张苏明—ICVD全程管理201105

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将制度链接到流程去执行①

将制度链接到流程去执行①
佚名
【期刊名称】《《计算机系统应用》》
【年(卷),期】2013(000)006
【摘要】系统地阐述了将制度链接到流程去执行的理念和方法,以政府部门建立质量管理体系为例进行了从制度分析到流程设计方法论的实证分析,并进而对将制度链接到流程去执行的载体—信息化应用平台的顶层设计与应用架构的规划进行了分析与建模,以期为政府部门建立质量管理体系、将制度链接到流程去执行提供一种过程方法和技术路线.
【总页数】8页(P6-13)
【正文语种】中文
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10.1007_s00253-010-2443-4

BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERINGEffects of biotic and abiotic elicitors on cell growth and tanshinone accumulation in Salvia miltiorrhiza cell culturesJiang-Lin Zhao &Li-Gang Zhou &Jian-Yong WuReceived:7September 2009/Revised:6January 2010/Accepted:6January 2010/Published online:2March 2010#Springer-Verlag 2010Abstract This study examined the effects of biotic and abiotic elicitors on the production of diterpenoid tanshi-nones in Salvia miltiorrhiza cell culture.Four classes of elicitors were tested,heavy metal ions (Co 2+,Ag +,Cd 2+),polysaccharides (yeast extract and chitosan),plant response-signaling compounds (salicylic acid and methyl jasmonate),and hyperosmotic stress (with sorbitol).Of these,Ag (silver nitrate),Cd (cadmium chloride),and polysaccharide from yeast extract (YE)were most effective to stimulate the tanshinone production,increasing the total tanshinone content of cell by more than ten-fold (2.3mg g -1versus 0.2mg g -1in control).The stimulating effect was concentration-dependent,most significant at 25μM of Ag and Cd and 100mg l -1(carbohydrate content)of YE.Of the three tanshinones detected,cryptotanshinone was stimulat-ed most dramatically by about 30-fold and tanshinones I and IIA by no more than 5-fold.Meanwhile,most of the elicitors suppressed cell growth,decreasing the biomass yield by about 50%(5.1–5.5g l -1versus 8.9g l -1in control).The elicitors also stimulated the phenylalanine ammonia lyase activity of cells and transient increases in the medium pH and conductivity.The results suggest that the elicitor-stimulated tanshinone accumulation was a stress response of the cells.Keywords Salvia miltiorrhiza .Cell culture .Tanshinones .Elicitors .Stress responseIntroductionSalvia miltiorrhiza Bunge (Lamiaceae),called Danshen in Chinese,is a well-known and important medicinal plant because its root is an effective herb for treatment of menstrual disorders and cardiovascular diseases and for the prevention of inflammation (Tang and Eisenbrand 1992).As its Chinese name refers,Danshen root is characterized by the abundance of red pigments which are mainly ascribed to numerous diterpene quinones generally known as tanshinones,e.g.,tanshinone I (T-I),tanshinone-IIA (T-IIA),and T-IIB,isotanshinone I and II,and cryptotanshinone (CT).Tanshinones constitute a major class of bioactive compounds in S .miltiorrhiza roots with proven therapeutic effects and pharmacological activities (Wang et al.2007).Danshen in combination with a few other Chinese herbs is an effective medicine widely used for the treatment of cardiovascular diseases and used as an emergency remedy for coronary artery disease and acute ischemic stroke.According to WHO statistics,cardiovas-cular diseases are and will continue to be the number one cause of death in the world (www.who.int/cardiovascular_diseases ).It is of significance to develop more efficient means for the production of Danshen and its active constituents.Although field cultivation is currently the major produc-tion means for Danshen and most other plant herbs,plant tissue cultures provide more well-controlled and sustainable systems for efficient production of desired bioactive compounds of the herb.Plant tissue cultures are the most useful and convenient experimental systems for examiningJ.-L.Zhao :L.-G.Zhou (*)Department of Plant Pathology,China Agricultural University,Beijing 100193,China email:lgzhou@J.-Y .Wu (*)Department of Applied Biology and Chemical Technology,The Hong Kong Polytechnic University,Hung Hom,Kowloon,Hong Kong email:bcjywu@.hkAppl Microbiol Biotechnol (2010)87:137–144DOI 10.1007/s00253-010-2443-4various factors on the biosynthesis of desired products and for exploring effective measures to enhance their produc-tion.The importance of Danshen for traditional and modern medicines has promoted the long-lasting research interest in the development of tiorrhiza tissue cultures for production of bioactive compounds for more than two decades.In an early study,Nakanishi et al.(1983)induced several cell lines from plant seedlings and screened out a cell line capable of producing significant amounts of CT and another diterpene,ferruginol.In later studies,the group performed a fuller evaluation and optimization of the medium for cell growth and CT production and,eventually,derived an effective production medium with a simpler composition(ten components)than the original Murashige and Skoog(MS) medium(about20components),achieving a high CT yield of 110mg l-1(Miyasaka et al.1987).Many recent studies have been focused on hairy root cultures of tiorrhiza transformed by Agrobacterium rhizogenes(Hu and Alfermann1993;Chen et al.2001)and by our group (Zhang et al.2004;Ge and Wu2005;Shi et al.2007).Most of the bioactive compounds in medicinal plants belong to secondary metabolites which are usually less abundant than primary metabolites in the plants.Since the accumulation of secondary metabolites in plants is a common response of plants to biotic and abiotic stresses, their accumulation can be stimulated by biotic and abiotic elicitors.Therefore,elicitation,treatment of plant tissue cultures with elicitors,is one of the most effective strategies for improving secondary metabolite production in plant tissue cultures(Chong et al.2005;Smetanska2008).The most common and effective elicitors used in previous studies include the components of microbial cells especially poly-and oligosaccharides(biotic)and heavy metal ions, hyperosmotic stress,and UV radiation(abiotic),and the signaling compounds in plant defense responses such as salicylic acid(SA)and methyl jasmonate(MJ;Zhou and Wu2006;Smetanska2008).Some of these elicitors,yeast extract(mainly the polysaccharide fraction),silver ion Ag+, and hyperosmotic stress(by an osmoticum)have also been applied and shown effective to enhance the production of tanshinones in tiorrhiza hairy root cultures(Chen et al.2001;Zhang et al.2004;Shi et al.2007).To the best of our knowledge,only a few studies have been documented on the effects of elicitors,YE,SA,and MJ,on the secondary metabolite production in Agro-bacterium tumefaciens transformed tiorrhiza cell cultures from one research group(Chen and Chen1999, 2000)but not any study in normal cell cultures.The present study focuses on the effects of common biotic and abiotic elicitors including polysaccharides,heavy metal ions, SA and MJ,and osmotic stress(with sorbitol)on the growth and accumulation of three major tanshinones T-I, T-IIA,and CT in suspension culture of normal tior-rhiza cells.In addition to the effects of various elicitors on the total tanshinone content of cells,the study will examine the effects on different tanshinone species and the potential relationship to plant stress response.Material and methodsCallus induction and cell suspension cultureYoung stem explants of tiorrhiza Bunge were collected from