. The flavonoids in petals of all cultivars were detected, while carotenoids were not detected; meanwhile, every cultivar, except for ‘Shangnong Ruxiang’, contained anthocyanins in petals. Among five anthocyanincontained cultivars, the highest anthocyanin content was observed in ‘Shangnong Purple Rose’, followed by ‘Shangnong Chenghong’. Totally 6 anthocyanin components were detected in petals of these cultivars, including Delphinidin 3,5-O-glucoside, Petunidin 3,5-O-glucoside, Delphinidin 3-O-glucoside, Malvidin 3,5-O-glucoside, Petunidin 3-O-glucoside and Malvidin 3-O-glucoside. Three anthocyanin components were detected in petals of ‘Shangnong Zixueqing’ and ‘Shangnong Gongfen’, of which were majorly malvidin and petunidin derivated anthocyanins. Malvidin-anthocyaninswith single or double glucosides was present in petals of ‘Shangnong Zixueqing’, while all anthocyanins in petals of ‘Shangnong Gongfen’ bonded with double glucosides, such as Delphinidin 3,5-O-glucoside, Petunidin 3,5-O-glucoside and Malvidin 3,5-O-glucoside, major Petunidin 3-O-glucoside and minor Delphinidin 3-O-glucoside in petals of ‘Shangnong Purple Rose’. While only one anthocyanin component was detected in petals of ‘Shangnong Golden Queen’ and ‘Shangnong Chenghong’, which was Delphinidin 3,5-O-glucoside and Malvidin 3,5-O-glucoside, respectively.
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    Adventitious Bud Induction with Immature Embryo of Herbaceous Peony
    WEI Dong-Xia1,2,3;GAO Kai4;YUAN Yan-Bo1;TANG Zheng-Jiao1;YU Xiao-Nan1,2,3*
    Bulletin of Botanical Research    2016, 36 (2): 190-194.   DOI: 10.7525/j.issn.1673-5102.2016.02.006
    Abstract372)      PDF(pc) (1069KB)(231)       Save
    Culture of zygotic embryos is an effective way to accelerate the breeding process of herbaceous peony. With the zygotic embryos of three herbaceous peony( Paeonia lactiflora Pall.) cultivars(‘Fen Yu Nu’, ‘Zhu Sha Pan’ and ‘Tuan Ye Hong’), we studied the affection of different embryo age, explant types, sterilization methods and starting medium formula on root initiation in vitro. The suitable development period were 50 and 70 d after flowering for embryo-culture of ‘Zhu Sha Pan’ and ‘Tuan Ye Hong’, respectively. We got better results when using excised zygotic embryos rather than keeping part of endosperm reserved. The optimal disinfection method for explant was that firstly treating carpel for 30-60 s with 70% alcohol, then for 20 min with 2% NaClO, and finally for 10 min with 2% NaClO. The optimal initial medium was 1/2MS+1.0 mg·L -1 IAA+0.5 mg·L -1 GA 3.
