种植年限对樱桃果园土壤微生物区系的影响毕业论文

分类号密级编号本科生毕业论文（设计）题目:种植年限对樱桃果园土壤微生物区系的影响学院:生物科学与化学学院专业：生物科学班级:09生科一班姓名:Xx学号：292010108指导教师:xx提交日期:2013年5月20日-13- 原创性声明本人郑重声明：所呈交的论文，是本人在指导教师的指导下独立进行研究所取得的成果。除文中已经注明引用的内容外，不包含任何其他个人或集体已经发表或撰写过的科研成果。本声明的法律责任由本人承担。论文作者签名：xx2013年5月21日论文指导教师签名：-13- 目录摘要4关键词41引言42材料与方法62.1试验材料62.2试验试剂及仪器……………………………………………………………….............62.2.1试验试剂62.2.2试验主要仪器设备62.3研究方法62.3.1研究区概况62.3.2土样的采集72.3.3土壤微生物的分离、统计72.3.3.1培养基配制72.3.3.2配制土壤悬液72.3.3.3土壤悬液接种72.3.3.4培养72.3.3.5数据统计................................................................................................83结果与分析3.1种植年限对樱桃果园土壤细菌数量的影响83.2种植年限对樱桃果园土壤放线菌数量的影响93.3种植年限对樱桃果园土壤真菌数量的影响94讨论10参考文献11致谢12-13- 种植年限对樱桃果园土壤微生物区系的影响（天水师范学院生命科学与化学学院甘肃天水741001）摘要：以时空互代法采集的天水市不同年限樱桃果园的土壤为试验材料，以8个不同树龄樱桃园的土壤为对象，研究了种植年限对樱桃果园土壤微生物区系（真菌、细菌、放线菌）的影响。结果表明，在4a～10a的种植年限樱桃土壤微生物数量，细菌＞放线菌＞真菌，细菌数量在土壤微生物中处于优势地位，次之为放线菌，真菌对土壤的作用强度和影响最低。结果还表明，随着年限的增加，微生物区系呈现动态变化。关键词：樱桃;土壤;真菌;细菌;放线菌OfplantinglifeofcherryorchardsoilmicrobialfloraAuthor:FanYaoInstructor:WangJing(TianshuiCollegeofLifeScienceandChemistry,Tianshui,Gansu741001)Summary:AcquisitiontimeandspaceonbehalfofeachmethodTianshuiCity,ofdiff-erentagescherryorchardsoilastestmaterialtoeightdifferentoldcherryorchardsoil,plantingyearsoncherryorchardsoilmicrobialflora(Fungi,Bacteria,Actinomycetesc).Theresultsshowedthat4ato10aofyearsofplantingcherrynumberofsoilmicrobes,Bacteria>Actinomycetes>Fungi,thenumberofbacteriainthesoilmicroorganismsinadominantposition,followedbyactinomycetes,fungiontheroleofsoilstrengthandminimalimpact.Theresultsalsoshowthatwiththeincreaseofyears,themicrobialfloraaredynamic.Keywords:Cherry;Soil;Fungi;Bacteria;Actinomycetesc1引言樱桃属于蔷薇科（Rosaceae）李属（PrunusL.）樱桃亚属（Cerasus.Pers.）落叶乔木果树。樱桃是上市较早的一种乔木果实，号称“百果第一枝”[1]。樱桃因其具有上市早、单位面积产值高、市场需求量大的优势,近年来成为我国发展最快的果树之一,各适栽地区均把甜樱桃列为果树生产的重要树种[2]。由于天水地处秦巴山区西秦岭北部黄土梁峁沟壑区，山地多，平地少易种植果树，自2000年以来，天水地区农民将大量农田改为樱桃园，使当地樱桃种植面积迅速增加，目前在天水市已种植0.43万hm2以上[3]-13- 。但是多年种植造成果园土壤质量下降问题已成为威胁樱桃生产的一个重要问题,部分樱桃园也面临更新，因此对不同种植年限樱桃果园土壤质量的研究尤为必要。土壤微生物是土壤生态系统的重要组成部分，在物质与能量循环、土壤结构、土壤的肥力和土壤营养元素的转化以及土壤微生态平衡保持等方面发挥着重要作用，而且对于进入土壤中的农药及其他有机污染物的自净、有毒金属及其化合物在土壤环境中的迁移转化等都起着极为重要的作用。