当前位置:文档之家› Transcriptomic responses to aluminum stress in soybean roots

Transcriptomic responses to aluminum stress in soybean roots

Transcriptomic responses to aluminum stress in soybean roots
Transcriptomic responses to aluminum stress in soybean roots

Abstract:Aluminum (Al)toxicity is the primary limitation to crop production and plant growth in acid soils.Soybean has multiple mechanisms of Al resistance including the complexing and exclusion of Al in root apices by Al-induced citrate se-cretion.Microarray analysis is available for the identification of genes in soybean.In the present study,Affymetrix soybean genome array was used to identify the Al-induced differentially expressed genes in Al-resistant genotype Jiyu 70.With a cutoff of >2.0-fold (p <0.05)between non Al-treated and Al-treated root apices,561genes were upregulated and 78genes were downregulated when roots were exposed to 30μmol/L AlCl 3for 4h.Quantitative real-time PCR was used to test the microarray data.The analysis showed that nearly half of the Al-responsive genes were of unknown biological function.A higher proportion of genes related to transcription regulation and cell wall processes were observed in Al-induced up-and downregulated genes,respectively.Some genes homologous to the citrate transporter MATE family gene or C 2H 2family transcription factor gene,STOP1,were detected in our analysis.Some genes related to lignin deposition were upregulated,which might be related to Al-induced root elongation inhibition.Key words:aluminium,microarray,gene expression,acid soil.

Résumé:La toxicitéde l ’aluminium (Al)est la plus importante contrainte àla production végétale et àla croissance des plantes dans les sols acides.Le soya possède plusieurs mécanismes de résistance àla toxicitéde l ’Al incluant la complexa-tion et l ’exclusion de l ’Al dans les apex racinaires par sécrétion de citrate induite par l ’Al.Des puces àADN sont disponi-bles pour identifier les gènes chez le soya.Dans ce travail,les auteurs ont employéune puce soya Affymetrix pour

identifier les gènes induits par l ’Al chez le génotype résistant àl ’Al Jiyu 70.En employant des seuils de 2×pour l ’ampleur des changements d ’expression (p <0,05)entre les apex racinaires exposés ou non àl ’Al,561gènes ont étésur-exprimés et 78gènes sous-exprimés suite àun traitement de 4h en présence de 30μmol/L d ’AlCl 3.Des analyses PCR en temps réel (qRT-PCR)ont étéemployées pour valider les données sur puce.Cette analyse a montréque,pour près de la moitiédes gè-nes dont l ’expression était changée en présence d ’Al,leur fonction est inconnue.Une surreprésentation de gènes liés àla ré-gulation transcriptionnelle et àla paroi cellulaire,respectivement,a étéobservée parmi les gènes sur-et sous-exprimés.Certains gènes homologues àla famille MATE des transporteurs de citrate ou au gène STOP1de la famille des facteurs de transcription C 2H 2ont étédétectés lors de cette analyse.Certains gènes impliqués dans la déposition de la lignine étaient sur-exprimés ce qui pourrait être en lien avec l ’inhibition de l ’élongation des racines qui est induite par l ’Al.Mots ‐clés :aluminium,puce àADN,expression des gènes,sol acide.[Traduit par la Rédaction]

Introduction

Acid soils (pH <5.5)are the most important limitation to the yield production in many crops,which cover about 30%of the world ’s arable soils and up to 70%of potentially arable land (von Uexküll and Mutert 1995).Soil acidification is an increasing problem in the world because of acid rain,the re-moval of the natural plant coverage from large production areas,and the use of ammonium-based fertilizers (Pannatier et al.2005).A recent report has shown the rapid decline of soil pH in China within the past 20years was associated with intensive agriculture and the excessive utilization of ni-trogen fertilizers (Guo et al.2010).Aluminum (Al)toxicity is considered to be one of the most important limiting factors to crop production in acid soils.Under acidic conditions,toxic forms of Al will mobilize into the soil solution and rapidly inhibit plant root growth,subsequently decreasing nutrient and water uptake,and result in poor crop growth and produc-tivity (Kochian et al.2004).

To deal with Al stress,many plant species have evolved a series of strategies to adapt to the acidic soil environment.Progress has been made in the previous few years to

better

923

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

understand these strategies.Al-induced organic acid anions efflux has been considered as the most important mechanism of Al resistance in a wide range of plant species,such as wheat,buckwheat,maize,soybean,common bean,Cassia tora ,and rye,by complexing and detoxifying Al in the rhizo-sphere (Kochian et al.2004).Malate and citrate are the gen-eral Al-induced organic acid anions in plants,which are suggested to be easy to synthesis and abundant within cells (Ryan and Delhaize 2010).Ma (2000)proposed that organic acid anion secretion can be classified into two patterns.In pattern I,Al might activate a preexisting transporter (anion channel)in the plasma membrane to initiate organic anion ef-flux,and the induction of novel protein synthesis is not re-quired.Whereas in pattern II,the induction of genes and synthesis of proteins involved in organic acid metabolism (bio-synthesis and decomposition)or transport (anion channel on plasma membrane and (or)tonoplast,or transport of organic acids from mitochondria)is required.

Soybean is considered as a pioneering crop and is widely grown in acid soils,including Southeast USA,Cerrados in Brazil,and South China.Citrate exudation from the root tips is crucial for Al resistance in soybean (Yang et al.2000,2001b ;Silva et al.2001).The citrate secretion from soybean has been shown to require at least 4h of Al exposure (Yang et al.2001b ).Furthermore,Al-induced citrate secretion from soybean roots was inhibited by the nonspecific protein syn-thesis inhibitor cycloheximide (You et al.2010).All the char-acteristics are consistent with pattern II proposed by Ma (2000),and thus it suggests that gene induction and novel protein synthesis are essential for Al-induced citrate efflux from soybean roots.However,the genes that confer citrate ef-flux to soybean roots have not been well established.

Besides citrate efflux,soybean appears to display additive mechanisms of Al resistance or tolerance.According to the report of Nian et al.(2005),the amounts of Al-induced cit-rate exudation was not well correlated with Al tolerance in a wide range of soybean genotypes,indicating that mechanisms other than citrate efflux may also play important roles.The hypothesis that multiple mechanisms of Al resistance existed in soybean is also supported by genetic studies (Bianchi-Hall et al.1998,2000)that suggest Al tolerance in soybean is a multigenetic trait.Thus an integrative molecular study ap-pears to be necessary to further classify the mechanisms of Al toxicity and tolerance.By the techniques,such as micro-array or suppression subtractive library,the transcriptional expression in response to Al stress has been studied on Ara-bidopsis (Kumari et al.2008),maize (Maron et al.2008),wheat (Guo et al.2007),Medicago truncatula (Chandran et al.2008),and common bean (Eticha et al.2010),and a series of Al-responsive genes were identified.Recently,a series of Al-tolerance genes were also studied by microarray in Al-resistant soybean genotype PI 416937(Duressa et al.2010a ).Microarray was also used to study the mechanisms of magnesium-alleviating Al toxicity in soybean at the tran-scriptional expression level (Duressa et al.2010b ).In our present study,Affymetrix soybean genome array was used to study the transcriptional profile of soybean under Al stress and to find the genes related to Al toxicity and resist-ance,particularly the genes that confer to Al-induced citrate efflux.

Materials and methods

Plant materials and culture conditions

Soybean seeds (cultivar Jiyu70)were surface sterilized in 1.0%(v /v )sodium hypochlorite for 5min,washed 3–4times with tap water,embedded in peat moss,and germinated for 3days at 25°C in the dark.After germination,the uniform seedlings were selected and cultured in 1L simplified nu-trient solution containing 0.5mmol/L CaCl 2solution at pH 4.5for 1day.Then,the seedlings were subjected to 0.5mmol/L CaCl 2solution without (control)or with AlCl 3(10,30,50m mol/L)for 4,8,12,and 24h.The root length was measured before and after the treatment.Relative root elongation (%)was calculated as follows:(root elongation in the treatment /root elongation in the control)×100.

Three-day-old seedlings were cultured in 1L plastic pots filled with complete nutrient solution as described by Horst et al.(1992)containing 750μmol/L KNO 3,250μmol/L Ca(NO 3)2,325μmol/L MgSO 4,10μmol/L KH 2PO 4,20μmol/L Fe-EDTA,8μmol/L H 3BO 3,0.2μmol/L CuSO 4,0.2μmol/L ZnSO 4,0.2μmol/L MnCl 2,and 0.2μmol/L (NH 4)6Mo 7O 24.The pH of the solution was adjusted to 4.5with 0.1mol/L HCl/KOH frequently and renewed every other day.After 5days culture,the seedlings were trans-ferred into 0.5mmol/L CaCl 2solution overnight.Then,seedlings were separated into two groups:(i )plants grown in 0.5mmol/L CaCl 2solution (pH 4.5);(ii )plants grown in 0.5mmol/L CaCl 2solution containing 30m mol/L AlCl 3(pH 4.5).After 4h treatment,the 1cm root tips were ex-cised and frozen immediately in liquid nitrogen for RNA isolation.All experiments were carried out in a controlled growth chamber at 25°C day :22°C night temperatures,70%constant relative humidity,14h light :10h dark cycles,300μmol m –2s –1of light intensity during the day,and aerated continuously.

RNA extraction and microarray hybridization

Total RNA of the 1cm root tips was extracted using the RNeasy Plant mini kit (QIAGEN,Valencia,California)and treated with RNase-free DNase (QIAGEN)following the manufacturer ’s instructions.cDNA and then cRNA were pre-pared,fragmented,hybridized,stained,and scanned accord-ing to the manufacturer ’s recommended protocols (Affymetrix,Santa Clara,California).Raw intensities were background corrected and normalized.The Affymetrix soy-bean genome array interrogated approximately 37500soy-bean transcripts with sequence information included in GeneBank and dbEST.The description of the GeneChip soy-bean genome array is available at the manufacturer ’s Web site.

Data analysis

Data analysis was performed by pair wise comparisons of GeneChips using dCHIP (Li and Wong 2001).In a compari-son analysis,we applied a paired t test to identify signifi-cantly differentially expressed genes between the control (no Al treatment)and Al treatment groups.Statistically signifi-cant differentially expressed genes were represented by a fold change (+Al/–Al treatment)of gene expression >2.0and p value <0.05.The sequence of each differentially ex-pressed gene was BLASTed in the Web site of Affymetrix

924Genome,Vol.54,2011

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

and manually annotated by comparison with the GenBank database using the online Basic Local Alignment Search Tool (BLAST)program (https://www.doczj.com/doc/e316156493.html,/Blast.cgi).

Quantitative real-time (qRT)-PCR

To validate the microarray results,RNA from three addi-tional independent replicates were extracted from the control and Al treatment groups,and cDNA was synthesized from 2μg of RNA using a high capacity RNA-to-cDNA kit (Ap-plied Biosystems,Foster City,California).DNAman 5.0soft-ware (Lynnon Corporation)was used to design gene-specific primers (see Supplementary data,1Table S1).The PCR pri-mer sets were designed to produce PCR products of about 120–140bp in length.For all the genes studied here,the op-timal primer concentration was 200nmol/L.qRT-PCR was performed using 0.5μL of above reverse transcription pro-duction in a 25μL reaction volume with SYBR Premix Ex Taq ?(TaKaRa Bio Inc.,Japan)on an ABIPRISM 7500Se-quence Detection System (Applied Biosystems).qRT-PCR conditions were as follows:95°C (10s);then 45cycles of 95°C (5s),58°C (20s),and 72°C (34s).The products were further analyzed by a dissociation curve program at 95°C (15s)to 60°C (1min)and then 95°C (15s).In each real time-PCR experiment,each gene was run in tripli-cate with different cDNAs synthesized from three biological replicates.Relative fold changes of gene expression were cal-culated using the comparative DD C t method (Livak and Schmittgen 2001),and b -tubulin was used as an internal standard.For microarray validation,the 2àDD C t values were calculated for each gene in each sample,log 2transformed,and plotted against its corresponding means data from the microarray.

