Table of Content

20 November 2014, Volume 34 Issue 6

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A New Species of Melampsora

XUE Yu；SHAO Li-Ping

2014, 34(6):  721-722.  doi:10.7525/j.issn.1673-5102.2014.06.001

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A new species of the genus Melampsora, M.salicis-polyadeniae Y.Xue et L.P.Shao, on leaves of Salix polyadenia is described from Jilin province. Latin and Chinese diagnosis are given.Type specimen is deposited in the Northeast Forestry University,Harbin.

Micro-morphological Characters of Leaf Epidermis of Ten Species in Genus Paphiopedilum

LI Zong-Yan；WU Yu-Lan；PENG Kun

2014, 34(6):  723-729.  doi:10.7525/j.issn.1673-5102.2014.06.002

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By observing leaf epidermis of ten species in Paphiopedilum under the light microscope, we compared their cell shapes, anticlinal walls, stomatal types, and subsidiary cell shape and measured eleven quantity indexes, including size of epidermis cell, size of stomata, stomatal density and its index, to study species' difference. The epidermis cell was polygon or rectangular, anticlinal wall was circular, nearly circular, straightarched and straight, and that stomata was mainly surrounded by tetracytic. By principal component analysis on 15 indexes, the quantity indexes related to ecological adaptation (size of epidermis cell and stomatal density) were greatly contributed to species variation, which reflected that there was significant adaptation differentiation to water and light in Paphiopedilum species from different habitation. Moreover, the types of anticlinal wall and stomata, the shapes of epidermis cell could be applied to divide its subgenus. The clustering result based on leaf epidermis characters supported treatments on interspecific relationship and division of subgenus by rDNA ITS evidence.

Anatomical Study on Leaf of Salvia miltiorrhiza from Different Areas

WANG Tao；LIU Shi-Yong；JIANG Yuan-Yuan；ZHANG Li*

2014, 34(6):  730-735.  doi:10.7525/j.issn.1673-5102.2014.06.003

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In order to provide scientific basis for the identification and improved varieties of Salvia miltiorrhiza, the anatomy characteristics of S.miltiorrhiza of different habitats were compared. The lamina and petiole of S.miltiorrhiza in 8 places were studied with paraffin sectioning, and their microstructures were observed. Moreover, the epidermis, xylems, phloems, etc. were measured. The data were studied by principal components analysis method. Leaf of S.miltiorrhiza is odd-pinnately compound leaf, bifacial leaf, villous. The upper epidermis thickness 11.87-26.47 μm, lower epidermis thickness 7.40-24.57 μm, number of palisade tissue 1-2, thickness of palisade tissue 42.93-58.13 μm, thickness of spongy tissue 57.17-102.67 μm, palisade tissue-spongy tissue ratio of leaf 0.55-0.87 etc. Leaf anatomical structure of S.miltiorrhiza has relationship with plant type and effective components. In summary, the anatomy characteristics of S.miltiorrhiza were distinguished from different origins. These conclusions can provide valuable reference for S.miltiorrhiza breeding.

Embryo Development in Bolbitis angustipinna(Hayata) H. Ito

ZHAO Er-Lan1；XU Yong-Xing2；GUAN Wei-Yuan2；LIU Bao-Dong1*

2014, 34(6):  736-740.  doi:10.7525/j.issn.1673-5102.2014.06.004

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We studied the development of embryo of Bolbitis angustipinna(Hayata) H. Ito by artificial cultivation and paraffin. The first division plane of the zygote is perpendicular to the prothallus, and parallel to the longitudinal axis of the archegonium neck. The plane is parallel to the prothallus and perpendicular to the longitudinal axis of the archegonium neck at the second zygote division. At the third division, the plane is parallel to the longitudinal axis of the prothallus and perpendicular to the archegonium neck. As the embryo reaches to the spherical phase, the upper and lower parts of the outside of embryo produce the apical cell of the first leaf and the apical cell of the first root respectively at the same time. These two apical cells develop into the first leaf primordium and the first root primordium. Subsequently, the first leaf primordium rapidly divides, breaks the cap-like structure and becomes the first young leaf. The first root primordium divides relatively slower than the first leaf primordium, and then breaks the cap-like structure, and forms the first root. We also discussed the phylogeny of embryo.

