隆线溞孤雌卵胚胎发育及其两性溞EST的初步研究
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摘要
隆线溞(Daphnia carinata)属节肢动物门(Arthropoda)、甲壳纲(Crustacea)、鳃足亚纲(Branchiopoda)、双甲目(Diplostraca)、枝角亚目(Cladocera)、溞科(Daphniidae)、溞属(Daphnia),是上海地区常见的淡水枝角类,采集方便,易于实验室培养。本研究以隆线溞为实验材料,采用组织学方法对其孤雌卵(夏卵)胚胎发育的全过程进行了详细地研究,以期为枝角类的规模化培养提供理论基础,并为深入研究枝角类胚胎发育的机理积累基础生物学资料。此外,运用分子生物学技术构建了隆线溞两性溞的cDNA文库,以该文库为材料进行了大规模的5'端EST测序,并将其与隆线溞孤雌溞文库进行比对,进行不同生殖状态下基因表达差异的研究,以期获取与生殖转化调控相关的基因,从分子角度初步探索枝角类生殖转化的内在规律和机理,为实现枝角类生殖转化的人为调控奠定基础,并为枝角类生殖生物学的深入研究提供线索。本研究的主要结果和结论如下:
1隆线溞孤雌卵胚胎发育的形态学研究
采用组织学方法较为系统地研究了隆线溞孤雌卵(夏卵)胚胎发育的全过程。隆线溞夏卵为中黄卵,室温24℃下,整个胚胎发育过程需45小时左右。根据隆线溞胚胎内部结构特征及外部形态学变化,将其胚胎发育分为卵裂期、囊胚期、原肠期、前无节幼体期、后无节幼体期、复眼色素期、准备孵化期七个时期。卵排至孵育囊约40 min后开始表面卵裂。卵裂至256细胞时,胚胎发育进入囊胚阶段,在卵表形成一薄层细胞层,囊胚腔则全被卵黄颗粒所充塞。囊胚后期,囊胚层细胞分裂加快,相互挤压向囊胚腔中移入形成原肠胚。随后,胚胎外部形态开始出现变化。首先在胚胎前端出现头部的三对附肢原基(两对触角原基及一对大颚原基),胚胎发育进入前无节幼体期。随后胸节分化,胚胎发育进入后无节幼体期,并形成胸肢、壳瓣、肠道等结构。复眼在复眼色素期的基础上,逐渐发育形成完整的复眼结构,同时其他各组织器官也不断发育完善。至准备孵化期的胚胎结构与幼体已基本相同。研究结果可为深入研究枝角类胚胎发育的机理积累基础生物学资料,同时为水产养殖业规模化培养枝角类提供基本生物学知识。
2隆线溞两性溞cDNA文库的构建和EST序列的初步分析
本研究构建了隆线溞两性种cDNA文库,并从中随机挑取3456个克隆测序。经去除低质量序列、载体序列及小于100bp的序列后,最终用于聚类拼接的EST数目为3367个(平均长度522bp,最小长度102bp,最大长度720bp),成功率97%。这3367个EST经聚类和拼接后得到Unique sequences为1576条,其中contigs445条,singletons 1131条。进行同源性比对后发现961条unique sequences(61%)与已知基因有较高同源性。由于本实验室同时构建了隆线溞孤雌溞的EST,我们对两种EST进行了表达差异分析,以期获取与生殖转化相关的功能基因。结果显示经IDEG.6软件分析后,得到12个基因在表达上有显著差异(p＜=0.01),其中,2个基因在孤雌溞中为相对高表达,10个基因在两性溞中为相对高表达。这12个基因经序列比对后,得到8个基因与已知基因有较高同源性,其中5个有功能注释。本文库及EST分析结果是帮助我们更深入的探究枝角类生殖转化机制的基础,也丰富了甲壳类动物发育生物学的研究内容,并为甲壳动物发育生物学的研究提供了新的研究思路。
Cladocera,generally speaking named daphnia or water fleas,are planktonic crustaceans that belong to the Branchiopoda.They appear in high concentrations in pools, ponds,lakes,ditches,slow-moving streams and swamps where organic material is decomposing,and become especially abundant with suitable conditions in a short period of time.Daphnia carinata is a kind of familiar freshwater Cladocera in Shanghai belonging to the genus Daphnia,the family Daphniidae.It is convenient to gather and culture this species in the lab condition.In this paper,we chose D.carinata as the experimental animal.
Recently,many researches have been concerntrated on mechanism of reproductive conversion,but the main researches were focused on induction of some environmental factors in reproductive conversion.As to the genes and proteins related to the reproductive conversion in Cladocera,few reports had been elucidated so far.Under the background,we hope that the analysis of express sequence tag on D.carinata may have both practical and theoretic values.
In this paper,we made a preliminary study on embryonic development of the parthenogenetic egg in D.carinata.And,we constructed a cDNA library from sexual female of D.carinata to find the genes and proteins related to the reproductive conversion in Cladocera.The findings will contribute basic data regarding the artificial control of reproductive conversion,and will provide fundamental and significant information for future studies of Cladocerans' reproductive biology.The results and conclusions were listed as following:
1.The morphological study on embryonic development of the parthenogenetic egg in Daphnia carinata
The embryonic development of the parthenogenetic egg(PE) in Daphnia carinata has been study morphologically in the present study.The PE is centrolecithal and its embryogenesis lasts about 45 hours at 24℃.According to the morphological and histological changes,the embryonic development of D.carinata could be divided into seven major stages,i.e.,the cleavage stage,blastula stage,gastrula stage,egg-nauplius stage,egg-metanauplius stage,embryo with eye pigments,and prepare-hatchling stage. The superficial cleavage of PE did not happen until approximately 40 minutes after egg laying.At the end of cleavage,the embryo consisted of 256 cells and reached the blastula stage at which a large part of the blastocoele was filled with yolk granules,accompanying by the formation of a thin sheet of divided cells.The cell division rate was accelerated and gastrulation took place by means of immigration of blastodermal cells at the end of this stage.Afterwards,the morphological changes in embryo development became remarkable.The egg-nauplius stage was characterized by the appearance of the rudiments of two antennae and a mandible in the head region of the embryo,while the egg-metanauplius stage characterized by the formation of the thoracic segments,thoracic limbs,carpace,and guts.At the eye pigments stage,the embryo was found presenting the eye pigmentation in the well-developed compound eyes.Simultaneously,other physiologically important tissues and organs started to develop.During the last prepare-hatching stage,the organelle structure in the embryo could be quite similar to that in the larval.The results could accumulate basic biological information for further research on embryonic development of Cladocera.
