Water sources of dominant species in three alpine ecosystems on the Tibetan Plateau, China

Plant water sources were estimated by two or three compartment linear mixing models using hydrogen and oxygen isotope （δD and δ^18O） values of different components such as plant xylem water, precipitation and river water as well as soil water on the Tibetan Plateau in the summer of 2005. Four dominant species （Quercus aquifolioides, Pinus tabulaeformis, Salix rehderiana and Nitraria tangutorum） in three typical ecosystems （forest, shrub and desert） were investigated in this study. Stable isotope ratios of the summer precipitations and the soil water presented variations in spatial and temporal scales. δ^18O values of N. tangutorum xylem water were constant in the whole growth season and very similar to those of deep soil water. Water sources for all of the plants came from both precipitations and soil water. Plants switched rapidly among different water sources when environmental water conditions changed. Rainwater had different contributions to the plants, which was influenced by amounts of precipitation. The percentage of plant xylem water derived from rainwater rose with an increase in precipitation. Water sources for broad-leaved and coniferous species were different although they grew in the same environmental conditions. For example, the broad-leaved species Q. aquifolioides used mainly the water from deep soil, while 92.5% of xylem water of the coniferous species P. tabulaeformis was derived from rainwater during the growth season. The study will be helpful for us to fully understand responses of species on the Tibetan Plateau to changes in precipitation patterns, and to assess accurately changes of vegetation distribution in the future.     

Recycling Isolation of Plant DNA,A Novel Method

DNA is one of the most basic and essential genetic materials in the field of molecular biology.To date,isolation of sufficient and good-quality DNA is still a challenge for many plant species,though various DNA extraction methods have been published.In the present paper,a recycling DNA extraction method was proposed.The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times,and correspondingly four DNA precipitations(termed as the 1st,2nd,3rd and 4th DNA sample, respectively) were conducted.This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method.This modified CTAB method was tested in eight plant species,wheat,sorghum,barley,corn,rice,Brachypodium distachyon,Miscanthus sinensis and tung tree.The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species.The DNA samples were good templates for PCR amplification of both ISSR and SSR markers.The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods,and thus is an effective and universal DNA isolation method.     

Localization and Dynamic Change of Saponin in Vegetative Organs of Polygala tenuifolia

Anatomical, histochemical and phytochemical methods were used to investigate the structure, localization and dynamic changes of total saponin and senegenin of vegetative organs in Polygala tenuifolia Willd. Histochemical localization results showed that saponin accumulated mainly in parenchyma cells of vegetative organs. The phytochemical results also showed that the saponin accumulated in the vegetative organs of P. tenuifolia, with higher content in roots and lower content in the aerial parts that included stems and leaves. The saponin content and dry weight of the vegetative organs of P. tenuifolia had dynamic variance at the developmental stages and all reached the highest level in the post-fruit period. Hence, the roots and aerial parts should be gathered in August to make full use of the plant. As the root is the main medicinal organ of P. tenuifolia, the content of total saponin and senegenin of different aged and different parts of the root were determined. The content of total saponin and senegenin exhibited a sustained decreasing trend with increasing root age; therefore, the annual roots had high quality. The content of total saponin and senegenin in different parts of the root showed obvious variation. The content in the ＂skin areas＂ was much higher than that of xylem. The results offer a theoretical basis for determining the appropriate harvesting stage and a reasonable harvest of P. tenuifolia.     