Results and discussion

Al tolerance and sensitivity of soybean used in the present study

We performed a microarray analysis to identify the Al-induced differentially expressed transcripts of soybean root tips.Inhibition of root elongation is the primary and most dramatic effect of Al toxicity,and root apex is the primary site of Al perception and response (Sivaguru and Horst 1998;Kollmeier et al.2000).According to our previous study,30μmol/L AlCl 3resulted in about 40%root elonga-tion inhibition of Jiyu 70(Hou et al.2010),which repre-sents a physiological stress responses,but not sever damage.Thus this optimal Al treatment concentration was used in this study.To select an appropriate duration of Al exposure for stress treatments,we conducted a time course response analysis for Al-induced root elongation inhibition.Figure 1shows that the root elongation was inhibited about 50%within 4h of Al treatment.Therefore,a 4h Al expo-sure period is needed before the onset of severe toxicity symptoms.Also,it has been reported that citrate efflux from soybean root tips was induced by Al treatment after a lag phase of 4h (Yang et al.2001b ).During the first 4h,gene induction was suggested to occur to facilitate citrate

efflux.Thus 4h was selected as the Al treatment duration in the present study.

Al-induced genes expression in root tips of soybean

Transcript levels of control (no Al treatment)and Al-treated soybean root apices were compared by microarray analysis.Figure 2depicts a global picture of gene expression during Al stress.In Fig.2A,the log 2(ratio)equal to zero and away from zero indicated nonsignificant and significant (p <0.05)change in gene expression,respectively.The majority of gene expression has no significant change under Al stress (Fig.2A).We focused our attention on transcripts that were significantly (p <0.05)regulated by >2.0-fold between Al-and non Al-treated root apices.As a result,a total of 639dis-tinct genes were identified as statistically significantly ex-pressed,of these,561genes were upregulated and 78genes were downregulated when roots were exposed to 4h Al stress.Figure 2B shows that there are genes with great tran-scriptional changes such as >30-fold,but most of the genes expression ranged from 2-to 4-fold.

Validation of microarray results by qRT-PCR

Microarray data were independently verified by qRT-PCR.Fourteen genes with different functions and expression levels were selected among all differentially regulated genes,and their transcript abundance was monitored by qRT-PCR.The results confirmed the gene expression observed in the micro-array results.Results from microarray and qRT-PCR methods show close correlation (R 2=0.8044)(Fig.3).

Functional categories of the up-and downregulated genes in response to Al

The transcriptional results were categorized based on their different functions,as shown in Fig.4,by searching from previously published Al-and other stress-related literatures.The Al-induced genes are involved in a wide range of func-tions.Nearly half of the Al-responsive genes (both up-and downregulated)were with unknown functions.More tran-scription factor-and cell-wall-related genes were found in Al-induced up-and downregulated genes,respectively (Fig.4).

The detailed information of genes with different functions is listed in the Supplementary data,Table S2.

Organic acid synthesis,metabolism,and transport-related genes

The genes involved in organic acid synthesis,metabolism,and transport processing may play important roles in Al-induced citrate exudation from root tips and thus Al resist-ance in soybean (Yang et al.2000,2001b ;Silva et al.2001).The role of organic acid metabolism in the root tis-sue in Al-induced citrate secretion are still not well defined (Ryan et al.2001).In soybean,citrate metabolism may play an important role in Al-induced citrate efflux.Silva et al.(2001)suggested that the Al-enhanced internal accumulation of citrate contributed to the enhanced citrate exudation.Our previous study (Xu et al.2010)revealed that the activities of mitochondrial malate dehydrogenase and citrate synthase increased and aconitase decreased with the increasing of Al

1Supplementary

data are available with the article through the journal Web site (https://www.doczj.com/doc/e316156493.html,/doi/suppl/10.1139/g11-060).

You et al.925

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

concentration and Al treatment duration,and the transcrip-tional level of the mitochondrial citrate synthase increased in soybean roots exposed to Al.Furthermore,present re-search failed to detect a significant change in the transcript abundance for any of the genes involved in the tricarboxylic acid (TCA)cycle.However,we found some genes related closely to the TCA cycle.

The expressions of four genes (Affymetrix gene ID GmaAffx.92463.1.S1_at,GmaAffx.74033.1.A1_s_at,Gma.7631.1.A1_at,and GmaAffx.74033.1.S1_s_at)with high similarity (different identity)to the sequence of NADP-dependent malic enzyme (NADP-ME; E.C. 1.1.1.40)gene were upregulated by Al treatment.NADP-ME catalyses the decarboxylation of L -malate to pyruvate,providing reductive power (NADPH)and pyruvate for biosynthesis or catabolism (Edwards and Andreo 1992).NADP-ME was suggested to contribute to the accumulation and secretion of citrate by its reaction product (pyruvate)supplying the carbon skeletons to the TCA cycle (Rangel et al.2010).

The transcription abundance of two putative mitochondrial dicarboxylate carrier (DIC)genes (Gma.3832.2.S1_a_at and Gma.3832.2.S1_x_at)significantly increased under Al stress.DICs were detected in rat heart and brain tissues and are sug-gested to fulfill the roles supplying substrates to the TCA cycle (Fiermonte et al.1998).Direct transport assay with re-combinant and reconstituted DIC proteins clearly demon-strated that they facilitate the transport of dicarboxylates,such as malate,oxaloacetate,and succinate at high rate (Pal-mieri et al.2008).According to their broad substrate spec-trum,DICs may play a role in importing dicarboxylates into mitochondria,which can be used to replenish the TCA cycle (Palmieri et al.2008).

It has been reported that the multidrug and toxic com-pound extrusion (MATE )family protein gene plays a major role in the regulation of Al-induced citrate efflux from plant root apices in barley (HvMATE ,Furukawa et al.2007),sor-ghum (SbMATE ,Magalhaes et al.2007),and Arabidopsis (AtMATE ,Liu et al.2009).In our previous study,we found Al increased the expression of Gm-AlCT (accession EU586179),a member of the MATE family identified in soy-bean (Xu et al.2010).In the present study,a gene (Gma.8768.1.A1_at)putatively belonging to the MATE fam-ily was found to be highly upregulated (approximately 23-fold change)by Al,which encodes a putative protein with 82%,76%,and 64%similarity to GmFRD3,AtFRD3,and ScFRDL1,respectively.The AtFRD3gene encodes a MATE protein expressed in cells surrounding the root vasculature where it facilitates the efflux of citrate into the xylem to ac-company iron movement to the shoots (Rogers and Guerinot 2002).Further study showed that Arabidopsis plants ectopi-cally expressing FRD3-GFP had significantly higher amounts of citrate in their root exudates and greater resistance to Al compared with the untransformed control plants (Durrett et al.2007).Two homologous genes ScFRDL1and ScFRDL2were isolated from rye,ScFRDL1was involved in the trans-port of citrate into the xylem for iron translocation from the roots to the shoots,and ScFRDL2was involved in Al-acti-vated citrate secretion from the roots of rye (Yokosho et al.2010).The expression of both GmFRD3a and GmFRD3b genes has been found to be induced by iron deficiency in a iron-efficient soybean cultivar (Rogers et al.2009).The role of the FDR gene in Al-induced citrate efflux from soybean roots is less known.

Recently,it has been reported that the induction of the Cys2His2(C2H2)zinc finger-type transcription factors ex-pression confers the tolerance to low pH and Al toxicity.For example,sensitive to proton rhizotoxicity 1(STOP1)in Ara-bidopsis (Iuchi et al.2007;Liu et al.2009)and Al resistance transcription factor 1(ART1)in rice (Yamaji et al.2009).STOP1was shown to be required for the AtMATE expression and Al-activated citrate exudation in Arabidopsis (Sawaki et al.2009;Liu et al.2009).In the present study,the

transcrip-Fig.1.Effect of Al treatment on the root growth of soybean (30μmol/L AlCl 3).Soybean seedlings (4-day-old)were exposed to 0or 30μmol/L AlCl 3in 0.5mmol/L CaCl 2solution (pH 4.5)for 4,8,12,and 24h.Root elongation was calculated as mentioned in the Materials and methods.Error bars represent SD (n =30).

926Genome,Vol.54,2011

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

tion levels of three genes (GmaAffx.58899.1.S1_at,GmaAffx.75412.1.S1_at,and Gma.4518.1.A1_at)with high similarity to STOP1sequence were increased under Al stress (around 2-fold),which might contribute to Al-induced citrate secretion in soybean.Furthermore,an additional four genes (Gma.235.1.S1_x_at,Gma.235.2.S1_a_at,GmaAffx.92932.1.S1_s_at,and GmaAffx.82571.1.S1_at)within the C2H2zinc finger protein family were also upregulated by Al.

The exudation of citrate from soybean root apices has been suggested to be accompanied by the increase of plasma mem-brane H +-ATPase activity (Shen et al.2005).They suggested that citrate release requires H +-ATPase activity and that Al treatment stimulates gene and protein expression of the H +-ATPase.Also,there are protein phosphorylation changes consistent with protein kinases that act upon threonine resi-dues.In rice bean it has beenproposed that the addition of magnesium to the toxic Al treatment helps maintain the tis-sue magnesium content and the activity of the plasma mem-brane H +-ATPase,which enhanced the Al-dependent citrate efflux (Yang et al.2007).In the present study,two genes (Gma.1047.1.S1_at and GmaAffx.83213.2.S1_at)with high similarity to the soybean plasma membrane Ca 2+

-ATPase

Fig.2.Numbers of Al-induced differentially expressed genes in soybean root apices by microarray analysis.(A)Volcano plot of normalized signal intensities of expressed transcripts on the microarray.The log 2(ratio)of transcript abundance of Al-treated root tips versus control was plotted against their corresponding –log 10(p value).(B)Distribution of genes in various fold-change categories based on the ratio of transcript abundance in Al-treated root tips compared with control.

You et al.927

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

(SCA1)and M.truncatula type IIB calcium ATPase (MCA2),respectively,were obviously upregulated at the transcription level (both around 5-fold)under Al stress,which might be coupled with the process of Al-dependent citrate efflux in soybean.Chandran et al.(2008)also found that Al stress increased the transcription level of Ca 2+-ATPase,which is suggested to be involved in maintaining Ca 2+ho-meostasis following Al exposure.

Kinases and phosphatases are key signal molecules in the perception of Al stress and appear to play importance roles in mediating stress responses.Osawa and Matsumoto (2001)suggested that protein kinase might be involved in the phos-phorylation of an anion channel leading to the modification of the channel and malate efflux.Their further study (Ko-bayashi et al.2007)showed that Al-inducible malate release was signi ?cantly reduced in plants pretreated with protein kinase inhibitor (K-252a,staurosporine,and calyculin A),and thusseveral levels were suggested at which reversible phosphorylation could regulate Al-inducible malate release:directly on the AtALMT1transporter to modulate transport activity by in ?uencing AtALMT1gene expression level,by either reducing or eliminating increases in AtALMT1expres-sion,or by some combination of both.In the present study,we found higher expression of five genes (GmaAffx.37908.1.A1_at,Gma.13830.3.S1_at,GmaAffx.23917.1.S1_at,Gma.8133.1.A1_at,and GmaAffx.83336.1.S1_at)with simi-lar sequence to serine-threonine kinase.In addition,we found the higher expression of genes (Gma.17453.1.S1_at and GmaAffx.16384.1.S1_s_at)possibly encoding calcium-dependent protein kinase (CDPK1)and mitogen-activated protein kinase 1(MAPK1),respectively.They all have the potential to couple with Al-induced citrate secretion from soybean.

