Bulletin of Botanical Research

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Comparative Structural Observation of Stems of 15 Species in Pottiaceae from the Glacier Retreat

AIFEIRE Abuduaini, CHEN Qiu-Yan, WANG Hong, GENG Ruo-Nan, WU Yu-Huan

Bulletin of Botanical Research 2019, 39 (1): 1-9. DOI: 10.7525/j.issn.1673-5102.2019.01.001

Abstract （3340）

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We used paraffin sections and scanning electron microscopy(SEM) to observe the structure and surface micromorphological characteristics of the stems of 15 species of Pottiaceae that grown from the retreat of Glacier No.1. The stem of Pottiaceae was divided into two types with central axis and no central axes. The cell walls were thickened to different degrees. The stems of central axis were divided into three parts of epidermal, cortex and central axis. Stem epidermis cells are short with one layer, most of the cell wall protrudes outward, the surface is rough, and the surface decoration is mostly granular. The cortex is the largest area, most of which have differentiation of the inner and outer skin. The cell walls of most species gradually become thinner from the outside to the inside, and the cells are orderly arranged from small to large. The area of the central axis is also different, the cell wall corner is thickened. For no central axes, the cell walls were thickened to the same extent.

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Photosynthetic Characteristics and Leaf Anatomical Structure of Eight Tree Species

GANG Hui-Xin, WANG Jia-Qi, HUANG Chun-Yan, WANG Xiu-Wei

Bulletin of Botanical Research 2019, 39 (1): 10-16. DOI: 10.7525/j.issn.1673-5102.2019.01.002

Abstract （3755）

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Urban greening not only includes the beautification of landscaping, but also has important ecological functions, and its ecological function is realized through the physiological activities of plants. Photosynthetic capacity often varies appreciably among species and genotypes. Such variations are usually related to basic differences in metabolism and/or leaf anatomy. The experiment was conducted to study the influence of leaf anatomical structure on photosynthetic characteristics in eight tree species in Harbin. Photosynthetic rate( P _n), respiratory rate( R _d), transpiration rate( T _r), stomatal conductance( G _s) and intercellular CO ₂ concentration( C _i) were measured by a portable photosynthesis system(Model Li-Cor 6400). The leaf anatomical structure, including leaf thickness, epidermal thickness, palisade tissue thickness and sponge tissue thickness were measured by micrometer microscope. The maximum photosynthetic rate, G _s, C _i, T _r and light saturation point( LSP) were significantly different( P<0.05) in the eight species. The difference of epidermal thickness, palisade tissue thickness, upper epidermal stomatal density and lower epidermal stomatal density was significant in the eight trees( P<0.05). Although there was significant difference in photosynthetic characteristics and leaf anatomical structure among the eight species, we found that there was a correlation between them. There was a significant positive correlation between light saturation point and leaf epidermal thickness in the eight species( P<0.01), and the correlation coefficient was 0.78. The intercellular CO ₂ concentration was significantly negatively correlated with the upper epidermal stomata density( P<0.05), and the correlation coefficient was -0.65. The maximum photosynthetic rate, R _d, T _r and LSP were not significantly correlated with epidermal thickness, palisade tissue thickness, sponge tissue thickness, upper epidermal stomatal density and epidermis stomatal density( P>0.05). There was no significant correlation between intercellular CO ₂ concentration and epidermal thickness, palisade tissue thickness, sponge tissue thickness and epidermis stomatal density( P>0.05). There was no significant correlation between light saturation point and palisade tissue thickness, sponge tissue thickness, upper epidermal stomatal density and epidermis stomatal density( P>0.05). Although there is still a great deal to learn how the leaf anatomical structure effect on physiological process, the measurements of leaf anatomical structure have great potential to provide a better understanding of physiological process.

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Pollen Morphology of 15 Cultivars of Freesia hybrida

SUN Yi, DING Su-Qin, SHI Yi-Min, TANG Dong-Qin

Bulletin of Botanical Research 2019, 39 (1): 17-26. DOI: 10.7525/j.issn.1673-5102.2019.01.003

Abstract （976）

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Pollen morphology of 15 cultivars of Freesia hybrida was measured using scanning electron microscopy(SEM), and then the similarity and differences among cultivars were evaluated by the shape, size, exine ornamentation and aperture of pollen. The pollens of the 15 cultivars of F.hybrida are all single and zygomorphic with far polar single germinal furrow. All pollen grains are elliptical in shape and boat-like or heart-like in polar view while long spheroidal shape in equatorial view of all investigated cultivars except Castor. The exine of Freesia was interspersed with spininess, and the pollen surface of most cultivars has circular holes and circular markings. Furthermore, the 15 cultivars of Freesia were classed into three groups based on these pollen characteristics by cluster analysis. Taken all together, pollen morphology of Freesia plants was studied in the present study at the first time, and we also found the moderate differences of pollen morphology possessed among cultivars, especially in the characters of exine ornamentation and aperture of pollen, which demonstrated the genetic diversity and divergence in Freesia. Our findings will be valuable to analyze the phylogenetic relationship among the Freesia accessions in future.

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Cytological Features of Gametophyte Development and Zygotic Formation in Senecio scandens Buch.-Ham. ex D.Don(Compositae)

XIE Xin, QIAN Qiu-Bo-Yan, WANG Lei, WANG Qian-Xing, PING Jun-Jiao, HE Li-Fang, QIAN Gang

Bulletin of Botanical Research 2019, 39 (1): 27-34. DOI: 10.7525/j.issn.1673-5102.2019.01.004

Abstract （690）

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Senecio scandens Buch.-Ham. ex D.Don(Compositae) is a crucial plant source of Chinese traditional medicine with antibacterial properties. To obtain basic knowledge for an insight into its cytology mechanism and further practical breeding application in this species, we focus on characterizing progress on gametophyte developments and zygote formation in this paper. Based on a chronological description of the male gametic development, two dimorphic sperm cells release from their separate position in the male germ unit. Moreover, mature embryo sac exhibits an ovoid to pyriform shape locating at wider micropylar extreme, by following the monosporic pattern on development of embryo sac in the current species. It is therefore reasonable to deduce that the origin of apico-basal polarity of zygotic embryo may be related to an unevenly spaced distribution of cytoplasm in zygotic embryo which results from fertilization between the dimorphic sperm cells and the polar ovule. As the distinguishable characteristics, these morphological features will be usefulness for the taxonomy studies in other Compositae species, including dimorphic sperm cells, polar embryo sac and zygotic polarity.

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Component Analysis of Volatile Organic Compounds Released by Branch and Leaf of Quercus variabilis and Response to Mechanical Injury in Beijing Pinggu

ZHANG Yu-Ting, WANG Xiao-Fei, NI Yan-Yan, LIU Jian-Feng, JIANG Ze-Ping

Bulletin of Botanical Research 2019, 39 (1): 35-44. DOI: 10.7525/j.issn.1673-5102.2019.01.005

Abstract （881）

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The experiment was conducted to study the component of VOCs released by branch and leaf of Quercus variabilis and response to mechanical injury in Beijing Pinggu. The plant volatile organic compounds(BVOCs) from branches and leaves of Q.variabilis were collected and analyzed with GC-MS. Eight kinds of volatile components, such as esters, alkanes, terpenes, aromatic hydrocarbons, aldehydes and ketones, alcohols, alkenes, as well as other N-containing, S-containing and Cl-containing compound were detected. Esters and alkanes were the main components of Q.variabilis, and account for 30.32%-41.60% and 30.02%-33.14% of the total plant volatile organic compounds content. The comparison of undamaged and mechanically-damaged leaves of Q.variabilis indicated that they had similar VOCs, while the components and contents of volatile substances changed. Dibutyl phthalate, 1-Hexanol, 2-ethyl-, Tetradecane, 2,6,10-trimethyl-, Cyclohexane, 2-ethenyl-1, 1-dimethyl-3-methylene-, Tetradecane, Nonadecane, 9-Octadecenamide,(Z)-, Nonanal, Decanal, etc. were the main components. Mechanical damage brings different degree effects on components and relative content of volatile organic compounds of Q.variabilis branches and leaves. Naphthalene, d-Limonene, 1,3,6-Octatriene, 3,7-dimethyl-,(Z)-,4-Hexen-1-ol, acetate, Dibutyl phthalate, Acetic acid, hexyl ester, Benzoic acid, butyl ester, Cyclohexane, 2-ethenyl-1,1-dimethyl-3-methylene-could be induced and their content increased obviously after mechanical damage. Esters and alkanes were the main VOCs components of Q.variabilis. The contents of naphthalene, d-Limonene, 4-Hexen-1-ol, acetate, Dibutyl phthalate were increased obviously after mechanical damage and these substances had certain ecological function and effect on the anti-herbivory. This may be a chemical defense strategy of Q.variabilis in response to herbivory or against external stress. In addition, many components also changed at different degree or small amplitude after mechanical damage. It's to be further studied on their change mechanism and ecological function.