the botanical garden at the Institute of Medicinal Plant Development,Chinese Academy of Med-ical Sciences,Beijing,China,in May2005.The fresh explants were washed with tap water,surface-sterilized with 75%ethanol for1min,and then soaked in0.1%mercuric chloride for10min and rinsed thoroughly with sterilized water.The clean and sterilized explants were cut into∼0.5-cm segments and placed on solid MS medium(Murashige and Skoog1962)supplemented with sucrose(30g l-1),2,4-D(2mg l-1)and6-BA(2mg l-1)to induce callus formation. The callus culture of tiorrhiza was maintained on a solid,hormone-free MS medium with8g l-1agar and 30g l-1sucrose at25°C in the dark and subcultured every 4weeks.The culture was deposited in Lab Y1210at The Hong Kong Polytechnic University with a collection number of Danshen cell-1.All experiments in this study were performed in suspension culture of tiorrhiza cells in a liquid medium of the same composition as for the solid culture but excluding agar.The cell suspension culture was maintained in shake-flasks,i.e.,125-ml Erlenmeyer flasks on an orbital shaker operated at110–120rpm,at 25°C in the dark.Each of the flasks was filled with25ml medium and inoculated with0.3g fresh cells from18–21-day-old shake–flask culture.Elicitor preparation and administrationEight elicitors were tested,each at three concentrations,in the initial elicitation experiments(Table1).These are representative of the four major classes of elicitors for the induction of plant responses and the stimulation of secondary metabolite production in plant tissue cultures (Zhou and Wu2006;Smetanska2008).All elicitors except MJ were prepared as a concentrated stock solution in water and autoclaved at121°C for15min,and stored at4°C in a refrigerator prior to use.Yeast elicitor(YE)was the polysaccharide fraction of yeast extract(Y4250,Sigma, St.Louis,MO,USA)prepared by ethanol precipitation as described previously(Hahn and Albersheim1978;Ge and Wu2005).In brief,yeast extract was dissolved in distilled water(20g/100ml)and then mixed with400ml of ethanol and allowed to precipitate for4days at4°C in arefrigerator.The precipitate was redissolved in100ml of distilled water and subjected to another round of ethanol precipitation.The final gummy precipitate was dissolved in 50ml of distilled water and stored at4°C before use.The concentration of YE was represented by total carbohydrate content which was determined by the Anthrone test using sucrose as a reference.Chitosan solution was prepared by dissolving0.5g crab shell chitosan(C3646,Sigma)in1ml glacial acetic acid at55–60°C for15min,and then the final volume was adjusted to50ml with distilled water and the pH adjusted to5.8with NaOH(Prakash and Srivastava 2008).MJ(Cat.39,270-7,Sigma-Aldrich)was dissolved in 95%ethanol and sterilized by filtering through a microfilter (0.2µm).SA(10,591-0,Sigma-Aldrich),sorbitol(S3755, Sigma),and the salts of heavy metals including cobalt chloride(C8661,Sigma-Aldrich),silver nitrate(S7276, Sigma-Aldrich),and cadmium chloride(C5081,Sigma-Aldrich)were dissolved in distilled water to the desired concentrations and adjusted to pH5.8.Elicitor treatment was administered to the shake–flask culture of tiorrhiza cell on day18,which was about 2–3days before reaching the stationary phase.This time point is usually favorable for elicitation when the biomass concentration is high(compared with earlier days of growth),and the cell metabolism is still active(compared with that during or after stationary phase;Buitelaar et al. 1992;Cheng et al.2006).Each of the elicitor solutions was added into the culture medium with a micropipette at the desired concentration.After the elicitor addition,the shake–flask culture of cells was maintained for another7days and then harvested for analysis.All treatments were performed in triplicate,and the results were averaged.After the initial experiments on the eight elicitors,the three most effective ones,Ag(25µM),Cd(25µM),and YE(100mg l-1)were applied in the following experiments on the time courses of elicitor-treated cell growth and tanshinone accumulation in the tiorrhiza cell culture.Measurement of cell weight,sucrose concentration, medium pH,and conductivityThe cells were separated from the liquid medium by filtration.The cell mass on the filter paper was rinsed thoroughly with water and filtered again,and blotted dry by paper towels and then dried at50°C in an oven to attain the dry weight.Sucrose concentration in the liquid medium was determined by the Anthrone test using sucrose as a reference(Ebell1969),and the medium pH and conduc-tivity were measured with the respective electrodes on an Orion720A+pH meter(Thermo Fisher Scientific,Inc., Beverly,MA,USA)and a CD-4303conductivity meter (Lutron,Taiwan),respectively.Measurement of PAL activityPhenylalanine ammonia lyase(PAL)was extracted from fresh tiorrhiza cells with borate buffer(pH8.8).The cells were ground in the buffer(0.15g ml-1)for2min with a pestle and mortar on ice,and then centrifuged at10,000rpm and4°C for20min to obtain a solid-free extract.The PAL activity was determined based on the conversion of L-phenylalanine to cinnamic acid as described by Wu and Lin(2002).Analysis of tanshinone contentsThe cell mass from culture was dried and ground into powder and extracted with methanol/dichloromethane(4:1, v/v,10mg ml-1)under sonication for60min.After removal of the solid,the liquid extract was evaporated to dryness and redissolved in methanol/dichloromethane(9:1,v/v). Tanshinone content was determined by high performance liquid chromatography(HPLC)on a HP1100system using C18column,acetonitrile/water(55:45,v/v)as the mobile phase,and UV detection at275nm as described previously (Shi et al.2007).Three tanshinone species CT,T-I,and T-IIA were detected and quantified with authentic standards obtained from the Institute for Identification of Pharmaceu-tical and Biological Products(Beijing,China).Total tanshinone content is the total content of the three tanshinones in the cells.Tanshinone content in the culture medium was negligible and not determined.ResultsCell growth and tanshinone accumulation in tiorrhiza cell cultureThe time course of tiorrhiza cell growth exhibited a lag phase or slow growth period in the first3–6days, a rapid,linear growth period between day9–18,and aTable1Elicitors and concentrations tested in the initial experiments Elicitors Unit ConcentrationC1C2C3Cobalt chloride(Co)µM 5.02550 Silver nitrate(Ag)µM 5.02550 Cadmium chloride(Cd)µM 5.02550 Salicylic acid(SA)µM1050100 Methyl jasmonate(MJ)µM1050100 Yeast elicitor(YE)mg l-150100200 Chitosan(CH)mg l-150100200 Sorbitol(SO)g l-152550stationary or declining phase in the later days,reaching the maximum biomass concentration (8.1g l -1)around day 21.The total tanshinone content of cells remained at a very low level from days 1–12and then increased steadily from days 12–27to a maximum of 0.16mg g -1.A significant portion (65%)of the tanshinone accumulation or content increase occurred during the stationary phase from days 21–27(Fig.1a ),which is characteristic of secondary metabolite production in a batch culture process.The time course of sugar (sucrose)concentration (Fig.1b )was nearly sym-metrical to that of cell growth,indicating a direct correlation of the