2.Construction of cDNA library from sexual female of Daphnia carinata and primary analysis of its ESTs
A high quality cDNA library was constructed from sexual female of D.carinata according to classical technique.The cDNA library contained 2.89×10~5pfu/ml.From this cDNA library,3456 EST sequences were obtained by randomly sequencing.A set of 1495 clusters was generated which contained 445 contigs and 1131 singletons.The nucleic acid and deduced amino acid sequences were compared with known GenBank sequences. Functional annotation found that 961 clusters showed significant homology with known genes involved in a broad range of metabolism,translation,development and reproduction.As our laboratory has obtained 3072 ESTs from parthenogenetic of D. carinata,we used IDEG6 to identify differentially expressed genes between sexual and parthenogenetic females of D.carinata.The results showed that there were 12 genes had differentially expressed.Among them,2 genes were highly expressed in parthenogenetic D.carinata,and the others were highly expressed in sexual D.carinata.Functional annotation found that 8 genes showed significant homology with known genes and 5 genes had function annotation.
1隆线溞孤雌卵胚胎发育的形态学研究
采用组织学方法较为系统地研究了隆线溞孤雌卵(夏卵)胚胎发育的全过程。隆线溞夏卵为中黄卵,室温24℃下,整个胚胎发育过程需45小时左右。根据隆线溞胚胎内部结构特征及外部形态学变化,将其胚胎发育分为卵裂期、囊胚期、原肠期、前无节幼体期、后无节幼体期、复眼色素期、准备孵化期七个时期。卵排至孵育囊约40 min后开始表面卵裂。卵裂至256细胞时,胚胎发育进入囊胚阶段,在卵表形成一薄层细胞层,囊胚腔则全被卵黄颗粒所充塞。囊胚后期,囊胚层细胞分裂加快,相互挤压向囊胚腔中移入形成原肠胚。随后,胚胎外部形态开始出现变化。首先在胚胎前端出现头部的三对附肢原基(两对触角原基及一对大颚原基),胚胎发育进入前无节幼体期。随后胸节分化,胚胎发育进入后无节幼体期,并形成胸肢、壳瓣、肠道等结构。复眼在复眼色素期的基础上,逐渐发育形成完整的复眼结构,同时其他各组织器官也不断发育完善。至准备孵化期的胚胎结构与幼体已基本相同。研究结果可为深入研究枝角类胚胎发育的机理积累基础生物学资料,同时为水产养殖业规模化培养枝角类提供基本生物学知识。
2隆线溞两性溞cDNA文库的构建和EST序列的初步分析
本研究构建了隆线溞两性种cDNA文库,并从中随机挑取3456个克隆测序。经去除低质量序列、载体序列及小于100bp的序列后,最终用于聚类拼接的EST数目为3367个(平均长度522bp,最小长度102bp,最大长度720bp),成功率97%。这3367个EST经聚类和拼接后得到Unique sequences为1576条,其中contigs445条,singletons 1131条。进行同源性比对后发现961条unique sequences(61%)与已知基因有较高同源性。由于本实验室同时构建了隆线溞孤雌溞的EST,我们对两种EST进行了表达差异分析,以期获取与生殖转化相关的功能基因。结果显示经IDEG.6软件分析后,得到12个基因在表达上有显著差异(p＜=0.01),其中,2个基因在孤雌溞中为相对高表达,10个基因在两性溞中为相对高表达。这12个基因经序列比对后,得到8个基因与已知基因有较高同源性,其中5个有功能注释。本文库及EST分析结果是帮助我们更深入的探究枝角类生殖转化机制的基础,也丰富了甲壳类动物发育生物学的研究内容,并为甲壳动物发育生物学的研究提供了新的研究思路。
Cladocera,generally speaking named daphnia or water fleas,are planktonic crustaceans that belong to the Branchiopoda.They appear in high concentrations in pools, ponds,lakes,ditches,slow-moving streams and swamps where organic material is decomposing,and become especially abundant with suitable conditions in a short period of time.Daphnia carinata is a kind of familiar freshwater Cladocera in Shanghai belonging to the genus Daphnia,the family Daphniidae.It is convenient to gather and culture this species in the lab condition.In this paper,we chose D.carinata as the experimental animal.
Recently,many researches have been concerntrated on mechanism of reproductive conversion,but the main researches were focused on induction of some environmental factors in reproductive conversion.As to the genes and proteins related to the reproductive conversion in Cladocera,few reports had been elucidated so far.Under the background,we hope that the analysis of express sequence tag on D.carinata may have both practical and theoretic values.