Leaf Fall Patterns of 12 Evergreen Woody Species in Subtropical Evergreen Forest in Tiantong Mountain of Zhejiang Province, China

LI Xin-Cheng1；MENG Feng-Qun2；LI Ming-Hong1；YANG Dong-Mei1*

2014, 34(6):  741-750.  doi:10.7525/j.issn.1673-5102.2014.06.005

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Leaf fall patterns can reflect the physiological ecology adapting of leaf to environment, and are the basis for the ecosystem process and function. In order to reveal the diversity of leaf fall patterns, we recorded the dynamics of perennial leaves (one-year old or older) number and current-year leaves number of the same twigs during leaf flushing period for twelve evergreen woody species in subtropical evergreen broad-leaved forest of Tiantong Mountain in Zhejiang Province, China. According to the peaks of leaf shedding and temporal distribution, we divided them into seasonal leaf fall peak and successively leaf fall patterns. Of twelve species, four species showed a spring leaf fall peak, two species showed a summer leaf fall peak, one species showed an autumn leaf fall peak, one species showed a spring and autumn bimodal leaf fall peaks, and four species shed their leaves successively round the year without a remarkable leaf fall peak. The sun-adapted species (including canopy trees and gap shrubs) are most seasonal leaf fall peaks patterns, while shade-adapted species are successively leaf fall patterns, and the former have larger percentage of total leaf fall amounts than the latter, which indicates the faster leaf turnover rate of the sun-adapted species. Combined with the climate data and the leaf flushing phenology, we discussed the impact factors (including leaf production, typhoon disturbance and water balance) influencing the leaf fall patterns. The leaf fall patterns in subtropical evergreen forest may help us to further explore the strategy of plant adaptation to environments.

SEM Observation on the Pollen Morphology of Eleven Ornamental Crabapple Varieties

WANG Yan-Xiu；CHEN Bai-Hong*；WANG Shu-Hua；HU Zi-Jing；DANG Zhao-Xia；WEI Xiao-Yan

2014, 34(6):  751-757.  doi:10.7525/j.issn.1673-5102.2014.06.006

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We observed pollen morphology of 11 Malus species ornamental crabapple cultivars by scanning electron microscope, and studied the relationships and groups by average (UPGMA) cluster analysis method according to the pollen shapes, ornamentation strips and hole-frequency. All of the 11 varieties’ pollen shapes in side view were oblongoval, and in polar view triangular or nearly round, which belonged to the N3P4C5 type pollen. The exine surface was covered with strips. There were significant variations among varieties in pollen shape, strips shape, decoration, and pore patterns. By UPGMA cluster analysis, 11 cultivars could be classified into three groups at similarity distance 3.30 level. ‘Hongshuang’, ‘Hongsha’ and ‘Hongbo’ could fall into the first group as the three varieties with long pollen stripe bundle’s branch, and sparse and regular ridge. ‘Hongfen’, ‘Hongliang’, ‘Xuelai’, ‘Xueqiu’, ‘Hongyan’ and ‘Xueqian’ could fall into the second category with sparse ridges, parallel to overall pollen, little sub-fork, and medium pore frequency. ‘Dorge’ and ‘Xuekai’ were in the third group with irregular ridges, smallest pollen grains and smaller pore diameter, and they were located in the top of evolution.

Responses of Photosynthesis,Chlorophyll Fluorescence of Poplar Leaf and Bark Chlorenchyma to Elevated Temperature

