Ecological functions of uncultured microorganisms in the cobaltrich ferromanganese crust of a seamount in the central Pacific are elucidated by fosmid sequencing

HUO Yingyi CHENG Hong Anton F. Post WANG Chunsheng JIANG Xiawei PAN Jie WU Min XU Xuewei

霍颖异, 程虹, AntonF.Post, 王春生, 江夏薇, 潘杰, 吴敏, 许学伟. 基于宏基因组测序研究中太平洋海山富钴结壳区非培养微生物生态功能[J]. 海洋学报英文版, 2015, 34(4): 92-113. doi: 10.1007/s13131-015-0650-7
引用本文: 霍颖异, 程虹, AntonF.Post, 王春生, 江夏薇, 潘杰, 吴敏, 许学伟. 基于宏基因组测序研究中太平洋海山富钴结壳区非培养微生物生态功能[J]. 海洋学报英文版, 2015, 34(4): 92-113. doi: 10.1007/s13131-015-0650-7
HUO Yingyi, CHENG Hong, Anton F. Post, WANG Chunsheng, JIANG Xiawei, PAN Jie, WU Min, XU Xuewei. Ecological functions of uncultured microorganisms in the cobaltrich ferromanganese crust of a seamount in the central Pacific are elucidated by fosmid sequencing[J]. Acta Oceanologica Sinica, 2015, 34(4): 92-113. doi: 10.1007/s13131-015-0650-7
Citation: HUO Yingyi, CHENG Hong, Anton F. Post, WANG Chunsheng, JIANG Xiawei, PAN Jie, WU Min, XU Xuewei. Ecological functions of uncultured microorganisms in the cobaltrich ferromanganese crust of a seamount in the central Pacific are elucidated by fosmid sequencing[J]. Acta Oceanologica Sinica, 2015, 34(4): 92-113. doi: 10.1007/s13131-015-0650-7

基于宏基因组测序研究中太平洋海山富钴结壳区非培养微生物生态功能

doi: 10.1007/s13131-015-0650-7

Ecological functions of uncultured microorganisms in the cobaltrich ferromanganese crust of a seamount in the central Pacific are elucidated by fosmid sequencing

  • 摘要: 富钴结壳是一种重要的海底矿产资源.富钴结壳是生物地球化学和微生物学的研究热点,但目前我们对富钴结壳区的微生物群落结构仍缺乏认识.本研究针对中太平洋海山富钴结壳区沉积物,采用非培养方法获得微生物基因组信息并分析其生态学功能.采用PCR的方法筛选得到78个带有16S rRNA基因的克隆.变形杆菌门和奇古菌门MGI类群分别为古菌和细菌的主要类群.测序并分析了9个带有16S rRNA 基因的fosmid克隆.这些克隆中含有较多的代谢相关和重金属抗性相关基因,表明多样的代谢和抗逆途径在微生物适应富钴结壳环境中发挥了重要作用.代谢途径分析表明,氮循环是富钴结壳区一种重要的生物地球化学过程.此外,发现多个水平基因转移事件,这些基因大部分转移自细菌,少量转移自古菌或植物.比较基因组分析表明,属于奇古菌门MGI类群的克隆W4-93包含一段共线性的基因簇.以上结果说明水平基因转移和基因组分歧进化在微生物适应深海环境过程中发挥了重要作用.