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Vertical Distribution of Hypopterygiaceae and Their Relationship with Environmental Factors in Karst Tiankeng

WU Jin, LI Cheng-Yi, LI Xiao-Fang, ZHANG Zhao-Hui

Bulletin of Botanical Research 2019, 39 (1): 45-53. DOI: 10.7525/j.issn.1673-5102.2019.01.006

Abstract （954）

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The karst Tiankeng is an unique negative landform with a huge and enclosed space, which forms a special habitat suitable for bryophyte growth and development. In order to study the relationship between environmental factors and distribution of Hypopterygiaceae, and their vertical distribution characteristics in Monkey-Ear Tiankeng(diameter of 300 m, and depth of 280 m) in Guizhou Province, 12 sampling sites in the Tiankeng were investigated in our fieldwork, and laboratory analysis and Canonical Correspondence Analysis(CCA) were used, and compared with the bryoflora of other sampling sites like karst rocky desertificated zone and Tiankeng marginal zone. The results showed that:(1)There were three genera and nine species of Hypopterygiaceae in the Tiankeng, accounting for 47.37% of the total species of this family in China, and accounting for 69.23% of the total species of this family in Guizhou, of which the Hypopterygiumflavo-limbatum C.Muell was more common species in six spots with a frequency of occurrence 50%; (2)Air humidity, light, canopy density, and human disturbance were the main environmental factors affecting the distribution of Hypopterygiaceae in the Monkey-Ear Tiankeng. The descending order in the influence of environmental factors was soil water content, soil pH, gradient, and air temperature; (3)The descending order in species richness of Hypopterygiaceae was karst Tiankeng,Tiankeng marginal zone,and desertification area; (4)In the vertical distribution pattern, the Hypopterygiaceae showed that the bottom of the Tiankengwas the center of biodiversity, and the distribution from the crater to the bottom was gradually increasing. It is a natural refuge for karst rocky desertification.

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Effects of Different Nitrogen Level on Growths of Moringa oleifera Seedling under Northern Facilities of China

XU Hai-Jun, CHENG Xin-Yu, WANG Xiao-Fei, GUAN Xiang-Jun

Bulletin of Botanical Research 2019, 39 (1): 54-60. DOI: 10.7525/j.issn.1673-5102.2019.01.007

Abstract （776）

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In order to clarify the rule of Moringa oleifera for nitrogen demand on fast-growing period, the experiment was carried out under five nitrogen level gradients applied on Moringa seedling, and to study the difference of characteristics on the growth of height, ground diameter, the contents of N, P, K in leaf, the contents of blade pigment and photosynthetic characteristics. The result showed that the height, ground diameter, N, P, chla, total pigment, carotenoid in leaf and photo-quantum efficiency rose first and descended later accompanied with nitrogen level increasing, while the light compensation point showed the contrary. Plant height growth was significantly correlated with leaf P and chla, and ground diameter was significantly correlated with dark respiration rate and leaf K content. When the nitrogen level was in 30 g·ind. ^-1(N ₃), M.oleifera seedling growth was the fastest, the plant nutrient status was better, and the utilization rate of light was the best.

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Relationship between Soil Moisture Dynamics and Vegetation Community in the Loess Area of Western Shanxi Province

QIANG Fang-Fang, WEI Tian-Xing, LIU Wei

Bulletin of Botanical Research 2019, 39 (1): 61-68. DOI: 10.7525/j.issn.1673-5102.2019.01.008

Abstract （816）

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Water availability is the most limiting factor for plants in forest ecosystems of semi-arid regions. To determine the spatiotemporal dynamics of soil moisture under different vegetation community in loess hilly region, by the field test, three typical vegetation communities( Pinus tabuliformis forest, Robinia pseudoacacia forest, and natural secondary forest) were investigated. By analyzing the temporal and spatial variation characteristics of soil water and the relationship between soil moisture and precipitation, temperature and soil nutrient, the results showed that:(1)In the past ten years, the rainfall distribution in the study area was extremely uneven, and characterized by insufficient pre-rainfall period and concentrated in the later stage. The natural secondary forest soil had the largest soil moisture, and R.pseudoacacia forest was the smallest, and there were significant differences between them, both of them were not significant difference with P.tabuliformis forest; (2)There were differences in the distribution of roots in the 0-60 cm soil layer of the three vegetation communities in the study area. Among them, the distribution of roots in different soil layers of the same planting community was significantly different. In different vegetation communities, the root distribution of natural secondary forests was significantly different with P.tabuliformis and R.pseudoacacia forests in the 20-60 cm soil layer; (3)The soil moisture of the vegetation community in the study area had obvious stratification, the soil moisture in the topsoil(0-40 cm) varies greatly, while the 40-100 cm soil layer had less fluctuation and basically maintained at 10%-15%. The soil moisture change in topsoil(0-40 cm) was related to the distribution of vegetation roots, and significantly negatively correlated with soil moisture; (4)Through analyzing the relationship of soil moisture and rainfall, temperature, organic carbon, total nitrogen, total phosphorus, total potassium in topsoil, soil moisture is significantly positively correlated with rainfall, and the correlation coefficient of P.tabuliformis forest is the highest. Temperature is only positively correlated with the soil moisture of the secondary forest. Soil moisture is positively correlated with organic carbon, total nitrogen, total phosphorus and total potassium, and the correlation with total phosphorus is the highest.

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Genetic Diversity of Ampelocalamus luodianensis in Different Microhabitats

LIU Ji-Ming, GUAN Rui-Ting, WANG Min, CHEN Jing-Zhong, TONG Bing-Li, WU Meng-Yao

Bulletin of Botanical Research 2019, 39 (1): 69-77. DOI: 10.7525/j.issn.1673-5102.2019.01.009

Abstract （3043）

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By random amplified polymorphic DNA(RAPD) markers analysis, 5 kinds of karst microhabitats of Ampelocalamus luodianensis was conducted to study genetic diversity and genetic structure. Through 8 RAPD primers, 87 discernible DNA fragments in all weighted from 200-3 000 bp were cloned, and 68 in those were polymorphic loci, with percentage of polymorphic loci at the species level, PPL=78.16%, Nei's index H=0.291 3, and Shannon's Information Index I=0.431 0. The A.luodianensis grew on soil surface(TM) possessed the highest level of genetic diversity( PPL=47.13%, H=0.195 3 and I=0.283 4), while the A.luodianensis in stone crevice exhibited the lowest levels of genetic diversity( PPL=39.08%, H=0.147 4 and I=0.219 2). Coefficient of genetic differentiation ( G _st) was 0.415 1(from the POPGENE32), and the gene flows( N _m) was 0.704 5. Among the 5 microhabitats, genetic identity varied from 0.777 3-0.999 4, genetic distance was different from 0.000 6-0.251 9, and the mean of them were 0.865 6-0.148 3 severally. Genetic diversity in niches at mean value was with PPL of 43.91%. Coefficient of genetic variation in niches was with G _st of 0.4054. The results indicated that differences of microhabitats increased the genetic diversity of A.luodianensis.

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Effects of Human Disturbance on Species Diversity and Soil Physical and Chemical Properties of Revetment Forest in Wenjiang Section of the Jinma River

ZHANG Xiao-Yue, QI Jin-Qiu, ZHANG Liu-Hua, WANG Mei-Zhen, LI Ting-Ting, YU Shun-Yao, HAO Jian-Feng

Bulletin of Botanical Research 2019, 39 (1): 78-86. DOI: 10.7525/j.issn.1673-5102.2019.01.010

Abstract （779）

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By the typical plot method, the interference theory and species diversity analysis method, and redundant analysis(RDA), we studied the relationship between 11 environmental factors and the diversity of revetment forest species, and explored the different disturbances of human disturbance to the Wenjiang section of the Jinma River. The impact of species diversity and soil physical and chemical properties on revetment forests. The results showed that:(1)Among the 264 different levels, 122 species of plants were recorded, belonging to 65 families and 102 genera, of which the herbaceous layer had the largest number of species, with 89 species, accounting for 73% of the total records, with the increase of human disturbance intensity. The number of species is gradually decreasing; (2)Human disturbance has a negative impact on the species diversity level of the Jinma River revetment community. With the increase of disturbance intensity, the Shannon-Wiener diversity index( H) and species richness index( D) of the community are gradually reduced, and on the arbor layer the species composition is single, resulting in a decline in community environmental function and stability; (3)The pH value of the soil in the community was alkaline, and the soil organic matter content was high. The physical and chemical properties of the soil under different disturbance intensity were significantly different( P<0.05); (4)Soil water content, available potassium and organic matter content are the most important factors affecting the species diversity of revetment forests.