REN Jie；WANG Hui-Mei*；WANG Wen-Jie；QU Dan；WANG Qiong；ZHONG Zhao-Liang

2014, 34(6):  758-764.  doi:10.7525/j.issn.1673-5102.2014.06.007

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Leaf and bark chlorenchyma are two kinds of important photosynthetic tissues for poplar. Their responses to elevated temperature in photosynthetic parameters will be useful for photosynthetic physiological changes of poplar during global warming. We studied the changes of photosynthesis and chlorophyll florescence in leaf and bark chlorenchyma of poplar with three temperature treatments of 23℃/18℃, 28℃/23℃ and 33℃/28℃(light/dark period). The chlorophyll content of bark chlorenchyma and leaf were hardly affected in different temperature treatments. However, the values of chlorophyll a/b in the bark chlorenchyma decreased significantly by increasing the temperature up to 33℃. The rising temperature produced important influences on photosynthesis in leaf and bark chlorenchyma. The net photosynthetic rate(P_n), photosynthetic efficiency(PE), stomata conductance(G_s) and transpiration rate(T_r) of leaf and bark chlorenchyma significantly enhanced, but water use efficiency obviously declined with rising temperature. Leaf and bark chlorenchyma chlorophyll fluorescence showed similar response to elevated temperature. Except for the maximum photochemical efficiency(F_v/F_m), actual photochemical efficiency(Φ_PSⅡ), photochemical quenching(qP), nonphotochemical quenching(NPQ) and photosynthetic electron transport rate(ETR) obviously decreased. Our results showed that stomatal factors mainly resulted in P_n increase of poplar bark chlorenchyma and leaf with elevated temperature.

Photosynthetic Physiological Responses of Five Woody Plants to NaHCO3 Stress

LIU Qiang1；WANG Qing-Cheng2；WANG Zhan-Wu1

2014, 34(6):  765-769.  doi:10.7525/j.issn.1673-5102.2014.06.008

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We investigated the effectiveness of photosynthetic characteristics as an evaluation index of salt tolerance in five trees differing in tolerance to salt stress. The salt tolerance rank of the five trees is Lycium dasystemum>Ulmus pumila>Celastrus orbiculatus>Syringa microphylla>Pinus sylvestris var. mongolica. Five woody plants responsed identically to NaHCO₃ stress, that is, net photosynthetic rate (P_n), transpiration rate (T_r), stomatal conductance (G_s), intercellular space CO₂ concentration (C_i) and stomata limitation (L_s) all decline obviously. P_n decreasing scale of five trees was P.sylvestris var. mongolica>S.microphylla>C.orbiculatus>U.pumila>L.dasystemum. T_r decreasing scale of five trees was P.sylvestris var. mongolica>S.microphylla>L.dasystemum>C.orbiculatus>U.pumila. G_s decreasing scale of five trees was P.sylvestris var. mongolica>L.dasystemum>Ulmus pumila>S.microphylla>C.orbiculatus. According to the changes of photosynthesis parameters of five seedlings, under lower stress intensity, the main factor leading to decreasing in photosynthesis rate of U.pumila and L.dasystemum was stomatal limitation, while there was non-stomatal limitation under higher stress intensity. The main factor leading to decreasing in photosynthesis rate of C.orbiculatus and P.sylvestris var. mongolica was stomatal limitation. Non-stomatal limitation was the main factor leading to decreasing in photosynthesis rate of S.microphylla. The more salt-tolerant U.pumila and L.dasystemum had a less decreasing scale of photosynthesis rate than that of S.microphylla and P.sylvestris var. mongolica.

Photosynthetic Characteristics and Its Micro-environmental Limiting Factors of Two Main Oil Peony

ZHANG Zhong-Hua1,2；TANG Zhong-Hua1；YANG Feng-Jian1*；WANG Hua3；ZHANG Jun1；ZU Yuan-Gang1

2014, 34(6):  770-775.  doi:10.7525/j.issn.1673-5102.2014.06.009

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It is important to study photosynthetic characteristics and micro-environmental constraints of Paeonia ostii and Paeonia rockii for the cultivation and promotion of oil peony. We selected P.ostii(Tongling, Anhui) and P.rockii(Lanzhou, Gansu) to measure the photosynthetic characteristics and micro-environmental factors, and analyzed photosynthesis limiting factors. At Phoenix Mountain (Tongling, Anhui), leaf temperature was the main limiting factor of photosynthesis in high temperature and drought seasons. When the temperature reached 31℃, the respiration rate was inhibited, and when the temperature reached 32℃, stomatal conductance and photosynthetic rate were reversed, and when the temperature reached 33 degrees, physiological damage appeared. At Gansu zhongchuan peony garden, the relative humidity was the most important influencing factor of photosynthesis characteristics, which determined the light saturation point, stomatal conductance, transpiration rate and water use efficiency of P.rockii. An appropriate shade habitat of the forest edge or forest gap is the most suitable environment for the growth of P.ostii and P.rockii.