cell growth (or biomass production)to sugar consumption.As the major carbon source,sugar was required for the S .miltiorrhiza cell growth,and when it was depleted (around day 21),the cell growth stopped,and the biomass concentration began to drop.As seen from Fig.1b ,the medium pH showed a notable drop in the first 3days (due to consumption of NH 4+and release of protons)and a gradual increase after day 6(due to consumption of nitrate NO 3-)(Morard et al.1998).Effects of various elicitors on cell growth and tanshinone productionFigure 2shows the effects of elicitor treatments on the cell growth and tanshinone accumulation in S .miltiorrhiza cell cultures,which were dependent both on the elicitor species and elicitor dose.As seen from Fig.2a ,most of the elicitor treatments except Co 2+and sorbitol at lower concentrations suppressed the cell growth to a lower biomass concentra-tion than that of the untreated control culture,and the growth suppression was more severe at a high elicitor dose.On the other hand,most of the elicitor treatments except Co 2+,sorbitol,SA,and MJ at lower concentrations increased the total tanshinone content of cell to a higher level than in the control (Fig.2b ).Overall the results indicated that the enhancement of tanshinone accumulation by an elicitor treatment concurred with a notable suppres-sion of cell growth or biomass production.Nevertheless,some of the elicitors had a much stronger stimulating effect on the tanshinone accumulation than the suppressing effect on the cell growth.In particular,Ag and Cd both at 25μM,and YE at 100mg l -1increased the total tanshinone content to 2.30mg g -1,about 11.5-fold versus that of the control (0.20mg g -1),but decreased the biomass production by no more than 50%(5.1–5.5g l -1versus 8.9g l -1).Another three elicitors,SA,MJ (both at 50μM),and sorbitol (50g l -1)increased the total tanshinone content by 2–3-fold but decreased the biomass by 30–45%compared with the control.The stimulating effect of chitosan on tanshinone accumulation (about 6-fold)was stronger than SA,MJ,and sorbitol but much weaker than Ag,Cd,and YE,while its suppressing effect on the cell growth was as severe as Ag,Cd,and YE.In summary,the results indicate that Ag,Cd,YE are the most favorable elicitors for the tanshinone production in S .miltiorrhiza cell culture and were used in the following experiments.Figure 3shows the time courses of cell growth and tanshinone production after treatment with the three most effective elicitors Ag (25μM),Cd (25μM),and YE (100mg l -1)and the control culture.All three elicitor treatments caused a steady decline of biomass concentration from initially 8.5g l -1to 5.3g l -1on day 6while biomass in00.040.080.120.160.20246810TT content (mg g -1)C e l l b i o m a s s (g d w l -1)dw TTa4.85.1 5.45.76.001020304036912151821242730p HS u c r o s e (g l -1)Culture time (d)bSucrosepHFig.1Time courses of biomass and total tanshinone content (a ),residue sugar (sucrose)and medium pH (b )in S .miltiorrhiza cell cultures (error bars for standard deviations,n =3)246810C e l l b i o m a s s (g l -1)0.00.51.01.52.02.5Control AgCdSAMJYECH SOT T c o n t e n t (m g g -1)Elicitor treatmentCo Fig.2Effects of various elicitors on biomass growth (a )andtanshinone production (b )in S .miltiorrhiza cell cultures (elicitors added to cultures on day 18at three concentrations C1,C2,and C3as shown in Table 1,and cultures harvested on day 25;error bars for standard deviations,n =3)the control culture was increased during this period (Fig.3a ).In the meantime,the tanshinone content of cells in the three elicitor-treated cultures increased sharply and most rapidly by Ag (from 0.14to 1.98mg g -1),while that of control increased slightly (from 0.14to 0.21mg g -1;Fig.3b ).The volumetric total tanshinone yields (the products of total tanshinone content and cell dry weight)were 1.9mg l -1in the control,and 9.2mg l -1,10.7mg l -1and 11.7mg l -1in cultures treated with 100mg l -1YE,25μM Cd,and Ag,respectively (on day 6).Another test was performed on the effects of two and three elicitors in combinations in the S .miltiorrhiza cell culture.As shown in Fig.4,the tanshinone content was increased about 20%with either two elicitors and about 40%with all three elicitors in combination compared with that with a single elicitor.The results suggest an additive or synergistic effect of these elicitors on the tanshinone accumulation in the cells.However,the combined use of two or three elicitors also suppressed the cell growth (biomass)more severely than with a single elicitor.Effects of elicitor treatments on different tanshinone species Of the three tanshinone species detected,CT was stimulated most significantly by all elicitors without exception;T-IIA was stimulated by most elicitors,and T-I was stimulated significantly only by chitosan but slightly stimulated or suppressed by other elicitors (Table 2).The highest CT content was about 2mg g -1(1,854–2,011μg g -1)in cellcultures treated with 25μM Ag and Cd,and 100mg l -1YE,about 31–34fold of the control level (60μg g -1),the highest T-I content 0.27mg g -1with 100mg l -1chitosan (3.4-fold of the control 80μg g -1)and the highest T-IIA content 0.37mg g -1with 25μM Cd (6-fold of the control 60μg g -1).As seen from the HPLC chromatograms (Fig.5),the cultures treated with the three different elicitors exhibited a similar profile with virtually identical major peaks.The experimental results do not suggest any specificity of particular tanshinone species to the type of elicitors,YE and chitosan as biotic polysaccharides,Cd and Ag as abiotic heavy metals,or SA and MJ as plant stress signaling pared with that of control,the HPLC profiles of elicitor-treated cultures also had three new unknown peaks appearing before the CT peak,between 10.0–11.5min and a high peak at 11.1min,which0.00.51.01.52.02.5123456T T c o n t e n t (m g g -1)Time after treatment (d)b4681012C e l l b i o m a s s (g l -1)Control Ag 25Cd 25YE 100aFig.3Time courses of biomass (a )and total tanshinone content (b )in S .miltiorrhiza cell cultures after treatment with Ag (25µM),Cd (25µM),and YE (100mg l -1;error bars for standard deviations,n =3)24681012345Cell dry weight (g l -1)T T c o n t e n t (m g g -1)Elicitor treatmentTTdwFig.4Effects of single and combined elicitors on S .miltiorrhiza cell growth and tanshinone accumulation (elicitors added to cell cultures on day 18at the same concentrations as in Fig.3,and cultures harvested on day 25;error bars for standard deviations,n =3)Table 2Effects of various elicitors on the accumulation of three tanshinones in S .miltiorrhiza cells Treatment aContent,μg/g (fold of content control)CTT-IT-IIA Control 59.9(1)81.6(1)57.6(1)Co-50263.7(4.4)67.5(0.83)55.5(0.96)Ag-251,817.5(30)71.0(0.87)225.8(3.9)Cd-251,854.0(31)80.3(0.98)369.0(6.4)SA-100390.0(6.5)78.5(0.96)72.8(1.3)MJ-100299.8(5.0)109.5(1.3)82.6(1.4)YE-1002,011.4(34)90.3(1.1)190.3(3.3)CH-100597.2(10)276.0(3.4)98.8(1.7)SO-50584.6(9.8)56.9(0.70)83.0(1.4)CT cryptotanshinone,T-I tanshinone I,T-IIA tanshinone-IIAaNumber after each elicitor symbol represents the elicitor concentra-tion as shown in Table 1may be ascribed to tanshinone relatives of higher polarity than CT induced by the elicitors.PAL activity,pH,and conductivity changes induced by