In this paper,we made a preliminary study on embryonic development of the parthenogenetic egg in D.carinata.And,we constructed a cDNA library from sexual female of D.carinata to find the genes and proteins related to the reproductive conversion in Cladocera.The findings will contribute basic data regarding the artificial control of reproductive conversion,and will provide fundamental and significant information for future studies of Cladocerans' reproductive biology.The results and conclusions were listed as following:
1.The morphological study on embryonic development of the parthenogenetic egg in Daphnia carinata
The embryonic development of the parthenogenetic egg(PE) in Daphnia carinata has been study morphologically in the present study.The PE is centrolecithal and its embryogenesis lasts about 45 hours at 24℃.According to the morphological and histological changes,the embryonic development of D.carinata could be divided into seven major stages,i.e.,the cleavage stage,blastula stage,gastrula stage,egg-nauplius stage,egg-metanauplius stage,embryo with eye pigments,and prepare-hatchling stage. The superficial cleavage of PE did not happen until approximately 40 minutes after egg laying.At the end of cleavage,the embryo consisted of 256 cells and reached the blastula stage at which a large part of the blastocoele was filled with yolk granules,accompanying by the formation of a thin sheet of divided cells.The cell division rate was accelerated and gastrulation took place by means of immigration of blastodermal cells at the end of this stage.Afterwards,the morphological changes in embryo development became remarkable.The egg-nauplius stage was characterized by the appearance of the rudiments of two antennae and a mandible in the head region of the embryo,while the egg-metanauplius stage characterized by the formation of the thoracic segments,thoracic limbs,carpace,and guts.At the eye pigments stage,the embryo was found presenting the eye pigmentation in the well-developed compound eyes.Simultaneously,other physiologically important tissues and organs started to develop.During the last prepare-hatching stage,the organelle structure in the embryo could be quite similar to that in the larval.The results could accumulate basic biological information for further research on embryonic development of Cladocera.
2.Construction of cDNA library from sexual female of Daphnia carinata and primary analysis of its ESTs
A high quality cDNA library was constructed from sexual female of D.carinata according to classical technique.The cDNA library contained 2.89×10~5pfu/ml.From this cDNA library,3456 EST sequences were obtained by randomly sequencing.A set of 1495 clusters was generated which contained 445 contigs and 1131 singletons.The nucleic acid and deduced amino acid sequences were compared with known GenBank sequences. Functional annotation found that 961 clusters showed significant homology with known genes involved in a broad range of metabolism,translation,development and reproduction.As our laboratory has obtained 3072 ESTs from parthenogenetic of D. carinata,we used IDEG6 to identify differentially expressed genes between sexual and parthenogenetic females of D.carinata.The results showed that there were 12 genes had differentially expressed.Among them,2 genes were highly expressed in parthenogenetic D.carinata,and the others were highly expressed in sexual D.carinata.Functional annotation found that 8 genes showed significant homology with known genes and 5 genes had function annotation.
引文
1、蒋燮治,堵南山.中国动物志:节肢动物门,甲壳纲,淡水枝角类[M].北京:科学出版社,1979,35-40.
2、郑重,曹文清.海洋枝角类生物学[M].厦门:厦门大学出版社,1987:74-77.
3、Green J.Growth,size and reproduction in Daphnia(Crustacea:Cladocera)[J].Proc.zool.Soc.Lond,1956,126:173-204.
4、Murugan N,Sivaramakrishnan K G..The biology of Simocephalus acutirostratus King(Cladocera,Daphnidae)-Laboratory studies of life span,instar duration,egg production,growth and stages in embryonic development[J].Freshwater Biology,1973,3(1):77-83.
5、Murugan N,Sivaramakrishnan K G..Laboratory studies on the longevity,instar duration,growth,reproduction and embryonic development in Scapholeberis kingi Sars(1903)(Cladocera:Daphnidae)[J].Hydrobiologia,1976,50(1):75-80.
6、施心路,史新柏.微齿喜马拉雅低额溞的生殖与发育[J].动物学报,1996,42(3):287-296.
7、曾错.多刺裸腹溞(Moina macrocopa)生殖腺发育和胚胎发育的形态学研究[D].上海:华东师范大学,2006,34-52.
8、黄祥飞.温度对近亲裸腹溞发育、生长和卵的生产量的影响[J].水生生物学集刊,1983,8(1):105-112.
9、郑又雄.发头裸腹溞卵和胚胎发育形态研究[J].动物学杂志,1993,28(6):5-8.
10、Kotov A A,Boikova O S.Comparative analysis of the late embryogenesis of Sida crystallina(O.F.M(u|¨)ller,1776) and Diaphanosoma brachyurum(Lievin,1848)(Crustacea:Branchiopoda:Ctenopoda)[J].Hydrobiologia,1998,380:103-125.
11、Kotov A A,Boikova O S.Study of the late embryogenesis of Daphnia(Anomopoda,'Cladocera',Branchiopoda) and a comparison of development in Anomopoda and Ctenopoda[J].Hydrobiologia,2001,442:127-143.
12、Murugan N,Venkataraman K.Study of the in vitro development of the parthenogenetic egg of Daphnia carinata King(Cladocera:Daphnidae)[J]. Hydrobiologia,1977,52(2-3):129-134.
13、张明凤,赵云龙,赵艳民,等.隆线溞夏卵胚胎发育过程中形态及蛋白质、氨基酸组成和含量的变化[J].水产学报,2005,29(6):783-790.
14、Grebelnyi S D.Influence of parthenogenetic reproduction on the genotypic constitution and evolutionary success of population and species[J].Hydrobiologia,1996,320(1-3):55-61.
15、Carvalho G R,Hughes R N.The effect of food availability,female culture-density and photoperiod on ephippia production in Daphnia magna Straus(Crustacea:Cladocera)[J].Freshwater Biology,1983,13:37-46.
16、Banta A M.Studies on the physiology,genetics and evolution of some Cladocera[M].Publ Carneg Inst,1939:513.
17、Green J.Seasonal variation in eggs productivity by Cladocera[J].J Anim Ecolo,1965,35:77-104.
18、Bunner H C,Halcrow K.Experimental induction of the production of ephippia by Daphnia magna Straus(Cladocera)[J].Crusteacea,1977,32(1):77-86.1
9、陆开宏,钱云霞,丁士芳.隆线潘冬卵的诱发试验[J].浙江水产学院学报,1992,6(11):9-15.