Cell wall-related genes

Several studies have suggested that the cell wall plays an important role in mechanisms of Al toxicity and resistance

(Tabuchi and Matsumoto 2001;Ma et al.2004).Some genes related to cell wall properties were induced by Al in this study,such as xyloglucan endotransglycosylase/hydrolase (XTH),pectin esterase,pectin acetyl esterase,and expansion.The polysaccharide xyloglucan is one major component of the primary cell wall of dicotyledons.Xyloglucan binds non-covalently to cellulose,coating and cross linking adjacent cellulose microfibrils.The cleavage of load-bearing xyloglu-can chains by hydrolytic enzymes,such as XTH,may cause a rapid wall loosening,which is necessary for root elongation.A consensus was reached that the member of XET/EXGT genes/proteins should be termed as XTH (Rose et al.2002).Thus,in present study,we found altogether nine genes puta-tively encoding XTH that were regulated by Al treatment,two (Gma.16471.1.S1_at and Gma.8236.1.S1_at)were upregu-lated and seven (GmaAffx.12832.1.S1_at,Gma.15814.1.A1_at,Gma.10791.2.S1_a_at,Gma.4216.2.S1_s_at,Gma.4216.1.S1_a_at,GmaAffx.91763.1.S1_s_at,and GmaAffx.91763.1.S1_x_at)of them were downregulated,which might be re-lated to the Al-induced root elongation inhibition of soy-bean.The diversity of their expression patterns might reflect the existence of different mechanisms of gene regu-lation within the same class of genes.

Glycosyl transferases (GTs)catalyze the transfer of sugar moieties from activated donor molecules to specific acceptor molecules,thereby forming glycosidic bonds (Coutinho et al.2003).The activity of GTs is necessary in the cell wall syn-thesis and may affect the thickening and rigidification of cell walls,which is related to the root elongation inhibition under Al stress (Sasaki et al.1996).The transcript abundance of four genes (GmaAffx.14473.1.S1_at,GmaAffx.42111.1.A1_at,GmaAffx.90870.1.S1_s_at,and GmaAffx.92715.1.S1_s_at)with putative glucosyl transferase sequence were upregu-lated in the present study,which might also be related to Al toxicity response in soybean.

Pectin esterase (PE)and pectinacetylesterase (PAE)cata-lyze the deesterification and deacetylation of pectin,respec-tively,which might play an important role in determining the extent to which pectin is accessible to degradation by cell wall hydrolyzing enzymes.Deacetylation of pectin and combing with the activation of cell wall hydrolyzing enzymes involved in cell expansionresults in root elongation.RNAi-in-duced gene silencing of PAE in M.truncatula decreased Al resistance,and PAE was suggested to be required for cell wall loosening and thus Al-induced cell elongation inhibition (Chandran et al.2008).In this study,higher transcription lev-els of putative pectinesterase gene (GmaAffx.91012.1.S1_s_at,GmaAffx.6824.2.S1_at,and GmaAffx.82617.1.S1_at)and putative pectinacetylesterase gene (Gma.4027.1.S1_at and Gma.3917.1.S1_at)in soybean roots under Al stress might contribute to its enhanced tolerance to Al stress.The role of PA and PAE in Al tolerance of higher plants needs fur-ther clarification.

Lignin deposition may also play a role in Al toxicity as a potential cause of root growth inhibition.Sasaki et al.(1996)reported that lignification in the elongation region coincides with the extent of inhibition of root growth by Al in wheat.Our array revealed a series of genes related to lignin biosyn-thesis that were upregulated under Al stress.Caffeoyl-CoA 3-O -methyltransferase (CCOMT)(Martz et al.1998;Zhang and Chiang 1997)and 4-coumarate:CoA ligase play

impor-Fig.3.Quantitative RT-PCR confirmation of microarray results for selected genes.PCR (qRT-PCR)analysis was performed for 10genes in soybean root apices under the same conditions used for microarray analysis (4h exposure to 0or 30μmol/L AlCl 3in 0.5mmol/L CaCl 2solution (pH 4.5)).

928

Genome,Vol.54,2011

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

tant roles in the regulation of cell wall lignification as both enzymes are involved in the biosynthesis of lignin.In the present study,we found three genes (GmaAffx.92100.1.S1_s_at,GmaAffx.92827.1.S1_at,and Gma.7423.2.S1_a_at)putatively encoding 4-coumarate:CoA ligase and one gene (GmaAffx.69410.1.S1_at)putatively encoding CCOMT that were highly significantly upregulated (17-,15-,and 9-fold,respectively)by Al.Chalcone synthase (ICHS1)and NADPH:isoflavone reductase (IFS)genes are key enzymes in the phenylpropanoid pathway and are also involved in lignin biosynthesis (Dixon et al.2002).We found the ex-pression of one gene (Gma.17605.1.S1_at)putatively encod-ing ICHS1and three genes (Gma.2686.1.S1_s_at,Gma.2686.2.S1_s_at,and Gma.2308.1.S1_s_at)putatively encoding IFS that were significantly upregulated.Addition-ally,as mentioned previously,we found four genes puta-tively encoding NADP-ME that were upregulated in this study,which may provide NADPH for lignin and flavonoid biosynthesis.The upregulation of lignin biosynthesis-related genes may be also related to the Al-induced inhibition of root elongation.

Plant peroxidases are bifunctional enzymes with diverse and even antagonistic functions involved in the cell-wall-related physiological processes such as lignin formation,cross-link of cell wall components,cell wall loosening,and cell elongation (Passardi et al.2004).The diversity of func-tions derives in part from two possible catalytic cycles of peroxidases involved in the consumption or release of H 2O 2and reactive oxygen species (ROS)(e.g.,O 2.–,H 2O 2,·OH).On one hand,peroxidases are involved in the cross-link of phenolic monomers in the formation of suberin and the ox-idative coupling of lignin subunits as part of lignin biosyn-thesis,which are related to the secondary cell wall formation and also the reduction of wall extensibility and thus the inhibition of root elongation (Tabuchi and Matsu-moto 2001).On the other hand,peroxidases can favor cell elongation by generating oxygen radicals (hydroxylic cycle)or by regulating the local concentration of H 2O 2.In the present study,Al enhanced the expressions of three genes (Gma.4305.3.S1_a_at,Gma.4305.2.S1_a_at,and Gma.4305.1.S1_a_at)putatively encoding peroxidase basic isozyme (monomer)and two genes (GmaAffx.91141.1.S1_at and Gma.5629.2.S1_a_at)putatively encoding peroxi-dase 1precursor (FBP1)and peroxidase 4precursor (FBP4).However,the transcription levels of two genes (GmaAffx.78664.1.S1_at and Gma.3804.1.A1_at)possibly en-coding peroxidase precursor were downregulated under Al stress.The diversity of peroxidases gene expression were also found in the transcriptional profile of maize (Maron et al.2008),and they contribute the diversity of expressions to the diversity of roles.

ATP binding cassette (ABC)transporter family genes seem to play important roles in conferring Al resistance of higher plants and have drawn a lot of attention in Al tolerance

study.

Fig.4.Functional categories of the Al-responsive genes in soybean roots.The number of genes found is included and,for clarity,genes classified as function unknown were not included in the figure.

You et al.

929

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

Huang et al.(2009)reported that two genes,sensitive to Al rhizotoxicity 1(STAR1)and STAR2encoded the nucleotide binding domain and the transmembrane domain of a bacte-rial-type ABC transporter,respectively,were responsible for Al tolerance in rice.STAR1interacted with STAR2to form a complex that localized to the vesicle membranes of root cells.They showed that the STAR –STAR2complex had efflux transport activity for UDP-glucose and hypothesized that UDP-glucose or glycosides derived from UDP-glucose mod-ify the cell walls to mask potential Al-binding sites in the apoplast,resulting in higher Al resistance in rice.STAR1and STAR2form the complex that functions as an ABC trans-porter,which is required for detoxification of Al in rice and transport of UDP-glucose,which may be used to modify the cell wall.In the present study,one gene (Gma.8365.1.S1_at)with a similar sequence to the rice STAR2was significantly upregulated (4-fold)by Al treatment.Moreover,we found Al increased the expression of two genes (GmaAffx.47590.1.S1_at and Gma.5478.1.S1_at)with putative UDP-glucosyltransferase sequence.The upregulation of STAR2and UDP-glucosyl-transferase genes may enhance the Al resistance by modify-ing the cell wall in soybean.

Hormones and signal transduction

Perception and transmission of stress signals could be the most important aspects of plant response to Al toxicity.Hor-mones play important roles in signaling.In our study,we found that Al induced expression of several hormone-related genes in soybean.

Abscisic acid (ABA)plays an important role in both plant development and the adaptation to abiotic stresses,such as drought,salinity,and low temperature (Xiong et al.2002;Shinozaki et al.2003).Our previous study proposed that ABA-mediated signal process might be involved in the regu-lation of Al resistance in soybean.Al treatment accelerated the ABA transport and induced more ABA accumulation in soybean,and ABA tended to distribute in the Al-exposed root part by split-root experiment (Hou et al.2010).In the present study,our array showed that a gene (GmaAffx.82642.2.S1_at)highly similar to ABA 8′-hydroxylase,a major ABA catabolic enzyme which converts ABA to phaseic acid (Saito et al.2004),was significantly downregulated by Al treatment,this could be related to the Al-enhanced ABA accumulation in the root tips of soybean (Hou et al.2010).We also found a gene related to the ABA transport in our microarray results.A gene (Gma.5574.2.S1_at)simi-lar to soybean PDR-like ABC-transporter was upregulated under Al stress.PDR-type ABC transporter has been identi-fied to be induced by salicylic acid and involved inABA transport (Eichhorn et al.2006;Kang et al.2010).Phospha-tidic acid triggers early signal transduction events that lead to responses to ABA during seed germination.Our array showed that Al induced the upregulation of two genes (Gma.4984.1.A1_at and Gma.4984.1.A1_x_at)putatively encoding phosphatidic acid phosphatase that catalyzes the conversion of phosphatidic acid to diacylglycerol and func-tions as a negative regulator upstream of ABI4,an AP2-type transcription factor (Katagiri et al.2005).Our array also revealed that Al increased the expression of several genes putatively encoding protein phosphatase 2C (GmaAffx.21217.3.S1_at,GmaAffx.82748.1.S1_at,and

GmaAffx.50980.1.S1_at).The plant protein phosphatase 2C

function as regulators of various signal transduction path-ways.In Arabidopsis,protein phosphatase 2C were proved to be central components in ABA signal transduction and are part of a negative control mechanism exerted by a postulated repressor of ABA-signal transfer (see review Ro-driguez 1998;Grill and Himmelbach 1998).

1-Aminocyclopropane-1-carboxylic acid oxidase (ACC-oxidase)involved in ethylene biosynthetic process and is proposed to be a typically stress-induced gene.Sun et al.(2007)reported that increased gene expression and enzyme activity of ACC-oxidase lead to increased ethylene produc-tion in Lotus japonicus and M.truncatula under Al stress,suggesting that ethylene is involved in the Al-induced inhibition of root elongation.The higher expression of ACC-oxidase gene was also identified as an Al toxicity re-sponse in the transcriptional profile studies of common bean (Eticha et al.2010)and M.truncatula (Chandran et al.2008).In the present study,Al significantly increased the expression of three genes (Gma.5064.1.S1_at,Gma.2776.1.S1_at,and Gma.2776.2.S1_s_at)putatively en-coding ACC-oxidase,two genes (GmaAffx.15015.1.S1_at and GmaAffx.92676.1.S1_s_at)putatively encoding two eth-ylene response factor,and two genes (Gma.3990.1.S1_s_at and Gma.3990.1.S1_at)putatively encoding ethylene-upregulated protein ER1.The expression of these genes could be related to the Al-induced inhibition of root elongation in soybean.

There are few studies concerning the role of jasmonic acid in Al tolerance of plants.A crucial step in the biosynthesis of jasmonic acid is the formation of its stereoisomeric precursor,cis -(+)-12-oxophytodienoic acid (OPDA),which is catalyzed by allene oxide cyclase (AOC;EC 5.3.99.6).In the present study,a gene (GmaAffx.22250.1.S1_at)putatively encoding for allene oxide cyclase was upregulated (2.2-fold)by Al treatment.However,we cannot conclude the role of allene oxide cyclase in soybean under Al stress.It has been sug-gested that the synthesis of both ethylene and jasmonic acid is positively controlled by stress-activated mitogen-activated protein kinases (MAPKs)(see review Schweighofer and Me-skiene 2008).Our array revealed that Al increased the tran-scription abundance of MAPK (GmaAffx.16384.1.S1_s_at),which might be related to the synthesis of ethylene and jas-monic acid.