Effect of Light Intensity on Growth, Photosynthetic and Fluorescence Characteristics of Polygonatum cyrtonema Hua

LI Ying-Chun；YANG Qing-Ping；CHEN Shuang-Lin*；GUO Zi-Wu；LI Yin；ZHUANG Ming-Hao

2014, 34(6):  776-781.  doi:10.7525/j.issn.1673-5102.2014.06.010

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We evaluated the ecological adaptability of Polygonatum cyrtonema Hua to different light conditions and determined the photosynthetic, fluorescence characteristics and growth performances of P.cyrtonema Hua. The survival rate was 100%, when the light intensity was equal or greater than 75% of full light, and the survival rate of P.cyrtonema Hua significantly reduced with the decreasing of the light intensity. Leaf area, basal diameter, and height of individual of P.cyrtonema Hua reached peak value at 45%-75% of full light, and specific leaf weight was the highest under 75% of light transmittance. When decreasing the light intensity, the contents of Chl a, Chl b and Chl contents increased firstly and then decreased, while the rate of Chl（a/b）and Car content changed in the inverse trend. Light had significant effect on T_r of P.cyrtonema Hua, while it showed no significant effect on P_n diurnal progress. When light intensity increased, Pmax decreased firstly and then increased. LCP and LSP decreased, while AQY changed insignificantly. Light intensity had significant effect on fluorescence parameters, while light intensity showed insignificant effect on F_v/F_m. Low light intensity influenced fluorescence parameters more obviously than that of high light intensity. Therefore, P.cyrtonema Hua is tolerant to low light environments which had high utilization efficiency in blue light, and it can grow well under 45%-75% of full light.

Distribution and Accumulating Regularity of Secondary Metabolites in Helianthus annuus L.

GAO Yin-Xiang1,2；ZU Yuan-Gang1；YANG Feng-Jian1；YU Xue-Ying1；YAO Li-Ping1*

2014, 34(6):  782-786.  doi:10.7525/j.issn.1673-5102.2014.06.011

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Helianthus annuus L. is one of the most famous ornamental plants and the oil crops in the world, its secondary metabolites have various biological and medical activities. We studied the distribution and accumulating regularity of four secondary metabolites-chlorogenic acid, polysaccharides, flavonoids and polyphenols in various growing stages of H.annuus L.. The higest chlorogenic acid content of 2.64 mg·g^-1 was in faceplates on flowering stage, the highest polysaccharide content of 52.2 mg·g^-1 was in faceplates on maturity stage, and the highest flavonoids and polyphenols contents of 20.94 and 11.75 mg·g^-1, respectively, were in leaves on seedling stage. Considering the distribution and accumulating regularity of these four secondary metabolites, the faceplates on flowering stage and leaves on budding stage have the most important medicinal value in H.annuus.

Dynamic Relations between Structural and Stilbene Glucoside Accumulating in Vegetative Organs of Polygonum multiflorum Thunb.

WANG Gui-Qin；ZHENG Yu-Hua；WANG Lu

2014, 34(6):  787-791.  doi:10.7525/j.issn.1673-5102.2014.06.012

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We studied the relationships between the anatomical structure and the content of stilbene glucoside in the vegetative organs of Polygonum multiflorum Thunb. by histochemical localization, light microscope and quantitative analysis. The stilbene glucoside is accumulated in every vegetative organs including root tube, rhizome, stem and leaf of P.multiflorum and the content of stilbene glucoside is arranged in the order of rhizomes>root tube>leave>stem. These vegetative organs (except for stem) at different growth stages all have different content of stilbene glucoside. The accumulation of stilbene glucoside in the root tuber has a linear growth trend (except on April) and achieves a peak value at the end of October. The stilbene glucoside content in rhizome and leave varies in a consistent trend and both accumulates to a maximum value at the end of August. The content of stilbene glucoside in the stem is very low and has no obvious changes at all growth stages. The root tube belongs to the tertiary structure and the stilbene glucoside mainly accumulates in phelloderm secondary phloem, tertiary phloem and vascular ray in the root tube. For the rhizome, the stilbene glucoside accumulates in the phelloderm, the cortex and the secondary phloem and vascular ray, and that in leaf accumulates mainly in the palisade tissue.