  • Adachi O, Ano Y, Toyama H, et al. 2007. Biooxidation with PQQ- and FAD-dependent dehydrogenases. In: Schmid R D, Urlacher V B, eds. Modern Biooxidation: Enzymes, Reactions and Applications. Hoboken, NJ: John Wiley & Sons, Inc, 41
    Béjà O, Koonin E V, Aravind L, et al. 2002. Comparative genomic analysis of archaeal genotypic variants in a single population and in two different oceanic provinces. Applied and Environmental Microbiology, 68(1): 335-345
    Beman J M, Popp B N, Alford S E. 2012. Quantification of ammonia oxidation rates and ammonia-oxidizing archaea and bacteria at high resolution in the Gulf of California and eastern tropical North Pacific Ocean. Limnology and Oceanography, 57(3): 711-726
    Blainey P C, Mosier A C, Potanina A, et al. 2011. Genome of a low-salinity ammonia-oxidizing archaeon determined by single-cell and metagenomic analysis. PLoS One, 6(2): e16626
    Candela T, Fouet A. 2006. Poly-gamma-glutamate in bacteria. Molecular Microbiology, 60(5): 1091-1098
    Clark M R, Rowden A A, Schlacher T, et al. 2010. The ecology of seamounts: structure, function, and human impacts. Ann Rev Mar Sci, 2: 253-278
    Craig J D, Andrews J E, Meylan M A. 1982. Ferromanganese deposits in the Hawaiian Archipelago. Marine Geology, 45(1-2): 127-157
    Delaney M L. 1998. Phosphorus accumulation in marine sediments and the oceanic phosphorus cycle. Global Biogeochemical Cycles, 12(4): 563-572
    Dell'Anno A, Danovaro R. 2005. Extracellular DNA plays a key role in deep-sea ecosystem functioning. Science, 309(5744): 2179