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    Effects of Soil Fauna on C Dynamics in Different Sized Forest Gaps in Pinus massoniana Plantation
    LI Xun1;LIU Yang1,2,3;ZHANG Yan1;LIU Hua1;YANG Wan-Qin1,2,3;ZHANG Jian1,2,3*
    Bulletin of Botanical Research    2016, 36 (2): 195-203.   DOI: 10.7525/j.issn.1673-5102.2016.02.007
    Abstract492)      PDF(pc) (1862KB)(314)       Save
    Soil fauna has an important effect on litter nutrient release, and the effect could vary with litter types and be controlled by environmental factors such as temperature and precipitation. We to characterized the effect of soil fauna on C dynamics in different sized forest gaps (G1: 100 m 2, G2: 400 m 2, G3: 900 m 2, G4: 1 600 m 2) during litter decomposition of a Pinus massoniana plantation in a hilly area of the upper reaches of Yangtze River. A field litterbag experiment was conducted in P.massoniana plantation of Sichuan, China, from January 2014 to November 2014. Samples of airdried leaf litter for P.massoniana( Pm) and Cinnamomum camphora(Cc) were placed in nylon litterbags (20 cm×20 cm, 10 g per bag) of two different mesh sizes (3.00 and 0.04 mm). The amount of C released were investigated in different sized forest gaps during the first year of decomposition, i.e., Spring, Summer, Autumn and Winter. There was a significant difference in the amount of litter C released through soil fauna among different sized gaps: medium sized gap (G3: 900 m 2) promoted the C release of Cc and Pm. The content of C displayed a declining trend in Spring and Summer, but with an obvious enrichment process in Winter and Autumn. Soil fauna imposed a positive effect on C release, the contribution of soil fauna to the C loss rate displayed as Cc> Pm. Based on the above results, soil fauna imposed a positive effect on C release in P.massoniana plantation. When we reconstruct the low-function forest, we can take advantage of medium sized gap and introduce native broadleaf tree species Cc which is easier to be decomposed by soil fauna to accelerate nutrient cycling and maintains the soil fertility of P.massoniana plantation.
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    Effects of Water Extraction from Roots of Carya illinoensis on Seed Germination and Seedling Growth of Two Crops
    ZHANG Quan1;YAO Xiao-Hua1*;TENG Jian-Hua2;SHAO Wei-Zhong3;FU Song-Ling4
    Bulletin of Botanical Research    2016, 36 (2): 204-210.   DOI: 10.7525/j.issn.1673-5102.2016.02.008
    Abstract460)      PDF(pc) (1439KB)(205)       Save
    In order to probe the impact of water extraction from roots of Carya illinoensis for Triticum aestivum L. and Glycine max(L.) Merr, we studied the effects of different levels(0.025, 0.05 and 0.1 g·mL -1) of water extraction from roots of C.illinoensis on the seeds and seedling growth of T.aestivum L. and G.max(L.) Merr with indoor bio-assay. The water extract prominently inhibited the seed germination and seedling growth of T.aestivum L. and G.max(L.) Merr with concentration effect(weak inhibition in low concentration, high inhibition in high concentration). The descending order of inhibition of water extract for germination rate, germination energy, germination index and vigor index of T.aestivum and G.max(L.) Merr were vigor index, germination energy, germination index, germination rate, and vigor index, germination energy, germination rate, germination index, respectively. The descending order of allelopathic effects of water extract on shoot height, root length, fresh weight and dry weight of T.aestivum and G.max(L.) Merr were both root length, shoot height, fresh weight, dry weight. The water extract prominently decreased the seedling root length, but had different impacts on the dry and fresh weight of receptor due to the different species. The order of the synthesis effects of water extraction from roots of C.illinoensis on two crops is T.aestivum L.> G.max(L.) Merr. The activity of SOD in the 7 d seedings was decreased, but the content of MDA, the activity of POD and CAT were increased with the increasing of concentration. Comprehensively, the inhibition of the water extraction from roots of C.illinoensis is higher for T.aestivum L. than for G.max(L.) Merr.
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    Determining the Main Organic Compounds in Ammodendron bifolium Seeds
    ZHU Yan-Lei1,2;AN Deng-Di2;SHE Xiao-Ping1*
    Bulletin of Botanical Research    2016, 36 (2): 211-215.   DOI: 10.7525/j.issn.1673-5102.2016.02.009
    Abstract444)      PDF(pc) (1026KB)(312)       Save
    Ammodendron bifolium(Pall.) Yakovl. is a rare psammophyte distributed in Xinjiang of China. In order to study the adaptation of the species to desert environment, the contents of main organic compounds in its seeds were determined. Carbohydrate, soluble protein and crude fat accounted for 16.8%, 23.7% and 15.5% of seed dry weight respectively. Soluble carbohydrate accounted for 10.1% of seed dry weight, and sucrose accounted for 43.4% of soluble carbohydrate content. Contents of amino acids in proteins and free amino acids accounted for 26.5% and 0.4% of seed dry weight, respectively. Unsaturated fatty acid was more than 90% of total fat content, and oleic acid and linoleic acid were its main component.