有些土壤中的微生物对农业有害，如反硝化细菌，能把硝酸盐还原成氨散失到大气中，降低土壤肥力。但多数是对农业有益的，对农业生产和环境保护有着不可忽视的影响。土壤微生物与植物果树的生长关系特别密切，不同的土壤微生物由于其生理活性和代谢产物的不同，对土壤肥力和植物营养产生积极或消极的作用，因此，土壤微生物群落结构及土壤微生物量能够间接反映土壤的健康状况[4]。土壤微生物中细菌最多，作用强度和影响最大，放线菌和真菌类次之，藻类和原生动物等数量较少，影响也小[5]。下面分别介绍一下细菌、真菌和放线菌，细菌：土壤中细菌可占土壤微生物总量的70％～90％，它们数量大、个体小，与土壤接触的表面积特别大，是土壤中最大的生命活动面，也是土壤中最活跃的生物因素，推动着土壤中的各种物质循环。土壤细菌有许多不同的生理类群，如固氮细菌、氨化细菌，纤维分解细菌等在土壤中都有存在。细菌积极参与着有机物的分解、腐殖质的合成和各种矿质元素的转化。放线菌：土壤中放线菌的数量仅次于细菌，它们以分枝丝状营养体缠绕于有机物或土粒表面，并伸展于土壤孔隙中。1g土壤中的放线菌孢子可达105～107个，占土壤微生物总数的5％～30％。由于单个放线菌菌丝体的生物量较单个细菌大得多，因此尽管其数量上少些，但放线菌总生物量与细菌的总生物量相当。土壤中放线菌的种类十分繁多，其中主要是链霉菌。目前已知的放线菌种大多是分离自土壤。放线菌主要分布于耕作层中，随土壤深度增加而数量、种类减少[6]。真菌：真菌是土壤中第三大类微生物，广泛分布于土壤耕作层，1g土壤中可含103～104 个真菌。真菌中霉菌的菌丝体像放线菌一样，缠绕在有机物碎片和土粒表面，向四周伸展，蔓延于土壤孔隙中，并形成有性或无性孢子。土壤霉菌为好氧性微生物，一般分布于土壤表层，深层较少发育。且较耐酸，在pH=5.0左右的土壤中，由于细菌和放线菌的发育受到限制而土壤真菌在土壤微生物总量中占有较高的比例。-13- 由于樱桃是多年生植物，经栽培后即在原地生长多年，所以会造成果园土壤质量下降，以致樱桃产品质量也受到一定的影响。对樱桃土壤的问题国内外均有报告，如于翠[7]等对大青叶樱桃根际微生物种群结构及其变化动态的研究，分析了樱桃不同物候期根际微生物种群结构的变化，并得出落叶期根际细菌种群结构最丰富而萌芽期最少这一结论。吕德国[8]等对樱桃根际土壤的基本特征的研究，揭示了樱桃根系适应性差，并为栽植几年后根际土壤易发生恶化状态的生理生化机制提供了参考依据。再如周晓康[9]对樱桃土壤有效化学元素硼的含量测定。张弢[10]对不同种植年限樱桃树根际土壤酶（过氧化氢酶、脲酶、蔗糖酶）活性的研究，反映了不同种植年限樱桃土壤的生态环境。就目前来看国内外针对不同种植年限蔬菜大棚及玉米，稻子等农作物土壤状况研究较多，对不同种植年限樱桃果园土壤状况的系统研究报道很少[11～13]。本文将对不同种植年限樱桃果园土壤微生物区系进行研究,分析随着种植年限的增加樱桃果园土壤微生物（主要指真菌、细菌、放线菌）区系的变化情况，旨在为进一步研究樱桃果园土壤管理提供科学依据，以及对提高樱桃产量品质提供理论基础。2材料与方法2.1试验材料以不同年限的樱桃果园土壤为试验材料，试样来自天水市樱桃果园。2.2试验试剂及仪器2.2.1试验试剂牛肉膏，蛋白胨，可溶性淀粉，孟加拉红，KNO3，NaCl，K2HPO4，MgSO4，FeSO4·7H2O,KH2PO4,MgSO4·7H2O，NaOH，琼脂，蒸馏水等。2.2.2试验主要仪器设备电子称，振荡仪，灭菌锅，移液抢，涂布棒，接菌台等。2.3研究方法2.3.1研究区概况天水市位于甘肃省东南部，地处秦巴山区西秦岭北部黄土梁峁沟壑区。属大陆性半高寒半湿润气候，年平均降水量530mm，年均气温10.72℃，无霜期170d左右。自2000年以来，天水地区农民将大量农田改为樱桃园，使当地樱桃种植面积迅速增加，肥料投入量大，远超过高产农田，这为本研究提供了可靠的研究对象。-13- 2.3.2土样的采集采用时空互代的方法(在一定的时间尺度内，以不同地点上选取不同种植年限的樱桃果园来代替同一地点上生长的不同年限的樱桃果园)，在天水市选择不同年限的樱桃果园，每个果园选择3株具有代表性的果树作为采样对象，在每株果树周围距树干120cm处，避开施肥点用土钻取土，取样深度40cm，每株果树周围取3钻，4℃保存。2.3.3土壤微生物分离、统计2.3.3.1培养基配制细菌采用牛肉膏蛋白胨培养基（牛肉膏3g，蛋白胨5g，NaCll5g，琼脂15g～18g，蒸馏水1000ml，用浓度为1mol/L的HCl或NaOH调pH至7.0～7.2，121℃高压灭菌30min）；真菌采用马丁氏培养基(孟加拉红，琼脂15g，蒸馏水1000ml，pH自然，121℃高压灭菌30min）；放线菌采用高氏一号培养基（可溶性淀粉10g，KCl0.5g，K2HPO41g，MgSO4·7H2O0.5g，FeSO4·7H2O0.01g，(NH4)2SO42g，琼脂5g，蒸馏水1000ml，pH7.6～7.8，121℃灭菌30min）[14、15]。