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Genetic Diversity Characteristics in Different Dominance Hierarchies of Pinus yunnanensis Franch. Trees

CAI Nian-Hui, XU Yu-Lan, WANG Ya-Nan, CHEN Shi, WANG Meng-Ting, LI Gen-Qian

Bulletin of Botanical Research 2019, 39 (1): 87-95. DOI: 10.7525/j.issn.1673-5102.2019.01.011

Abstract （897）

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The trees were divided into the dominant trees, the intermediate trees and the suppressed trees according to the height and diameter(ground diameter) parameters. The variation of genetic diversity characteristics in different dominance hierarchies of Pinus yunnanensis Franch. population was studied by SSR molecular markers. In forest canopy layer, the mean number of effective alleles, Shannon's information index, observed heterozygosity and expected heterozygosity of dominant trees were 2.083, 0.762, 0.290 and 0.423, respectively, for the dominant trees, which slightly exceeded the intermediate and suppressed trees. In regeneration layer mean number of effective alleles, Shannon's information index, observed heterozygosity and expected heterozygosity were 2.063, 0.774, 0.272 and 0.410, respectively, for the dominant tree, which slightly exceeded the intermediate and suppressed trees except for observed heterozygosity. There were no significant differences among the different dominance hierarchy trees for the genetic diversity by the difference significance test of statistics method.

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Cloning and Expression Analysis of BpSPL2 Promoter from Betula platyphylla

WANG Sheng-Yu, ZHANG Qi, ZHANG Zheng-Yi, HU Xiao-Qing, TIAN Jing, ZHANG Yong, LIU Xue-Mei

Bulletin of Botanical Research 2019, 39 (1): 96-103. DOI: 10.7525/j.issn.1673-5102.2019.01.012

Abstract （944）

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The SPL(SQUAMOSA promoter-binding protein-like) is a plant specific transcription factor, and the study showed that it plays an important role in the transformation of developmental stage, flower and fruit development. A 1 960 bp promoter sequence of BpSPL2 gene was cloned from Betula platyphylla genomic DNA using the method of PCR. The cis-regulatory elements were analyzed by PLACE and PlantCARE web tools. Multiple flowering elements, abiotic stress response elements and hormone-responsive elements were predicted in the promoter region. It indicated that it played an important role in plant growth and development and stress response.Further, the BpSPL2 promoter was inserted to pBI121 vector under control of the 35S promoter to generate the pBI121- BpSPL2 promoter:: GUS recombinant construct, which was transient expressed in B.platyphylla and Arabidopsis thaliana seedlings by Agrobacterium tumefaciens mediated method, then investigated the expression pattern via histechemical GUS staining. BpSPL2 promoter could drive the expression of GUS gene in B.platyphylla and A.thaliana, and expressed in high level in leaves, buds, and roots of B.platyphylla while in high level in anthers, pistils, and leaves of A.thaliana. This study will provide a reference for further studies on the expression regulation and functional analysis of BpSPL2 gene in B.platyphylla.

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Relationship between H ₂S and ABA Signaling in Arabidopsis thaliana under Drought Stress

ZHAO Min, WANG Yue-Xuan, XU Yun-Fei, ZHAO Qi-An, LIU Bo, YANG Ning

Bulletin of Botanical Research 2019, 39 (1): 104-112. DOI: 10.7525/j.issn.1673-5102.2019.01.013

Abstract （1397）

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With the seedlings of wild type Arabidopsis thaliana(WT), hydrogen sulfide(H ₂S) synthase deletion mutant lcd, abscisic acid(ABA) deletion mutant aba1, 0.3 mol·L ^-1 mannitol simulated drought stress, we studied the effects of drought stress on ABA content and H ₂S content, analyzed their roles and signal relationships in drought tolerance. Drought stress significantly increased the LCD and ABA1 gene relative expressions, contents of H ₂S and ABA; Drought stress significantly inhibited the seed germination of lcd and aba1 mutants; Application of NaHS and ABA significantly increased the endogenous H ₂S content in WT under drought stress; Application of NaHS could promote the endogenous H ₂S production in WT, lcd and aba1, the relative expression of LCD and ABA1 also up-regulated under drought stress. While the application of ABA increased the H ₂S content and the LCD, ABA1 relative expressions in WT and aba1 under drought stress, but had no significant effect on the H ₂S content and LCD relative expression in lcd. The results showed that the signal molecules H ₂S and ABA played a vital role response to the drought stress in Arabidopsis, and H ₂S is located at downstream of ABA to participated in the regulation of signaling process.

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Cloning and Expression Analysis of S-Adenosyl-L-Methionine Synthetase( ThSAMS) Gene from Tamarix hispida

ZHANG Yue, ZHAO Xin, HOU Zheng, WANG Yan-Min, WANG Yu-Cheng, WANG Chao

Bulletin of Botanical Research 2019, 39 (1): 113-122. DOI: 10.7525/j.issn.1673-5102.2019.01.014

Abstract （739）

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A full length cDNA of a S-Adenosyl-L-Methionine Synthetase gene(named ThSAMS) was isolated from the transcriptome cDNA librarys of Tamarix hispida. The open reading frame of ThSAMS cDNA was 1 185 bp encoding 394 amino acids. The relative molecular weight of ThSAMS protein was 97.85 kDa, with isoelectric point of 5.02. Multiple alignment analysis based on the amino acids indicated that ThSAMS had high homology to SAMS proteins from different species, and the highest similarity of 95% to the Ziziphus jujuba SAMS. Quantitative real-time PCR(qRT-PCR) assay revealed that ThSAMS gene can respond to NaCl, PEG and ABA stress, suggesting that it may be involved in salt and drought stresses in T.hispida. This study provided theoretical groundwork for further exploration into the function of stress tolerance of SAMS gene in plants.

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Antisense CCoAOMT Gene Regulates Lignin Biosynthesis in Betula platyphylla

YAO Lian-Mei, HU Xiao-Qing, ZHOU Fei, ZHENG Yao-Qiang, WANG Guo-Dong

Bulletin of Botanical Research 2019, 39 (1): 123-130. DOI: 10.7525/j.issn.1673-5102.2019.01.015

Abstract （734）

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Betula platyphylla is an important forestation tree species in northern China, but its high lignin content severely restricts its development and utilization as a papermaking resource plant. In this paper, the full-length ORF sequence of CCoAOMT(Caffeoyl-coenzyme A-3-O-methyltransferase) gene was obtained using RACE technology. The antisense CCoAOMT expression vector was constructed and transformed into B.platyphylla by Agrobacterium infection method. The molecular detection indicated that the CCoAOMT antisense gene had been successfully integrated into B.platyphylla. By semi-quantitative PCR analysis of the transgenic plants, the CCoAOMT gene expression of the transgenic lines was decreased. Compared to the wild type, by the Wiesner staining, the lignin content of the transgenic plants was decreased. Chemical compositions of seven-year-old transgenic and wild-type birch were analyzed. The phenyl alcohol extracts and Klason lignin of transgenic birch were significantly reduced, and the pentosan content was increased. The BpCCoAOMT gene plays an important role in the synthesis of lignin, which lays a foundation for the cultivation of a new species of birch with low lignin content.

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Variation Analysis of Phenotypic Traits in F ₁ Population of Rosa spp

ZHOU Li-Jun, YU Chao, CHANG Xiao, WAN Hui-Hua, LUO Le, PAN Hui-Tang, ZHANG Qi-Xiang

Bulletin of Botanical Research 2019, 39 (1): 131-138. DOI: 10.7525/j.issn.1673-5102.2019.01.016

Abstract （3294）

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Heredity of some characters of flower and leaf in F ₁ individuals of mother( Rosa ‘Yunzheng Xiawei’) and father( R. ‘Sun City’) were measured and analyzed with variance, coefficient of variation, inheritance and correlation. The coefficient of variation in the hybrid population was 7.33%-68.08%, indicating a significant phenotypic variation. Among them, the number of petals in the population showed the highest degree of variation. All the traits measured had high degree of dispersion and came out new phenotypic traits, different from parents. The analysis of the genetic characteristics and correlation of phenotypic traits will benefit for selecting breeding and assisting breeding for dominant genes in controlling phenotypic traits.