Genetic Diversity and Relationships of Endangered Sinowilsonia fienryi Hemsl by ISSR

HE Yan-Ni；JI Wen-Li*；ZHOU Lin-Yue；MA Jun-Ren；JI Xin-Miao；LI Hou-Hua

2014, 34(6):  792-797.  doi:10.7525/j.issn.1673-5102.2014.06.013

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ISSR-PCR was used to assess the genetic diversity of six wild populations Sinowilsonia fienryi Hemsl, along with two cultivated populations including 75 test materials. We detected 85 polymorphic bands from total 89 bands with 10 primers, and the percentage of polymorphic bands (PPB) was 95.5% indicating that S.fienryi Hemsl had a high level of genetic diversity at species level. Nei’s gene diversity (H), Shannon’s information index (I), the coefficient of gene differentiation (G_st) and the percentage of total variance among and within populations were 0.2833, 0.403 4, 0.350 7, 25.49% and 74.51%, respectively, indicating that there was a significant genetic differentiation both within and between the populations, and genetic diversity was mainly derived from the differentiation within populations. With UPGMA dendrogram, at the specify threshold of 0.87, S.fienryi Hemsl species were separated into three clusters, and most individuals from same habitat were clustered, indicating that there might be a rule between clustering analysis and geographical distribution.

Identification of Sex Related SRAP Molecular Marker in Humulus scandens

DENG Chuan-Liang；REN Ying-Xue；ZHANG Wei-Li；LI Shu-Fen；GAO Wu-Jun；LU Long-Dou

2014, 34(6):  798-802.  doi:10.7525/j.issn.1673-5102.2014.06.014

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Humulus scandens is a model material for studying the molecular mechamisms of sex differentiation in dioecious plants. We used SRAP (sequence related amplified polymorphism) to screen male or female specific fragments in H.scandensby using male and female gene pool as template. Fifty pairs of primers showed a good amplification pattern after screening 256 pairs of primers. Polyacrylamide gel electrophoresis amplification patterns of SRAP showed that 671 bands were amplified using 50 primer combinations, of which 247 bands were polymorphic and the polymorphic ratio is 36.81%. With five single male plants and five single female plants as materials, we validated the amplification patterns of primer combinations me15+em11, me14+em16, me13+em12 and me7+em10 again by agarose gel electrophoresis. Although the amplification patterns were more or less different between polyacrylamide gel electrophoresis and agarose gel electrophoresis, male or female specific bands were relatively stable.If cloning, sequencing and chromosomal locating the obtained sex-related DNA sequences on chromosome of H.scandens by using fluorescence in situ hybridization technique, it will be contributed to studying the molecular mechanisms of sex chromosome evolution in H.scandens.

AFLP Analysis of Genetic Differences among Alnus with Different Ploidies

RAO Long-Bing1；YANG Han-Bo1,2；GUO Hong-Ying3；DUAN Hong-Ping2；CHEN Yi-Tai1

2014, 34(6):  803-809.  doi:10.7525/j.issn.1673-5102.2014.06.015

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We studied the genetic difference between species in Alnus with different ploidies by using AFLP. Five AFLP primers produced 301 bands, of which 282 were polymorphic, accounting for 93.7%. The percentage of polymorphic bands of four population was 35.2% 72.4%, respectively, with an average percentage of 54.6%. The total population of genetic diversity Nei’s was 0.3043, and total Shannon polymorphism information was 0.4578. Among and within populations all had abundant genetic diversity, and the genetic difference interspecific was higher than that of intraspecific. By NJ cluster and PCOA, the closely related populations are geographically restricted and occur in proximity to each other.