Water sources of Populus euphratica and Tamarix ramosissima in Ejina Delta,the lower reaches of the Heihe River China北大核心CSCD

Aims: We aim to evaluate the water sources of typical riparian arbor species (Populus euphratica) and shrub species (Tamarix ramosissima), and analyze the spatial and temporal dynamics of plant water source in Ejina Delta, the lower reaches of the Heihe River. China. Methods: Eight sampling sites were selected in the riparian zones along the East River and West River in Ejina. The plant xylem water, soil moisture, rainwater, stream water and groundwater were taken and pretreated during the growing season in 2015-2016, and the stable oxygen isotope ratio (δ18O) for each water sample was measured. The δ18O of plant xylem water and soil moisture were compared to estimate the dominant depth of root water uptake, and the linear-mixed model called "IsoSource" were applied to determine plant water sources and quantify their proportions. Important findings: This study indicated that the main recharge sources for P. euphratica and T. ramosissima were stream water and groundwater. The contributions of rain water to them was negligible due to the limited amount and the shallow infiltration depth of local rainfall. As affected by groundwater level fluctuation, soil physical properties, as well as lateral and vertical recharge of stream water on soil moisture, the dominant depth of root water uptake spatially varied. However, the relative contributions of stream water or groundwater to plant water sources did not change significantly across space. Populus euphratica used more stream water (68%), while T. ramosissima used more groundwater (65%). Plant water sources were sensitive to environmental flow controls. The contributions of stream water to the water sources of the two species went up to 84% and 48% for P. euphratica and T. ramosissima respectively during the discharge period, but dropped to 63% and 30% during the non-discharge period. On the other hand, the contributions of groundwater decreased to 16% and 52% during the discharge period, but increased to 37% and 70% during non-discharge period. It is noteworthy that the high similarity of δ18O between stream water and groundwater due to extensive water exchange in the riparian zone made increase the uncertain in quantifying plant water sources.     

The Ca2+-dependent DNases are Involved in Secondary Xylem Development in Eucommia ulmoides

Secondary xylem development has long been recognized as a typical case of programmed cell death (PCD) in plants. During PCD, the degradation of genomic DNA is catalyzed by endonucleases. However, to date, no endonuclease has been shown to participate in secondary xylem development. Two novel Ca 2+ -dependent DNase genes, EuCaN1 and EuCaN2, were identified from the differentiating secondary xylem of the tree Eucommia ulmoides Oliv., their functions were studied by DNase activity assay, in situ hybridization, protein immunolocalization and virus-induced gene silencing experiments. Full-length cDNAs of EuCaN1 and EuCaN2 contained an open reading frame of 987 bp, encoding two proteins of 328 amino acids with SNase-like functional domains. The genomic DNA sequence for EuCaN1 had no introns, while EuCaN2 had 8 introns. EuCaN1 and EuCaN2 digested ssDNA and dsDNA with Ca 2+ -dependence at neutral pH. Their expression was confined to differentiating secondary xylem cells and the proteins were localized in the nucleus. Their activity dynamics was closely correlated with secondary xylem development. Secondary xylem cell differentiation is influenced by RNAi of endonuclease genes. The results provide evidence that the Ca 2+ -dependent DNases are involved in secondary xylem development.     

Feature selection for the prediction of translation initiation sites

Translation initiation sites （TISs） are important signals in cDNA sequences. In many previous attempts to predict TISs in cDNA sequences, three major factors affect the prediction performance： the nature of the cDNA sequence sets, the relevant features selected. and the classification methods used. In this paper, we examine different approaches to select and integrate relevant features for TIS prediction. The top selected significant features include the features from the position weight matrix and the propensity matrix, the number of nucleotide C in the sequence downstream ATG, the number of downstream stop codons. the number of upstream ATGs, and the number of some amino acids, such as amino acids A and D. With the numerical data generated from these features, different classification methods, including decision tree. naive Bayes, and support vector machine, were applied to three independent sequence sets. The identified significant features were found to be biologically meaningful. while the experiments showed promising results.     