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Cloning and Expression Analysis of Two PLT Transcription Factors Genes in Fraxinus mandshurica

DING Yi-Wei, ZHAN Ya-Guang, ZHANG Jia-Wei, HE Li-Ming

Bulletin of Botanical Research 2019, 39 (1): 139-147. DOI: 10.7525/j.issn.1673-5102.2019.01.017

Abstract （1264）

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In order to investigate the function and mechanism of the PLT transcription factor in the root development of Fraxinus mandshurica, we cloned two PLT transcription factors from F.mandshurica, and named them FmPLT2 and FmPLT3 by homologous alignment, encoding 532 and 497 amino acid residues, respectively. By bioinformatics analysis, the relative molecular masses of FmPLT2 and FmPLT3 were 59 and 55 kDa, respectively, and the isoelectric points were 5.98 and 5.79, respectively. They were all hydrophilic unstable proteins, and both contained two AP2 conserved domains. By phylogenetic analysis, FmPLT2 and FmPLT3 had the highest homology with the OePLT2 and OeOLT3 genes of the Oleaceae, respectively. Subcellular localization predictions showed that both FmPLT2 and FmPLT3 proteins were mainly concentrated in the nucleus. The expression of FmPLT2 and FmPLT3 in different tissues and roots of different developmental stages was analyzed by qRT-PCR. The tissue-specific expression of FmPLT2 and FmPLT3 was similar and they all have the highest expression in roots and the lowest expression in leaves. The expression level of FmPLT2 in roots at different developmental stages is extremely significant. Especially, the expression level of FmPLT2 on the 21st day of root development is 32 times as high as that at the 0 day, but the expression of FmPLT3 was not significant, indicating that FmPLT2 not only plays an important role in root development, but also may participate in multiple developmental pathways such as cell proliferation and growth.

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Characteristics of Microsatellite in Camellia saluenensis by High-throughput Sequencing

WANG Da-Wei, ZHOU Fan, SHEN Bing-Qi, WANG Lian-Chun

Bulletin of Botanical Research 2019, 39 (1): 148-155. DOI: 10.7525/j.issn.1673-5102.2019.01.018

Abstract （847）

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Leaves sample of Camellia saluenensis Stapf ex Bean was sequenced with high-throughput sequencing technology(Illumina Hiseq 2000). In total 140 996 transcripts of C.saluenensis were screened using MISA software. The 32 696 SSRs were identified, and the frequency of SSR was 23.2% and mean distribution density was dinucleotide 32.02 kB. The di-nucleotide repeats were the highest, followed by tri-nucleotide and mono-nucleotide repeat types. The tetra-nucleotide, penta-nucleotide and hexa-nucleotide were all less than 1%. Among the mononucleotide repeats, the A/T repeats motifs were the highest frequency(10.92%). AG/CT was the most frequent repeat motifs in di-nucleotide repeats(49.72%), followed by AT/AT and AC/GT. CG/CG were minimum. AAG/CTT repeats motifs was the highest in tri-nucleotide followed by ACC/GGT, ATC/ATG and AGG/CCT, while CCG/GGC, ACT/AGT and ACG/CGT were lowest(<1%). The repeat motifs were very few in tetra-nucleotide, petra-nucleotide and hexa-nucleotide and had no obvious differences. The microsatellite number in C.saluenensis decreased with the increased number of repeats and length of repeats.

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Sequencing and Assembly of Populus simonii× P.nigra Genome and SSR Analysis

ZHOU Yu-Min, WANG Sui, LIU Yi, LI Kai-Long, YOU Xiang-Ling

Bulletin of Botanical Research 2019, 39 (1): 156-160. DOI: 10.7525/j.issn.1673-5102.2019.01.019

Abstract （839）

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Populus simonii× P.nigra, which is the hybrid crossed by P.simonii and P.nigra, inherits the advantages of fast growth and strong resistance from their parents. In this study, the genome sequence of Populus simonii× P.nigra was preliminarily sequenced and assembled by the next generation sequencing(NGS) technology. The SSR sequences were identified and analyzed. We hope this study can provide a reference for classification and phenotypic correlation of Populus simonii× P.nigra. The results show that we total assembled 368.96 Mbp genome sequence, which contain 366 876 contigs. SSR analysis was performed on 21 788 non-redundant contigs which were not less than 2 000 bp. In total, 18 111 SSR sequences were identified, and most of them are one, two or three nucleosides acid repeat motif. The primers were designed for the obtained SSR sequences, and a total of 12 838 primers were obtained for future experiments.

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Characteristics of Leaf Epidermis of Meconopsis integrifolia under Different Altitudes in the Qinghai-Tibet Plateau

TANG Nan, LI Miao-Miao, TANG Dao-Cheng

Bulletin of Botanical Research 2019, 39 (2): 161-168. DOI: 10.7525/j.issn.1673-5102.2019.02.001

Abstract （1459）

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Characteristics of leaf epidermis of Meconopsis integrifolia were observed under optical microscope, including trichome, stomata and epidermis cell. The relationship between epidermis trichome, stomata, epidermis cell of leaf and altitude was discussed. The trichome density, stomata density and epidermis cell density of both upper and lower leaf epidermis increased gradually as the altitude increased. While the length, width and area of both stomata and epidermis cell decreased. Morphology of epidermis cell changed from irregular to polygonal. The shape of anticlinal wall altered from corrugated to arched or straight. These variation of characteristics probably be a general response of plants which living under plateau environment in a long-term, providing theoretical support for further study on alpine plant adaptation.

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Cuticle Micromorphology of Podocarpus from China and Its Taxonomic Significance

SUN Tong-Xing, DONG Yun-Qiu, HOU Xue-Liang

Bulletin of Botanical Research 2019, 39 (2): 169-180. DOI: 10.7525/j.issn.1673-5102.2019.02.002

Abstract （1136）

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We studied the cuticle micromorphology of leaves from 8 species and 2 varieties of Podocarpus with scanning electron microscopy. The leaf epidermal structures of Podocarpus revealed remarkable consistency. Epidermal cells are rectangular or irregular in shape, the outlines of epidermal cells are slightly undulating with prominent butresses. The stomata are oriented parallel to the long axis of the leaf. Floring rings and stomatal plugs are usually present. The guard cells have prominent polar extension and stomata usually have two to four subsidiary cells, often with polar subsidiary cells lacking. However, there are more or less discrepancies in the arrangement of stomatal apparatus, the shape of epidermal cells within these species. In P.falcatus and P.gracilior, stomatal apparatus distributes on both surfaces, cuticle flanges on epidermal cells are straight and without buttress. In other taxa, stomatal apparatus is only restricted to abaxial surfaces. On both adaxial and abaxial surfaces, the cuticle flanges on epidermal cells in P.henkelii are thick and with prominent buttresses. In P.wangii, the epidermal cells of adaxial surfaces are fairly regular, most cells are tetragonum and perpendicular to the long axis of leaf vein, the cuticle flanges are more prominent and extending to the hypodermis. In P.costalis, the adaxial epidermal cells are irregular and polygonous. In P.forrestii, the stomatal intervals are less and the adaxial epidermal cells are shorter, the cuticle flanges are less pronounced. In P.annamiensis, the stomatal intervals are relatively wide, the stomata exhibit broad elliptic, the epidermal cells of adaxial surfaces are thin rectangle in outline. In addition, the shape of epidermal cells on both surfaces and buttress characteristics in P.macrophyllus var. maki and P.macrophyllus var. angustifolius are distinctly different from those in P.macrophyllus. The epidermal cells of adaxial surfaces in P.macrophyllus var. maki are irregular and sinuate in outline, the flange between epidermal cells are fairly thick. In P.macrophyllus var. angustifolius, the epidermal cells of adaxial surfaces are square to rectangular, in outline, often shorter than those in P.macrophyllus, and the flange between epidermal cells are thicker than that in P.macrophyllus. Therefore, these differences of cuticle micromorphology may be used to distinguish species or variety in Podocarpus.