Characterization of EST-SSR Among Six Genera of Rosaceae

ZHANG De-Fang1；LI Shu-Xian2；XIA Tao2*

2014, 34(6):  810-815.  doi:10.7525/j.issn.1673-5102.2014.06.016

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EST-SSR is a molecular marker based on functional genomics, had been widely used because of its evenly distributed among the genome, better universality between species or genera and characters associated with the phenotypic traits. We downloaded the EST sequences of six genera in Rosaceae (Rosa, Malus, Pyrus, Prunus, Fragaria and Rubus) to study the distributed characteristic of SSR in Rosaceae by bioinformatics methods. There was significant difference of EST-SSR distribution frequency among genera, from 10.94%(Rubus) to 43.49%(Pyrus). The dinucleotide and trinucleotide repeats were the major types, AG/CT, GA/TC and AT/TA were the preponderant dinucleotide motifs, the trinucleotide repeats showed significant difference among genera, and the dominant type included AGA/TCT, GAA/TTC, AAG/CTT, CAC/GTG and ACA/TGT. Our results provided a research platform for plant molecular phylogeny and evolution, germplasm identification, genetic diversity and genetic mapping of Rosaceae.

Cloning and Bioinformation Analysis of PLA Gene Family in Lycium barbarum L.

CHEN Hua-Feng1；CHEN A-Li2；SHANG Jie3*；ZHANG Feng-Jiao4

2014, 34(6):  816-823.  doi:10.7525/j.issn.1673-5102.2014.06.017

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Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is a key regulatory enzyme that link primary and secondary metabolism in plants by catalyzing the conversion of L-phenylalanine to cinnamic acid. Four partial sequences of PAL in Lycium barbarum L. were successfully cloned by RT-PCR using a sequence homology strategy, and the bioinformation analysis were carried out. The cDNA fragment of PAL gene family was 1 662-1 683 bp in length, which encodes proteins of 554-561 amino acids with the predicted molecular mass of 60.681-61.250 kD. The amino acid sequence variation between the PAL gene families is 0.005%-0.336%. The estimated isoelectric point (pI) of the putative protein is from 7.02 to 7.71. According to the amino acid sequence and structural analysis, it showed that this protein family contained one conserved domain, namely the shielding domain, and one active site which is phenylalanine/histidine ammonia-lyases. Subcellular localizations of LyPAL1-3 proteins were likely in the mitochondrion and LyPAL4 in the cytoplasm. The secondary structures and tertiary structures of LyPAL1 were abundant in alpha helixs (306) and random coils (179), while were less in beta turns (30) and extended strains (46). Phylogenetic analysis showed that the genes of LyPAL1-3 in L.barbarum L. were closely related to PAL in Capsicum annuum, while LyPAL4 was closely related to PAL in Petunia Juss. (Petunia axillaris, Petunia x hybrid, P.exserta). These four sequences have been registered in GenBank with the accession numbers KC75915-KC759156. Four LyPAL were cloned by molecular biology technology, which would contribute to the molecular mechanisms of the formation of flavonoids and other secondary metabolites in L.barbarum L..

Fine Root Biomass Vertical Distribution Character of Main Afforestation Tree Species in Horqin Sandy Land

WANG Kai1；SONG Li-Ning2*；Lü Lin-You3；ZHANG Liang1；QIN Zheng-Yuan1

2014, 34(6):  824-828.  doi:10.7525/j.issn.1673-5102.2014.06.018

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We studied the fine root vertical distribution characters of Populus simonii, Ulmus pumila, Pinus tabulaeformis and Pinus sylvestris var. mongolica in Horqin sandy land, and discussed the relationships with soil water and organic matter. The distribution extinction coefficient of fine root was 0.968-0.979 for main afforestation tree species in Horqin sandy land. There was no significant difference in the total fine root biomass between the same age class tree species (P>0.05), while the value of P.sylvestris var. mongolica was significantly higher at 32 years old than that at the other age (P<0.05). The correlation of the fine root biomass was higher with soil water content than with soil organic matter content except 22-ear old P.sylvestris var. mongolica. There were significant positive linear correlations between the fine root biomass and soil water content for 10-, 22- and 42-year P.sylvestris var. mongolica, 44-year P.tabulaeformis, and 42-year U.pumila(P<0.05), but the relationships were significant negatively correlated for 32 years old P.sylvestris var. mongolica and 13 years old P.simonii(P<0.05). The fine root biomass vertical distribution was deeper for main afforestation tree species in Horqin sandy land with significant dependence on the soil water content.