    DeLong E F. 1992. Archaea in coastal marine environments. Proceedings of the National Academy of Sciences of the United States of America, 89(12): 5685-5689
    Duffy E J. 2008. "Seamount". CenSeam: a global census of marine life on seamounts content partner and national oceanic and atmospheric administration content source. In: Cleveland C J, ed. Encycleopedia of Earth. Washington DC: Environmental Information Coalition, National Council for Science and the Environment
    Ehrhardt C J, Haymon R M, Lamontagne M G, et al. 2007. Evidence for hydrothermal Archaea within the basaltic flanks of the East Pacific Rise. Environmental Microbiology, 9(4): 900-912
    Emerson D, Moyer C L. 2010. Microbiology of seamounts: common patterns observed in community structure. Oceanography, 23: 148-163
    Finkel S E. 2006. Long-term survival during stationary phase: evolution and the GASP phenotype. Nat Rev Microbiol, 4(2): 113-120
    Finkel S E, Kolter R. 2001. DNA as a nutrient: novel role for bacterial competence gene homologs. Journal of Bacteriology, 183(21): 6288-6293
    Fu Yazhou, Peng Jiantang, Qu Wenjun, et al. 2005. Os isotopic compositions of a cobalt-rich ferromanganese crust profile in Central Pacific. Chin Sci Bull, 50(18): 2106-2112
    Gadd G M. 2007. Geomycology: biogeochemical transformations of rocks, minerals, metals and radionuclides by fungi, bioweathering and bioremediation. Mycological Research, 111(Pt 1): 3-49