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Effects of Exogenous Hydrogen Peroxide on Growth and Resistance Physiology of Naked Oat Seedlings under Saline-alkali Mixed Stress

LIU Jian-Xin, OU Xiao-Bin, WANG Jin-Cheng

Bulletin of Botanical Research 2019, 39 (2): 181-191. DOI: 10.7525/j.issn.1673-5102.2019.02.003

Abstract （1028）

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The experiment was conducted to study whether exogenous hydrogen peroxide(H ₂O ₂) could improve the saline-alkali tolerance of naked oat( Avena nuda) seedlings and its physiological mechanism. Seedlings of naked oat(‘Dingyou No.6’) at three-leaf period cultured with perlite in a solar greenhouse were irrigated with the Hoagland nutrient solution containing 75 mmol·L ^-1 saline-alkali mixed solution(NaCl∶Na ₂SO ₄∶NaHCO ₃∶Na ₂CO ₃=12∶8∶9∶1) combination with dimethylthiourea(DMTU, a quencher of H ₂O ₂), and the leaves were sprayed with 0.01 mmol·L ^-1 H ₂O ₂. Seedlings were irrigated with Hoagland nutrient solution, and the leaves were sprayed with water served as the control. Seedlings growth, photosynthetic pigment content, active oxygen metabolism, and osmotic regulatory substance accumulation in leaves were measured. The results showed that exogenous H ₂O ₂ could alleviate the inhibition effects of saline-alkali mixed stress on seedlings growth of naked oat. Spraying H ₂O ₂ significantly increased root length, plant height, dry weight, the leaf chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents, and the superoxide dismutase, peroxidase, catalase, ascorbate peroxidase activities of naked oat seedlings under saline-alkali mixed stress, but decreased the contents of superoxide anion, H ₂O ₂, malondialdehyde, ascorbic acid, glutathione, and free amino acids. Treatments with spraying H ₂O ₂ also increased the contents of antioxidants(such as flavonoids, total phenols, procyanidins) and osmotic regulatory substance(such as soluble protein, soluble sugars, and proline) in leaves. The above effects of exogenous H ₂O ₂ were partially or completely reversed by DMTU. A comprehensive evaluation based on subordinate function analyses revealed that foliar spraying with H ₂O ₂ could improve the saline-alkali resistance of naked oat seedlings, and the promoting effect of H ₂O ₂ could be completely reversed by DMTU. Therefore, the exogenous H ₂O ₂ supply can increase saline-alkali adaptability in naked oat seedlings by regulating active oxygen metabolism and osmotic regulatory substance accumulation in leaves of naked oat plants under saline-alkali mixed stress.

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Nutritional Effect and Rhythm of Spring and Summer Flowering in Magnolia soulangeana‘Changchun’

JIANG Zheng, SUN Li-Yong, LIU Xu, LIU Chen-Ni, YIN Zeng-Fang

Bulletin of Botanical Research 2019, 39 (2): 192-199. DOI: 10.7525/j.issn.1673-5102.2019.02.004

Abstract （1337）

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In order to systematically master the phenological rhythm of spring and summer flowering, and discuss its relationship with nutrients in Magnolia soulangeana ‘Changchun’, a six-year-old M.soulangeana ‘Changchun’ tree was taken as material, the annual growth and developmental rhythm, phenological characteristic of spring and summer flowering were observed, and the changes of nutrient content in the stem were tested. The results were: (1)The dormant period of M.soulangeana ‘Changchun’ was from December of this year to late February of the next year, and then its flower buds began to burst in the late February. The spring flowering lasted for 20 days in March. The vegetation growth began in April. The flower bud differentiation completed in May, and burst again at the end of May partly, followed by the summer flowering in June. The summer flowering period lasted for nearly 20 days. The developmental lag period of unexpanded flower buds was from July to September. In addition, a small amount of lateral buds at the base of flowers bloomed in summer were differentiated into flower buds. From October to December, as the leaves fallen off, the tree entered the dormant period gradually. (2)The flower buds of M.soulangeana ‘Changchun’ that differentiated after vegetative growth had ability to flowering twice in spring and summer. The spring flowering was characterized by anthesis posterior leaf, and the rate of flowering synchronization was high. The flowering rate of spring flowering was about 100%. Furthermore, the flowers were fertile with normal pistils and stamens. The summer flowering had the characteristic of flowering and leafing at the same time, but flowering synchronism was lower. The flowering rate of summer flowering was about 30%. Expressly, the flowers were sterile with abnormal pistils and stamens. (3)Soluble sugar and soluble protein were shown with a trend of descending among spring flowering, while starch content was decreased at the late flowering stage. And the contents of soluble sugar and starch were shown with a trend of first descending then rising, while the soluble protein content continued to decline during the summer flowering. Therefore, there were some differences in the spring and summer flowering pattern of M.soulangeana ‘Changchun’, which was related to changes of nutrients content. It was suggested that low levels of soluble sugar and high levels of starch and soluble protein were beneficial to the initiation of spring flowering, while low levels of soluble protein and high levels of soluble sugar and starch were contributed to the realization of summer flowering.

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Effects of Clonal Integration on Biomass Allocation and Leaf Structure of Glechoma longituba(Nakai) Kupr in Different Water Availability

XIANG Yun-Rong, ZHANG Fang, DUAN Jing, HUANG Hui-Min, HE Dan-Ni, LIU Yuan, TAO Jian-Ping

Bulletin of Botanical Research 2019, 39 (2): 200-207. DOI: 10.7525/j.issn.1673-5102.2019.02.005

Abstract （1034）

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Because of discontinuous soil coverage, poor soil fertility and soil water scarcity, the high heterogeneity of karst habitats seriously restricts the growth and distribution of local plants. To clarify the adaptive strategies of clonal plants in karst area, two connected ramets with a single internode of Glechoma longituba were planted in yellow calcareous soil. Then, we cut off the half of the internode and made the ramets disconnected, supplied different amounts of water to clarify the effects of the clonal integration on biomass accumulation, distribution, stomata and tissue characteristics of leaves under different treatments. The results showed that clonal integration significantly promoted the biomass accumulation of G.longituba, and increased biomass allocation to roots and leaves. The stomatal conductance were increased and the stomatal index were decreased by clonal integration of leaves. There was less affected on leaves sponge tissue, but the palisade tissue and the ratio of palisade to spongy tissue showed that the non-clonal integration ramets were higher than the integrated ramets. The clonal integration could increase the investment of roots and leaves of G.longituba, and further promoted clonal plant survival and adaptation in karst habitats with better leaves stomata and tissue structure.

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Relationship between Anatomical Structures and Heat Resistance of Davidia involucrata Natural Populations in Sichuan Province

ZHANG Teng-Ju, CHEN Xiao-Hong, LIU Jing, KANG Xi-Kun

Bulletin of Botanical Research 2019, 39 (2): 208-221. DOI: 10.7525/j.issn.1673-5102.2019.02.006

Abstract （4042）

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In order to understand the heat resistance mechanism and the level of heat resistance of Dove tree( Davidia involucrata) populations in Sichuan Province, 16 anatomical structures of 15 representative natural populations in Sichuan Province were investigated by paraffin section and NaClO. Moreover, coefficient of variation, correlation analysis, and cluster analysis were used to screen representative structures. The 16 anatomical structure indexes all reached a significant level difference among 15 natural populations( P<0.01), thickness ratio of palisade and spongy tissue, thickness of palisade tissue, stoma density, cuticle thickness of lower epidermis, protuberant degree of main vein, cuticle thickness of upper epidermis, cuticle thickness of upper epidermis and thickness of lower epidermis were the main factors related to the heat resistance, while other indexes, such as leaf thickness, sponge tissue thickness, thickness of main vein, thickness of epidemic cell, thickness of vascular bundle in main vein stoma length, stoma width, tightness of leaf structure and looseness of leaf structures did not show much effect on heat resistance. There were some differences among 15 populations in heat resistance, and the descending order was Hanyuan, Junlian, Leibo, Muchuan, Pingshan, Yingjing, Tianquan, Meigu, Pingwu, Bifengxia, Beichuan, Hongya, Baoxing, Dayi, Qionglai. According to the heat hardiness, these populations could be divided into four groups, Hanyuan; Jun Lian, Leibo and Muchuan; Pingshan, Yingjing, Tianquan, Meigu, Pingwu, Bifengxia, Beichuan LR; and Hongya, Baoxing, Dayi, Qionglai.