Transcription factors

The majority of Al-responsive transcription factors belong to WRKY,NAC,C2H2,and myelloblastosis (MYB)fami-lies.WRKY transcription factors are typically associated with stress responses.In the present study,we identified 17putative WRKY-type transcription factors induced by Al stress,of these,all were upregulated,except the GmWRKY10(GmaAffx.37060.1.S1_at)gene which was downregulated.The involvement of NAC transcription factors in the regula-tion of Al stress of soybean was found in the present study.Al increased the expression of the putative genes GmNAC6(GmaAffx.13806.1.S1_at),GmNAC32(GmaAffx.50673.2.S1_s_at,GmaAffx.50673.1.S1_at,Gma.5519.3.S1_a_at,and Gma.5519.2.S1_at),GmNAC27(Gma.5331.1.S1_at),and GmNAC4(Gma.5331.1.S1_a_at).Besides the above dis-cussed STOP1gene,another four genes (Gma.235.1.S1_x_at,

930Genome,Vol.54,2011

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

Gma.235.2.S1_a_at,GmaAffx.92932.1.S1_s_at,and GmaAffx.82571.1.S1_at)within the C2H2zinc finger protein family were also found to be upregulated by Al treatment.Three genes (GmaAffx.74289.1.S1_at,GmaAffx.84604.1.S1_at,and GmaAffx.57525.3.S1_at)with similarity to GmMYB124,GmMYB127,and GmMYB29B2were upre-gulated (7.2-,3.2-,and 2.5-fold,respectively)by Al treat-ment.

Oxidative stress response genes

Oxidative stress is commonly presumed to be a major component of Al stress.The activated antioxidative systems are beneficial for plant growth under Al stress,because ad-equate capacity of antioxidative enzymes and other antioxi-dant metabolites may help in the removal of excess ROS and inhibit lipid peroxidation (Mittler 2002).In the present study,we observed the increased transcript abundance of several genes,glutathione S -transferases (GSTs),peroxidases,lipoxy-genase (LOX),and glutaredoxin,all of which are related to oxidative stress.The complex regulation of GSTs by environ-mental stimuli implies that they have important protective functions (see review Edwards et al.2000).GSTs may be in-volved in either detoxification of xenobiotics or the protec-tion of cells from lipid oxidation resulting from oxidative stress (Edwards et al.2000;Yang et al.2001a ).Plant GSTs attach glutathione to electropholic xenobiotics,which tags them for vacuolar sequestration.Al significantly upregulated the expressions of genes (Gma.2593.1.S1_s_at,Gma.9013.1.S1_at,Gma.8518.1.S1_at,and GmaAffx.26859.1.S1_s_at)puta-tively encoding GSTs,which might represent an Al resist-ance response.As discussed previously,a series of probes (Gma.5725.1.S1_s_at,GmaAffx.91141.1.S1_at,Gma.4305.3.S1_a_at,Gma.4305.2.S1_a_at,and Gma.4305.1.S1_at)with similarity to genes encoding peroxidase were also upregulated (about 2-to 3-fold).Peroxides are considered as bifunc-tional enzymes that can oxidize various substrates in the presence of H 2O 2but also produce ROS.Their involvement in many physiological and developmental processes can be detected in plants from germination to senescence.Additionally,genes (GmaAffx.24359.1.A1_at and GmaAffx.65360.1.S1_at)with putative lipoxygenase-9(LOX9)sequence were upregulated by Al in the present study.LOX (EC 1.13.11.12)is a ubiquitously occurring en-zyme that catalyzes the peroxidation of unsaturated fatty acids of biomembranes to produce hydroperoxides and oxy-free radicals.The increase of LOX activity may increase the formation of oxidation products (Axelrod et al.1981;Gard-ner 1991).The upregulation of LOX enzyme genes might be associated with Al toxicity response.

Concluding remarks

When grown in acid soils,plants generally encountered multistresses associated with soil acidity (low pH,low cal-cium,high Al,and high manganese),which limited proper root growth and development of the plants.As a member of the Fabaceae family,soybean is able to establish a symbiosis with N 2fixing bacteria of the Rhizobia family,which in-volves a complex molecular dialogue leading to the develop-ment of root nodules.Root tissue response to Al in the case of soybean is only a part of plant reaction in the soil.Hydro-

ponic culture permitted to address the sole Al toxicity re-sponse.

The present study identified many Al-responsive genes in-volved in either Al resistance or Al toxicity of soybean.This study will further facilitate our understanding of Al-induced root elongation inhibition and provide a molecular study ba-sis of Al resistance in soybean.However,many genes with unknown biological function were found,increasing or de-creasing at a high level in response to Al stress.These un-known genes may represent novel mechanisms of Al toxicity and resistance and thus need to be further clarified.

Acknowledgement

The authors thank Z.Yang for revising the text.Financial support for this research was provided by Specialized Research Fund for the Doctoral Program of Higher Education (2007018163),the National Natural Science Foundation of China (No.40903029),the Fundamental Research Funds for the Central Universities,and the 211Project of Jilin University.

References

Axelrod,B.,Cheesbrough,T.M.,and Laakso,S.1981.Lipoxygenase from soybeans.Methods Enzymol.71:441–451.doi:10.1016/0076-6879(81)71055-3.

Bianchi-Hall,C.M.,Carter,T.E.,Rufty,T.W.,Arellano,C.,Boerma,H.R.,and Ashley,D.A.1998.Heritability and resource allocation of aluminum tolerance derived from soybean PI 416937.Crop Sci.38(2):513–522.doi:10.2135/cropsci1998.0011183X003800020040x.Bianchi-Hall,C.M.,Carter,T.E.,Bailey,M.A.,Mian,M.A.R.,Rufty,T.W.,Ashley,D.A.,et al.2000.Aluminum tolerance associated with quantitative trait loci derived from soybean PI 416937in hydroponics.Crop Sci.40(2):538–544.doi:10.2135/cropsci2000.402538x.

Chandran,D.,Sharopova,N.,Ivashuta,S.,Gantt,J.S.,VandenBosch,K.A.,and Samac,D.A.2008.Transcriptome profiling identified novel genes associated with aluminum toxicity,resistance and tolerance in Medicago truncatula .Planta,228(1):151–166.doi:10.1007/s00425-008-0726-0.PMID:18351384.

Coutinho,P.M.,Deleury,E.,Davies,G.J.,and Henrissat,B.2003.An evolving hierarchical family classification for glycosyltransferases.J.Mol.Biol.328(2):307–317.doi:10.1016/S0022-2836(03)00307-3.PMID:12691742.

Dixon,R.A.,Achnine,L.,Kota,P.,Liu,C.-J.,Reddy,M.S.S.,and Wang,L.J.2002.The phenylpropanoid pathway and plant defence —a genomics perspective.Mol.Plant Pathol.3(5):371–390.doi:10.1046/j.1364-3703.2002.00131.x.PMID:20569344.

Duressa,D.,Soliman,K.,and Chen,D.Q.2010a .Identification of aluminum responsive genes in Al-tolerant soybean line PI 416937.Int.J.Plant Genomics.In press.doi:10.1155/2010/164862.

Duressa,D.,Soliman,K.M.,and Chen,D.Q.2010b .Mechanisms of magnesium amilioration of aluminum toxicity in soybean at the gene expression level.Genome,53(10):787–797.doi:10.1139/G10-069.PMID:20962885.

Durrett,T.P.,Gassmann,W.,and Rogers,E.E.2007.The FRD3-mediated efflux of citrate into the root vasculature is necessary for efficient iron translocation.Plant Physiol.144(1):197–205.doi:10.1104/pp.107.097162.PMID:17351051.

Edwards,G.E.,and Andreo,C.S.1992.NADP-malic enzyme from plants.Phytochemistry,31(6):1845–1857.doi:10.1016/0031-9422(92)80322-6.PMID:1368216.

Edwards,R.,Dixon,D.P.,and Walbot,V.2000.Plant glutathione S -transferases:enzymes with multiple functions in sickness and in

You et al.931

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

health.Trends Plant Sci.5(5):193–198.doi:10.1016/S1360-1385(00)01601-0.PMID:10785664.

Eichhorn,H.,Klinghammer,M.,Becht,P.,and Tenhaken,R.2006.Isolation of a novel ABC-transporter gene from soybean induced by salicylic acid.J.Exp.Bot.57(10):2193–2201.doi:10.1093/jxb/erj179.PMID:16720608.

Eticha,D.,Zahn,M.,Bremer,M.,Yang,Z.,Rangel,A.F.,Rao,I.M.,and Horst,W.J.2010.Transcriptomic analysis reveals differential gene expression in response to aluminum in common bean (Phaseolus vulgaris )genotypes.Ann.Bot.(Lond.),105(7):1119–1128.doi:10.1093/aob/mcq049.

Fiermonte,G.,Palmieri,L.,Dolce,V.,Lasorsa,F.M.,Palmieri,F.,Runswick,M.J.,and Walker,J.E.1998.The sequence,bacterial expression,and functional reconstitution of the rat mitochondrial dicarboxylate transporter cloned via distant homologs in yeast and Caenorhabditis elegans .J.Biol.Chem.273(38):24754–24759.doi:10.1074/jbc.273.38.24754.PMID:9733776.

Furukawa,J.,Yamaji,N.,Wang,H.,Mitani,N.,Murata,Y.,Sato,K.,et al.2007.An aluminum-activated citrate transporter in barley.Plant Cell Physiol.48(8):1081–1091.doi:10.1093/pcp/pcm091.PMID:17634181.

Gardner,H.W.1991.Recent investigation into the lipoxygenase pathway of plants.Biochim.Biophys.Acta,1084(3):221–239.doi:10.1016/0005-2760(91)90063-N.PMID:1909580.

Grill,E.,and Himmelbach,A.1998.ABA signal transduction.Curr.Opin.Plant Biol.1(5):412–418.doi:10.1016/S1369-5266(98)80265-3.PMID:10066625.

Guo,P.G.,Bai,G.H.,Carver,B.,Li,R.H.,Bernardo,A.,and Baum,M.2007.Transcriptional analysis between two wheat near-isogenic lines contrasting in aluminum tolerance under aluminum stress.Mol.Genet.Genomics,277(1):1–12.doi:10.1007/s00438-006-0169-x.PMID:17039377.

Guo,J.H.,Liu,X.J.,Zhang,Y.,Shen,J.L.,Han,W.X.,Zhang,W.F.,et al.2010.Significant acidification in major Chinese croplands.Science,327(5968):1008–1010.doi:10.1126/science.1182570.PMID:20150447.

Horst,W.J.,Asher,C.J.,Cakmak,I.,Szulkiewicz,P.,and Wissemeier,A.H.1992.Short-term responses of soybean roots to aluminium.J.Plant Physiol.140:174–178.

Hou,N.N.,You,J.F.,Pang,J.D.,Xu,M.Y.,Chen,G.,and Yang,Z.M.2010.The accumulation and transport of abscisic acid in soybean (Glycine max L.)under aluminum stress.Plant Soil,330(1–2):127–137.doi:10.1007/s11104-009-0184-x.

Huang,C.F.,Yamaji,N.,Mitani,N.,Yano,M.,Nagamura,Y.,and Ma,J.F.2009.A bacterial-type ABC transporter is involved in aluminum tolerance in rice.Plant Cell,21(2):655–667.doi:10.1105/tpc.108.064543.PMID:19244140.

Iuchi,S.,Koyama,H.,Iuchi,A.,Kobayashi,Y.,Kitabayashi,S.,Kobayashi,Y.,et al.2007.Zinc finger protein STOP1is critical for proton tolerance in Arabidopsis and co-regulates a key gene in aluminum tolerance.Proc.Natl.Acad.Sci.U.S.A.104(23):9900–9905.doi:10.1073/pnas.0700117104.PMID:17535918.