    García-Martínez J, Martínez-Murcia A, Antón A I, et al. 1996. Comparison of the small 16S to 23S intergenic spacer region (ISR) of the rRNA operons of some Escherichia coli strains of the ECOR collection and E. coli K-12. Journal of Bacteriology, 178(21): 6374-6377
    Ghiorse W C. 1984. Biology of iron- and manganese-depositing bacteria. Annu Rev Microbiol, 38: 515-550
    Gillan D C, Danis B. 2007. The archaebacterial communities in Antarctic bathypelagic sediments. Deep-Sea Research Part II: Topical Studies in Oceanography, 54(16-17): 1682-1690
    Guibaud G, van Hullebusch E, Bordas F, et al. 2009. Sorption of Cd(II) and Pb(II) by exopolymeric substances (EPS) extracted from activated sludges and pure bacterial strains: Modeling of the metal/ligand ratio effect and role of the mineral fraction. Bioresource Technology, 100(12): 2959-2968
    Heijs S K, Haese R R, van der Wielen P W, et al. 2007. Use of 16S rRNA gene based clone libraries to assess microbial communities potentially involved in anaerobic methane oxidation in a Mediterranean cold seep. Microbial Ecology, 53(3): 384-398
    Hillier J K, Watts A B. 2007. Global distribution of seamounts from ship-track bathymetry data. Geophysical Research Letters, 34(13): L13304
    Ito M, Tsunekawa M, Yamaguchi E, et al. 2008. Estimation of degree of liberation in a coarse crushed product of cobalt-rich甠獦?獲畲扯瑭楡汮楧獡??捳汥漠湣楲湵杳?愯湮摯?獵敬煥畳攠湡据敤???潳甠牧湲慡汶?潴晹??敥湰敡牲慡汴??楮挮爠潉扮楴潥汲潮条祴??????????????????扩牮????㈠???獣敥歳慳湩湮?听???渨椳琭琴攩氺???‰中愱搰愵氼楢杲 ̄呉??敲琠?愠汄???づと????椠癃攬爠獄楡瑳礠?