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    Effect of Shade Treatment in Summer on the Expression of Genes Related to Theanine Biosynthesis in Tea Plants( Camellia sinensis)
    CHEN Qi;YU Shu-Wei;JIANG Xue-Mei;ZHAO Ying;MENG Xiang-Yu;WAN Xiao-Chun*
    Bulletin of Botanical Research    2016, 36 (2): 216-223.   DOI: 10.7525/j.issn.1673-5102.2016.02.010
    Abstract466)      PDF(pc) (745KB)(328)       Save
    The shade can effectively improve the quality of tea( Camellia sinensis) by causing an increase in the concentration of theanine, the main compound that contributes to the fresh and brisk taste of tea leaves. We analyzed the influence of shade on theanine biosynthesis in relation to the expression of genes related to theanine synthesis in tea leaves during summer. Our data reveal that shade has a notable effect on the level of both theanine and total free amino acids in young leaves. The results of qPCR and western blot analysis suggest that these effects are mainly brought about via an increase in the mRNA expression of TS and promotion of TS protein synthesis. The qPCR results also suggest that the mRNA expression of all the genes related to the theanine pathway in the leaves, including glutamine synthetase( GS), glutamate dehydrogenase( GDH), glutamine 2-oxoglutarate amino-transferase( GOGAT), nitrite reductase( NiR), arginine decarboxylase( ADC), are significantly affected by shade. However, from two different expression patterns, the GS/GOGAT pathway is triggered by shade and members of this pathway show higher expression in old leaves, which reassimilate more NH + 4 to supply more amides to young leaves. The gene expression of NiR and ADC was markedly lower in both the old and young leaves from the shaded plants, which suggests that nitric nitrogen metabolism might be inhibited by shade treatment. Thus, shade treatment can increase the content of free amino acids especially theanine in tea leaves and improve the effective utilization of summer tea.
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    Effects of Complex Saline-alkali Stress on Seed Germination and Stress-resistance Physiological Characteristics of Naked Oat Seedlings
    LIU Jian-Xin1,2;WANG Jin-Cheng1,2;WANG Rui-Jian1,2;JIA Hai-Yan1,2
    Bulletin of Botanical Research    2016, 36 (2): 224-231.   DOI: 10.7525/j.issn.1673-5102.2016.02.011
    Abstract688)      PDF(pc) (1653KB)(246)       Save
    We studied the effects of salt-alkaline mixed stress on seeds germination, seedlings growth and leaf physiological characteristics of stress-resistance for the physiological response of a new naked oat( Avena nude) cultivar ‘Dingyou No 6’ to complex saline-alkali stress. A.nude seeds and seedlings were cultivated in sands with different complex saline-alkali solution addition according to soil salinity composition of the planting areas. Germination rate, germination index, vigor index of seeds, water content and dry weight of seedlings were significantly decreased, while the contents of superoxide anion, hydrogen peroxide and malondialdehyde and relative plasma membrane permeability in leaves were increased evidently with increasing of the complex saline-alkaline stress. The complex saline-alkaline stress increased significantly Na + contents, but reduced K + contents and K +/Na +. The activities of plasma membrane H +-ATPase, peroxidase, ascorbate peroxidase and soluble protein content in leaves of the seedlings exhibited a humped-curve, while superoxide dismutase activities and the contents of glutathione, organic acid and proline were significantly increased. Catalase activity and ascorbic acid content were decreased evidently, free amino acid content first decreased and then increased, and soluble sugar content demonstrated a rose at first, and then dropped at last rose again. The imbalance of active oxygen metabolism and the disturbance of Na + and K + homeostasis in leaves resulted in the inhibition of naked oat seedlings growth, and the organic acid and proline are two possible osmotic adjusting substances of naked oats that are adapted to complex salt-alkaline stress.
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    Sequence Analysis of Nuclear DNA  JRD5680 for Determining Genetic Diversity and Genetic Structure Analysis of Common Walnut( Juglans regia L.)