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Physiological Response of Suspension Cells of Helianthus tuberosus to NaCl Stress

CAO Ming-Wu, LUO Rui, AN Hui, PANG Qiu-Ying

Bulletin of Botanical Research 2019, 39 (2): 222-228. DOI: 10.7525/j.issn.1673-5102.2019.02.007

Abstract （1058）

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To investigate the physiological response of Helianthus tuberosus suspension cells to salt stress, biomass, viability, activity of antioxidant enzyme, the contents of hydrogen peroxide, malonaldehyde, proline, soluble sugar, soluble protein, total phenols were measured, and 14 phenolic compounds of suspension cells were analyzed under the treatments of 0, 50, 100, 150 and 200 mmol·L ^-1 NaCl. The salt stress markedly weakened the H.tuberosus suspension cell viability, and the growth of suspension cells was obviously inhibited. Under the condition of 200 mmol·L ^-1NaCl treatment, suspension cells almost stopped growing. Salt stress induced the oxidative stress, the content of malonaldehyde significantly increased, the activity of ascorbate peroxidase, peroxidase, the content of total phenols and part phenolic compounds increased with the increasing concentration of NaCl. Both phenolic compounds and antioxidant enzyme system participated in antioxidative process. Proline played an important role in the osmoregulation of H.tuberosus suspension cells responding to NaCl osmotic stress, meanwhile, the soluble sugar had little effect.

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Species Diversity of Bryophytes in Miyagi Relics of Shangjing Longquanfu,Parhai State,China

CONG Ming-Yang, TANG Lu-Yan, LI Jin-Jiang, ZHANG Mei-Ping, CHEN Bao-Zheng, XU Yue-Yue

Bulletin of Botanical Research 2019, 39 (2): 229-238. DOI: 10.7525/j.issn.1673-5102.2019.02.008

Abstract （778）

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Relics of Shangjing Longquanfu of Parhai State is the largest site in northeast region in medieval times, which demonstrate the style and features of Tang Dynasty. However, researches about bryophyte diversity in special relics habitats remain rare. The miyagi relics is the core of protection, and it has irreplaceable scientific value. In order to fully understand the diversity of bryophytes, a total of 216 specimens were collected from 45 quadrats at 9 sampling sites. Species composition, habitat types, similarity coefficient, α diversity indexes and water ecological types of bryophytes were analyzed by adopting Excel 2010 and R 3.5.1. Main results were as follows: (1)the 35 species were recorded in total, including 1 in Hepaticae and 34(17 genera, 7 families) in Musi. Dominant families were Brachytheciaceae, Pottiaceae, Bryaceae and Entodontaceae. Dominant species were Entodon dolichocucullatus S.Okam., Brachythecium coreanum Card. and Didymodon fallax Hedw. etc. (2)Habitat types were wall(40.00%) and rock(28.89%), with soil(17.78%) and tree(13.33%) habitats took second place. Similarity coefficient was the highest between wall and rock(38.71%), while the value was zero between tree and other 3 habitats. (3)Values of Patrick, Shannon-Wiener and Simpson indexes were wall>rock>soil>tree. (4)Water ecological types were only xerophyte, mesophyte and phreatophyte, in which mesophyte(51.43%) was dominate. Above research findings have important significance and will provide basic data and scientific basis for effective protection of the miyagi site.

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Variation Analysis of Growth Traits in Different Growth Year of Half-sib Fraxinus mandshurica Families

LUAN Ke-Quan, ZHANG Heng, TIAN Yong-Gang, YANG Shu-Cheng, WANG Hong-Wu, WANG Lian-Kui, LI Yan-Long, LU Zhi-Min, ZHAO Xi-Yang

Bulletin of Botanical Research 2019, 39 (2): 239-245. DOI: 10.7525/j.issn.1673-5102.2019.02.009

Abstract （827）

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In order to select high-quality germplasm resources of Fraxinus mandshurica, with 75 half-sib families of 16-years F.mandshurica at Sanchazi Forestry Bureau, tree height, basal diameter(diameter at breast height) and volume were investigated in the 4th, 11th and 16th growth years. By ANOVA analysis, there was significant differences among different variation source( P<0.01). The variation coefficients of phenotypic and heritabilities of different traits ranged in 22.20%-63.30% and 0.796-0.981, respectively. There was significant positive correlation among different traits. Based on 10% selected rate, according to volume to evaluate the families, eight families were selected as elite families. The average volumes of elite family were higher than average values of all the families by 0.003 m ³, whose genetic gain were 36.03%. The elite families can supply basis materials for the improved variety register and the rebuilt seed orchard of F.mandshurica.

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Effects of EMS on Seed Germination of Anemarrhena asphodeloides in the Simulated Environment

YANG Nan, NIE Jiang-Li, XIN Wei, XIA Xu, LANG Yu-Jie, PEI Yi

Bulletin of Botanical Research 2019, 39 (2): 246-251. DOI: 10.7525/j.issn.1673-5102.2019.02.010

Abstract （1180）

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The seeds of Anemarrhena asphodeloides were treated with different contents and treatment times of EMS at 1%, 2%, 4%, 6% and 6, 8, 12, 24 h, respectively, in the constant temperature incubator in this experiment.We studied the effects of the different concentrations and treatment time of EMS on the seed germination characteristics of A.asphodeloides. The results of seed germination showed that the seed germination of A.asphodeloides reduced with theincreasings of the contents and the treatment time of EMS. With the relative germination rate reaching the semi-lethal concentration as the standard, the suitable condition for EMS treatment of A.asphodeloides seeds are the concentration of 6% and soaking seeds for 12 and 24 hours.

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Physiological Analysis of Three Anti-Browning Agents Affecting Somatic Embryogenesis in Fraxinus mandshurica

PENG Chun-Xue, CUI Xue-Mei, LIU Chun-Ping, YANG Ling, WANG Qiu-Shui, SHEN Hai-Long

Bulletin of Botanical Research 2019, 39 (2): 252-258. DOI: 10.7525/j.issn.1673-5102.2019.02.011

Abstract （944）

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The somatic embryogenesis of Fraxinus mandshurica Rupr. is often accompanied by browning, and most somatic embryos grow on browning explants.We analyzed the relationship between explant browning and somatic embryogenesis, by adding PVP(polyvinylpyrrolidone), L-Glu(L-glutamic acid) and AgNO ₃(silver nitrate) to the culture medium, and studied its effects on browning, somatic embryogenesis and physiological and biochemical characteristics of explants.The results showed that: (1)Low concentration(0.1 and 0.5 g·L ^-1) PVP, and 100 mg·L ^-1 L-Glu treatment exacerbated browning of explants, but significantly promoted somatic embryogenesis, somatic embryogenesis rate exceeded 60%(increased by 6.59%, 24.08% and 22.88% compared to the control,respectively). (2)The 200 mg·L ^-1 L-Glu treatment effectively reduced the browning of explants, the browning rate was 68.11%(down 5.83% compared with the control), but the somatic embryogenesis rate was reduced to 46.32%(down 22.8%compared with the control). (3)The activities of polyphenol oxidase(PPO) and superoxide dismutase(SOD) in explants were lower than those of control after treatment with three anti-browning agents, peroxidase(POD) activity and malondialdehyde(MDA) content were higher than the control.Therefore, it is believed that a low concentration of anti-browning agent can promote somatic embryogenesis, and in this process, an increase in POD activity and an increase in MDA content are involved. We analyzed the biological mechanism of somatic embryogenesis with explants browning and got the frequency of somatic embryogenesis of F.mandshurica.

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Phenotypic Genetic Diversity of Pinus elliottii× P.caribaea Morelet var. hondurensis Clones

GUO Wen-Li, LI Yi-Liang, ZHAO Fen-Cheng, TIE Jun, LIAO Fang-Yan, ZHONG Sui-Ying, LIN Chang-Ming, YE Wei-Fang

Bulletin of Botanical Research 2019, 39 (2): 259-266. DOI: 10.7525/j.issn.1673-5102.2019.02.012

Abstract （4872）

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Thirty-five Pinus elliottii× P.caribaea Morelet var. hondurensis clones were used as experimental materials, and correlation analysis, cluster analysis and principal component analysis were performed on eight phenotypic traits to explore genetic diversity level. The level of genetic diversity was discussed to provide scientific basis for the utilization of P.elliottii× P.caribaea Morelet var. hondurensis clones and breeding of improved varieties. The results showed that: (1)The coefficient of variation of growth traits ranged from 12.31%-29.28%, the mean value was 22.90%, and the diversity index ranged from 1.61-1.80 with an average of 1.69. The coefficient of variation of shape traits was between 13.34% and 47.25%, the mean value was 26.86, and the diversity index was between 1.50 and 1.94, with a mean value of 1.77. The P.elliottii×P.caribaea Morelet var. hondurensis clones was rich in genetic variation and high in genetic diversity. (2)Among the eight pairs of phenotypic traits, eleven pairs of trait correlations were extremely positively significant, and five pairs of them were positively significant. The selected volume was the main index for breeding selection of the material, followed by tree height. (3)When the Euclidean distance was 12, thirty-five clones were divided into six groups, and some of them come from the same source or the same male parent. (4)Three principal components were extracted from the principal component analysis of eight phenotypic traits, and the cumulative contribution rate reached 86.00%, which mainly represented six traits, so as to express the comprehensive variation of all the tested materials.