elicitorsFigure 6shows the changes of intracellular PAL activity and medium pH and conductivity in the S .miltiorrhiza cell culture after the treatment by Ag (25μM),Cd (25μM),and YE (100mg l -1).The PAL activity of cells was stimulated by all three elicitors to the similar level,from 1.4-to 1.9-fold of the control level over 6days (Fig.6a ).PAL is a key enzyme at the entrance step in the phenylpropanoid pathway in plants,and its activity increase stimulated by the elicitors is suggestive of an enhanced secondary metabolism in the plant cells (Taiz and Zeiger 2006).The pH and conductivity of culture medium were also increased (to higher levels than those of the control)by all three elicitors but more significantly by YE (Fig.6b,c ).Most significant increases (differences from the control level)in the medium pH and conductivity were shown in the very early period from day 0–1.The increase in medium conductivity in the early period was most probably attributed to the release of potassium K +ion from the cells or K +efflux across the cell membrane (Zhang et al.2004).Transient medium pH increase (alkalinization)and K +efflux across the cell membrane are early and important events in the elicitation of plant responses and phytoalexin production (Ebel and Mithöer 1994;Roos et al.1998).The conductivity decline in the later period after day 1of Ag +and Cd 2+-treated cultures and the control cultures can be attributed to the consumption of inorganic and mineral nutrients in the culture medium (Kinooka et al.1991).Overall,the results here provide further evidence forthe01234R e l a t i v e P A L a c t i v i t yControl Ag CdYEa5.05.45.86.26.6M e d i u m p H b2.03.04.05.06.00246M e d i u m c o n d u c t i v i t y (m S )Time after treatment (d)cFig.6Time courses of PAL activity (a ),medium pH (b ),and conductivity (c )of S .miltiorrhiza cell cultures after elicitor treatments in comparison with the control (error bars for standard deviation,n =3)elicitor activities of Ag,Cd,and YE in stimulating the stress responses and secondary metabolism of the S. miltiorrhiza cells.DiscussionThe effects of various elicitors on tanshinone accumulation found here in the normal tiorrhiza cell cultures are in general agreement with those found in transformed cell and hairy root cultures of tiorrhiza.In transformed cell cultures(Chen and Chen1999),the CT accumulation was also stimulated significantly by YE but not by SA or MJ alone,and YE also inhibited the cell growth.The tanshinone(mainly CT)production in hairy root cultures was also enhanced significantly(3–4fold)by Ag(Zhang et al.2004)and YE(Shi et al.2007).In all these culture systems,CT was the major tanshinone species stimulated by various elicitor treatments.CT has been identified as a phytoalexin in tiorrhiza plant which plays a defense role against pathogen invasion of the plant(Chen and Chen 2000).In this connection,the stimulated CT accumulation by the elicitors may be a defense or stress response of the cells.CT was also the major diterpenoid produced by a normal tiorrhiza cell line which was initially grown in the MS medium and then transferred to a production medium containing only about half of the nutrient compo-nents of the MS medium(Miyasaka et al.1987).It is very possible that the improvement of CT yield in this production medium was also attributed,at least partially, to the stress imposed by the nutrient deficiency which suppressed growth but stimulated secondary metabolite accumulation.MJ or its relative jasmonic acid has been shown effective for stimulating a variety of secondary metab-olites in plant tissue cultures such as hypericin in Hypericum perforatum L.(St.John’s Wort)cell cultures (Walker et al.2002),paclitaxol(diterpenoid)and related taxanes in various Taxus spp.and ginsenosides in Panax spp.(Zhong and Yue2005),and bilobalide and ginkgo-lides in Ginkgo biloba cell cultures(Kang et al.2006). However,MJ showed only a moderate or insignificant stimulating effect on tanshinone accumulation in normal and transformed tiorrhiza cell cultures.The discrep-ancy suggests that the effects of various elicitors on secondary metabolite production in plant tissue cultures are dependent on the specific secondary metabolites.This argument is also supported by the much stronger stimu-lation of CT than T-I and T-IIA by most elicitors found in our tiorrhiza cell cultures.In addition,the hairy roots appeared more tolerant to the elicitor stress,and the growth was less inhibited by the elicitors or even enhanced in some cases,e.g.,by YE(Chen et al.2001)and sorbitol(Shi et al.2007).Moreover,sorbitol as an osmotic agent significantly stimulated the tanshinone accumulation(3–4folds)in tiorrhiza hairy root cultures,but not so significantly in the cell cultures.This shows that the elicitor activities for the same metabolites can vary with the tissue culture systems.In conclusion,the polysaccharide fraction of yeast extract and two heavy metal ions Ag+and Cd2+were potent elicitors for stimulating the tanshinone production in tiorrhiza cell culture.The stimulated tanshinone production by most elicitors was associated with notable growth suppression.CT was more responsive to the elicitors and enhanced more dramatically than another two tanshinones,T-I and IIA.The results from this study in the tiorrhiza cell cultures and from previous studies in hairy root cultures suggest that the cell and hairy root cultures may be effective systems for CT production, provided with the elicitors.As most of the elicitor chemicals are commercially available or can be readily prepared in the laboratory and easily administered to the cell and root cultures,they are suitable for practical applications in the laboratory or large-scale production. Acknowledgements This work was supported by grants from The Hong Kong Polytechnic University(G-U502and1-BB80)and the China Hi-Tech Research and Development Program(2006AA10A209).ReferencesBuitelaar RM,Cesário MT,Tramper J(1992)Elicitation of thiophene production by hairy roots of Tagetes patula.Enzyme Microb Technol14:2–7Chen H,Chen F(1999)Effects of methyl jasmonate and salicylic acid on cell growth and cryptotanshinone formation in Ti transformed Salvia miltiorrhiza cell suspension cultures.Biotechnol Lett 21:803–807Chen H,Chen F(2000)Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia miltiorrhiza cell suspension cultures.Plant Cell Rep19:710–717Chen H,Chen F,Chiu FCK,Lo CMY(2001)The effect of yeast elicitor on the growth and secondary metabolism of hairy root cultures of Salvia miltiorrhiza.Enzyme Microb Technol28:100–105Cheng XY,Zhou HY,Cui X,Ni W,Liu CZ(2006)Improvement of phenylethanoid glycosides biosynthesis in Cistanche deserticola cell suspension cultures by chitosan elicitor.J Biotechnol 121:253–260Chong TM,Abdullah MA,Lai QM,Nor’Aini FM,Lajis NH(2005) Effective elicitation factors in Morinda elliptica cell suspension culture.Process Biochem40:3397–3405Ebel J,Mithöer A(1994)Early events in the elicitation of plant defence.Planta206:335–348Ebell LF(1969)Variation in total soluble sugars of conifer tissues with method of analysis.Phytochemistry8:227–233Ge XC,Wu JY(2005)Tanshinone production and isoprenoid pathways in Salvia miltiorrhiza hairy roots induced by Ag+and yeast elicitor.Plant Sci168:487–491。