2.3.3.2配制土壤悬液2.3.3.2.1称取10g鲜土样加入盛有90ml无菌水的三角瓶中，用振荡仪剧烈振荡15min，使土样均匀分散在稀释液中成为土壤悬浮液。2.3.3.2.2将上述土壤悬浮液静置10min后，用无菌移液枪吸取1ml土壤悬液到盛有9ml无菌水的试管中，依次按10倍法稀释到10–4。2.3.3.3土壤悬液接种根据待分离的微生物类型，分别吸取不同稀释倍数的土壤悬浊液(细菌10-3、10-4，放线菌10-3、10-4，真菌10-1、10-2)100ul，加入至预先制备好的平板培养基，用无菌涂布棒将土壤悬浊液均匀涂布在培养基上，每种处理设3次重复。2.3.3.4培养将培养基置于30℃的培养箱中进行培养，细菌培养2～3天，放线菌培养5～7天，真菌培养3～5天。2.3.3.5数据统计根据不同培养基中生长出菌落数统计真菌、细菌和放线菌数量，-13- 结合土壤样品的稀释倍数，按照公式：土壤微生物浓度(cfu/g土)=菌落平均数×稀释倍数，求得土壤中可培养真菌、细菌和放线菌总数[16]。3结果与分析土壤细菌、真菌和放线菌构成了土壤微生物的主要量，它们的组成和数量变化通常能反映土壤生物活性水平，显示土壤中物质代谢的旺盛程度，是土壤物理化学特性的综合体现，也是土壤肥力的一个重要指标[17]。有表1可知，在4a～10a的种植年限樱桃土壤微生物数量，细菌＞放线菌＞真菌，其中细菌占总数的79.41％～52.43％。由此可见，细菌数量在土壤微生物中处于优势地位，然而种植12a放线菌数量比细菌数量多52.2％，这可能由于在种植12a的果园土壤偏碱性且有机物含量丰富有利于放线菌的生长。总体来看微生物数量，尤其是细菌数量的多少在某种程度上可以反映土壤质量的变化。表1不同种植年限樱桃果园土壤微生物数量的变化Table1DifferentPlantingYearscherryorchardsoilmicrobialamount（×103cfu/g土）种植年限/a细菌放线菌真菌微生物总数4156400.44196.44585480.21133.2163122520.51564.517118880.31206.31865590.22124.2292671060.32373.32102161200.68336.68121592420.38401.383.1种植年限对樱桃果园土壤中细菌数量的影响由图1可知，樱桃果园土壤中细菌数量随种植年限的增长呈双抛物线的变化趋势。在种植年限为4a到6a期间，土壤中细菌总数开始增加，平均每年增加222％，种植6a时达到峰值,细菌数量为3.12×105（cfu/g土）；种植6a～8a间，随种植年限的增加，土壤中细菌数量呈减少的变化趋势，平均每年减少56％；种植年限为9a时，细菌总数再次升高，细菌总数比种植8a的增加311%；在种植年限9a～12a的樱桃果园土壤中细菌总数随种植年限的增加而又呈现下降趋势，每年平均下降23.1％。-13- 图1不同种植年限樱桃果园土壤中细菌总数的变化Figure1DifferentPlantingYearscherryorchardsoilinthetotalnumberofBacteria3.2种植年限对樱桃果园土壤中放线菌数量的影响由图2可见，4a～12a种植年限之间放线菌数量呈先增加后降低，之后再增加的变化趋势。种植年限在4a～6a期间，土壤中放线菌呈增加的变化趋势，平均每年增加223％，种植6a时放线菌数量增加到2.52×105（cfu/g土），达到峰值；6a～8a的樱桃果园土壤细菌数随种植年限的增加而逐渐减少,平均每年减少49％；种植8a～12a之间,随着种植年限的增加，放线菌数量以平均每年增加65％的趋势逐渐上升，种植12a再次达到峰值，放线菌数量为2.42×105（cfu/g土）。图2不同种植年限樱桃果园土壤中放线菌总数的变化Figure2DifferentPlantingYearscherryorchardsoilinthetotalnumberofActinomycetes3.3种植年限对樱桃果园土壤中真菌数量的影响由图3可知，樱桃果园土壤中真菌数量随种植年限的增长也呈现双抛物线的变化趋势。种植-13- 5a的真菌数量比种植4a的真菌数量减少52.3％；5a到6a土壤真菌总数增加143％；种植年限在6a～8a期间，随种植年限的增加，樱桃果园土壤真菌数逐渐减少，每年平均减少33.8％；8a～10a树龄的樱桃果园土壤中真菌数量随种植年限的增加，以每年平均增加69.7％的趋势上升，种植10a达到峰值，真菌数量为2.42×105（cfu/g土）；种植12a时真菌数量又下降到了0.38×103（cfu/g土）。