    ZHANG Tian;WANG Ma-Li;ZHAO Peng*
    Bulletin of Botanical Research    2016, 36 (2): 232-241.   DOI: 10.7525/j.issn.1673-5102.2016.02.012
    Abstract525)      PDF(pc) (2102KB)(469)       Save
    We studied the genetic diversity and population genetic structure of 265 individuals from different regions of China by using the roles of phenylalanine ammonia-lyase(PAL) nuclear gene JRD5680 DNA sequence of 40 common walnut( Juglans regia) and 5 out group plant species. The length of JRD5680 sequence was 809 bp, the content of G+C was 46.2% with 74 information sites. A total of 30 haplotypes, haplotype diversity variation showed low genetic diversity( H d=0.370, π=0.005 3). There is a clear geographical structure( N ST> G ST), which is a regional distribution pattern. The spatial genetic structure analysis showed that the correlation between geographical isolation and genetic distance was significant( r=0.263 2; P=0.032 4*), and there was obvious geographical isolation effect among different geographic populations. The results of comprehensive mismatch analysis and neutral test can be deduced from the recent history of the different regions of the population expansion event. From the genetic structure, the walnut population can be divided into two groups(southwest and other regions). By AMOVA, the genetic variation among the populations was mainly stored in the populations(56.54%), and the genetic differentiation among the populations was higher( F ST=0.885). The total of 5 species of Juglandaceae could be a single branch, and the JRD5680 sequence might be with high molecular identification radio in the members of the Juglandaceae.
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    Allelic Variation in 4-coumarate:CoA ligase(4CL) Associated with Wood Properties of Larix olgensis
    WANG Yan-Hong1,2;JIA Qing-Bin1;ZHANG Lei1;ZHANG Han-Guo1*
    Bulletin of Botanical Research    2016, 36 (2): 242-251.   DOI: 10.7525/j.issn.1673-5102.2016.02.013
    Abstract464)      PDF(pc) (1662KB)(220)       Save
    We used 10 provenances of Larix olgensis to study the effects of polymorphism of 4CL gene on wood quality and growth traits in L.olgensis. The polymorphism in 4CL gene was identified using DNA sequencing, 64 mean SNPs were identified,the median SNP frequency was one site per 31 bp,and the average nucleotide diversity for the sequenced regions was calculated to be π T =0.015 77 and θ W =0.008 78. The diversity level of synonymous nucleotide sbustituons( π syn=0.022 56) was 1.4 of nonsynoymous nucleotide substitutions( π nonsyn=0.015 75), suggesting that the gene misht be evolved under purifying selection at the synonoymous sites of the coding region in L.olgensis. The association between the 27 common SNPs(frequency>10%), haplotypes and wood quality and growth traits were analyzed respectively, indicating that 7 SNPs and 6 haplotypes were significantly associated with wood quality and growth traits, explaining between 0.09%-2.08% of the phenotypic variance. Ourstudy provided an important genetic foundation for molecular marker-assistant selection breeding programs with the goals of improving the wood quality and growth products in L.olgensis.
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    Sequence and Expression Analysis of MYB Family Genes in Betula platyphylla Suk.