中国信息技术服务标准(ITSS)白皮书第一版

《中国信息技术服务标准(ITSS)白皮书》（第一版）国家信息技术服务标准指导协调组组　长：陈　伟副组长：郭建兵　陈　英　高素梅　戴红　胡　燕　林　宁何小龙　侯建仁秘书长：尹洪涛成　员：任利华　何海林　姜广智　朱宗尧　陈少媚　池　宇陈建共国家信息技术服务标准工作组组　长：林　宁副组长：马洪杰　赵国祥　于　跃　张　帆　欧阳树生邱善勤　陈渌萍秘书长：周　平Ⅱ编写组王宝艾　高　林　周　平　潘纯峰左天祖　马洪杰　张　帆陈世林　高　巍　范　勇　欧阳树生　寸丹梅　廖　昕　崔　静　李　娜　王春涛　贺东锋　李慧敏子　毛立新　李　新　刘瑞慧　白　璐　王永华参加单位（排名不分先后）中国电子技术标准化研究所神州数码系统集成服务有限公司中国软件与技术服务股份有限公司东软集团股份有限公司上海翰纬信息管理咨询有限公司山东浪潮齐鲁软件产业股份有限公司南天电子信息产业股份有限公司快威科技集团有限公司成都勤智数码科技有限公司上海宝信软件股份有限公司广州市金禧信息技术服务有限公司ⅢCopyright © 2010 版权所有前言2009年4月15日，国务院正式发布《电子信息产业调整和振兴规划》（以下简称：规划），在强化自主创新能力建设方面明确提出“加快制定信息技术服务标准和规范”。