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Differential Expression of PodaPIN9 Gene in Tissues of Populus davidiana× P.alba var. pyramidlis Induced by Trichoderma

LI Shu-Hang, MIAO Rui, CHANG Yuan, LI Jun-Nan, YAN Xiao-Jie, LIU Zhao-Ying, ZHANG Rong-Shu

Bulletin of Botanical Research 2019, 39 (2): 267-275. DOI: 10.7525/j.issn.1673-5102.2019.02.013

Abstract （759）

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The PIN proteins contain multiple transmembrane domains, and are involved in the efflux of auxin, affecting many growth and development-related processes in higher plants. Trichoderma is a biocontrol agent that promotes the growth and improves the defense mechanisms of plants against various diseases. Therefore, it is important to study the effects of Trichoderma on the distribution of auxin in the woody species Populus davidiana×P.alba var. pyramidlis(Shanxin poplar). In this study, the gene PodaPIN9 was cloned from Shanxin poplar, and its nucleotide and protein sequences were analyzed. The constructed phylogenetic tree showed that PodaPIN9 has a high identity with 9 PIN genes from 6 species(>80%). By the qRT-PCR analysis, PodaPIN9 was expressed in the stem tips, mature leaves, and roots of the plant. The expression of PodaPIN9 was significantly low in the roots, but was significantly high in the stem tips and leaves(503 and 346 times higher than that in the roots, respectively). Following inoculation with Trichoderma, the expression of PodaPIN9 was significantly downregulated in the stem tips and leaves, whereas its expression was significantly upregulated in the roots. The expression of PodaPIN9 in the roots was 32.01 times higher than the control group. Additionally, we found that the levels of auxin in the stem tips, leaves, and roots of the plant were decreased at 48 h after inoculating the Trichoderma. It demonstrated that Trichoderma can affect auxin levels and the expression of PodaPIN9 in the stem tips, leaves, and roots of poplar. Furthermore, by Pearson correlation analysis, the correlation between the expression of PodaPIN9 and auxin levels differed in the stem tips, leaves, and roots.

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Transcriptome Analysis for Rare and Endangered Plants of Semiliquidambar cathayensis

YE Xing-Zhuang, LIU Dan, LUO Jia-Jia, FAN Hui-Hua, ZHANG Guo-Fang, LIU Bao, CHEN Shi-Pin

Bulletin of Botanical Research 2019, 39 (2): 276-286. DOI: 10.7525/j.issn.1673-5102.2019.02.014

Abstract （796）

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In order to strengthen the conservation and utilization of China's endangered plant, Semiliquidambar cathayensis, a high-throughput sequencing platform, Illumina HiSeq 2500, was used to sequence the transcriptome of the plants. The obtained data was filtered and assembled by de novo. Then, 77 629 unigenes were obtained through classification and redundancy removal. By comparison analysis, annotation of the 9 functional databases, 45 293 unigenes get annotation information, which was divided into 25 sub-categories by function in KOG with 25 253 functional annotation information obtained. The functional annotation of GO could be divided into such three parts as cell components, biological processes, and molecular functions, which is composed of 2 226, and 17 subclasses respectively (65 subclasses in total). With the comparison of KEGG database, 286 metabolic pathways are found, among which there are 177 pathways to biosynthesis of secondary metabolites, which may be related to the medicinal active components of S.cathayensis. Based on the assembling result, 1 547 unigenes encoding the transcription factors are predicted in 88 gene families. Thus, a family of transcription factors controlling pharmacokinetic synthesis are discovered. Based on the annotation results of S.cathayensis, 12 579 SNP polymorphic sites are detected and 57 671 CDS sites are predicted. This study is the first time to analyze the transcriptional sequences of S.cathayensis, providing a basic data for the in-depth study on molecular biology in S.cathayensis.

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Genetic Transformation and Resistance Analysis of BpBEE2 Gene from Betula platyphylla

YAN Bin, WU Dan-Yang, LI Hui-Yu

Bulletin of Botanical Research 2019, 39 (2): 287-293. DOI: 10.7525/j.issn.1673-5102.2019.02.015

Abstract （968）

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Brassinolide Enhanced Expression2( BEE2) genes belong to bHLH transcription factors family,and were involved in Brassinolide signal pathway.In this study, BpBEE2 gene was cloned using RT-PCR, and plant over-expression and inhibiting expression vector were constructed. Genetic transformation was operated through agrobacterium mediated, then mass growths of transgenic lines and non-transgenic line were analyzed under natural conditions and salt, drought stress. The length of 1 080 bp cDNA sequence was obtained, over-expression and inhibiting expression vectors were constructed successfully, and transgenic lines were obtained. Seedlings height of over-expression transgenetic line was higher than that of the control line, however, that of inhibiting expression lines was lower than that of the control line. BEE2 gene can ffect fresh weight of plants after salt and drought stress, and BpBEE2 is involved in the process of plant growth and development and plays a key role in drought and salt resistance response.

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Characteristic Analysis of Microsatellites in the Transcriptome of Phyllanthus emblica,an Important Edible and Medicinal Plant

LIU Xiong-Fang, LI Tai-Qiang, ZHANG Xu, LI Zheng-Hong, WAN You-Ming, AN Jing, LIU Xiu-Xian, MA Hong

Bulletin of Botanical Research 2019, 39 (2): 294-302. DOI: 10.7525/j.issn.1673-5102.2019.02.016

Abstract （899）

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In order to comprehensively understand the characteristics of distribution and variation of the SSR loci in the transcriptome of Phyllanthus emblica L., transcriptome sequencing was conducted on young leaves of P.emblica by using Illumina Hiseq 4000. The SSR loci were studied using MISA software from the obtained unigenes. A total of 9 991 SSRs distributed on 9 538 unigenes were detected, with an average density of one SSR per 5.49 kB. Mononucleotide and dinucleotide repeats were the main SSR types, accounting for 42.3% and 30.79% of all SSRs, respectively. There were 1 731 SSR loci distributed across the protein-coding regions, with an overall density of 0.039 SSRs/kB. The dominant repeat type was trinucleotide repeat type. Among all the 169 repeat motifs, A/T(42.10%) was the richest motif, followed in order by AG/CT(22.91%) and AAG/CTT(5.02%). The number of repeats ranged from 4 to 75, most of them were concentrated on four to 20. The SSRs with repeat fragment length ≥20 bp accounted for 21.20%. There was significant negative correlation( P<0.01) between the frequency of SSRs and the repeat fragment length, with correlation coefficient of -0.561. The SSR loci in the transcriptome of P.emblica showed high frequency and density of distribution, rich low-level repeat motifs, large repeat number and long fragments, most of which exhibited high potential of polymorphisms in our study. These SSR loci would lay an important foundation for further developing a large number of EST-SSRs and studying on population genetics of P.emblica, and they would also provide a reference for conservation and rational exploitation of genetic resources of the species.

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Effects of Different Growth Stages on Secondary Metabolites of Acanthopanax senticosus

XU Ming-Yuan, WANG Qian-Bo, GUO Sheng-Lei, WANG Ying-Wei, LI Feng-Xia, LIU Jia, TANG Zhong-Hua, WANG Zhen-Yue

Bulletin of Botanical Research 2019, 39 (2): 303-309. DOI: 10.7525/j.issn.1673-5102.2019.02.017

Abstract （851）

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The common influence of different cultivation years on the accumulation of various active components in the root and stem of Acanthopanax senticosus was evaluated. It was important for raising the cultivation and high value utilization of A.senticosus. It provide a theoretical basis for standardized planting and rational exploitation and utilization of A.senticosus. The A.senticosus of different growth stage was collected from the same origin, and the Ultra-Performance LC(Waters, Japan) was used to analyze different growth stages of six secondary metabolites in the A.senticosus roots and stems. The total score of six main active components in root and stem of A.senticosus was the highest in five years. Among them, quercetin has the highest content in three-year rooting and the highest content in five-year stem; rutin has the highest content in five year roots and stems, the lowest in three-years in root and lowest in nine-year stem; Hyperoside is the highest in five-year root and three-year stem. The content of isofraxidin in three year root and stem is the highest, and the content of root and stem is the lowest in five years. The content of syringin is the highest in nine-year root and in five-year stem. The highest content of eleutheroside e in the five-year rooting is the highest in the three-year stem. Different medicinal ingredients accumulated differently in roots and stems of A.senticosus in different cultivation years and targeted cultivation could select suitable harvest years according to the active components.