Kang,J.Y.,Hwang,J.U.,Lee,M.Y.,Kim,Y.Y.,Assmann,S.M.,Martinoia, E.,and Lee,Y.2010.PDR-type ABC transporter mediates cellular uptake of the phytohormone abscisic acid.Proc.Natl.Acad.Sci.U.S.A.107(5):2355–2360.doi:10.1073/pnas.0909222107.PMID:20133880.

Katagiri,T.,Ishiyama,K.,Kato,T.,Tabata,S.,Kobayashi,M.,and Shinozaki,K.2005.An important role of phosphatidic acid in ABA signaling during germination in Arabidiopsis thaliana .Plant J.43(1):107–117.doi:10.1111/j.1365-313X.2005.02431.x.PMID:15960620.

Kobayashi,Y.,Hoekenga,O.A.,Itoh,H.,Nakashima,M.,Saito,S.,Shaff,J.E.,et al.2007.Characterization of AtALMT1expression in

aluminum-inducible malate release and its role for rhizotoxic stress tolerance in Arabidopsis.Plant Physiol.145(3):843–852.doi:10.1104/pp.107.102335.PMID:17885092.

Kochian,L.V.,Hoekenga,O.A.,and Pi?eros,M.A.2004.How do crop plants tolerate acid soils?Mechanisms of aluminum tolerance and phosphorous efficiency.Annu.Rev.Plant Biol.55(1):459–493.doi:10.1146/annurev.arplant.55.031903.141655.PMID:15377228.

Kollmeier,M.,Felle,H.H.,and Horst,W.J.2000.Genotypical differences in aluminum resistance of maize are expressed in the distal part of the transition zone.Is reduced basipetal auxin flow involved in inhibition of root elongation by aluminum?Plant Physiol.122(3):945–956.doi:10.1104/pp.122.3.945.PMID:10712559.

Kumari,M.,Taylor,G.J.,and Deyholos,M.K.2008.Transcriptomic responses to aluminum stress in roots of Arabidopsis thaliana .Mol.Genet.Genomics,279(4):339–357.doi:10.1007/s00438-007-0316-z.PMID:18270741.

Li,C.,and Wong,W.H.2001.Model-based analysis of oligonucleo-tide arrays:expression index computation and outlier detection.Proc.Natl.Acad.Sci.U.S.A.98(1):31–36.doi:10.1073/pnas.011404098.PMID:11134512.

Liu,J.P.,Magalhaes,J.V.,Shaff,J.,and Kochian,L.V.2009.Aluminum-activated citrate and malate transporters from the MATE and ALMT families function independently to confer Arabidopsis aluminum tolerance.Plant J.57(3):389–399.doi:10.1111/j.1365-313X.2008.03696.x.PMID:18826429.

Livak,K.J.,and Schmittgen,T.D.2001.Analysis of relative gene expression data using real-time quantitative PCR and the 2àDD C t method.Methods,25(4):402–408.doi:10.1006/meth.2001.1262.PMID:11846609.

Ma,J.F.2000.Role of organic acids in detoxification of aluminum in higher plants.Plant Cell Physiol.41(4):383–390.PMID:10845450.

Ma,J.F.,Shen,R.F.,Nagao,S.,and Tanimoto,E.2004.Aluminum targets elongating cells by reducing cell wall extensibility in wheat roots.Plant Cell Physiol.45(5):583–589.doi:10.1093/pcp/pch060.PMID:15169940.

Magalhaes,J.V.,Liu,J.P.,Guimar?es,C.T.,Lana,U.G.P.,Alves,V.M.C.,Wang,Y.H.,et al.2007.A gene in the multidrug and toxic compound extrusion (MATE)family confers aluminum tolerance in sorghum.Nat.Genet.39(9):1156–1161.doi:10.1038/ng2074.PMID:17721535.

Maron,L.G.,Kirst,M.,Mao,C.,Milner,M.J.,Menossi,M.,and Kochian,L.V.2008.Transcriptional profiling of aluminum toxicity and tolerance responses in maize roots.New Phytol.179(1):116–128.doi:10.1111/j.1469-8137.2008.02440.x.PMID:18399934.

Martz,F.,Maury,S.,Pincon,G.,and Legrand,M.1998.cDNA cloning,substrate specificity and expression study of tobacco caffeoyl-CoA 3-O -methyltransferase,a lignin biosynthetic en-zyme.Plant Mol.Biol.36(3):427–437.doi:10.1023/A:1005969825070.PMID:9484483.

Mittler,R.2002.Oxidative stress,antioxidants and stress tolerance.Trends Plant Sci.7(9):405–410.doi:10.1016/S1360-1385(02)02312-9.PMID:12234732.

Nian,H.,Yang,Z.,Huang,H.,Yan,X.,and Matsumoto,H.2005.Citrate secretion induced by aluminum stress may not be a key mechanism responsible for differential aluminum tolerance of some soybean genotypes.J.Plant Nutr.27(11):2047–2066.doi:10.1081/PLN-200030112.

Osawa,H.,and Matsumoto,H.2001.Possible involement of protein phosphorylation in aluminum-responsive malate efflux from

932

Genome,Vol.54,2011

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

wheat root apex.Plant Physiol.126(1):411–420.doi:10.1104/pp.126.1.411.PMID:11351103.

Palmieri,L.G.,Picault,N.,Arrigoni,R.,Besin,E.,Palmieri,F.,and Hodges,M.2008.Molecular identification of three Arabidopsis thaliana mitochondrial dicarboxylated carrier isoforms:organ distribution,bacterial expression,reconstitution into liposomes and functional characterization.Biochem.J.410(3):621–629.doi:10.1042/BJ20070867.PMID:18039180.

Pannatier,G.E.,Luster,J.,Zimmermann,S.,and Blaser,P.2005.Monitoring of water chemistry in forest soils:an indicator for acidification.Chimia (Aarau),59(12):989.

Passardi, F.,Penel, C.,and Dunand, C.2004.Performing the paradoxical:how plant peroxidases modify the cell wall.Trends Plant Sci.9(11):534–540.doi:10.1016/j.tplants.2004.09.002.PMID:15501178.

Rangel, A.F.,Rao,I.M.,Braun,H.-P.,and Horst,W.J.2010.Aluminum resistance in common bean (Phaseolus vulgaris )involves induction and maintenance of citrate exudation from root apices.Physiol.Plant.138(2):176–190.doi:10.1111/j.1399-3054.2009.01303.x.PMID:20053183.

Rodriguez,P.L.1998.Protein phosphatase 2C (PP2C)function in higher plants.Plant Mol.Biol.38(6):919–927.doi:10.1023/A:1006054607850.PMID:9869399.

Rogers,E.E.,and Guerinot,M.L.2002.FRD3,a member of the multidrug and toxin efflux family,controls iron deficiency responses in Arabidopsis.Plant Cell,14(8):1787–1799.doi:10.1105/tpc.001495.PMID:12172022.

Rogers,E.E.,Wu,X.,Stacey,G.,and Nguyen,H.T.2009.Two MATE protein play a role in iron efficiency in soybean.J.Plant Physiol.166(13):1453–1459.doi:10.1016/j.jplph.2009.02.009.PMID:19342121.

Rose,J.K.C.,Braam,J.,Fry,S.C.,and Nishitani,K.2002.The XTH family of enzymes involved in xyloglucan endotransglucosylation and endohydrolysis:current perspectives and a new unifying nomenclature.Plant Cell Physiol.43(12):1421–1435.doi:10.1093/pcp/pcf171.PMID:12514239.

Ryan,P.R.,and Delhaize,E.2010.The convergent evolution of aluminum resistance in plants exploits a convenient currency.Funct.Plant Biol.37(4):275–284.doi:10.1071/FP09261.

Ryan,P.R.,Delhaize, E.,and Jones, D.L.2001.Function and mechanism of organic anion exudation from plant roots.Annu.Rev.Plant Physiol.Plant Mol.Biol.52(1):527–560.doi:10.1146/annurev.arplant.52.1.527.PMID:11337408.

Saito,S.,Hirai,N.,Matsumoto,C.,Ohigashi,H.,Ohta,D.,Sakata,K.,and Mizutani,M.2004.Arabidopsis CYP707A s encode (+)-abscisic acid 8′-hydroxylase,a key enzyme in the oxidative catabosism of abscisic acid.Plant Physiol.134(4):1439–1449.doi:10.1104/pp.103.037614.PMID:15064374.

Sasaki,M.,Yamamoto,Y.,and Matsumoto,H.1996.Lignin deposition induced by aluminum in wheat (Triticum aestivum )roots.Physiol.Plant.96(2):193–198.doi:10.1111/j.1399-3054.1996.tb00201.x.

Sawaki,Y.,Iuchi,S.,Kobayashi,Y.,Kobayashi,Y.,Ikka,T.,Sakurai,N.,et al.2009.STOP1regulates multiple genes that protect Arabidopsis from proton and aluminum toxicities.Plant Physiol.150(1):281–294.doi:10.1104/pp.108.134700.PMID:19321711.Schweighofer, A.,and Meskiene,I.2008.Regulation of stress hormones jasmonates and ethylene by MAPK pathways in plants.Mol.Biosyst.4(8):799–803.doi:10.1039/b718578m.PMID:18633480.

Shen,H.,He,L.F.,Sasaki,T.,Yamamoto,Y.,Zheng,S.J.,Ligaba,A.,et al.2005.Citrate secretion cupled with the modulation of soybean root tip under aluminum stress.Up-regulation of transcription,translation,and threonine-oriented phosphorylation

of plasma membrane H +-ATPase.Plant Physiol.138(1):287–296.doi:10.1104/pp.104.058065.PMID:15834009.

Shinozaki,K.,Yamaguchi-shinozaki,K.,and Seki,M.2003.Regulatory network of gene expression in the drought and cold stress responses.Curr.Opin.Plant Biol.6(5):410–417.doi:10.1016/S1369-5266(03)00092-X.PMID:12972040.

Silva,I.R.,Smyth,T.J.,Raper,C.D.,Carter,T.E.,and Rufty,T.W.2001.Differential aluminum tolerance in soybean:an evaluation of the role of organic acids.Physiol.Plant.112(2):200–210.doi:10.1034/j.1399-3054.2001.1120208.x.PMID:11454225.

Sivaguru,M.,and Horst,W.J.1998.The distal part of the transition zone is the most aluminum-sensitve apical root zone of maize.Plant Physiol.116(1):155–163.doi:10.1104/pp.116.1.155.

Sun,P.,Tian,Q.-Y.,Zhao,M.-G.,Dai,X.-Y.,Huang,J.-H.,Li,L.-H.,and Zhang,W.-H.2007.Aluminum-induced ethylene production is associated with inhibition of root elongation in Lotus japonicus L.Plant Cell Physiol.48(8):1229–1235.doi:10.1093/pcp/pcm077.PMID:17573361.

Tabuchi, A.,and Matsumoto,H.2001.Changes in cell-wall properties of wheat (Triticum aestivum )roots during aluminum-induced growth inhibition.Physiol.Plant.112(3):353–358.doi:10.1034/j.1399-3054.2001.1120308.x.PMID:11473692.

von Uexküll,H.R.,and Mutert,E.1995.Global extent,development and economic impact of acid soils.Plant Soil,171(1):1–15.doi:10.1007/BF00009558.

Xiong,L.,Schumaker,K.S.,and Zhu,J.K.2002.Cell signaling during cold,drought,and salt stress.Plant Cell,14(Suppl.):S165–S168.doi:10.1105/tpc.000596.PMID:12045276.