Graft‐accelerated virus‐induced gene silencing facilitates functional genomics in rose flowers

Rose has emerged as a model ornamental plant for studies of flower development, senescence, and morphology, as well as the metabolism of floral fragrances and colors.Virus-induced gene silencing(VIGS) has long been used in functional genomics studies of rose by vacuum infiltration of cuttings or seedlings with an Agrobacterium suspension carrying TRV-derived vectors. However, VIGS in rose flowers remains a challenge because of its low efficiency and long time to establish silencing. Here we present a novel and rapid VIGS method that can be used to analyze gene function in rose,called ‘graft-accelerated VIGS', where axil ary sprouts are cut from the rose plant and vacuum infiltrated with Agrobacterium. The inoculated scions are then grafted back onto the plants to flower and silencing phenotypes can be observed within 5 weeks, post-infiltration. Using this new method, we successfully silenced expression of the RhDFR_1, RhA G, and RhNUDX_1 in rose flowers, and affected their color, petal number, as well as fragrance, respectively. This grafting method will facilitate high-throughput functional analysis of genes in rose flowers. Importantly, it may also be applied to other woody species that are not currently amenable to VIGS by conventional leaf or plantlet/seedling infiltration methods.     

简单快速的DNA银染和胶保存方法

本文介绍了一套简单快速的DNA银染以及胶保存的方法,整个过程仅需10~15分钟,而且背景浅,条带清楚,灵敏度高,稳定性好。胶保存采用双层玻璃纸夹心法,可长久地保存胶显色时的原貌。以常规PAG胶检测和HLA的SSCP分型为例,利用该套方法进行了银染以及胶的保存,均得到了满意的结果。该方法具有推广价值。 Abstract:This paper introduced the simple and rapid methods of silver staining and gel preservation.It was taken only about 10 and 15 minutes to stain a gel.The background of gel was light,the bands were clear,the sensibility was high and the stabilization was well by the method of silver staining.The gel preservation adopted a method named two-layer transparent plastic paper "Sandwich" which could keep the gel with primitive colors for a long time.The methods were used on PAG checking and SSCP typing of HLA and the results were satisfactory.The set of methods are expected to be widely used in laboratories.     

Influence of light and habitat on predation of Culex quinquefasciatus （Diptera： Culicidae）larvae by the waterbugs （Hemiptera： Heteroptera）

The influence of light and habitat structure on the predation of Culex quinquefasciatus larvae by the common heteropteran water bug, Diplonychus （= Sphaerodema） annulatus, D. rusticus and Anisops bouvieri was assessed in the laboratory. It was revealed that water bugs predate more in presence of light than in dark conditions. While A. bouvieri consumed more prey in structured conditions, D. annulatus and D. rusticus consumed more prey in open conditions. The selection of prey size as well as the respective numbers varied between predators and treatments significantly. Prey vulnerability （PV）, an indicator of predatory efficiency, was highest for D. annulatus, moderate for D. rusticus and low for A. bouvieri. Prey consumption and PV values under different treatment conditions indicate that for belostomatid water bugs D. annulatus and D. rusticus, the order of prey consumption under different habitat and light/dark combinations is light open 〉 dark open 〉 light vegetated 〉 dark vegetated. In the case of the backswimmer, A. bouvieri, the order of prey consumption appeared to be light structured 〉 dark structured 〉 light open 〉 dark open. These findings were consistent with resource-partitioning by water bug species sharing the same guild. If the observed results are extended to natural settings the efficiency of these predators in controlling mosquito populations will vary with the structural complexity of habitats and the intensity of light.     

图位克隆技术在分离植物基因中的应用

本文综述图位克隆技术的原理及基本技术环节,并与其它基因克隆技术相比较说明图位克隆技术的优缺点。本文还对近年来图位克隆技术在分离不同的植物发育基因上的广泛应用进行了简要概述,并对其应用前景和近期的预期进展作出展望。 Abstract:In this review,the principle and basis steps of map-based gene cloning on plant were introduced in detail.At the same time,the advantages and disadvantage of the method were evaluated as compared with the other methods.The extensive application of the map-based gene cloning method on cloning genes of different plants in the past was also summarized in brief.The prospect and the expected progress in several years were proposed in this paper.     