Analysis on Microsatellites in Rhodiola algida Based on Solexa Sequencing

LEI Shu-Yun1,2；GAO Qing-Bo1；FU Peng-Cheng1；YANG Hui-Ling1*；CHEN Shi-Long1；ZHANG Fa-Qi1,3

2014, 34(6):  829-834.  doi:10.7525/j.issn.1673-5102.2014.06.019

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Solexa sequencing was used to analyze microsatellites in Rhodiola algida based on EST-assembled sequences, expecting to find out the genetic diversity and to develop microsatellite markers for Rhodiola. A total of 6552 EST sequences were obtained in the current study, trinucleotide repeats were the most with 41.5%, mononucleotide and dinucleotide repeats were almost similar with 27.76% and 24.76%, respectively. The distribution densities of dinucleotides repeats to hexanucleotides were directly proportional to the number of SSR number. In mononucleotides repeats, thymine(T) and adenine(A) accounted for 14.91% and 12.70%, respectively, while with less G and C repeats. In dinucleotide repeats, AG repeats were the dominated with 5.60%, followed by GA(4.75%) and TC(4.72%). In trinucleotides repeats, GAA were the most frequent, accounted for 1.85%, followed by GAT(1.79%), while TTC, TCT, TCA, GGA, GCT, GAG with similar composition. The tetranucleotide, pentanucleotide and hexanucleotide repeats were very few. Except for pentanucleotide and hexanucleotide repeats, the variation of repeats are inversely proportional to the length of repeated nucleotide, while on one type, the variation of repeats is in negative correlation.

Effects of Soil Moisture on Biomass of Vaccaria segetalis(Necr.) Gracke and Vaccarin Content

YUAN Xiao-Han1,2；ZHANG Yan-Dong1；GAO Xue-Jun2；GU Cheng-Bo1*

2014, 34(6):  835-839.  doi:10.7525/j.issn.1673-5102.2014.06.020

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We tested pot cultured Vaccaria segetalis(Necr.) Gracke to study the effects of soil moisture on biomass and vaccarin (VR) content of V.segetalis in four different kinds of soil moisture. We determined the biomass and VR content of different organs of V.segetalis after 15, 30 and 45 days in different soil water conditions. The biomass and VR content of V.segetalis are affected significantly by the followed soil water conditions of severe soil drought, moderate soil drought, and excessive soil moisture compared with the control group, and VR content in different organs of V.segetalis can be improved significantly in moderate soil drought condition. The best biomass and VR yield of V.segetalis are in soil moisture of 70% 75% of field capacity. Proper soil moisture was proposed to maintain to balance between yield and quality of this medicinal herb in production of V.segetalis.

Allelopathy of Gallic Acid on Microcystis aeruginosa

PEI Yi1；DU Xian1；NIE Jiang-Li1*；ZHAI Huan-Huan2；ZHANG Bao-Kun1

2014, 34(6):  840-844.  doi:10.7525/j.issn.1673-5102.2014.06.021

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We studied the inhibitory effect of gallic acid on Microcystis aeruginosaand the effect of Gallicacid on ultrastructure of M.aeruginosa. M.aeruginosa was stressed with different concentrations of Gallic acid. With the increase of both concentration and time of Gallic acid, the growth of M.aeruginosa was significantly reduced. We observed cellular ultrastructure by SEM and TEM.The cells ultrastructure of M.aeruginosa after 72 h showed the changes, compared with the control, the cells walls of M.aeruginosa became wrinkled, collapsed and ruptured. Cytoplasm thylakoids photosynthetic lamellae showed dispersion and fracture. The phycobilisomes attached to the thylakoids decreased or dispersed to the cytoplasm, the number of the lipid particles and the cyanophycin particles of cytoplasm emerged different degrees increasing, a large void appeared in the nuclear area until most of the algal cells died.Therefore, gallic acid with obvious allelopathy can effectively inhibit the growth of M.aeruginosa.