20、Tifnouti A,Pourriot R,Rougier C.The effect of temperature on the growth and reproduction of four species of planktonic Cladocera(Crustacea) reared with natural resources present[J].Annales de Limnologie,1993,29(1):3-13.
21、Gordo T,Lubian L M,Canavate J P.Influence of temperature on growth,reproduction and longevity of Moina salina(Daday,1888)(Cladocera:Moinidae)[J].Journal of Plankton Research,1994,16(11):1513-1523.
22、Suzanne E M.Intersex and male development in Daphnia magna[J].Hydrobiologia,2001,442:145-156.
23、D'Abramo L R.Ingestion rate decrease as the stimulus for sexuality in populations of Moina macrocopa[J].Limnology and Oceanography,1980,25(3):422-429.
24、Frey D G.Contrasting strategies of gamogenesis in Northern and Southern population of Cladocera[J].Ecology,1982,63(1):223-241.
25、Korpelainen H.The effects of temperature and photoperiod on life history parameters of Daphnia magna (Crustecea:Cladocera)[J].Freshwater biology,1980,16:615-620.
26、Baer K N,Owens K D.Evaluation of selected endocrine disrupting compounds on sex determination in Daphnia magna using reduced photoperiod and different feeding rates[J].Bull Environ Contam Toxicol,1999,62:214-221.
27、Stross R G Photoperiod control of diapause in Daphnia Ⅱ:Induction of winter diapause in the Artic[J].Biol Bull,1969,136:264-273.
28、Stross R G Photoperiod control of diapause in Daphnia Ⅲ:Two-stimulus control of long-day,short-day induction[J].Biol Bull,1969,137:359-374.
29、Moghraby A I.A study on diapause of zooplankton in a tropical river-The Blue Nile[J].Freshwater Biology,1977,7:207-212.
30、Arnold D E.Ingestion assimition,surivival and reproduction by Daphnia pulex fed seven species of blue-green Algae[J].Limon Oceanogr,1971,16(6):906-922.
31、Banta A M,Brown L A.Control of sex in Cladocera I:Crowding the mothers as a means of controlling male production[J].Physiological Zoology,1929,2(1):80-92.
32、Terao A,Tanaka T.Influence of population density upon the rate of reproduction in the water-flea,Moina macrocopa Straus[J].Proc Imp Acad,1928,(4):556-558.
33、Zaffagnini F.Formation of an ovarian follicle and its function in genesis of parthenogenetic eggs in Daphnia magna[J].Rend 1st Sci Univ Camerino,1961,14(2):231-243.
34、Bacci G,Cognetti G,Vaccari A.Endomeiosis and sex determination in Daphnia pulex[J].Experientia,1961,17:505-506.
35、Zaffagnini F.Some cytological and cytochemical observations on the maturation of the testes inDaphnia magna[J].Caryologia,1965,18(1):1-13.
36、Berard J J.Ultrastructural study of spermatogenesis and spermatozoon in Daphnia magna Straus (Entomostraca,Branchiopoda,Cladocera)[J].Bulletin De La Societe Zoologique De France -Evolution Et Zoologie,1974,99(4):723-730.
37、Delavaul R,Berard J J.Ultrastructural study of spermatogenesis in Daphnia magna Straus (Entomostracese,Branchiopoda,Cladocerae) [J].Comptes Rendus Hebdomadaires Des Seances De L Academie Des Sciences Serie D,1974,278(12):1589-1592.
38、Beaton J J,Hebert P D N.Variation in chromosome numbers in Daphnia[J].Hereditas,1994,120:275-279.
39、Trentini M.Chromosome numbers of nine species of Daphniidae(Crustacea,Cladocera)[J].Genetica,1980,54:221-223.
40、曹文清,王永聪,林元烧.多刺裸腹溞染色体实验研究[J].台湾海峡,1995,14(3):284-287.
41、曹文清,林元烧,郭东晖,等.蒙古裸腹溞染色体组型研究[J].台湾海峡,1999,18(1):71-75.
42、Allen E,Banta A M.Growth and maturation in the parthenogenetic and sexual eggs of Monia macrocopa[J].J Morph,1929,48:123-151.
43、Ojima Y.A cytologyical study on the development and maturation of the parthenogenetic and sexual eggs of Daphnia pulex(Crustacea:Cladocera)[J].Kwansei Gakuin Univ Annual Studies,1958,6:123-176.
44、Zaffagnini F,Sabelli B.Karyologic observations on the maturation of the summer and winter eggs of Daphnia pulex and Daphnia middendorffiana[J].Chromosoma,1972,36(2):193-203.
45、Dehn M von.Experimentelle.Untersuchungen uber den Generationswechsel der Cladoceren[J].Chromosoma(Berl),1948,3:167-193.
46、Banta A M.A thelytokous race of Cladocera in which pseudosexual reproduction occurs[J].Z induct Abstamm-u Vererb-L,1925,40:28-41.
47、Schrader F.The cytology of pseudosexual eggs in a species of Daphnia[J].Z indukt Abstamm-u Vererb-L,1925,40:1-36.
48、Banta A M,Brown L A.Control of sex in cladocera-locatization of the critical periodical for control of sex[J].Proc Natal Acd Sci,1929,15:71-81.
49、Baker D M,Herbert P D N.Secondary sex ratio of the cyclic parthenogene Daphnia magna in the Candsi in Arctic[J].Canadian Journal of Zoology,1986,64:1137-1143.
50、王金秋,张佳忆,黄博.生态因子的波动对多刺裸腹溞雄体发生的影响[J].应用 生态学报,2000,11(1):135-137.
51、Sciacchitano I.Cyclical reproduction and variation of some Sardinian Cladocera[J].Internat Rev Ges,Hydrobiol u Hydrogr,1926,15(5-6):335-346.
52、张明凤,赵云龙,曾错.隆线溞孤雌溞和两性雌溞的蛋白质差异表达[J].动物学报,2006,52(5):916-923.