愬渠摥?愠扡畬渮搠愲渰挱攲?漠晓?慡敭牯潵扮楴捳—慣湨摡?慡湣慴敥牲潩扳楴捩?浳攬琠桦慯湲敭?潴硩楯摮椬稠敭物獮?慲瑡?琠桤敥??慳慩歴潳渠??潤猠扢祩??畩摶?噲潳汩捴慹渮漠???慬牯敧湩瑣獡?十散慴???瀱瀰氨椳攩携?愲渹搵??渰瘸椼牢潲渾浊敩湡瑮慧氠??楡捷牥潩戬椠潘汵漠杘祵?????ㄠえ?????????????戠牡??愠爲琰??金???湥??畩慦摩牣慡摴潩?????????????灴敥穲??慡牴捩??㈠???慮?偶???汥扳慴??????攠瑦?慯汭??㈠つづ????敥瑡愠杳敥湤潩浭楥据獴?潭晥?瑡桧敥?摯敭敥瀮??敲摣楨瑩敶牥牳愠湯敦愠湍??慲?睢慩牯浬?执慹琬栠礱瀹攴氨愳朩椺挠′栰愷戭椲琱愴琼??倾?漦匣′伴渶攻????????攠?????扡牲??慁爠瑅椬渠??甠慬摡爠慔摯潲??????刬漠摥牴椠条畬攮稠?嘰愰氵攮爠慉?????潩牯敮椠牯慦????敡瑵?慯汴???とど???業湭摯獮楩条栭瑯?楩湤?瑺桩敮?爠敭污慲瑩楮癥攠?慲扣畨湡摥慯湮挮攠??浴敵瑲慥戬漠水椳挷?瀷漰琵攸温琺椠愵水″愭渵搴?朼敢湲漾测敡?摥票湩慳浡椠捍猬?潇景?畯渠捓甮氠琲椰瘰愰琮攠摋?浇慇爺椠湫敹?慴牯挠桥慮散慹?晬牯潰浥?捩潡洠灯慦爠慧瑥楮癥敳?浡敮瑤愠杧敥湮潯浭楥捳?愠湎慵汣祬獥敩獣?潁晣?扤慳琠桒祥灳敥污慲杣楨挬?瀲永愨渱欩琺漠渲?漭昳‰搼楢晲显敋牡整湯琠?漬挠敋慯湢楡捹?牳敨杩椠潃測猠??呫桥敧??卡????潥畴爠湡慬氮?′日?????????????戠牣??敭湵慮物摴???圠??ㄠ??????慩牬楩湣敡??敩潣汨漠杭祩?潲景?瑩桡敬?偭慡捴楳映楡捴??乥敥睰?女潥牡欠???捲??牨慥睲??楬氠汦?扥牬??椠牯潦猠桴湨楥挠桓敯湵歴潨?????楡灲灩敡????卲瑯慵捧歨攮戠牅慮湶摩瑲????敮瑴?慬氠???ひは????獯潧汹愬琠椱漱渨?愩渺搠′挰根愴爭愲挱琱攱爼楢穲愾瑋楡潴湯?潓昬?呙桡敮牡浧潡捷潡挠捋甬猠?獵楮扡業牵楲捡甠獍?猠灥??湡潬瘮?′昰爰漹洮?慁?坵敮獤瑡敮牣湥?卯楦戠敚牥楴慡?桲楯杴桥?瑢敡浣灴敥牲慩瑡甠牷敩?潨楩汮?牣敲獵敳牴癡潬椠牦???硤瑳爠敩浮漠灢桡楣汫攭獡??????????????扬爠??略楬??㈠???潴獨?匠?????敥楲湮???割???牡愠湔歲????攮琠?慮汶???の?????敬攠灍?獣敲慯??敯??湧?挬爠由猱琨猱′昩爺漠洳′琱栰攭″串漲爲琼桢敲愾獋瑯??瑨汩慮湳瑫楹挠?伬挠效慥湩??捊漠浒瀮漠猲椰琰椳漮渠?慰湴摡?牥攠獯潦甠牥捬敥?捥潮湴獳椠摦敲牯慭琠楳潥湡獷???慲爠楢湹攠??敲潲牯敭獡潮畧牡据敥獳????敵潳瑴敳挺栠湳潯汬潩杤礭?????ㄠ????っ??ぴ?扯牮?丠楡敮獤?????????㈠??剥散獩楡獴瑩慯湮挮攠?瑡潲?据慥搠浇楥畯浬??捹漬戠愱氹琸?″稭椴温挺?″愳渱搭″渵椱挼止敲氾?楩渠?浥楮捧爬漠扇敵猠??偤汯慮獧洮椠搲??社????????????扳牴?乵潣牴瑵潲湥???????污潮瑳穣??????卥瑳数楯湮???夠??攠瑡?慡汭????ぢ????潲浩灡氬攠瑁敏?本攠湡潮浤攠?獏敂焠畩敮渠捭敡?潧晲?乶楥琠牳潥獤潩獭灥楮牴愠?浩畣汲瑯楣景潳牭浳椠獡??慮湤?慤洠海潩湴楨愠?潭硭楯摮楩穵業渠条?扤愠据瑩整牲楩畴浥?映牁潰浰?瑩桥敤?獍潩楣汲?敢湩癯楬牯潧湹洠敡湮瑤???灯灴汥楣敨摮?慬湯摧??渠瘹椷爨漲渲洩攺渠琹愸氵??椹挸爷漴戼楢潲氾潌杩礠??????????????????㈠?扨牡?剧漠睘摩敡湯???????漠睡敬爮??‰???匠捖桥汲慴捩档敡牬?呤????敩瑢?慴汩??㈠は????偣慴牥慲摩楡杬洠獡?楤渠?獲散慨浡潥畡湬琠?