    LIU Hui-Zi;SUN Dan;YU Ying;ZHANG Nan;WANG Chao*
    Bulletin of Botanical Research    2016, 36 (2): 252-257.   DOI: 10.7525/j.issn.1673-5102.2016.02.014
    Abstract738)      PDF(pc) (1202KB)(294)       Save
    The MYB transcription factor family is one of the important plant transcription factor family members, and they have the key function on the growth, development, cell wall biosynthesis and stress response in plant. We obtained the total of 17 MYB genes, and by phylogenetic analysis, 17 MYB genes of birch belong to different subfamilies and groups. Among these genes, 10 MYBs belong to the 1R/4R subfamilies, and 7 MYBs belong to the 2R subfamily. BplMYB13 is homologous to the AtMYB46 which functions in the secondary cell wall biosynthesis. BplMYB15 and BplMYB26 belong to the group which involved in the stress response. BplMYB23 is homologous to the AtMYBs involved in the glucosinolate biosynthesis. BplMYB9, 21 and 22 have homologous with the AtMYBs functioned in phenylpropanoid biosynthesis. The expression patterns analysis among the development stages of the growth season and the tension wood and opposite wood bended for 6 h, and the normal wood were performed by real time RT-PCR. 17 MYB genes had the peak express levels from the middle of May to the middle of July, and the BplMYB13 maintained the high express levels among all the growth season in the cambium and xylem. The BplMYB13 may play the role in the cambium and xylem development. After 6 h bending, 14 MYBs were up-regulated in the tension wood compare with the opposite wood or normal wood, and 7 MYBs were increased expressed in the opposite wood than those in the normal wood. Therefore, these genes response to the artificial bending and play key roles in the physiology change during the xylem development response to the bending stress.
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    Cloning and expression analysis of GrCMS gene in Gentiana rigescens
    ZHANG Xiao-Dong1;LI Cai-Xia1;WANG Yuan-Zhong2*
    Bulletin of Botanical Research    2016, 36 (2): 258-265.   DOI: 10.7525/j.issn.1673-5102.2016.02.015
    Abstract508)      PDF(pc) (1894KB)(257)       Save
    2-C-methyl-D-erythritol 4-phosphatecytidyltransferase(CMS, EC is the third enzyme in methylerythritol phosphate(MEP) pathway. The Open Reading Frame(ORF) of GrCMS gene was cloned by RT-PCR technology from young leaves of Gentiana rigescens based on the transcriptome of G.rigescens and its prokaryotic and the tissue specific expression analysis were performed. The ORF of GrCMS gene(accession number: KJ917164) was 933 bp long coding for a protein of 310 amino acids, and the predicted relative molecular weight of GrCMS was 34.23 kD with its theoretical pI of 7.68. The results of GrCMS protein analysis showed that GrCMS which possessed the conserved domains of CMS proteins and may localize in chloroplast was a hydrophilic stable protein without signal peptide, and it was composed of mainly α-helix(26.45%) and random coils(54.84%). By phylogenetic analysis, GrCMS was close to CrCMS of Catharanthus roseus. By prokaryotic analysis, the recombinant protein of GrCMS gene in E.coli was approximately 60.23 kD(containing GST tag protein 26 kD), which was consistent with the anticipated size. By real-time PCR analysis, GrCMS gene was primarily expressed in leaf. Our results will provide reference for further functional researches of GrCMS gene and the biosynthetic pathway of gentiopicroside.
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    Response of Germination on Ten Chenopodiaceae Seeds to Drought Stress and Resistances Evaluation in Tarim Basin
    HAN Zhan-Jiang;CHENG Long;LI Zhi-Jun*
    Bulletin of Botanical Research    2016, 36 (2): 266-273.   DOI: 10.7525/j.issn.1673-5102.2016.02.016
    Abstract564)      PDF(pc) (1666KB)(232)       Save
    The experiments were conducted with ten Chenopodiaceae seeds as materials to examine responses of seed germination and evaluate their tolerance under drought stress by PEG-6000. The final germination rates and radicle length of seeds were inhibited while the mean germination times extended continuously in various degrees with the drought intensifying in 10 Chenopodiaceae species. Seeds of Suaeda glauca, Suaeda rigida, Suaeda heterophylla, Halocnermum strobilaceum, Halostachys caspica, Salicornia europaea and Halogeton arachnoideus obtained their germination recoveries after relieving drought stresses. The limit values of drought tolerance at seed germination stage from large to small were H.caspica, H.arachnoideus, S.heterophylla, S.rigida, S.europaea, Suaeda arcuata, H.strobilaceum, Suaeda stellatiflora, Chenopodium aristatum and S.glauca. Comprehensive evaluation of drought resistances of seed germination were conducted with fuzzy subordinate function analysis by selecting the final genmination rate, mean generation time, radicle length and germination recovery rate. The drought resistances at seed germination stage from large to small were H.caspica, H.arachnoideus, S.rigida, H.strobilaceum, S.heterophylla, S.stellatiflora, S.europaea, C.aristatum, S.glauca and S.arcuata. Seed germinations of H.caspica, H.arachnoideus, S.rigida and H.strobilaceum showed the best resistances under drought stresses between ten Chenopodiaceae species, and their weighted average values of subordinate function were 0.915, 0.793, 0.762 and 0.737, respectively.