53、Watanabe H,Tatarazako N,Oda S,et al.Analysis of expressed sequence tags of the water flea Daphnia magna[J].Genome,2005,48:606-609.
54、Venter J C,Adams M D,Eugene W M,et al.The sequence of the Human Genome[J].Science,2001,291(5507):1304-1351.
55、Goff S A,Ricke D,Lan T H,et al.A draft sequence of the rice genome(Oryza sativa L.ssp.japonica)[J].Science,2002,296:92-100.
56、Costanzo F.Cloning of several cDNA segments coding rot human liver proteins[J].EMBO J.,1983,2:57-60.
57、Adams M D,Kelley J M,Gocayne J D,et al.Complementary DNA sequencing:expressed sequence tags and human genome project[J].Science,1991,252:1651-1656.
58、Boguski M S,Lowe T M,Tolstoshev C M.dbEST—database for "expressed sequence tags"[J].Nature Genet,1993,4(4):332-333.
59、Bedell J A,Korf I,Gish W.MaskerAid:a performance enhancement to RepeatMasker[J].Bioinformatics,2000,16(11):1040-10411.
60、Schuler G D,Boguski M S,Stewart E A,et al.A gene map of the human genome[J].Science,1996,274(5287):540-546.
61、Huang X,Madan A.CAP3:A DNA sequence assembly program[J].Genome Res,1999,9:868-877.
62、Romualdi C,Bortoluzzi S,D'Alessi F,et al.IDEG6:a web tool for detection of differentially expressed genes in multiple tag sampling experiments[J].Physiological Genomics.2003,12(2):159-162.
63、Wixon J,Kell D.The Kyoto encyclopedia of genes and genomes-KEGG[J].Yeast.2000,17(1):48-55.
64、Arakawa K,Kono N,Yamada Y,et al.KEGG-based pathway visualization tool for complex omics data[J].In Silico Biology.2005,5(4):419-423.
65、Liu J,Hara C.Umeda M1 analysis of randomly isolated cDNA from developing endosperm of rice[J].Plant Molecular Biology.1995.29(4):685-689.
66、Epple P.EST Rreveal multigene family for plant in Arabidopsis Thalina[J].FEBS letter.1997,400(2):168-192.
67、Allona I,Quinn M,Shoop E.Analysis of xylem formation in pine by cDNA sequencing[J].Proc Natl Acad Sci USA.1998,5(16):9693-9689.
68、卢亚芳,2001.枝角类在水产养殖中的应用[J].台湾海峡,20:69-75.
69、Kleiven O T,Larsson P,Hobaek A.Sexual reproduction in Daphnia magna requires three stimuli[J].Oikos,1992,65(2):197-206.
70、曹文清,林元烧,郭东晖.若干生态因子对蒙古裸腹溞生殖方式转变效应的实验[J].台湾海峡,2001,20(增刊):190-195.
71、张明凤,赵云龙,杨志彪,等.隆线潘孤雌溞生殖系统的组织学[J].动物学杂志,2004,39(4):68-72.
72、张明凤,赵云龙,曾错,等.隆线溞雄性生殖系统的组织学研究[J].华东师范大学学报(自然科学版),2004,118(4):111-116.
73、张明凤,赵云龙,王群,等.隆线溞精子发生及成熟精子的超微结构[J].动物学报,2005,51(4):710-717.
74、Gulbrandsen J,Johhsen G H.Temperature-dependent development of parthenogenetic embryos in Daphnia pulex de Geer[J].J Plankton Res,1990,12(3):443-453.
75、赵文,张焕,徐宪仲,等.蒙古裸腹溞胚胎发育及两种裸腹溞滋养盘的形态观察[J].大连水产学院学报,2006,21(3):196-202.
76、堵南山,赖伟,邓雪怀,等.隆线溞(Daphnia carinata King)生长与生殖力[J].华东师范大学学报(自然科学版),1983,1:85-91.
77、孟凡丽,赵云龙,陈立侨,等.红螯螯虾胚胎发育的研究:Ⅱ.消化系统的发生[J].动物学研究,2001,22(5):383-387.
78、赵云龙,王群,赖伟,等.罗氏沼虾胚胎发育研究Ⅰ、胚胎外部结构形态发生[J].动物学报,1998,44(3):249-256.
79、堵南山,赵云龙,赖伟.中华绒螯蟹胚胎发育的研究[A].甲壳动物学会主编.甲壳动物学论文集(第三辑)[C].青岛:青岛海洋大学出版社,1992,128-135.
80、山东海洋学院,上海水产学院.水产动物胚胎学[M].北京:农业出版社,1961,61-67.
81、Kotov A A.Fate of the second maxilla during embryogenesis in some Anomopoda Crustacea(Branchiopoda)[J].Zoological Journal of the Linnean Society,1996,116:393-405.
82、李红,赵云龙,王群,等.日本沼虾胚胎发育不同阶段主要生化成分的变化[J].水产学报,2003,27(6):545-549.
83、王桂忠,汤鸿,李少菁,等.锯缘青蟹胚胎发育过程主要生化组成[J].台湾海峡,1995,14(3):280-283.
84、胡松年.基因表达序列标签(EST)数据分析手册[M].浙江:浙江大学出版社,2005:8.
85、晏慧君,黄兴奇,程在全.cDNA文库构建策略及其分析研究进展[J].云南农业大学学报,2006,21(1):7-12.
86、郑明刚,孙修勤,张进兴.皱纹盘鲍肝和肾cDNA文库的构建及免疫相关基因的初步分[J].高技术通讯,2007,17(3):319-324.
87、王兴友,陈杭薇,李兵.流感病毒A单抗杂交瘤细胞cDNA文库的构建[J].国际呼吸杂志,2007,27(15):1128-1130.