图3不同种植年限樱桃果园土壤中真菌总数的变化Figure3DifferentPlantingYearscherryorchardsoilinthetotalnumberofFungi4讨论本研究根据对不同种植年限樱桃果园土壤中不同类型微生物的统计及分析结果发现，在0～10ａ种植年限内，樱桃土壤的微生物数量分布为：细菌＞放线菌＞真菌，且细菌是优势类群。一般认为，细菌数量较多是土壤肥力较高的一种表现，对樱桃根系造成危害的主要是一些致病的真菌。而土壤中Bacillusspp.和Pseudomonasspp.等细菌类群对土壤致病真菌有抑制作用[18]。本研究表明，种植年限为6a时土壤中细菌数量和放线菌数量均达到峰值，其中细菌占总数的55.3％，放线菌占总数的44.6,％，真菌数量却只占到0.09％是4～12a的最小比例。真菌的峰值年限为10a，细菌的数量较少。因此可以推测，6a树龄的樱桃果园土壤微生物群落是较适于樱桃生长的系统。12a的樱桃树龄放线菌数量占总数60.3％，细菌却只占到39.6％，微生物区系发生变化。土壤管理、种植模式改变以及果树生长年限，都会引起土壤微生物生态系统的改变，进而导致整个果园土壤生态系统的改变。因此，在某种程度上土壤微生物区系的变化可以作为衡量土壤质量和土壤肥力水平高低的一项指标。-13- 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致谢在做实验和论文的写作过程中遇到了无数的困难和障碍，都在同学和指导老师的帮助下度过了。尤其要特别感谢我的论文指导老师—王静老师，她对我进行了无私的指导和帮助，王老师严谨的治学态度和忘我的工作精神使我受益非浅：不仅使我的专业能力得到了很大的提高，而且要特别感谢王老师在做人和做学问方面的真诚教诲。在此我向尊敬的王老师致以最衷心的感谢，谢谢！感谢这篇论文所涉及到的各位学者，本文引用了数位学者的研究文献，如果没有各位学者的研究成果的帮助和启发，我将很难完成本篇论文的写作。感谢我的实验小组和朋友，在我写论文的过程中提供的热情帮助。由于我的学术水平有限，所写论文难免有不足之处，恳请各位老师批评和指正！2013年5月21日9JWKffwvG#tYM*Jg&6a*CZ7H$dq8KqqfHVZFedswSyXTy#&QA9wkxFyeQ^!djs#XuyUP2kNXpR89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^G89AmUE9aQ@Gn8xp$R#͑Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z8vG#tYM*Jg&6a*CZ7H$dq8KqqfHVZFedswSyXTy#&QA9wkxFyeQ^!djs#XuyUP2kNXpRWXmA&UE9aQ@Gn8xp$R#͑Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmUE9aQ@Gn8xp$R#͑Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNuGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$U*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89Amv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$U*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz84!z89Amv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$U*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$U*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNuGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^GjqvadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5ux^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wEwZ#Qc@UE%&qYp@Eh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$U*3tnGK8!z89AmYWpazadNu##KN&MuWFA5uxY7JnD6YWRrWwc^vR9CpbK!zn%Mz849Gx^Gjqv^$UE9wE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