为了贯彻落实规划要求，2009年4月23日，工业和信息化部软件服务业司成立了信息技术服务标准工作组（以下简称：工作组），负责研究并建立信息技术服务标准体系，制定信息技术服务领域的相关标准。

该工作组的成立得到了国家标准化管理委员会、工业和信息化部运行监测协调局、科技司、电子信息司以及北京、上海、广东、江苏、湖北、重庆、成都、沈阳、杭州等省市工业和信息化主管部门的大力支持。

目前，工作组主要围绕信息系统建设、运行维护、服务管理、治理、外包等专业领域开展标准研究制定工作，并针对云计算服务新兴领域开展前期标准预研工作。

工作组形成的标准成果对修订我国《软件产业统计报表制度》、《国民经济行业分类》（GB/T 4754）发挥了重要的支持作用；同时，通过《软件产业统计报表制度》的实施，初步掌握了我国信息技术服务业的总体规模、增长速度及发展趋势；另外，标准在WTO服务贸易多边磋商、中欧服务贸易谈判中也得到了应用。

生产管理领域授课张老师简介

为今天工作成绩优异而努力学习，为明天事业腾飞培训学习以蓄能！是企业对员工培训的意愿，是学员参加学习培训的动力，亦是蓝草咨询孜孜不倦追求的目标。

蓝草咨询提供的训练培训课程以满足初级、中级、中高级的学员（含企业采购标的），通过蓝草精心准备的课程，学习达成当前岗位知识与技能；晋升岗位所需知识与技能；蓝草课程注意突出实战性、技能型领域的应用型课程；特别关注新技术、新渠道、新知识创新型知识课程。

蓝草咨询坚定认为，卓越的训练培训是获得知识的绝佳路径，但也应是学员快乐的旅程，蓝草企业的口号是：为快乐而培训为培训更快乐！蓝草咨询为实现上述目标，为培训机构、培训学员提供了多种形式的优惠和增值快乐的政策和手段，可以提供开具培训费的增值税专用发票。

张铭老师简介⏹ISO质量管理专家⏹华南理工大學MBA⏹ISO10012高级审核员⏹知识产权管理体系审核员⏹卓越绩效培训资深讲师⏹广东省名牌评价中心评审专家⏹湖北工业大学创业导师（教授）⏹ISO/TS16949质量管理及五大工具资深讲师⏹ISO90001、14001、OHSAS18001咨询师/培训师⏹测量管理体系高级审核员（编号：CMS-P-1899）、测量系统分析（MSA）资深讲师实战背景（长达22年工作经验，活跃于ISO系列体系的管理和咨询工作）曾任：联合利华品质部经理/管理者（1994年至2004年）曾任：珠海功控玻璃纤维有限公司管理者（2004年至2006年）曾任：广东蓉胜超微线材有限公司（汽配件）品质经理（2006年至2008年）曾任：深圳XXX管理顾问有限公司高级顾问（2008年至2011年）曾任：中启计量体系认证中心广东分中心（2011年至2015年）资历优势:●组织过多家著名企业的管理体系一体化整合，对质量、环境、职业健康安全管理体系和测量管理体系的保持和持续改进，以及管理体系整合有丰富的经验。

●卓越绩效培训资深讲师，对企业文化、目标管理、系统设计及优化有独到的见解。

执行力塑造和提升

夜很深了，刘军看着桌面上一个个拖延的计划，感觉无限的无奈……
2020/3/3
9
在你的公司或团队中是否有这样一些现象
➢ 经常计划同最终的实际有很大的差别 ➢ ➢ 会议上总有些员工一言不发，而你不知道他们在想什么 ➢ ➢ 经理会议上常常为找出到底应由谁负责而争吵不休 ➢ ➢ 有多少次你的会议制定了实施计划？ ➢ ➢ 为辞退一些业绩不理想的员工而烦恼
每个企业通常都不缺少目标，然而，他们却总是难以实现目标，为什么那么多的公司都实现不了目标呢?
2020/3/3
29
1.设定目标的原则执行力领导者设立的目标，一定是可以转化为各项工
作的。
目标的SMART原则
• S——SPECIFIC • M—— • A——ATTAINABLE • R——REALISTIC • T——TIME
……
小组讨论：结合实际工作，你对此寓言及GE的做法有何感触？
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三.执行力是企业经理层和员工层的共同提高
做职业化经理人做职业化员工
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四.中国企业的执行问题分析及出路
（一）中国企业过去成功在哪里？ 1.第一代企业家：技术引进型 2.第二代企业家：市场政治家（靠谋略、靠勇气） 3.第三代企业家：战略管理型
4
从两则故事谈起：
西点军校的故事 GE成功的故事
2020/3/3
5
主要讲解哪些问题
第一篇整体把握企业的执行力第一章.执行力决定企业的成败第二章.企业执行力的总体境界
第二篇经理人的执行力塑造第三章.执行力之根—掌握4R执行管理模式第四章.执行力之干—有效激励员工第五章.执行力之枝—企业文化的变革第六章.执行力之叶—员工甄选与任用