敯捭潭汵潮杩祴??晳愠捡瑬??晧椠捤瑩楳潣湲?慴湥搠?晡畹瑥畲牳攠???慡爠楤湥敥??捳潥污漠杣祯?????卤畩灭灥?????????ㄠ?扴爠?卨慥渠瑅敡汳汴椠??????佣爠捒畩瑳瑥???丱?…?愱渷渶椻湎朩????整瑲?慭汯???ぬづ????戲用渴搩愺渠挵攷″愭渵搸‵搼楢癲放牌獩楡瑯礠?漬映?浵椠捘爠潗戬椠慊汩?汮楧映敘?楗測?潥捴攠慡湬?挠爲田猱琱??乍慩瑣畲牯敢?????????????????????戭牳?卡愠湳瑥敤汩汭楥??????坭攠扴扨?匠?????潴栭湲慩汣歨漠癣慲??????数瑯?慩汴???で?????楮瘠整牨獥椠瑐祡?潩晦??渠?潣硥楡摮攮猠?灅牍潓搠畍捩散摲?扢祩??湯?????潯硬楯摧楹稬椠渷朸?昳甩渺朠椵?‵?攵漸挵格楢浲椾捌慯?敥瑺??潩獡浺漠捉栬椠浃楬捡慲??挠瑓愬??????ぬ??????金?㈱????戠牳?卯捉桉慄甠敏牰?割???楯敦渠桂潡汣摩????刀愀洀攀琀琀攀????攀琀?愀氀???  ????愀挀琀攀爀椀愀氀?搀椀瘀攀爀猀椀琀礀?愀渀搀?戀椀漀最攀漀最爀愀瀀栀礀?椀渀?搀攀攀瀀?猀攀愀?猀甀爀昀愀挀攀?猀攀搀椀洀攀渀琀猀?漀昀?琀栀攀?匀漀甀琀栀??琀氀愀渀琀椀挀?伀挀攀愀渀??吀栀攀??匀????漀甀爀渀愀氀?????????????? ?戀爀?匀挀栀攀渀愀甀?匀?????攀??愀渀最攀???????  ???倀栀漀猀瀀栀漀爀甀猀?爀攀最攀渀攀爀愀琀椀漀渀?瘀猀??戀甀爀椀愀氀?椀渀?猀攀搀椀洀攀渀琀猀?漀昀?琀栀攀??爀愀戀椀愀渀?匀攀愀???愀爀椀渀攀??栀攀洀椀猀琀爀礀?????????? ??????戀爀?匀挀栀氀愀挀栀攀爀?吀????刀漀眀搀攀渀???????漀眀攀爀??????攀琀?愀氀??? ? ??匀攀愀洀漀甀渀琀?猀挀椀攀渀挀攀?猀挀愀氀攀猀?甀渀搀攀爀猀攀愀?洀漀甀渀琀愀椀渀猀??渀攀眀?爀攀猀攀愀爀挀栀?愀渀搀?漀甀琀氀漀漀欀???愀爀椀渀攀??挀漀氀漀最礀?????匀甀瀀瀀????????戀爀?匀琀愀甀搀椀最攀氀?????漀瀀瀀攀爀猀?????倀??倀氀愀渀欀?吀????攀琀?愀氀??? ? ??匀攀愀洀漀甀渀琀猀?椀渀?琀栀攀?猀甀戀搀甀挀琀椀漀渀?昀愀挀琀漀爀礀??伀挀攀愀渀漀最爀愀瀀栀礀?????????????????戀爀?吀愀洀甀爀愀????倀攀琀攀爀猀漀渀????倀攀琀攀爀猀漀渀?一??攀琀?愀氀??? ???????????洀漀氀攀挀甀氀愀爀?攀瘀漀氀甀琀椀漀渀愀爀礀?最攀渀攀琀椀挀猀?愀渀愀氀礀猀椀猀?甀猀椀渀最?洀愀砀椀洀甀洀?氀椀欀攀氀椀栀漀漀搀??攀瘀漀氀甀琀椀漀渀愀爀礀?搀椀猀琀愀渀挀攀??愀渀搀?洀愀砀椀洀甀洀?瀀愀爀猀椀洀漀渀礀?洀攀琀栀漀搀猀???漀氀攀挀甀氀愀爀??椀漀氀漀最礀?愀渀搀??瘀漀氀甀琀椀漀渀?????? ?????????????戀爀?吀愀琀甀猀漀瘀?刀?????愀氀瀀攀爀椀渀???夀??一愀琀愀氀攀??????攀琀?愀氀???   ??吀栀攀??伀??搀愀琀愀戀愀猀攀??愀?琀漀漀氀?昀漀爀?最攀渀漀洀攀?猀挀愀氀攀?愀渀愀氀礀猀椀猀?漀昀?瀀爀漀琀攀椀渀?昀甀渀挀琀椀漀渀猀?愀渀搀?攀瘀漀氀甀琀椀漀渀??一甀挀氀攀椀挀??挀椀搀猀?刀攀猀攀愀爀挀栀???????????????戀爀?吀攀戀漀???????愀爀最愀爀???刀???氀攀洀攀渀琀??????攀琀?愀氀???  ????椀漀最攀渀椀挀?洀愀渀最愀渀攀猀攀?漀砀椀搀攀猀??瀀爀漀瀀攀爀琀椀攀猀?愀渀搀?洀攀挀栀愀渀椀猀洀猀?漀昀?昀漀爀洀愀琀椀漀渀???渀渀甀愀氀?刀攀瘀椀攀眀?漀昀??愀爀琀栀?