88、Ainscough R,Bardill S,Barlow K,et al.Genome Sequence of the Nomatode C.elegans:A Platform for Investigating Biology[J].Science,1998,282(5396):2012-2018.
89、Vaisman B,Fibach E,Konijn A M.Utilization of intracellular ferritin iron for hemoglobin synthesis in developing human erythroid precursors[J].Blood,1997,90(2):831-838.
90、Hanazato T,Yasuno M,Hosomi M.Significance of a low oxygen layer for a Daphnia population in Lake Yunoko,Japan[J].Hydrobiologia,1989,185(1):19-27.
91、Kato Y,Tokishita S,Toshihiro O,et al.A vitellogenin chain containing a superoxide dismutase-like domain is the major component of yolk proteins in cladoceran crustacean Daphnia magna[J].Gene.2004,334:157-165.
92、王海燕,张敏.细胞周期检测点激酶1的研究进展[J].医学分子生物学杂志,2005,2(2):147-149.
93、Yong W,Marc W.Kirschner Identification of multiple CDH1 homologues in vertebrates conferring different substrate specificities.PNAS,2001,98(23):13066-13071.
94、郑青亮,盛清,张耀洲.Kazal型蛋白酶抑制剂结构与功能研究进展[J].生物工程学报,22(5):698-700.
2、郑重,曹文清.海洋枝角类生物学[M].厦门:厦门大学出版社,1987:74-77.
3、Green J.Growth,size and reproduction in Daphnia(Crustacea:Cladocera)[J].Proc.zool.Soc.Lond,1956,126:173-204.
4、Murugan N,Sivaramakrishnan K G..The biology of Simocephalus acutirostratus King(Cladocera,Daphnidae)-Laboratory studies of life span,instar duration,egg production,growth and stages in embryonic development[J].Freshwater Biology,1973,3(1):77-83.
5、Murugan N,Sivaramakrishnan K G..Laboratory studies on the longevity,instar duration,growth,reproduction and embryonic development in Scapholeberis kingi Sars(1903)(Cladocera:Daphnidae)[J].Hydrobiologia,1976,50(1):75-80.
6、施心路,史新柏.微齿喜马拉雅低额溞的生殖与发育[J].动物学报,1996,42(3):287-296.
7、曾错.多刺裸腹溞(Moina macrocopa)生殖腺发育和胚胎发育的形态学研究[D].上海:华东师范大学,2006,34-52.
8、黄祥飞.温度对近亲裸腹溞发育、生长和卵的生产量的影响[J].水生生物学集刊,1983,8(1):105-112.
9、郑又雄.发头裸腹溞卵和胚胎发育形态研究[J].动物学杂志,1993,28(6):5-8.
10、Kotov A A,Boikova O S.Comparative analysis of the late embryogenesis of Sida crystallina(O.F.M(u|¨)ller,1776) and Diaphanosoma brachyurum(Lievin,1848)(Crustacea:Branchiopoda:Ctenopoda)[J].Hydrobiologia,1998,380:103-125.
11、Kotov A A,Boikova O S.Study of the late embryogenesis of Daphnia(Anomopoda,'Cladocera',Branchiopoda) and a comparison of development in Anomopoda and Ctenopoda[J].Hydrobiologia,2001,442:127-143.
12、Murugan N,Venkataraman K.Study of the in vitro development of the parthenogenetic egg of Daphnia carinata King(Cladocera:Daphnidae)[J]. Hydrobiologia,1977,52(2-3):129-134.
13、张明凤,赵云龙,赵艳民,等.隆线溞夏卵胚胎发育过程中形态及蛋白质、氨基酸组成和含量的变化[J].水产学报,2005,29(6):783-790.
14、Grebelnyi S D.Influence of parthenogenetic reproduction on the genotypic constitution and evolutionary success of population and species[J].Hydrobiologia,1996,320(1-3):55-61.
15、Carvalho G R,Hughes R N.The effect of food availability,female culture-density and photoperiod on ephippia production in Daphnia magna Straus(Crustacea:Cladocera)[J].Freshwater Biology,1983,13:37-46.
16、Banta A M.Studies on the physiology,genetics and evolution of some Cladocera[M].Publ Carneg Inst,1939:513.
17、Green J.Seasonal variation in eggs productivity by Cladocera[J].J Anim Ecolo,1965,35:77-104.
18、Bunner H C,Halcrow K.Experimental induction of the production of ephippia by Daphnia magna Straus(Cladocera)[J].Crusteacea,1977,32(1):77-86.1
9、陆开宏,钱云霞,丁士芳.隆线潘冬卵的诱发试验[J].浙江水产学院学报,1992,6(11):9-15.
20、Tifnouti A,Pourriot R,Rougier C.The effect of temperature on the growth and reproduction of four species of planktonic Cladocera(Crustacea) reared with natural resources present[J].Annales de Limnologie,1993,29(1):3-13.
21、Gordo T,Lubian L M,Canavate J P.Influence of temperature on growth,reproduction and longevity of Moina salina(Daday,1888)(Cladocera:Moinidae)[J].Journal of Plankton Research,1994,16(11):1513-1523.
22、Suzanne E M.Intersex and male development in Daphnia magna[J].Hydrobiologia,2001,442:145-156.
23、D'Abramo L R.Ingestion rate decrease as the stimulus for sexuality in populations of Moina macrocopa[J].Limnology and Oceanography,1980,25(3):422-429.
24、Frey D G.Contrasting strategies of gamogenesis in Northern and Southern population of Cladocera[J].Ecology,1982,63(1):223-241.
25、Korpelainen H.The effects of temperature and photoperiod on life history parameters of Daphnia magna (Crustecea:Cladocera)[J].Freshwater biology,1980,16:615-620.
26、Baer K N,Owens K D.Evaluation of selected endocrine disrupting compounds on sex determination in Daphnia magna using reduced photoperiod and different feeding rates[J].Bull Environ Contam Toxicol,1999,62:214-221.