应用经济学硕士研究生培养方案

应用经济学硕士研究生培养方案(学科代码:0202)一、培养目标本学科致力于培养具有严谨求实的学术作风，德、智、体全面发展，具有坚定正确的政治方向，具有扎实的经济学理论基础、合理的知识结构和宽广的知识面，具有独立从事经济研究的能力，能胜任经济类课程的教学，能胜任实际经济工作。

较为熟练地掌握一门外语并能阅读本学科的外文资料;毕业后可承担本学科的教学、科研工作和中高层次的经济管理工作;具有健康的心理和体魄。

二、学科专业1、区域经济学2、数量经济学3、财政学(含税收学)4、产业经济学5、统计学三、学习年限及应修学分全日制硕士研究生的学习年限一般为3年。

在完成培养要求的前提下，对少数学业优秀、科研成果突出的硕士生，可申请提前毕业，提前期一般不超过1年。

如确需延长学习年限的，延长期一般不超过1年。

至少须修满35学分，其中，课程学习32学分，实践环节3学分。

四、课程设置及考核方式(具体见课程设置与教学计划表)实践环节由科研实践和教学实践组成，科研实践必须参加校内外相关学科学术会议1次，撰写心得体会一份(计1学分);选听学科前沿系列讲座1次，至少6学时;撰写相关文献综述一份(计1学分)。

教学实践必须听课30学时，讲课30学时，提交教学大纲一份(计1学分)。

科研实践和教学实践均由导师负责考核。

五、培养方式研究生由导师及导师小组全面负责培养，以导师指导和本学科教师集体培养相结合为原则，建立和完善有利于学术群体作用的培养机制。

课程学习和研究并重;专业课的学习采取系统讲授、重点辅导、讨论讲座以及任课教师制定参考文献、书目，学习阅读后写综述和评论等多种形式。

加强研究生的自学能力、表达能力、写作能力、实际工作能力等的训练和培养。

六、学位(毕业)论文研究生在修完全部学位课程和修满所要求的总学分后，要在导师的指导下，进行学位(毕业)论文的研撰，由硕士研究生独立完成，论文写作时间不少于一年。

论文选题必须经过充分调查研究，查阅相关的文献，了解国内外本领域的研究历史和现状，选择本学科内有重要学术价值和实用价值、研究基础较为薄弱的问题，或能为解决当前、当地经济和社会发展的热点、难点问题以及为政府决策提供借鉴的问题作为论文选题;研究生确定了论文选题后，在论文写作之前，必须撰写开题报告，开题报告应包括论文选题的理由或意义、国内外有关该论题研究的现状及趋势、本人的详细研究计划、写作提纲、主要参考文献等内容。

张锡民开课目录提纲表(管理及人力类)

二.目标管理的优缺点 1. 目标管理的优点 2. 目标管理的不足之处三.目标管理的组织和流程 1. 目标管理的组织 2. 各部门在目标管理中的作用 3. 目标管理的流程四.目标管理的方法与技巧 1 参考指标体系 2.关键业绩指标实例：小奇为什么不服气？？ 3.工作目标的设计方法 4.目标协议书的签订 5.目标执行的管理实例：丁雪为什么士气不高？？五.工作计划的制定 1. 公司/部门/员工年度工作计划方法 2. 公司/部门/员工季度工作计划方法 3. 公司/部门/员工月度工作计划方法 4. 日常工作计划方法六.案例: 某知名公司目标管理/工作计划项目过程及成果共享引子：面对“仇仔”的挑衅王经理怎么办？？ 4 绩效管理：绩效考核与关键业绩指标的设定与平衡计分卡方法的运用一.绩效管理的概念、作用与价值 1. 绩效管理的概念 2. 绩效管理的作用与价值二.企业绩效管理通常存在的问题 1. 只有绩效考核，没有绩效管理实例：小冯的工作业绩第二年仍不理想，为什么？？ 2. 不知如何定考核指标 3. 考核指标只注重短期效益，没考虑公司长期战略要求 4. 考核结果和奖惩不兑现 5. 等等三．绩效管理循环过程与绩效管理制度 1. 目标管理和绩效管理的关系 2. 绩效管理过程详解 3. 如何制定好的绩效管理制度四.绩效管理的组织和规程 1. 是谁负责公司的绩效管理？ 2. 人力资源部的作用 3. 各业务部门的作用实例：王经理的见解对吗？？ 4. 绩效管理的力度如何把握 6H 低中高层
序号
1
课程名称
组织设计与管理（含集团公司）
课程提纲（两至三层）
一•初步认识组织管理 1.什么是企业的组织？2.组织在管理中的价值 3.组织管理常见问题 4.组织结构的特征因素 5.组织结构的权变因素 6.管理平台与组织设计 –企业管理常见问题； –管理平台及其作用； –管理平台搭建过程。7.企业生命周期各阶段的组织特征二• 7 组织设计的原则与程序； 1.组织设计的基本原则 2.组织设计的程序三•组织框架设计； 1.组织系统表 2.组织系统表的构成要素四•管理幅度与管理层次设计； 1.管理幅度设计 2.管理层次设计 3.高长结构的优缺点 4.扁平结构的优缺点 5.组织变化的趋势五•集权与分权设计； 1.集权与分权的影响因素 2.经济责任中心 3.不同责任中心的权限 4.经济责任中心的选择六•不同组织形式及优缺点； 1.组织形式及特点 2.直线职能制示意 3.直线职能制优缺点 4. 事业部制示意 5.事业部制优缺点 5.矩阵制示意 6.矩阵制优缺点 7.多维立体组织结构七•高层组织设计 1.股东大会职权 2.董事会职权 3.总经理职权 4.监事会职权八.部门结构设计； 1.职权分解 2.横向协调设计小组讨论：结合公司组织管理状况，讨论组织设计与管理的改善思路九•组织评价及组织变革 1.对组织模式的评价标准 2.组织变革的内涵 3.组织变革的方式 4.知识经济对组织管理提出的挑战十.从组织设计到人力资源规划 1.组织设计是人力资源规划的前提 2.人力资源规划的方法