愀渀搀?倀氀愀渀攀琀愀爀礀?匀挀椀攀渀挀攀猀??????????????戀爀?嘀攀爀氀愀愀渀?倀???????????攀渀琀栀椀挀?爀攀挀爀甀椀琀洀攀渀琀?愀渀搀?洀愀渀最愀渀攀猀攀?挀爀甀猀琀?昀漀爀洀愀琀椀漀渀?漀渀?猀攀愀洀漀甀渀琀猀???愀爀椀渀攀??椀漀氀漀最礀??????????????????戀爀?圀愀氀欀攀爀??????搀攀?氀愀?吀漀爀爀攀???刀???氀漀琀稀??????攀琀?愀氀??? ? ??一椀琀爀漀猀漀瀀甀洀椀氀甀猀?洀愀爀椀琀椀洀甀猀?最攀渀漀洀攀?爀攀瘀攀愀氀猀?甀渀椀焀甀攀?洀攀挀栀愀渀椀猀洀猀?昀漀爀?渀椀琀爀椀昀椀挀愀琀椀漀渀?愀渀搀?愀甀琀漀琀爀漀瀀栀礀?椀渀?最氀漀戀愀氀氀礀?搀椀猀琀爀椀戀甀琀攀搀?洀愀爀椀渀攀?挀爀攀渀愀爀挀栀愀攀愀?倀爀漀挀攀攀搀椀渀最猀?漀昀?琀栀攀?一愀琀椀漀渀愀氀??挀愀搀攀洀礀?漀昀?匀挀椀攀渀挀攀猀?漀昀?琀栀攀?唀渀椀琀攀搀?匀琀愀琀攀猀?漀昀??洀攀爀椀挀愀??? ?????????????????戀爀?圀愀渀最?堀椀愀漀栀漀渀最????氀氀攀爀?圀???????  ????愀爀椀渀攀?戀椀漀洀椀渀攀爀愀氀猀??瀀攀爀猀瀀攀挀琀椀瘀攀猀?愀渀搀?挀栀愀氀氀攀渀最攀猀?昀漀爀?瀀漀氀礀洀攀琀愀氀氀椀挀?渀漀搀甀氀攀猀?愀渀搀?挀爀甀猀琀猀??吀爀攀渀搀猀?椀渀??椀漀琀攀挀栀渀漀氀漀最礀?????????????????戀爀?圀愀渀最?堀椀愀漀栀漀渀最??匀挀栀氀漀猀猀洀愀挀栀攀爀?唀??一愀琀愀氀椀漀????攀琀?愀氀???  ????瘀椀搀攀渀挀攀?昀漀爀?戀椀漀最攀渀椀挀?瀀爀漀挀攀猀猀攀猀?搀甀爀椀渀最?昀漀爀洀愀琀椀漀渀?漀昀?昀攀爀爀漀洀愀渀最愀渀攀猀攀?挀爀甀猀琀猀?昀爀漀洀?琀栀攀?倀愀挀椀昀椀挀?伀挀攀愀渀??椀洀瀀氀椀挀愀琀椀漀渀猀?漀昀?戀椀漀氀漀最椀挀愀氀氀礀?椀渀搀甀挀攀搀?洀椀渀攀爀愀氀椀稀愀琀椀漀渀???椀挀爀漀渀??? ???????????????戀爀?圀愀渀最?堀椀愀漀栀漀渀最??圀椀攀渀猀????匀挀栀爀??????搀攀爀????攀琀?愀氀??? ?????漀氀攀挀甀氀愀爀?戀椀漀洀椀渀攀爀愀氀椀稀愀琀椀漀渀??琀漀眀愀爀搀?愀渀?甀渀搀攀爀猀琀愀渀搀椀渀最?漀昀?琀栀攀?戀椀漀最攀渀椀挀?漀爀椀最椀渀?漀昀?瀀漀氀礀洀攀琀愀氀氀椀挀?渀漀搀甀氀攀猀??猀攀愀洀漀甀渀琀?挀爀甀猀琀猀??愀渀搀?栀礀搀爀漀琀栀攀爀洀愀氀?瘀攀渀琀猀???渀????氀氀攀爀?圀??????攀搀???漀氀攀挀甀氀愀爀??椀漀洀椀渀攀爀愀氀椀稀愀琀椀漀渀???攀爀氀椀渀??攀椀搀攀氀戀攀爀最??匀瀀爀椀渀最攀爀??????? ?戀爀?圀攀搀攀瀀漀栀氀????????????吀栀攀?挀漀洀瀀漀猀椀琀椀漀渀?漀昀?琀栀攀?挀漀渀琀椀渀攀渀琀愀氀?挀爀甀猀琀???攀漀挀栀椀洀椀挀愀?攀琀??漀猀洀漀挀栀椀洀椀挀愀??挀琀愀???????????????????戀爀?圀攀猀猀攀氀?倀??匀愀渀搀眀攀氀氀???吀???椀洀?匀?匀??? ? ??吀栀攀?最氀漀戀愀氀?猀攀愀洀漀甀渀琀?挀攀渀猀甀猀??伀挀攀愀渀漀最爀愀瀀栀礀???????????????戀爀?圀爀椀最栀琀???????漀渀眀愀爀???????愀氀氀愀洀?匀????? ?????椀挀爀漀戀椀愀氀?攀挀漀氀漀最礀?漀昀?攀砀瀀愀渀搀椀渀最?漀砀礀最攀渀?洀椀渀椀洀甀洀?稀漀渀攀猀??一愀琀?刀攀瘀??椀挀爀漀戀椀漀氀??? ?????????????戀爀?圀甀?夀甀攀栀漀渀最???椀愀漀??椀??圀愀渀最??栀甀渀猀栀攀渀最??