Analysis of the Caudal Vortices Evolvement around Flapping Foil

The viscous flow field around two-dimensional flapping （ heaving and pitching） foils was numerically computed. The structural characteristics of caudal vortices were investigated and the contour curves at different phase angles were obtained. The relationships between the structural characteristics of the vortices and the force acting on the foil and between the widths of the caudal vortex street and of the caudal flow field were analyzed. A method to determine the shedding frequency of the vortices was proposed.     

Na~ and Water Uptake in Relation to the Radial Reflection Coefficient of Root in Arrowleaf Saltbush Under Salt Stress

The response of halophyte arrowleaf saltbush(Atriplex triangularis Willd)plants to a gradient of salt stress were investigatedwith hydroponically cultured seedlings.Under salt stress,both the Na~ uptake into root xylem and negative pressures inxylem vessels increased with the elevation of salinity(up to 500 mol/m~3)in the root environment.However,the increment innegative pressures in root xylem far from matches the decrease in the osmotic potential of the root bathing solutions,evenwhen the osmotic potential of xylem sap is taken into consideration.The total water potential of xylem sap in arrowleafsaltbush roots was close to the osmotic potential of root bathing solutions when the salt stress was low,but a progressivelyincreased gap between the water potential of xylem sap and the osmotic potential of root bathing solutions was observedwhen the salinity in the root environment was enhanced.The maximum gap was 1.4 MPa at a salinity level of 500 mol/m~3without apparent dehydration of the tested plants.This discrepancy could not be explained with the current theories inplant physiology.The radial reflection coefficient of root in arrowleaf saltbush decreased with the enhanced salt stress wasand accompanied by an increase in the Na~ uptake into xylem sap.However,the relative Na~ in xylem exudates based onthe corresponding NaCl concentration in the root bathing solutions showed a tendency of decrease.The results showedthat the reduction in the radial reflection coefficient of roots in the arrowleaf saltbush did not lead to a mass influx of NaClinto xylem when the radial reflection coefficient of the root was considerably small;and that arrowleaf saltbush could usesmall xylem pressures to counterbalance the salt stresses,either with the uptake of large amounts of salt,or with thedevelopment of xylem pressures dangerously negative.This strategy could be one of the mechanisms behind the highresistance of arrowleaf saltbush plants to salt stress.     

Chemical Constituents of the Roots of Anemone altaica Fisch. ex C. A. Mey.

The roots of Anemone altaica Fisch. ex C. A. Mey. have been used in the treatment of epilepsia, neurasthenia, and arthritis in Chinese folk medicine for a long time. In order to find new and bioactive compounds, the chemical constituents of the roots of A. altaica were investigated and nine compounds were isolated from the EtOH extract of this plant. On the basis of spectroscopic methods, the structures of these compounds were elucidated as 4-（9H-β-carbolin- 1-yl）-4-oxo-butyric acid （1）, carboxymethyl isoferulate （2）, isoferulic acid （3）, cirsiumaldehyde （4）, 5-hydroxy-4-oxo-pentanoic acid （5）, triacontane （6）, palmic acid （7）, β-sitosterol （8）, and daucosterol （9）. Among them, 1 and 2 were new compounds, and 3 and 4 were obtained from this genus for the first time.     

Nitrogen released from different plant residues of the Loess Plateau and their additions on contents of microbial biomass carbon, nitrogen in soil