27、Stross R G Photoperiod control of diapause in Daphnia Ⅱ:Induction of winter diapause in the Artic[J].Biol Bull,1969,136:264-273.
28、Stross R G Photoperiod control of diapause in Daphnia Ⅲ:Two-stimulus control of long-day,short-day induction[J].Biol Bull,1969,137:359-374.
29、Moghraby A I.A study on diapause of zooplankton in a tropical river-The Blue Nile[J].Freshwater Biology,1977,7:207-212.
30、Arnold D E.Ingestion assimition,surivival and reproduction by Daphnia pulex fed seven species of blue-green Algae[J].Limon Oceanogr,1971,16(6):906-922.
31、Banta A M,Brown L A.Control of sex in Cladocera I:Crowding the mothers as a means of controlling male production[J].Physiological Zoology,1929,2(1):80-92.
32、Terao A,Tanaka T.Influence of population density upon the rate of reproduction in the water-flea,Moina macrocopa Straus[J].Proc Imp Acad,1928,(4):556-558.
33、Zaffagnini F.Formation of an ovarian follicle and its function in genesis of parthenogenetic eggs in Daphnia magna[J].Rend 1st Sci Univ Camerino,1961,14(2):231-243.
34、Bacci G,Cognetti G,Vaccari A.Endomeiosis and sex determination in Daphnia pulex[J].Experientia,1961,17:505-506.
35、Zaffagnini F.Some cytological and cytochemical observations on the maturation of the testes inDaphnia magna[J].Caryologia,1965,18(1):1-13.
36、Berard J J.Ultrastructural study of spermatogenesis and spermatozoon in Daphnia magna Straus (Entomostraca,Branchiopoda,Cladocera)[J].Bulletin De La Societe Zoologique De France -Evolution Et Zoologie,1974,99(4):723-730.
37、Delavaul R,Berard J J.Ultrastructural study of spermatogenesis in Daphnia magna Straus (Entomostracese,Branchiopoda,Cladocerae) [J].Comptes Rendus Hebdomadaires Des Seances De L Academie Des Sciences Serie D,1974,278(12):1589-1592.
38、Beaton J J,Hebert P D N.Variation in chromosome numbers in Daphnia[J].Hereditas,1994,120:275-279.
39、Trentini M.Chromosome numbers of nine species of Daphniidae(Crustacea,Cladocera)[J].Genetica,1980,54:221-223.
40、曹文清,王永聪,林元烧.多刺裸腹溞染色体实验研究[J].台湾海峡,1995,14(3):284-287.
41、曹文清,林元烧,郭东晖,等.蒙古裸腹溞染色体组型研究[J].台湾海峡,1999,18(1):71-75.
42、Allen E,Banta A M.Growth and maturation in the parthenogenetic and sexual eggs of Monia macrocopa[J].J Morph,1929,48:123-151.
43、Ojima Y.A cytologyical study on the development and maturation of the parthenogenetic and sexual eggs of Daphnia pulex(Crustacea:Cladocera)[J].Kwansei Gakuin Univ Annual Studies,1958,6:123-176.
44、Zaffagnini F,Sabelli B.Karyologic observations on the maturation of the summer and winter eggs of Daphnia pulex and Daphnia middendorffiana[J].Chromosoma,1972,36(2):193-203.
45、Dehn M von.Experimentelle.Untersuchungen uber den Generationswechsel der Cladoceren[J].Chromosoma(Berl),1948,3:167-193.
46、Banta A M.A thelytokous race of Cladocera in which pseudosexual reproduction occurs[J].Z induct Abstamm-u Vererb-L,1925,40:28-41.
47、Schrader F.The cytology of pseudosexual eggs in a species of Daphnia[J].Z indukt Abstamm-u Vererb-L,1925,40:1-36.
48、Banta A M,Brown L A.Control of sex in cladocera-locatization of the critical periodical for control of sex[J].Proc Natal Acd Sci,1929,15:71-81.
49、Baker D M,Herbert P D N.Secondary sex ratio of the cyclic parthenogene Daphnia magna in the Candsi in Arctic[J].Canadian Journal of Zoology,1986,64:1137-1143.
50、王金秋,张佳忆,黄博.生态因子的波动对多刺裸腹溞雄体发生的影响[J].应用 生态学报,2000,11(1):135-137.
51、Sciacchitano I.Cyclical reproduction and variation of some Sardinian Cladocera[J].Internat Rev Ges,Hydrobiol u Hydrogr,1926,15(5-6):335-346.
52、张明凤,赵云龙,曾错.隆线溞孤雌溞和两性雌溞的蛋白质差异表达[J].动物学报,2006,52(5):916-923.
53、Watanabe H,Tatarazako N,Oda S,et al.Analysis of expressed sequence tags of the water flea Daphnia magna[J].Genome,2005,48:606-609.
54、Venter J C,Adams M D,Eugene W M,et al.The sequence of the Human Genome[J].Science,2001,291(5507):1304-1351.
55、Goff S A,Ricke D,Lan T H,et al.A draft sequence of the rice genome(Oryza sativa L.ssp.japonica)[J].Science,2002,296:92-100.
56、Costanzo F.Cloning of several cDNA segments coding rot human liver proteins[J].EMBO J.,1983,2:57-60.
57、Adams M D,Kelley J M,Gocayne J D,et al.Complementary DNA sequencing:expressed sequence tags and human genome project[J].Science,1991,252:1651-1656.
58、Boguski M S,Lowe T M,Tolstoshev C M.dbEST—database for "expressed sequence tags"[J].Nature Genet,1993,4(4):332-333.
59、Bedell J A,Korf I,Gish W.MaskerAid:a performance enhancement to RepeatMasker[J].Bioinformatics,2000,16(11):1040-10411.