Xu,M.Y.,You,J.F.,Hou,N.N.,Zhang,H.M.,Chen,G.,and Yang,Z.M.2010.Mitochondrial enzymes and citrate transporter contribute to the aluminium-induced citrate secretion from soybean (Glycine max )roots.Funct.Plant Biol.37(4):285–295.doi:10.1071/FP09223.Yamaji,N.,Huang,C.F.,Nagao,S.,Yano,M.,Sato,Y.,Nagamura,Y.,and Ma,J.F.2009.A zinc finger transcription factor ART1regulates multiple genes implicated in aluminum tolerance in rice.Plant Cell,21(10):3339–33349.doi:10.1105/tpc.109.070771.Yang,Z.M.,Sivaguru,M.,Horst,W.J.,and Matsumoto,H.2000.Aluminium tolerance is achieved by exudation of citric acid from roots of soybean (Glycine max ).Physiol.Plant.110(1):72–77.doi:10.1034/j.1399-3054.2000.110110.x.

Yang,Y.S.,Cheng,J.Z.,Singhal,S.S.,Saini,M.,Pandya,U.,Awasthi,S.,and Awasthi,Y.C.2001a .Role of glutathione S -transferases in protection against lipid peroxidation.J.Biol.Chem.276(22):19220–19230.PMID:11279091.

Yang,Z.M.,Nian,H.,Sivaguru,M.,Tanakamaru,S.,and Matsumoto,H.2001b .Characterization of aluminium-induced citrate secretion in aluminium-tolerant soybean (Glycine max )plants.Physiol.Plant.113(1):64–71.doi:10.1034/j.1399-3054.2001.1130109.x.

Yang,J.L.,You,J.F.,Wu,P.,and Zheng,S.J.2007.Magnesium enhances aluminum-induced citrate secretion in rice bean roots (Vigna umbellata )by restoring plasma membrane H +-ATPase activity.Plant Cell Physiol.48:66(1):66–73.PMID:17132634.Yokosho,K.G.,Yamaji,N.,and Ma,J.F.2010.Isolation and characterization of two MATE genes in rye.Funct.Plant Biol.37(4):296–303.doi:10.1071/FP09265.

You,J.F.,Hou,N.N.,Xu,M.Y.,Zhang,H.M.,and Yang,Z.M.2010.Citrate transporters play an important role in regulating alumium-induced citrate secretion in Glycine max .Biol.Plant.54(4):766–768.doi:10.1007/s10535-010-0138-4.

Zhang,X.H.,and Chiang,V.L.1997.Molecular cloning of 4-coumarate:coenzyme A ligase in loblolly pine and the roles of this enzyme in the biosynthesis of lignin in compression wood.Plant Physiol.113(1):65–74.doi:10.1104/pp.113.1.65.PMID:9008388.

You et al.

933

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

This article has been cited by:

1.Mireille Cabane, Dany Afif, Simon HawkinsLignins and Abiotic Stresses 61, 219-26

2. [CrossRef ]

G e n o m e D o w n l o a d e d f r o m w w w .n r c r e s e a r c h p r e s s .c o m b y O h i o S t a t e U n i v e r s i t y o n 12/29/13F o r p e r s o n a l u s e o n l y .

最新The_Monster课文翻译

Deems Taylor: The Monster 怪才他身材矮小,头却很大,与他的身材很不相称——是个满脸病容的矮子。他神经兮兮,有皮肤病,贴身穿比丝绸粗糙一点的任何衣服都会使他痛苦不堪。而且他还是个夸大妄想狂。他是个极其自负的怪人。除非事情与自己有关,否则他从来不屑对世界或世人瞧上一眼。对他来说,他不仅是世界上最重要的人物,而且在他眼里,他是惟一活在世界上的人。他认为自己是世界上最伟大的戏剧家之一、最伟大的思想家之一、最伟大的作曲家之一。听听他的谈话,仿佛他就是集莎士比亚、贝多芬、柏拉图三人于一身。想要听到他的高论十分容易,他是世上最能使人筋疲力竭的健谈者之一。同他度过一个夜晚,就是听他一个人滔滔不绝地说上一晚。有时,他才华横溢;有时,他又令人极其厌烦。但无论是妙趣横生还是枯燥无味,他的谈话只有一个主题:他自己,他自己的所思所为。他狂妄地认为自己总是正确的。任何人在最无足轻重的问题上露出丝毫的异议,都会激得他的强烈谴责。他可能会一连好几个小时滔滔不绝,千方百计地证明自己如何如何正确。有了这种使人耗尽心力的雄辩本事,听者最后都被他弄得头昏脑涨,耳朵发聋,为了图个清静,只好同意他的说法。他从来不会觉得,对于跟他接触的人来说,他和他的所作所为并不是使人产生强烈兴趣而为之倾倒的事情。他几乎对世间的任何领域都有自己的理

论,包括素食主义、戏剧、政治以及音乐。为了证实这些理论,他写小册子、写信、写书……文字成千上万,连篇累牍。他不仅写了,还出版了这些东西——所需费用通常由别人支付——而他会坐下来大声读给朋友和家人听,一读就是好几个小时。他写歌剧,但往往是刚有个故事梗概,他就邀请——或者更确切说是召集——一群朋友到家里,高声念给大家听。不是为了获得批评,而是为了获得称赞。整部剧的歌词写好后,朋友们还得再去听他高声朗读全剧。然后他就拿去发表,有时几年后才为歌词谱曲。他也像作曲家一样弹钢琴,但要多糟有多糟。然而,他却要坐在钢琴前,面对包括他那个时代最杰出的钢琴家在内的聚会人群,一小时接一小时地给他们演奏,不用说,都是他自己的作品。他有一副作曲家的嗓子,但他会把著名的歌唱家请到自己家里,为他们演唱自己的作品,还要扮演剧中所有的角色。他的情绪犹如六岁儿童,极易波动。心情不好时,他要么用力跺脚,口出狂言,要么陷入极度的忧郁,阴沉地说要去东方当和尚,了此残生。十分钟后,假如有什么事情使他高兴了,他就会冲出门去,绕着花园跑个不停,或者在沙发上跳上跳下或拿大顶。他会因爱犬死了而极度悲痛,也会残忍无情到使罗马皇帝也不寒而栗。他几乎没有丝毫责任感。他似乎不仅没有养活自己的能力,也从没想到过有这个义务。他深信这个世界应该给他一条活路。为了支持这一信念,他

新版人教版高中语文课本的目录。

必修一阅读鉴赏第一单元1.*沁园春?长沙……………………………………毛泽东3 2.诗两首雨巷…………………………………………戴望舒6 再别康桥………………………………………徐志摩8 3.大堰河--我的保姆………………………………艾青10 第二单元4.烛之武退秦师………………………………….《左传》16 5.荆轲刺秦王………………………………….《战国策》18 6.*鸿门宴……………………………………..司马迁22 第三单元7.记念刘和珍君……………………………………鲁迅27 8.小狗包弟……………………………………….巴金32 9.*记梁任公先生的一次演讲…………………………梁实秋36 第四单元10.短新闻两篇别了,“不列颠尼亚”…………………………周婷杨兴39 奥斯维辛没有什么新闻…………………………罗森塔尔41 11.包身工………………………………………..夏衍44 12.*飞向太空的航程……………………….贾永曹智白瑞雪52 必修二阅读鉴赏第一单元1.荷塘月色…………………………………..朱自清2.故都的秋…………………………………..郁达夫3.*囚绿记…………………………………..陆蠡第二单元4.《诗经》两首氓采薇5.离骚………………………………………屈原6.*《孔雀东南飞》(并序) 7.*诗三首涉江采芙蓉《古诗十九首》短歌行……………………………………曹操归园田居(其一)…………………………..陶渊明第三单元8.兰亭集序……………………………………王羲之9.赤壁赋……………………………………..苏轼10.*游褒禅山记………………………………王安石第四单元11.就任北京大学校长之演说……………………..蔡元培12.我有一个梦想………………………………马丁?路德?金1 3.*在马克思墓前的讲话…………………………恩格斯第三册阅读鉴赏第一单元1.林黛玉进贾府………………………………….曹雪芹2.祝福………………………………………..鲁迅3. *老人与海…………………………………….海明威第二单元4.蜀道难……………………………………….李白5.杜甫诗三首秋兴八首(其一) 咏怀古迹(其三) 登高6.琵琶行(并序)………………………………..白居易7.*李商隐诗两首锦瑟马嵬(其二) 第三单元8.寡人之于国也…………………………………《孟子》9.劝学……………………………………….《荀子》10.*过秦论…………………………………….贾谊11.*师说………………………………………韩愈第四单元12.动物游戏之谜………………………………..周立明13.宇宙的边疆………………………………….卡尔?萨根14.*凤蝶外传……………………………………董纯才15.*一名物理学家的教育历程……………………….加来道雄第四册阅读鉴赏第一单元1.窦娥冤………………………………………..关汉卿2.雷雨………………………………………….曹禹3.*哈姆莱特……………………………………莎士比亚第二单元4.柳永词两首望海潮(东南形胜) 雨霖铃(寒蝉凄切) 5.苏轼词两首念奴娇?赤壁怀古定风波(莫听穿林打叶声) 6.辛弃疾词两首水龙吟?登建康赏心亭永遇乐?京口北固亭怀古7.*李清照词两首醉花阴(薄雾浓云愁永昼) 声声慢(寻寻觅觅) 第三单元8.拿来主义……………………………………….鲁迅9.父母与孩子之间的爱……………………………..弗罗姆10.*短文三篇热爱生

高中外研社英语选修六Module5课文Frankenstein's Monster

Frankenstein's Monster Part 1 The story of Frankenstein Frankenstein is a young scientist/ from Geneva, in Switzerland. While studying at university, he discovers the secret of how to give life/ to lifeless matter. Using bones from dead bodies, he creates a creature/ that resembles a human being/ and gives it life. The creature, which is unusually large/ and strong, is extremely ugly, and terrifies all those/ who see it. However, the monster, who has learnt to speak, is intelligent/ and has human emotions. Lonely and unhappy, he begins to hate his creator, Frankenstein. When Frankenstein refuses to create a wife/ for him, the monster murders Frankenstein's brother, his best friend Clerval, and finally, Frankenstein's wife Elizabeth. The scientist chases the creature/ to the Arctic/ in order to destroy him, but he dies there. At the end of the story, the monster disappears into the ice/ and snow/ to end his own life. Part 2 Extract from Frankenstein It was on a cold November night/ that I saw my creation/ for the first time. Feeling very anxious, I prepared the equipment/ that would give life/ to the thing/ that lay at my feet. It was already one/ in the morning/ and the rain/ fell against the window. My candle was almost burnt out when, by its tiny light,I saw the yellow eye of the creature open. It breathed hard, and moved its arms and legs. How can I describe my emotions/ when I saw this happen? How can I describe the monster who I had worked/ so hard/ to create? I had tried to make him beautiful. Beautiful! He was the ugliest thing/ I had ever seen! You could see the veins/ beneath his yellow skin. His hair was black/ and his teeth were white. But these things contrasted horribly with his yellow eyes, his wrinkled yellow skin and black lips. I had worked/ for nearly two years/ with one aim only, to give life to a lifeless body. For this/ I had not slept, I had destroyed my health. I had wanted it more than anything/ in the world. But now/ I had finished, the beauty of the dream vanished, and horror and disgust/ filled my heart. Now/ my only thoughts were, "I wish I had not created this creature, I wish I was on the other side of the world, I wish I could disappear!” When he turned to look at me, I felt unable to stay in the same room as him. I rushed out, and /for a long time/ I walked up and down my bedroom. At last/ I threw myself on the bed/ in my clothes, trying to find a few moments of sleep. But although I slept, I had terrible dreams. I dreamt I saw my fiancée/ walking in the streets of our town. She looked well/ and happy/ but as I kissed her lips,they became pale, as if she were dead. Her face changed and I thought/ I held the body of my dead mother/ in my arms. I woke, shaking with fear. At that same moment,I saw the creature/ that I had created. He was standing/by my bed/ and watching me. His