一次成功的组织蜕变

一次成功的组织蜕变
赵春明
【期刊名称】《企业管理》
【年(卷),期】2007(000)006
【摘要】组织设计为企业发展所需的企业改革提供方案，科学且务实的组织设计对企业发展影响重大。

上世纪30年代，斯隆在通用公司采用事业部制的组织模式变革企业，是组织设计成功促进企业发展的典范；同样，不佳的组织设计也为企业带来灾难性的损失，较近例子中如实达和康佳两公司，2000年前后，因为采用了不正确的组织设计方案，导致了巨额的损失。

【总页数】8页(P62-69)
【作者】赵春明
【作者单位】上海复斯管理咨询公司
【正文语种】中文
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物业管理书籍名称

物业管理书籍名称,物业管理?拿来即用?案头手册(10条)覃朝晖编著/2010年03月/中国经济出版社规范的管理标准、流程与工作范本实用的工作表单工具化繁为简，拿来即用，高效务实,24.30,38.00 折扣:64折,物业管理(修订第二版)(0条)季如进主编/2008年01月/首都经济贸易大学出版社本书作者为清华大学房地产研究所副所长，对物业管理方面有多年的理论研究和实践经验。

因此，本书既阐述了物业管理的一些基本理论，同时又介绍了物业管理的具体方法，如如何签订物业服务委托合同、物业..,22.8035.00 折扣:65折 ,,实用小区物业管理表格o (10条)《实用小区物业管理表格》编写组编/2006年05月/机械工业出版社本书是一本物业管理的工具民收。

它以表格的形式对小区物业管理的各个环节进行了设计，具有实用性和创新性。

内容包括:招标投标与接管入信管理表格，公共事务管理表格，设备设施管理表格，消防、安全管...,21.00,28.00 折扣:75折create "safe and civilized model works fine in Zhuhai city"; Section II quality management system in order to strengthen the quality control of project construction site, according to the project management handbook (Second Edition), the quality management system manual and procedures specifies the control plan. Applies to all construction quality management in construction work of this project, this project in project management will fully implement the requirements of the quality management system standard. This quality management system is theproject of implementing quality assurance in mechanical and electrical installation engineering construction one of the basic requirements, relating to the construction of the project staff will ... Fruit Visual should is following standard: flush qualified standard equipment speed (r/min) filter network specifications (accounts number) qualifiedstandard ? 6000 200 Visual filter network, each square centimeters range within remaining of dirt real not than 3 star <6000 100 2, and water pipeline system disinfection: drinking water pipeline system in official delivered using Qian needed used each rose water in the containing 20~mg of free chlorine of water irrigation full pipeline for disinfection. Chlorine-containing water in the tube should be let stand for 24 hours or more. After disinfection, rinse with potable water piping, and sampling by the health and epidemic prevention departments after passing inspection, turn off all valves and closed all entrances, preventforeign bodies from entering. 5.4.2.1 drainage system debugging debugging: debugging sanitary appliances, drain pump debugging, debugging,物业管理风险防范与服务案例o (2条)邵小云，王高翔等编/2011年01月/化学工业出版社本书以物业管理者需要规避风险为出发点，依据最新的法律法规以案例形式介绍了物业公司与业主委员会沟通的案例，小区防范的案例，与业主纠纷的案例，停车场管理的案例，业主投诉案例等内容，可操作性较..,23.90,38.00 折扣:63折,物业管理员—职业培训计划培训大纲o (0条)中华人民共和国劳动和社会保障部培训就业司组织制定/2007年01 月/中国劳动社会保障出版社为进一步贯彻《民办教育促进法》，更好地规范职业培训机构的办学行为，提高职业培训质量，劳动和社会保障部组织有关专家编制了《物业管理员职业培训计划培训大纲》(以下简称《培训计划培训大纲》...,6.00,8.00 折扣:75折,物业设施设备管理与维修(第2版)(物业管理?物业设施管理专业通o (2条)刘薇，张喜明，孙萍主编/2010年06月/清华大学出版社本书共分十二章，紧密结合物业设施设备的范围、物业电气设施设备管理与维护维修工作内容、给排水设施设备管理与维护维修工作内容、通风设施设备与维护维修工作内容、供暖设施设备与维护维修工作内容、...,24.80,33.00 折扣:75折create "safe and civilized model works fine in Zhuhai city"; Section II quality management system in order to strengthen the quality control of project construction site, according to the project management handbook (Second Edition), the quality management system manual and procedures specifies the control plan. Applies to all construction quality management in construction work of this project, this project in project management will fully implement the requirements of the quality management system standard. This quality management system is theproject of implementing quality assurance in mechanical and electrical installation engineering construction one of the basic requirements, relating to the construction of the project staff will ... Fruit Visual should is following standard: flush qualified standard equipment speed (r/min) filter network specifications (accounts number) qualified standard ? 6000 200 Visual filter network, each square centimeters range within remaining of dirt real not than 3 star <6000 100 2, and water pipeline system disinfection: drinking water pipeline system in official delivered using Qian needed used each rose water in the containing 20~mg of free chlorine of water irrigation full pipeline for disinfection. Chlorine-containing water in the tube should be let stand for 24 hours or more. After disinfection, rinse with potable water piping, and sampling by the health and epidemic prevention departments after passing inspection, turn off all valves and closed all entrances, preventforeign bodies from entering. 5.4.2.1 drainage system debuggingdebugging: debugging sanitary appliances, drain pump debugging, debugging当当网图书,物业管理服务常见问题100例o (3条)王比刚，王寿华编著/2010年07月/中国建筑工业出版社本书根据《物权法》、《物业管理条例》，详细阐述了物业服务工作中常见的一些矛盾和纠纷，以及如何完善和做好物业管理服务工作。