60、Schuler G D,Boguski M S,Stewart E A,et al.A gene map of the human genome[J].Science,1996,274(5287):540-546.
61、Huang X,Madan A.CAP3:A DNA sequence assembly program[J].Genome Res,1999,9:868-877.
62、Romualdi C,Bortoluzzi S,D'Alessi F,et al.IDEG6:a web tool for detection of differentially expressed genes in multiple tag sampling experiments[J].Physiological Genomics.2003,12(2):159-162.
63、Wixon J,Kell D.The Kyoto encyclopedia of genes and genomes-KEGG[J].Yeast.2000,17(1):48-55.
64、Arakawa K,Kono N,Yamada Y,et al.KEGG-based pathway visualization tool for complex omics data[J].In Silico Biology.2005,5(4):419-423.
65、Liu J,Hara C.Umeda M1 analysis of randomly isolated cDNA from developing endosperm of rice[J].Plant Molecular Biology.1995.29(4):685-689.
66、Epple P.EST Rreveal multigene family for plant in Arabidopsis Thalina[J].FEBS letter.1997,400(2):168-192.
67、Allona I,Quinn M,Shoop E.Analysis of xylem formation in pine by cDNA sequencing[J].Proc Natl Acad Sci USA.1998,5(16):9693-9689.
68、卢亚芳,2001.枝角类在水产养殖中的应用[J].台湾海峡,20:69-75.
69、Kleiven O T,Larsson P,Hobaek A.Sexual reproduction in Daphnia magna requires three stimuli[J].Oikos,1992,65(2):197-206.
70、曹文清,林元烧,郭东晖.若干生态因子对蒙古裸腹溞生殖方式转变效应的实验[J].台湾海峡,2001,20(增刊):190-195.
71、张明凤,赵云龙,杨志彪,等.隆线潘孤雌溞生殖系统的组织学[J].动物学杂志,2004,39(4):68-72.
72、张明凤,赵云龙,曾错,等.隆线溞雄性生殖系统的组织学研究[J].华东师范大学学报(自然科学版),2004,118(4):111-116.
73、张明凤,赵云龙,王群,等.隆线溞精子发生及成熟精子的超微结构[J].动物学报,2005,51(4):710-717.
74、Gulbrandsen J,Johhsen G H.Temperature-dependent development of parthenogenetic embryos in Daphnia pulex de Geer[J].J Plankton Res,1990,12(3):443-453.
75、赵文,张焕,徐宪仲,等.蒙古裸腹溞胚胎发育及两种裸腹溞滋养盘的形态观察[J].大连水产学院学报,2006,21(3):196-202.
76、堵南山,赖伟,邓雪怀,等.隆线溞(Daphnia carinata King)生长与生殖力[J].华东师范大学学报(自然科学版),1983,1:85-91.
77、孟凡丽,赵云龙,陈立侨,等.红螯螯虾胚胎发育的研究:Ⅱ.消化系统的发生[J].动物学研究,2001,22(5):383-387.
78、赵云龙,王群,赖伟,等.罗氏沼虾胚胎发育研究Ⅰ、胚胎外部结构形态发生[J].动物学报,1998,44(3):249-256.
79、堵南山,赵云龙,赖伟.中华绒螯蟹胚胎发育的研究[A].甲壳动物学会主编.甲壳动物学论文集(第三辑)[C].青岛:青岛海洋大学出版社,1992,128-135.
80、山东海洋学院,上海水产学院.水产动物胚胎学[M].北京:农业出版社,1961,61-67.
81、Kotov A A.Fate of the second maxilla during embryogenesis in some Anomopoda Crustacea(Branchiopoda)[J].Zoological Journal of the Linnean Society,1996,116:393-405.
82、李红,赵云龙,王群,等.日本沼虾胚胎发育不同阶段主要生化成分的变化[J].水产学报,2003,27(6):545-549.
83、王桂忠,汤鸿,李少菁,等.锯缘青蟹胚胎发育过程主要生化组成[J].台湾海峡,1995,14(3):280-283.
84、胡松年.基因表达序列标签(EST)数据分析手册[M].浙江:浙江大学出版社,2005:8.
85、晏慧君,黄兴奇,程在全.cDNA文库构建策略及其分析研究进展[J].云南农业大学学报,2006,21(1):7-12.
86、郑明刚,孙修勤,张进兴.皱纹盘鲍肝和肾cDNA文库的构建及免疫相关基因的初步分[J].高技术通讯,2007,17(3):319-324.
87、王兴友,陈杭薇,李兵.流感病毒A单抗杂交瘤细胞cDNA文库的构建[J].国际呼吸杂志,2007,27(15):1128-1130.
88、Ainscough R,Bardill S,Barlow K,et al.Genome Sequence of the Nomatode C.elegans:A Platform for Investigating Biology[J].Science,1998,282(5396):2012-2018.
89、Vaisman B,Fibach E,Konijn A M.Utilization of intracellular ferritin iron for hemoglobin synthesis in developing human erythroid precursors[J].Blood,1997,90(2):831-838.
90、Hanazato T,Yasuno M,Hosomi M.Significance of a low oxygen layer for a Daphnia population in Lake Yunoko,Japan[J].Hydrobiologia,1989,185(1):19-27.
91、Kato Y,Tokishita S,Toshihiro O,et al.A vitellogenin chain containing a superoxide dismutase-like domain is the major component of yolk proteins in cladoceran crustacean Daphnia magna[J].Gene.2004,334:157-165.
92、王海燕,张敏.细胞周期检测点激酶1的研究进展[J].医学分子生物学杂志,2005,2(2):147-149.
93、Yong W,Marc W.Kirschner Identification of multiple CDH1 homologues in vertebrates conferring different substrate specificities.PNAS,2001,98(23):13066-13071.
94、郑青亮,盛清,张耀洲.Kazal型蛋白酶抑制剂结构与功能研究进展[J].生物工程学报,22(5):698-700.