人教版高中语文必修必背课文精编WORD版

人教版高中语文必修必背课文精编W O R D版 IBM system office room 【A0816H-A0912AAAHH-GX8Q8-GNTHHJ8】

必修1 沁园春·长沙(全文)毛泽东 独立寒秋, 湘江北去, 橘子洲头。 看万山红遍, 层林尽染, 漫江碧透, 百舸争流。 鹰击长空, 鱼翔浅底, 万类霜天竞自由。 怅寥廓, 问苍茫大地, 谁主沉浮。 携来百侣曾游, 忆往昔峥嵘岁月稠。

恰同学少年, 风华正茂, 书生意气, 挥斥方遒。 指点江山, 激扬文字, 粪土当年万户侯。 曾记否, 到中流击水, 浪遏飞舟。 雨巷(全文)戴望舒撑着油纸伞,独自 彷徨在悠长、悠长 又寂寥的雨巷, 我希望逢着 一个丁香一样地

结着愁怨的姑娘。 她是有 丁香一样的颜色, 丁香一样的芬芳, 丁香一样的忧愁, 在雨中哀怨, 哀怨又彷徨; 她彷徨在这寂寥的雨巷,撑着油纸伞 像我一样, 像我一样地 默默彳亍着 冷漠、凄清,又惆怅。她默默地走近, 走近,又投出 太息一般的眼光

她飘过 像梦一般地, 像梦一般地凄婉迷茫。像梦中飘过 一枝丁香地, 我身旁飘过这个女郎;她默默地远了,远了,到了颓圮的篱墙, 走尽这雨巷。 在雨的哀曲里, 消了她的颜色, 散了她的芬芳, 消散了,甚至她的 太息般的眼光 丁香般的惆怅。 撑着油纸伞,独自

彷徨在悠长、悠长 又寂寥的雨巷, 我希望飘过 一个丁香一样地 结着愁怨的姑娘。 再别康桥(全文)徐志摩 轻轻的我走了,正如我轻轻的来; 我轻轻的招手,作别西天的云彩。 那河畔的金柳,是夕阳中的新娘; 波光里的艳影,在我的心头荡漾。 软泥上的青荇,油油的在水底招摇; 在康河的柔波里,我甘心做一条水草! 那榆荫下的一潭,不是清泉, 是天上虹揉碎在浮藻间,沉淀着彩虹似的梦。寻梦?撑一支长篙,向青草更青处漫溯, 满载一船星辉,在星辉斑斓里放歌。

集团客户综合金融服务方案及营销要点

集团客户综合金融服务方案及营销要点集团客户综合金融服务方案及营销要点集团客户部童罗兴2011年1月1 目录集团业务经营现状和形势1对集团客户金融服务需求的理解2服务方案的设计思路3服务方案的具体内容及营销要点42 集团业务经营现状及形势13 集团客户对建设银行的重要意义??集团客户贷款余额27179亿元,占全行对公贷款40818亿元的7><67%,不良率为0.93% ;??存款余额12259亿元,占全行对公存款的25%,其中活期存款占62%;??表外业务余额103<67亿元,垫款余额仅1.61亿元,垫款率不到0.02%;??2010年上半年,我行共为总行级战略性客户发行中期票据6期、共232亿元,发行短期融资券23期、共1174亿元,分别占承销总量的51%和90%。??截至三季度末,总行级战略性客户毛收入为134亿元,税前利润82 亿元,EVA 为27 亿元,年化资本回报率为19.46。??基础性作用??高附加值4??低风险 强者的握手——建设银行与集团客户中国建设银行集团客户??规模化、多元化经营趋势;??强大的品牌优势;??个性化金融服务需求趋势;??机构与客户经理队伍;??融资渠道拓宽,议价能力提??境内外、综合化经营平台;升;??风险管理体系;??风险因素集聚,监控和防范难??业务流程的差别化问题突出。度加大,而破坏性也加大;机遇与挑战并存:在做好客户甄别与风险控制的前提下,为集团客户提供更为专业化和个性化的金融服务方案。5

对集团客户金融服务需求的理解26 “黑海”-成熟期“红海”-稳定期??集团经营管理高度成熟,脱媒现象严重;??集团管理相对成熟,已初步完成系统构建;??合作银行已相对稳定;??对银行服务仍有个性化需求;??综合收益明显,竞争惨烈。??营销和维护成本极高。??电力、石油行业的战略性客户??某些外资集团:GE“黄海”-发展期“蓝海”-成长期??经营管理体制正在完善,一定程度上依托银行;??集团正处于发展初期,需求多且旺盛;??标准产品匹配个性化服务,若能在此阶段想方设??银行通过引导客户,有可能取得先导期的超法解决其“核心关切”,将利于提高客户黏性;额收益;??前景看好,竞争加剧,但传统合作银行仍有一定??竞争压力较小,客户甄别是关键。优势。??某些转型期集团企业??中小型集团客户7 对不同发展阶段集团客户的服务策略黑海-综合分析建设银行当前利益和长远利益,考量财务收益与社会收益;-若仍“入不敷出”,则应选择不再新增投入。红海-综合算账,提升整体收益,积极介入新需求,效率取胜;-积累产品和客户关系优势,提升客户黏性,有策略地争取业务回报;-例:中国中铁现金管理业务黄海-基于当前和长远收益,积极拓展;-特别关注关键业务对维护客户关系的关键性作用,效率和创新是关键;-例:中冶保障性住房业务需求蓝海-加强贴身服务,引导客户需求;-提升客户依赖度,在控制风险的基础上,可适度争取高额回报。-例:产品覆盖度的重要性8 服务方案的设计思路39 方案设计思路投融资中心拓宽集团融资渠道提高资金使用效率资金管理中心配合集团发展战略为集团创造价值战略协同中心10

(完整版)Unit7TheMonster课文翻译综合教程四

Unit 7 The Monster Deems Taylor 1He was an undersized little man, with a head too big for his body ― a sickly little man. His nerves were bad. He had skin trouble. It was agony for him to wear anything next to his skin coarser than silk. And he had delusions of grandeur. 2He was a monster of conceit. Never for one minute did he look at the world or at people, except in relation to himself. He believed himself to be one of the greatest dramatists in the world, one of the greatest thinkers, and one of the greatest composers. To hear him talk, he was Shakespeare, and Beethoven, and Plato, rolled into one. He was one of the most exhausting conversationalists that ever lived. Sometimes he was brilliant; sometimes he was maddeningly tiresome. But whether he was being brilliant or dull, he had one sole topic of conversation: himself. What he thought and what he did. 3He had a mania for being in the right. The slightest hint of disagreement, from anyone, on the most trivial point, was enough to set him off on a harangue that might last for hours, in which he proved himself right in so many ways, and with such exhausting volubility, that in the end his hearer, stunned and deafened, would agree with him, for the sake of peace. 4It never occurred to him that he and his doing were not of the most intense and fascinating interest to anyone with whom he came in contact. He had theories about almost any subject under the sun, including vegetarianism, the drama, politics, and music; and in support of these theories he wrote pamphlets, letters, books ... thousands upon thousands of words, hundreds and hundreds of pages. He not only wrote these things, and published them ― usually at somebody else’s expense ― but he would sit and read them aloud, for hours, to his friends, and his family. 5He had the emotional stability of a six-year-old child. When he felt out of sorts, he would rave and stamp, or sink into suicidal gloom and talk darkly of going to the East to end his days as a Buddhist monk. Ten minutes later, when something pleased him he would rush out of doors and run around the garden, or jump up and down off the sofa, or stand on his head. He could be grief-stricken over the death of a pet dog, and could be callous and heartless to a degree that would have made a Roman emperor shudder. 6He was almost innocent of any sense of responsibility. He was convinced that

人教版高中语文必修一背诵篇目

高中语文必修一背诵篇目 1、《沁园春长沙》毛泽东 独立寒秋,湘江北去,橘子洲头。 看万山红遍,层林尽染;漫江碧透,百舸争流。 鹰击长空,鱼翔浅底,万类霜天竞自由。 怅寥廓,问苍茫大地,谁主沉浮? 携来百侣曾游,忆往昔峥嵘岁月稠。 恰同学少年,风华正茂;书生意气,挥斥方遒。 指点江山,激扬文字,粪土当年万户侯。 曾记否,到中流击水,浪遏飞舟? 2、《诗两首》 (1)、《雨巷》戴望舒 撑着油纸伞,独自 /彷徨在悠长、悠长/又寂寥的雨巷, 我希望逢着 /一个丁香一样的 /结着愁怨的姑娘。 她是有 /丁香一样的颜色,/丁香一样的芬芳, /丁香一样的忧愁, 在雨中哀怨, /哀怨又彷徨; /她彷徨在这寂寥的雨巷, 撑着油纸伞 /像我一样, /像我一样地 /默默彳亍着 冷漠、凄清,又惆怅。 /她静默地走近/走近,又投出 太息一般的眼光,/她飘过 /像梦一般地, /像梦一般地凄婉迷茫。 像梦中飘过 /一枝丁香的, /我身旁飘过这女郎; 她静默地远了,远了,/到了颓圮的篱墙, /走尽这雨巷。 在雨的哀曲里, /消了她的颜色, /散了她的芬芳, /消散了,甚至她的 太息般的眼光, /丁香般的惆怅/撑着油纸伞,独自 /彷徨在悠长,悠长 又寂寥的雨巷, /我希望飘过 /一个丁香一样的 /结着愁怨的姑娘。 (2)、《再别康桥》徐志摩 轻轻的我走了, /正如我轻轻的来; /我轻轻的招手, /作别西天的云彩。 那河畔的金柳, /是夕阳中的新娘; /波光里的艳影, /在我的心头荡漾。 软泥上的青荇, /油油的在水底招摇; /在康河的柔波里, /我甘心做一条水草!那榆阴下的一潭, /不是清泉,是天上虹 /揉碎在浮藻间, /沉淀着彩虹似的梦。寻梦?撑一支长篙, /向青草更青处漫溯, /满载一船星辉, /在星辉斑斓里放歌。但我不能放歌, /悄悄是别离的笙箫; /夏虫也为我沉默, / 沉默是今晚的康桥!悄悄的我走了, /正如我悄悄的来;/我挥一挥衣袖, /不带走一片云彩。 4、《荆轲刺秦王》 太子及宾客知其事者,皆白衣冠以送之。至易水上,既祖,取道。高渐离击筑,荆轲和而歌,为变徵之声,士皆垂泪涕泣。又前而为歌曰:“风萧萧兮易水寒,壮士一去兮不复还!”复为慷慨羽声,士皆瞋目,发尽上指冠。于是荆轲遂就车而去,终已不顾。 5、《记念刘和珍君》鲁迅 (1)、真的猛士,敢于直面惨淡的人生,敢于正视淋漓的鲜血。这是怎样的哀痛者和幸福者?然而造化又常常为庸人设计,以时间的流驶,来洗涤旧迹,仅使留下淡红的血色和微漠的悲哀。在这淡红的血色和微漠的悲哀中,又给人暂得偷生,维持着这似人非人的世界。我不知道这样的世界何时是一个尽头!