Identification of Kiwifruit Ploidy Using Flow Cytometry

SHI Chun-Hui1,2；XU Lan-Lan3；WANG Xiao-Qing1,2；LUO Jun1,2*

2014, 34(6):  845-849.  doi:10.7525/j.issn.1673-5102.2014.06.022

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The contents of nuclear DNA in different ploidy kiwifruit cultivars were measured by flow cytometry, a simple, rapid method for the identification of kiwifruits ploidy was established. Diploid “Red Sun” Kiwi was used as control, ‘81-5’, ‘Wan Chui’ Kiwi as material, the ploidy was identified by screening the optimal leaf area, improving dissociation liquid formulations, improving membrane mesh and the number of filters, increasing the number of centrifugation. The results showed that open conditions of field cultivation, the early exhibition leaf under the stem tip is the best part of ploidy identification; twice centrifugation; dissociation solution 2% PVP-30; 500 mesh membrane filtration twice can effectively resolve the blocking of tube, effectively remove impurities and obtain a clear peak sample peak. The optimized technology enables the simultaneous determination of two or more kiwifruit mixed samples.

Discriminant Predictive Linear Regression Analysis Method for Astragalus

CHEN Yuan-Yuan1,2；ZHANG Xiao-Qin2；QIN Xue-Mei1；LI Zhen-Yu1*

2014, 34(6):  850-855.  doi:10.7525/j.issn.1673-5102.2014.06.023

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In this paper, the unary linear regression model between diameter of the xylem and cross section diameter was respectively set up for Astragalus membranaceus(Fisch.) Bge var. mongholicus(Bge.) Hsiao and A.membranaceus(Fisch) Bge.. Regression analysis and the classification of samples were conducted. Through several extracting random samples, correct classification rate was got. The results showed that the classification effect of A.membranaceus(Fisch.) Bge var. mongholicus(Bge.) Hsiao model was very good. In addition, the homogeneity of variance t test on samples of different growth years was carried on the samples of A.membranaceus(Fisch.) Bge var. mongholicus(Bge.) Hsiao. Through t test, it gave the confidence interval of the ratio between xylem and cross section diameter of samples of 2 years and above 2 years. Results showed that the interval of 2 years and above 2 years samples did not cross and could be obviously apart so that the growth years of Astragalus can be deduced. The study provides a simple and fast method for determining the varieties and growth years of Astragalus herbs. Hence, Astragalus samples can simply be classified and the growth years of a certain type of Astragalus can be determined.

Ultrasonic-assisted Extraction of Total Flavonoids from Peony Tree Seed Coat and Its Anticoagulation Activity

WANG Hong-Zheng1,2；LIU Wei1,2；ZHANG Lin1,2；CONG Ying1,2；ZAN Peng1,2；WANG Hua1,2,3；ZU Yuan-Gang1,2；CHEN Hua-Feng1,2*；ZHAO Xiu-Hua1,2

2014, 34(6):  856-860.  doi:10.7525/j.issn.1673-5102.2014.06.024

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We used ultrasonic-assisted extraction method to extract total flavonoids from seed coat of peony tree, and studied the extraction conditions including volume fraction of ethanol, ratio of liquid to solid (mL·g^-1), ultrasonic time and power which influenced the extraction yield of total flavonoids. The results of orthogonal test indicated the optimum extraction conditions of total flavonoids of seed coat were: volume fraction of 55% as solvent, liquid to solid ratio of 20∶1, ultrasonic time of 40 min and ultrasonic power of 140 W. Under these conditions, the total flavonoids yield of seed coat was 46.50‰. After column chromatography for flavonoids purification of seed coat extract, it contained 203.5 mg·g^-1 quercetin and 23.2 mg·g^-1 luteolin. Seed coat flavonoids of peony tree showed high activity of anticoagulation activity in experiments of in vivo rat.

Aqueous Extraction Technology on Main Active Component from Fresh Eucommia Bark

LI Yuan-Yuan；XIANG Feng-Ying；GAO Yue；ZU Yuan-Gang*；LU Qi；ZHANG Ying

2014, 34(6):  861-865.  doi:10.7525/j.issn.1673-5102.2014.06.025

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We investigated the total solid and the active component yields from fresh Eucommia bark by singlefactor experiments of aqueous extraction under the liquid ratio of 1∶20 to 1∶60 g·mL^-1, extraction time from 10-120 min, and extraction temperature from 30℃ to 90℃. Under the optimum extraction processes of singlefactor experiment, the total solid yield was 11.61%, and the extraction yields of chlorogenic acid, aucubin, geniposide and pinoresinol diglucoside were 0.073%, 0.704%, 0.122% and 0.108%, respectively. Aqueous extraction has advantages of simple process without environmental pollution and solvent recovery.