An incubation method was used to investigate the nitrogen release characteristics from the residue of ten plant species which commonly grow in the northern part of the Loess Plateau. The effect of the residue on soil microbial biomass carbon (SMBC) and soil microbial biomass nitrogen (SMBN) was also determined. There were significant differences in the total N content and the C/N ratios among the different types of plant residue. The total N content of the residues ranged from 6.61 to 32.78 g kg^?1. The C/N ratio of the residue ranged from 14 to 65. There was an immediate increase in soil N after alfalfa, erect milkvetch, and korshinsk peashrub residue was added to the soil. In contrast, soil N decreased after elm, sea buckthorn, and wild peach residue was added to the soil. The soil N content remained relatively low for 14–34 days and then increased. This indicated that N immobilization occurred during the early portion of the incubation period when elm, sea buckthorn and wild peach residue was added to the soil. Soil N levels were low during the entire incubation period when simon poplar, locust, Stipa bungeana, and old world bluestem residue were added to the soil. The addition of plant residue significantly increased SMBC and SMBN in all treatments. The SMBC and SMBN values were greatest in treatments containing plant residue with high total N content and low C/N ratios. The C/N ratios of korshinsk peashrub, sea buckthorn, and wild peach residues were similar, but the amount of N released from these residues and the effects of the residue on SMBC and SMBN in soil were significantly different. This indicates that not only the C/N ratio but also the chemical composition of the plant residue affected decomposition. It is important to consider C and N release characteristics from plant residue in order to adjust the C and N balance of soil when revegetating degraded ecosystems.     

Apoptosis of breast cancer cell MCF-7 induced by morcantharidin(NCTD)

To study the cytotoxic effects and antitumour mechanism of norcantharidin(NCTD) on human breast cancer cell, MCF-7 cells were exposed to culture medium with NCTD in different doses and hours. The cell growth inhibition curves of NCTD on MCF-7 cells were acquired by detecting the growth ratio of MCF-7 cells with MTT methods. The results indicated that NCTD had inhibition effects on growth of MCF-7 cells depending on the doses and times of treatment. When the concentrations of NCTD used was lower than 5 μg/ml, the cells grew as same as the control cells. With increasing doses and times of treatment, the inhibition effects of NCTD on the cells' growth increased, for example, when exposed to 10 μg/ml NCTD     

Nitrogen released from different plant residues of the Loess Plateau and their additions on contents of microbial biomass carbon, nitrogen in soil

An incubation method was used to investigate the nitrogen release characteristics from the residue of ten plant species which commonly grow in the northern part of the Loess Plateau. The effect of the residue on soil microbial biomass carbon (SMBC) and soil microbial biomass nitrogen (SMBN) was also determined. There were significant differences in the total N content and the C/N ratios among the different types of plant residue. The total N content of the residues ranged from 6.61 to 32.78 g kg^?1. The C/N ratio of the residue ranged from 14 to 65. There was an immediate increase in soil N after alfalfa, erect milkvetch, and korshinsk peashrub residue was added to the soil. In contrast, soil N decreased after elm, sea buckthorn, and wild peach residue was added to the soil. The soil N content remained relatively low for 14–34 days and then increased. This indicated that N immobilization occurred during the early portion of the incubation period when elm, sea buckthorn and wild peach residue was added to the soil. Soil N levels were low during the entire incubation period when simon poplar, locust, Stipa bungeana, and old world bluestem residue were added to the soil. The addition of plant residue significantly increased SMBC and SMBN in all treatments. The SMBC and SMBN values were greatest in treatments containing plant residue with high total N content and low C/N ratios. The C/N ratios of korshinsk peashrub, sea buckthorn, and wild peach residues were similar, but the amount of N released from these residues and the effects of the residue on SMBC and SMBN in soil were significantly different. This indicates that not only the C/N ratio but also the chemical composition of the plant residue affected decomposition. It is important to consider C and N release characteristics from plant residue in order to adjust the C and N balance of soil when revegetating degraded ecosystems.     

小红参的抗癌环己肽甙和乔木萜烷型三萜新成分

The roots of Rubia yunnanensis Diels.known as"Xiao Hong Sheng",are used asantitumor drug in Yunnan.From methanol extract of the plant,a newglycocyclohexapeptide and a cyclohexapeptide(deoxybouvadin)with high antitumor ac-tivity,together,with ten arborane type triterpenoids Including seven new ones.were iso-lated and their structures were elucidated on the basis of spectral and chemical methods.     