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Correction of Typographical Errors in the Protologue of Eleven Taxa(Dicotyledon) in China

LIN Yun, BI Hai-Yan, LI Chao, YUN Ying-Xia

Bulletin of Botanical Research 2019, 39 (2): 310-320. DOI: 10.7525/j.issn.1673-5102.2019.02.018

Abstract （895）

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We correct the typographical errors in the protologues of eleven taxa(Dicotyledon) in China. The type of Castanopsis yanshanensis Hu(Fagaceae), incorrectly cited as C.W.Wang 84116 in the protologue, is actually C.W.Wang 84416; the former collection belongs to Inuna helianthus-aquatica C.Y.Wu ex Ling(Asteraceae). The type of Pasania longinux Hu(Fagaceae), incorrectly cited as K.M.Feng 13012 in the protologue, is actually K.M.Feng 13102; the former collection belongs to Ilex triflora Bl.(Aquifoliaceae). The holotype of Laportea fujianensis C.J.Chen(Urticaceae), incorrectly cited as C.J.Chen & Z.Y.Li 109 in the protologue, is actually C.J.Chen & Z.Y.Li 103; the former collection belongs to a species of Oreocnide frutescens(Thunb.) Miq.(Urticaceae). The holotype of Oreocnide integrifolia(Gaudich.) Miq. subsp. subglabra C.J.Chen(Urticaceae), incorrectly cited as N.K.Chun 44099 in the protologue, is actually N.K.Chun 44033; the former collection belongs to Lyonia ovalifolia(Wallich) Drude var. rubrovenia(Merr.) Judd.(Ericaceae). The holotype of Clematis austrogansuensis W.T.Wang & L.Q.Li(Ranunculaceae), incorrectly cited as Baishuijiang Exped. 4490 in the protologue, is actually Baishuijiang Exped. 4990; the former collection belongs to Euonymus alatus(Thunb.) Sieb.(Celastraceae). The type of Delphinium pachycentrum Hemsl. var. humilius W.T.Wang(Ranunculaceae) was wrongly cited as Sichuan Veg. Exped. 3137in the protologue, but actually is Sichuan Veg. Exped. 3173; the former collection belongs to Gentiana conduplicata T.N.Ho(Gentianaceae).The holotype of Astragalus chengkangensis Podlech & L.R.Xu(Fabaceae), incorrectly cited as T.T.Yu 17255 in the protologue, is actually T.T.Yu 17225; the former collection belongs to Scirpus lushanensis Ohwi(Cyperaceae). The type of Oxytropis latialata P.C.Li(Fabaceae), incorrectly cited as Qinghai-Xizang Comp. Exped. 9484 in the protologue, is actually Qinghai-Xizang Comp. Exped. 9485; the former collection belongs to Arenaria kansuensis Maxim.(Caryophyllaceae). The type of Oxytropis reniformis P.C.Li(Fabaceae), incorrectly cited as Qinghai-Xizang Comp. Exped.3650 in the protologue, is actually Qinghai-Xizang Comp. Exped.3605; the former collection belongs to a species of Ephedra gerardiana Wall. ex Mey.(Ephedraceae). The type of Acer longipes Franch. ex Rehd. var. hunanense W.P.Fang & W.K.Hu(Aceraceae), incorrectly cited as Z.T.Lee 2944 in the protologue, is actually Z.T.Lee 2994; the former collection belongs to Pieris formosa D.Don(Ericaceae). The type of Berchemia omeiensis W.P.Fang ex Y.L.Chen & P.K.Chou(Rhamnaceae), incorrectly cited as G.H.Yang 54729 in the protologue, is actually G.H.Yang 54723; the former collection belongs to Helwingia chinensis Batalin.(Cornaceae).

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A Study on Breeding Characteristics and Ecological Adaptability of the Ferula ferulaeoides

ZHAO Xiao-Yi, WANG Zhong-Ke, LI Gui-Fang, LÜ Xin-Hua, ZHAO Wen-Qin, ZHUANG Li

Bulletin of Botanical Research 2019, 39 (3): 321-328. DOI: 10.7525/j.issn.1673-5102.2019.03.001

Abstract （686）

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Ferula ferulaeoides is a endemic species of ephemeroid plant in Xinjiang. It has great ecological function and potential economic value. In this experiment, we studied the floral morphological characteristics, breeding system and pollination insects of F.ferulaeoides. To explain the mechanism about the adaptation of environment. The results showed that:(1)The F.ferulaeoides inflorescence anthesis lasted for 10-15 d and the single flower florescence was 7 days. Herkogamy and dichogamy were found in the hermaphroditic flower; (2)The type of breeding system was out-crossing mixed with self-pollination and the fruiting rate was 72.24% under natural conditions; (3)Many species of insects visited the flowers and most of them were Diptera and Hymenoptera; (4)The pollen viability on the first day of anthesis was highest at 12:00, the stigma receptivity was strongest after the second day of completely disseminating pollen.

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Floral Morphology and Flowering Process of Acer yangjuechi, the Extremely Endangered Plant

CHEN Xiang-Bo, LÜ Xiu-Li, LIU Yang, ZHAO Ming-Shui, CUI Xin-Hong, ZHANG Dong-Mei

Bulletin of Botanical Research 2019, 39 (3): 329-337. DOI: 10.7525/j.issn.1673-5102.2019.03.002

Abstract （1014）

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To reveal the reproduction mechanism of low embryo development rate, the floral morphology and flowering process of Acer yangjuechi Fang et P.L.Chiu, the extremely endangered plants in Chinawas determined by testing trees cultivated in National Nature Reserve of Tianmu Mountain, Zhejiang Province.The results show that:(1)Bisexual flowers and maleflowers are inserted on a same apical inflorescence, and the bisexual flowers are always on the top of rachis while maleflowers on lateral rachis top or bottom. The quantity ratio of maleflowers to bisexual flowers is 6.89 of 2.6 times exceeding to A.buergerianum, which indicates less bisexual flowers of A.yangjuechi than A.buergerianum. (2)Two type of maleflowers were observed. Type Ⅰmaleflowers have tiny ovary, the stigma shortened extremely to a black spot and with flat flower disc while Type Ⅱmaleflowers have biforked stigma outstanding upon ovary degenerated into white fluff and with thick and radial sunken flower disc. Bisexual flowers possess of outstanding oblate spheroid and shorter filament compared with maleflowers. (3)The flowering process of individual tree is of bisexual flowers flowering within 3 days previous to maleflowers while only under seven percent of maleflowers were on pollination stage at the same time. (4)For population flowering process, the flowering stage between bisexual flowers and maleflowers present an obvious interval and bisexual flowers flowering intensively previous to maleflowers. Therefore, the main cause of the high embryo abortion, which leads to endangered situation of A.yangjuechi, was discussed.

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Embryonic Callus Induction and Plant Regeneration of Lilium leucanthum

ZHANG Lu, PAN Yuan-Zhi, LIU Shi-Liang, CHEN Yuan, SHEN Li-Jia

Bulletin of Botanical Research 2019, 39 (3): 338-346. DOI: 10.7525/j.issn.1673-5102.2019.03.003

Abstract （1052）

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With Lilium leucanthum Bakeri bulbs as explants, L ₉(3 ⁴) orthogonal test was used to study the effects of different hormones and their concentrations on its callus induction, bulblet differentiation and rooting culture. Histological observation of embryogenic callus was carried out by paraffin sectioning technique. The optimal medium combinations for inducing callus of L.leucanthum under light and dark conditions were MS+TDZ 0.5 mg·L ^-1+NAA 1.0 mg·L ^-1+2, 4-D 0.05 mg·L ^-1 and MS+TDZ 0.5 mg·L ^-1+NAA 1.5 mg·L ^-1+2, 4-D 0.1 mg·L ^-1, with the induction rates of 62.690 and 59.423, respectively. The medium suitable for bulblet rooting was 1/2MS+NAA 0.2 mg·L ^-1+1 g AC. After 60-day culture, the seedlings were transferred to peat:vermiculite:perlite=1:1:1 in a medium and cultured in a light incubator for 35 days with the survival rate of 90%. Our experiment successfully established the regeneration system of squamous callus in L.leucanthum, which laid a foundation for the research of tissue culture transgenic seedling and the innovation of germplasm resources.

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