集团客户大客户业务改革创新年措施

集团客户业务业务改革创新提升年 工作实施细则 “天与水违行,君子以做事谋始”德州分公司集团客户部,认真学习省公司下发的《山东广电网络有限公司“改革创新提升年“实施意见》,深刻理解“改革创新提升年“在公司发展过程中的重要影响,领会省公司党委对于2020年度工作的部署和战略发展意图,与省公司整体同向、同心、同力,切实的把思想与行动统一到省公司党委的要求上来。 集团客户部将在理解领会“改革创新提升年“实施意见的前提下,结合自身发展实际情况、本地发展特点,在其指导思想下,确定原则、指定实施细则,认真部署谋划,扎实推进。 健全完善集客业务发展机制。改革创新年的集客业务改革创新,需要激发人才的创新创造活力,这就需要相应的机制制度来激发鼓励。部门内强化集客业务发展流程推进制度,确保用合理的制度利用现有业务资源进行拓展拔高。同时,放宽业务激励奖励至全员,调动全员积极性共同助力集客业务的全面发展,鼓励全员营销全员发展集客业务。在业务开展过程中,即重收入也重利润。进一步完善集客项目的立项审批,进行精细化管理,加强成本控制与利润核算,确保集客业务的收入、利润双增双高。最后项目到人、责任到人、落实到人。保证集客业务项目的落地与推进,并且提供良好的售前、售中、售后服

务,提高集团客户的好感度与忠诚度,以促进集客业务稳定永续的发展。 强化项目引领,塑造集客品牌。强化品牌的力量、利用榜样的力量。大力着重建设集客业务的品牌项目、明星项目。如此可以提高对我公司的认可度,便于以后业务的进一步推广和发展;对品牌项目、明星项目的复制,能够快速部署到各区县范围内,缩小集客业务发展的不平衡,提高整体集客业务发展的最短板。其中,着重发力县级荣媒体建设,将鲜鸡肉跟媒体中心与新时代文明时间中心、乡村文化振兴统筹发展,与智慧党建融合互通,利用我们自身的行业优势,切入宣传文化部门核心工作任务,提供出我们广电网络的解决方案。

人教版高中语文必修必背课文

必修1 沁园春·长沙(全文)毛泽东 独立寒秋, 湘江北去, 橘子洲头。 看万山红遍, 层林尽染, 漫江碧透, 百舸争流。 鹰击长空, 鱼翔浅底, 万类霜天竞自由。 怅寥廓, 问苍茫大地, 谁主沉浮。 携来百侣曾游, 忆往昔峥嵘岁月稠。 恰同学少年, 风华正茂, 书生意气, 挥斥方遒。 指点江山, 激扬文字, 粪土当年万户侯。 曾记否, 到中流击水, 浪遏飞舟。 雨巷(全文)戴望舒 撑着油纸伞,独自 彷徨在悠长、悠长 又寂寥的雨巷, 我希望逢着 一个丁香一样地 结着愁怨的姑娘。 她是有 丁香一样的颜色, 丁香一样的芬芳, 丁香一样的忧愁, 在雨中哀怨, 哀怨又彷徨;

她彷徨在这寂寥的雨巷, 撑着油纸伞 像我一样, 像我一样地 默默彳亍着 冷漠、凄清,又惆怅。 她默默地走近, 走近,又投出 太息一般的眼光 她飘过 像梦一般地, 像梦一般地凄婉迷茫。 像梦中飘过 一枝丁香地, 我身旁飘过这个女郎; 她默默地远了,远了, 到了颓圮的篱墙, 走尽这雨巷。 在雨的哀曲里, 消了她的颜色, 散了她的芬芳, 消散了,甚至她的 太息般的眼光 丁香般的惆怅。 撑着油纸伞,独自 彷徨在悠长、悠长 又寂寥的雨巷, 我希望飘过 一个丁香一样地 结着愁怨的姑娘。 再别康桥(全文)徐志摩 轻轻的我走了,正如我轻轻的来;我轻轻的招手,作别西天的云彩。 那河畔的金柳,是夕阳中的新娘;波光里的艳影,在我的心头荡漾。 软泥上的青荇,油油的在水底招摇;

在康河的柔波里,我甘心做一条水草! 那榆荫下的一潭,不是清泉, 是天上虹揉碎在浮藻间,沉淀着彩虹似的梦。 寻梦?撑一支长篙,向青草更青处漫溯, 满载一船星辉,在星辉斑斓里放歌。 但我不能放歌,悄悄是别离的笙箫; 夏虫也为我沉默,沉默是今晚的康桥。 悄悄的我走了,正如我悄悄的来; 我挥一挥衣袖,不带走一片云彩。 记念刘和珍君(二、四节)鲁迅 二 真的猛士,敢于直面惨淡的人生,敢于正视淋漓的鲜血。这是怎样的哀痛者和幸福者?然而造化又常常为庸人设计,以时间的流驶,来洗涤旧迹,仅使留下淡红的血色和微漠的悲哀。在这淡红的血色和微漠的悲哀中,又给人暂得偷生,维持着这似人非人的世界。我不知道这样的世界何时是一个尽头! 我们还在这样的世上活着;我也早觉得有写一点东西的必要了。离三月十八日也已有两星期,忘却的救主快要降临了罢,我正有写一点东西的必要了。 四 我在十八日早晨,才知道上午有群众向执政府请愿的事;下午便得到噩耗,说卫队居然开枪,死伤至数百人,而刘和珍君即在遇害者之列。但我对于这些传说,竟至于颇为怀疑。我向来是不惮以最坏的恶意,来推测中国人的,然而我还不料,也不信竟会下劣凶残到这地步。况且始终微笑着的和蔼的刘和珍君,更何至于无端在府门前喋血呢? 然而即日证明是事实了,作证的便是她自己的尸骸。还有一具,是杨德群君的。而且又证明着这不但是杀害,简直是虐杀,因为身体上还有棍棒的伤痕。 但段政府就有令,说她们是“暴徒”! 但接着就有流言,说她们是受人利用的。 惨象,已使我目不忍视了;流言,尤使我耳不忍闻。我还有什么话可说呢?我懂得衰亡民族之所以默无声息的缘由了。沉默啊,沉默啊!不在沉默中爆发,就在沉默中灭亡。

(客户管理)中国移动集团客户营销体系

(客户管理)中国移动集团客户营销体系

中国移动集团客户营销体系 从移动通信市场运营经验探索的意义上解析小灵通,它对中国电信的意义就绝不仅仅限于于“3G’之前找到壹个新增点,解决中国电信收入增长率的问题。中国电信的运营者,也提前意识到小灵通的战略意义,因此,于壹些小灵通发展较早的电信公司,于完成“用户规模积累”阶段后,开始于小灵通客户群的服务体系等方面进行尝试:例如成均电信,于小灵通用户规模达到130万户后,成立了专门的小灵通客户部,负责用户尤其是高端用户的服务和保有;浙江电信则全省统壹于10000号建立专门的小灵通班组。 从这些刚刚开始进行的探索中,中国电信已经感受到对移动客户尤其是高值移动客户关注的重要性。以成均电信为例,2005年,实行客户经理负责的近8万小灵通高端客户,全年流失率仅为2%,大大低于小灵通客户平均30%的流失率水平。 中国电信和其从头开始探索移动客户群的服务之路,不如“站于领先者的肩上”,研究壹下中国移动于客户服务尤其是高值客户服务方面的发展历程,作为中国电信思考的参照。 基于这壹初衷,我们于收集大量关联资料的基础上,对中国移动集团客户体系近年来的发展历程及思路进行了梳理,且提出了结合中国电信调整和改善市场营销体系的建议。 中国移动集团客户体系的发展沿革及简介 2000年,中国移动从中国电信剥离后,这壹批当初的电信人很快意识到移动客户群的运营和服务和固定客户群有着迥然不同的特征,其中最明显的区别于于移动客户强烈的个人性。我们能够见到,

中国移动早期正是针对移动客户个人性的特点,对不同客户群进行分级服务,且大力打造俩大服务品牌:针对普通客户服务的1860品牌和针对高端客户的全球通VIP服务品牌。 可是,值得重视的是,从2002年以后,中国移动开始低调但快速地把客户服务体系的重心转向另壹方面:集团客户。和中国电信的大客户体系相比,移动的集团客户体系能够说是后来者,而且由于移动客户的个人性,集团体系的建设面临许多的困难。为此,中国移动不得不借用电力系统“网格化”管理的方面,通过网格化管理,逐步收集和建立集团客户档案。经过四年的努力,中国移动的集团客户的覆盖率已经达到近30%,集团客户对于高值用户群的稳定、移动数据及增值业务的推广、行业应用等方面,已经具有相当的战略地位。中国移动是如何于短短四年内建立和健全这壹体系的?我们首先简单回顾移动集团客户体系的发展历程。 2002年是中国移动集团客户体系建设的试点推广年。这壹年移动主要于以下方面进行了初步的探索:成立项目组、产品模块化、典型项目推广、建立工作流程。 2003年是移动集团客户体系建设的全面促进年。他们明确提出“集团客户工作不能再是业余爱好,要打正规战役。”这壹年他们于集团客户体系建设方面的主要进展是:建立工作体系、健全客户资料、建设平台支撑系统、大力推广标准化应用产品、创新商务模式、建立营销渠道。为此,各级移动主要开展了四项工程:摸底工程,主要是地毯式清查,健全数据;圈地工程,以四川移动为例,他们要求当年

Unit5THEMONSTER课文翻译大学英语六

Unit 5 THE MONSTER He was an undersized little man, with a head too big for his body -- a sickly little man. His nerves were had. He had skin trouble. It was agony for him to wear anything next to his skin coarser than silk. And he had seclusions of grandeur. He was a monster of conceit.Never for one minute did he look at the world or at people, except in relation to himself. He was not only the most important person in the world,to himself;in his own eyes he was the only person who existed. He believed himself to be one of the greatest dramatists in the world, one of the greatest thinkers, and one of the greatest composers. To hear him talk, he was Shakespeare, and Beethoven, and Plato, rolled into one. And you would have had no difficulty in hearing him talk. He was one of the most exhausting conversationalists that ever lived. An evening with him was an evening spent in listening to a monologue. Sometimes he was brilliant; sometimes he was maddeningly tiresome. But whether he was being brilliant or dull, he had one sole topic of conversation: himself. What he thought and what he did. He had a mania for being in the right.The slightest hint of disagreement,from anyone, on the most trivial point, was enough to set him off on a harangue that might last for house, in which he proved himself right in so many ways, and with such exhausting volubility, that in the end his hearer, stunned and deafened, would agree with him, for the sake of peace. It never occurred to him that he and his doing were not of the most intense and fascinating interest to anyone with whom he came in contact.He had theories about almost any subject under the sun, including vegetarianism, the drama, politics, and music; and in support of these theories he wrote pamphlets,le tters, books? thousands upon thousands of words, hundreds and hundreds of pages. He not only wrote these things, and published them -- usually at somebody else's expense-- but he would sit and read them aloud, for hours, to his friends and his family. He wrote operas,and no sooner did he have the synopsis of a story, but he would invite -- or rather summon -- a crowed of his friends to his house, and read it aloud to them. Not for criticism. For applause. When the complete poem was written, the friends had to come again,and hear that read aloud.Then he would publish the poem, sometimes years before the music that went with it was written. He played the piano like a composer, in the worst sense of what that implies, and he would sit down at the piano before parties that included some of the finest pianists of his time, and play for them, by the hour, his own music, needless to say. He had a composer's voice. And he would invite eminent vocalists to his house and sing them his operas, taking all the parts.

Unit7TheMonster课文翻译综合优质教程四.docx

最新资料欢迎阅读 Unit 7 The Monster课文翻译综合教程四 Unit 7 The Monster Deems Taylor 1 He was an undersized little man, with a head too big for his body ― a sickly little man. His nerves were bad. He had skin trouble. It was agony for him to wear anything next to his skin coarser than silk. And he had delusions of grandeur. 2 He was a monster of conceit. Never for one minute did he look at the world or at people, except in relation to himself. He believed himself to be one of the greatest dramatists in the world, one of the greatest thinkers, and one of the greatest composers. To hear him talk, he was Shakespeare, and Beethoven, and Plato, rolled into one. He was one of the most exhausting conversationalists that ever lived. Sometimes he was brilliant;sometimes he was maddeningly tiresome. But whether he was being brilliant or dull, he had one sole topic of conversation: himself. What he thought and what he did. 3 He had a mania for being in the right. The slightest hint of disagreement, from anyone, on the most trivial point, was enough to set him off on a harangue that might last for hours, in which he proved himself right in so many ways, and with such exhausting volubility, that in the end his hearer, stunned and deafened, would agree with him,for the sake of peace. 4 It never occurred to him that he and his doing were not of the most intense and fascinating interest to anyone with whomhe came in contact. He had theories about almost any subject under the sun,including vegetarianism, the drama, politics, and music; and in support of these theories he wrote pamphlets, letters,books ... thousands upon thousands

相关主题
相关文档 最新文档