Distributions of arsenic and essential elements in pinna ofarsenic hyperaccumulator Pteris vittata L.

The distributions of arsenic and 6 essential elements in the pinna of As hyperac-cumulator, Pteris vittata L., were studied using synchrotron radiation X-ray fluorescence (SRXRF). Significant correlation between the distribution and mobility of the elements revealed that SRXRF study on the elemental distribution was feasible to inspect the transportations of elements in plants. The distribution of As in the pinna showed that As had great abilities to be transported in xylem vessels and from xylem to mesophyll. The distribution of K, one of the most mobile elements in plants, was similar to that of As, whereas the distributions of Fe and Ca with less mobility in plants were almost opposite to that of As in the pinna.     

DNA Extraction Protocol for Biological Ingredient Analysis of Liuwei Dihuang Wan

Traditional Chinese medicine(TCM) preparations are widely used for healthcare and clinical practice. So far, the methods commonly used for quality evaluation of TCM preparations mainly focused on chemical ingredients. The biological ingredient analysis of TCM preparations is also important because TCM preparations usually contain both plant and animal ingredients,which often include some mis-identified herbal materials, adulterants or even some biological contaminants.For biological ingredient analysis, the efficiency of DNA extraction is an important factor which might affect the accuracy and reliability of identification. The component complexity in TCM preparations is high, and DNA might be destroyed or degraded in different degrees after a series of processing procedures. Therefore, it is necessary to establish an effective protocol for DNA extraction from TCM preparations. In this study, we chose a classical TCM preparation,Liuwei Dihuang Wan(LDW), as an example to develop a TCM-specific DNA extraction method.An optimized cetyl trimethyl ammonium bromide(CTAB) method(TCM-CTAB) and three commonlyused extraction kits were tested for extraction of DNA from LDW samples. Experimental results indicated that DNA with the highest purity and concentration was obtained by using TCM-CTAB. To further evaluate the different extraction methods, amplification of the second internal transcribed spacer(ITS2) and the chloroplast genome trnL intron was carried out.The results have shown that PCR amplification was successful only with template of DNA extracted by using TCM-CTAB. Moreover, we performed high-throughput 454 sequencing using DNA extracted by TCM-CTAB. Data analysis showed that 3–4 out of 6 prescribed species were detected from LDW samples, while up to 5 contaminating species were detected, suggesting     

Bacterial diversity in deep-sea sediment from northeastern Pacific Ocean

16S rDNA sequencing method is one of the effectively used culture-independent techniques in recent years. In this study, 16S rDNA sequencing method was used to investigate the bacterial diversity in deep-sea sediment from northeastern Pacific polymetallic nodule province. Total DNAs were extracted by using 2 different methods (chemical method and DNA extracting kit method). After purification, genomic DNA was amplified by using 2 universal primers (27F and 1492R). Clones were selected and sequenced randomly. After the sequences were checked by using the Chimera Check Program of the RDP database, a bacterial 16S rRNA gene library of 79 clones was established. Phylogenetic analysis indicated that 79 clones could be divided into 11 phylotypes. Gamma Proteobacteria (22.8%) and alpha Proteobacteria (16.5%) were the dominant components of the sediment bacterial community, followed by Planctomycetacia (7.6%), delta Proteobacteria (6.3%), Nitrospira (6.3%), Actinobacteria (6.3%), beta Proteobacteria (5%), Acidobacteria (5.1%), Sphingobacteria (3.8%), Firmicutes (2.5%) and uncultured bacteria (17.7%). Gamma Proteobacteria also dominated at slices 0–2 cm and 4–6 cm. Different slices had different types of bacteria, alpha Proteobacteria, gamma Proteobacteria, delta Proteobacteria, Planctomycetacia, Nitrospira, Actinobacteria and Acidobacteria, however, appeared in all slices. Pseudomonas is common in many different deep-sea environments. In this study, it accounted for 22.2% of the total gamma Proteobacteria.