攀琀?愀氀??? ??????挀漀洀瀀愀爀椀猀漀渀?漀昀?洀椀挀爀漀戀椀愀氀?挀漀洀洀甀渀椀琀椀攀猀?椀渀?搀攀攀瀀?猀攀愀?瀀漀氀礀洀攀琀愀氀氀椀挀?渀漀搀甀氀攀猀?愀渀搀?琀栀攀?猀甀爀爀漀甀渀搀椀渀最?猀攀搀椀洀攀渀琀猀?椀渀?琀栀攀?倀愀挀椀昀椀挀?伀挀攀愀渀???攀攀瀀?匀攀愀?刀攀猀攀愀爀挀栀?倀愀爀琀????伀挀攀愀渀漀最爀愀瀀栀椀挀?刀攀猀攀愀爀挀栀?倀愀瀀攀爀猀??????? ????戀爀?堀甀??攀椀砀椀愀渀最??圀愀渀最?倀攀渀最??圀愀渀最??攀渀最瀀椀渀最??攀琀?愀氀???  ????椀挀爀漀戀椀愀氀?搀椀瘀攀爀猀椀琀礀?愀琀?愀?搀攀攀瀀?猀攀愀?猀琀愀琀椀漀渀?漀昀?琀栀攀?倀愀挀椀昀椀挀?渀漀搀甀氀攀?瀀爀漀瘀椀渀挀攀???椀漀搀椀瘀攀爀猀椀琀礀????漀渀猀攀爀瘀愀琀椀漀渀?????????????????? ?戀爀?夀攀猀猀漀渀?????氀愀爀欀???刀??吀愀礀氀漀爀??????攀琀?愀氀??? ????吀栀攀?最氀漀戀愀氀?搀椀猀琀爀椀戀甀琀椀漀渀?漀昀?猀攀愀洀漀甀渀琀猀?戀愀猀攀搀?漀渀?? ?愀爀挀?猀攀挀漀渀搀猀?戀愀琀栀礀洀攀琀爀礀?搀愀琀愀???攀攀瀀?匀攀愀?刀攀猀攀愀爀挀栀?倀愀爀琀????伀挀攀愀渀漀最爀愀瀀栀椀挀?刀攀猀攀愀爀挀栀?倀愀瀀攀爀猀?????????????????戀爀?娀栀愀渀最??甀椀??匀攀欀椀最甀挀栀椀?夀???愀渀愀搀愀?匀??攀琀?愀氀???  ????攀洀洀愀琀椀洀漀渀愀猀?愀甀爀愀渀琀椀愀挀愀?最攀渀??渀漀瘀???猀瀀??渀漀瘀???愀??爀愀洀?渀攀最愀琀椀瘀攀??愀攀爀漀戀椀挀??瀀漀氀礀瀀栀漀猀瀀栀愀琀攀??愀挀挀甀洀甀氀愀琀椀渀最?洀椀挀爀漀?漀爀最愀渀椀猀洀??琀栀攀?昀椀爀猀琀?挀甀氀琀甀爀攀搀?爀攀瀀爀攀猀攀渀琀愀琀椀瘀攀?漀昀?琀栀攀?渀攀眀?戀愀挀琀攀爀椀愀氀?瀀栀礀氀甀洀??攀洀洀愀琀椀洀漀渀愀搀攀琀攀猀?瀀栀礀氀??渀漀瘀???渀琀攀爀渀愀琀椀漀渀愀氀??漀甀爀渀愀氀?漀昀?匀礀猀琀攀洀愀琀椀挀?愀渀搀??瘀漀氀甀琀椀漀渀愀爀礀??椀挀爀漀戀椀漀氀漀最礀???????????????????戀爀?娀栀愀渀最??甀礀甀愀渀??娀栀愀渀最?圀攀椀礀愀渀??娀栀甀??攀挀栀愀漀??攀琀?愀氀???  ????椀猀琀爀椀戀甀琀椀漀渀?挀栀愀爀愀挀琀攀爀椀猀琀椀挀猀?漀昀?挀漀戀愀氀琀?爀椀挀栀?昀攀爀爀漀洀愀渀最愀渀攀猀攀?挀爀甀猀琀?爀攀猀漀甀爀挀攀猀?漀渀?猀甀戀洀愀爀椀渀攀?猀攀愀洀漀甀渀琀猀?椀渀?琀栀攀?圀攀猀琀攀爀渀?倀愀挀椀昀椀挀???挀琀愀??攀漀氀漀最椀挀愀?匀椀渀椀挀愀?????????????? ??戀爀?娀栀愀漀?儀椀礀甀愀渀????????伀挀攀愀渀??攀漀挀栀攀洀椀猀琀爀礀???攀椀樀椀渀最??吀栀攀??攀漀氀漀最椀挀愀氀?倀甀戀氀椀猀栀椀渀最??漀甀猀攀?戀爀?娀栀甀?圀攀渀栀愀渀???漀洀猀愀搀稀攀?????漀爀漀搀漀瘀猀欀礀????? ? ???戀?椀渀椀琀椀漀?最攀渀攀?椀搀攀渀琀椀昀椀挀愀琀椀漀渀?椀渀?洀攀琀愀最攀渀漀洀椀挀?猀攀焀甀攀渀挀攀猀??一甀挀氀攀椀挀??挀椀搀猀?刀攀猀攀愀爀挀栀??????????攀???
  • 加载中
计量
  • 文章访问数:  1833
  • HTML全文浏览量:  44
  • PDF下载量:  1178
  • 被引次数: 0
出版历程
  • 收稿日期:  2014-01-03
  • 修回日期:  2014-07-05

目录

    /

    返回文章
    返回