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    Influence of Different Vegetation Types on the Soil Inorganic Nitrogen at Small Watershed in Northern Zhejiang Province
    WANG Qing-Bing1;WU Hao1;ZHANG Jian-Feng1*;CHEN Guang-Cai1;LI Ze-Bo1;WANG Li1,2;YANG Quan-Quan1
    Bulletin of Botanical Research    2016, 36 (2): 274-282.   DOI: 10.7525/j.issn.1673-5102.2016.02.017
    Abstract457)      PDF(pc) (1668KB)(242)       Save
    We studied the contents of inorganic nitrogen in surface and subsurface soil within six different vegetation types at Fushi Reservoir watershed in Anji County of northern Zhejiang Province, and the influence of different vegetation types on soil inorganic nitrogen distribution. There is significant difference( P<0.05) between the contents of soil NO - 3-N and NH + 4-N at Tianjia mountain and Rihui mountain watershed, and the maximum differences of soil NO - 3-N and NH + 4-N are 10.09 and 11.45 mg·kg -1 in plant non-growing season with more drastic fluctuations when compared to the plant growing season(9.6 and 2.72 mg·kg -1). Soil organic nitrogen accounts for more than 97.0% of the total nitrogen at six vegetation types in two seasons. By the correlation analysis, there is a close relationship between NO - 3-N and NH + 4-N of 0-20 cm soil with the physical and chemical properties of soil itself for six vegetation types. The total nitrogen input to reservoir in the plant nongrowing season is higher than that in the growing season, and that of Rihui mountain watershed is significantly greater than that of Tianjia mountain watershed, 1.49 times and 1.55 times more than that of Tianjia mountain watershed in two plant seasons, respectively. The plant non-growing season shows a greater risk of nitrogen loss in this region, and it is also the key stage for the nitrogen pollution control, Metasequoia glyptostroboides forest and Echinochloa crusgalli(L.) Beauv show a larger capacity for storing and retaining nitrogen, which are suitable for large-scale construction to protect the water quality of Fushi Reservoir watershed.
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    Construction of Plant Expression Vector and Genetic Transformation Analysis of Arabidopsis thaliana CYCD2;1 Gene in Nicotiana tabacum
    DAI Li-Juan;ZHENG Tang-Chun;LIU Cai-Xia;LI Kai-Long;QU Guan-Zheng*
    Bulletin of Botanical Research    2016, 36 (2): 283-290.   DOI: 10.7525/j.issn.1673-5102.2016.02.018
    Abstract518)      PDF(pc) (1917KB)(385)       Save
    Cell division is one of the basic characteristics of organisms, which plays an extremely important role in the plant growth and development. CYCD2;1, as a regulatory factor, has been identified in regulation of cell cycle. In our study, CYCD2;1 was amplified from Arabidopsis inflorescences cDNA by PCR. Plant overexpression vector (pROKII- AtCYCD2;1) was constructed and transformed into wild-type tobacco by Agrobacterium-mediated leaf disc transformation. The transgenic plants were confirmed by PCR, and the result showed AtCYCD2;1 gene was integrated into tobacco genome. The result of qRT-PCR detection of T2 generation plants showed the mRNA of AtCYCD2;1 was transcribed in transgenic tobaccos. There were significant differences in the flower between the transgenic and wild type plant, such as the width of the corolla, the lengths of the petal and sepal, and increased volume of seed pod, which suggesting overexpression of AtCYCD2;1 has an obvious influence on the development of flowers.
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