胶州湾浮游植物rbcL基因分子遗传多样性研究

摘要/Abstract

摘要： 利用聚合酶链式反应扩增胶州湾表层海水浮游植物核酮糖1,5-二磷酸羧化/氧化酶大亚基基因(rbcL)片段,建立了该基因片段变异类型文库.随机测定了28个rbcL片段序列,依此初步分析了胶州湾表层海水浮游植物rbcL基因分子遗传多样性.结果表明,春季胶州湾表层海水浮游植物优势种群为D类rbcL代表的浮游植物,其中隐藻占28.6%、Stramenopies占32.1%、定鞭藻占28.6%、红藻占3.6%.B类rbcL代表的浮游植物为绿藻,占7.1%.根据各操作分类单元丰度计算的分子遗传多样性指数为2.85,根据逆翻译成的氨基酸序列计算的序列多样性为0.20.

关键词: 浮游植物, 核酮糖1,5-二磷酸羧化/氧化酶大亚基基因, 分子遗传多样性

Abstract: The variants of a 500 base pair fragment of RubisCo large subunit gene(rbcL)from the phytoplanktonic DNA of Jiaozhou Bay surface seawater were amplified by using polymerase chain reaction and cloned.Twenty-eight clones were randomly selected and sequenced,which were further used to determine the molecular genetic diversity of the phytoplankton of Jiaozhou Bay surface seawater.Systematic analysis showed that the clones representing cryptophyta counted for 28.6%,Stramenopiles 32.1%,Haptophyta 28.6%,Rhodophyta 3.6% and Chlorophyta 7.1%.The sequences from Cryptophyta,Stramenopiles,Haptophyta and Rhodophyta belonged to type D of Form I rbcL and Chlorophyta to type B,indicating that the dominant phytoplankton were those represented by type D rbcL.The genetic diversity index and the reversely translated amino acid sequence diversity of Jiaozhou Bay phytoplankton were 2.85 and 0.20,which were determined by the abundances of operational taxonomy units and the reversely translated amino acid sequences respectively.

Key words: Phytoplankton, Ribulose-1, 5-bisphosphate carboxynase/oxygenase large subunit gene(rbcL), Molecular genetic diversity

中图分类号:

Q344
Q943.2

刘永健, 杨官品, 管晓菁. 胶州湾浮游植物rbcL基因分子遗传多样性研究[J]. 应用生态学报, 2004, (9): 1626-1632.

LIU Yongjian, YANG Guanpin, GUAN Xiaojing. Molecular genetic diversity of phytoplankton rbcL gene in Jiaozhou Bay[J]. Chinese Journal of Applied Ecology, 2004, (9): 1626-1632.

参考文献

[1] Amann RI, Ludwig W, Schleifer KH. 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev, 59:143～ 169
[2] Andersen RA, Bailey JC. 2002. Phylogenetic analysis of 32 strains of Vaucheria (Xanthophyceae) using the rbcL gene and ITS two flanking space regions. J Phycol, 38: 583 ～ 592
[3] Chernykh NA. 1995. Detection of the ribulose bisphosphate carboxylase gene in natural samples by polymerase chain reaction.Mikrobiologiya, 64: 792 ～ 796
[4] Clegg MT. 1992. Chloroplast gene sequences and the study of plant evolution. Proc Natl Acad Sci USA, 89: 5685～ 5689
[5] Daugbjerg N, Andersen RA. 1997. A molecule phylogeny of the Heterokont algae based on analysis of chloroplast-encoded rbcL sequences data. J Phycol, 33 :1031～1041
[6] Daugbjerg N, Henriksen P. 2001. Pigment composition and rbcL sequence data from the silicoflagellate dictyocha speculum: A herterokont algae with pigments similar to some haptophytes. J Phycol,37:1110～ 1120
[7] Daugbjerg N, Andersen RA. 1997. Phylogenetic analysis of the rbcL sequences form Haptophytes and Heterokont algae suggest their chloroplasts are unrelated. Mol Biol Evol, 14 ( 12): 1242 ～1251
[8] Debashish B, Medlin L. 1998. Algal phylogeny and the origin of land plants. Plant Physiol,116:9～15
[9] Elsaied H, Naganuma T. 2001. Phylogenetic diversity of ribulose-1,5-bisphosphate carboxylase/oxygenase large-subunit genes from deep-sea microorganisms. Appl Environ Microbiol, 67(4): 1751～ 1765
[10] Freshwater DW, Fredericq S, Hommersand MH. 1995. A molecular phylogeny of the Gelidiales(Rhodophyta)based on analysis of plastid rbcL nucleotide sequences. J Phycol, 31: 616～ 632
[11] Freshwater DW, Frendericq S, Butler, BS, et al. 1994. A gene phylogeny of the red algae(Rhodophyta)based on plastid rbcL.Proc Natl Acad USA, 91:7281～7285
[12] Fuhrman JA, McCallum K, Davis AA. 1993. Phylogenetic diversity of subsurface marine microbial communities from Atlantic and Pacific Oceans. Appl Environ Microbiol, 59:1294 ～ 1302
[13] Fujiwara S, Tsuzuki M, Kawachi M, et al. 2001. Molecular phylogeny of the Haptophyta based on the rbcL gene and sequence variation in the spacer region of the RubisCo operon. J Phycol, 37:121～ 129
[14] Giovannoni SJ, Britschgi TB, Moyer CL, et al. 1990. Genetic diversity in Sargasso Sea bacterioplankton. Nature, 345: 60～ 63
[15] Han X-R(韩秀荣),Wang X-L(王修林),Sun X(孙霞).2003.Nutrient distribution and its relationship with occurrence of red tide in coastal area of East China Sea.Chin Appl Ecol(应用生态学报),14(7):1097～1101(in Chinese)
[16] Hwang SR, Tabita R. 1991. Constraption, deduced primer structure, and expression of the chloroplast-encoded rbcL and rbcS genes of the marine diatom Cylindrotheca sp. strain N1. J Biol C hem,266: 6271 ～ 6279
[17] Kitano K, Maeda N, Fukui T, et al. 2001. Crystal structure of a novel-type archaeal RubisCo with pentagonal symmetry. Structure,59: 473 ～ 481
[18] Liu D-Y(刘东艳),Sun J(孙军),Zhang L-Y(张利永).2002.Structural characteristics of phytoplankton community during harmful algae bloomin Jiaozhou Bay.Chin J Appl Ecol(应用生态学报),14(11):1963～1966(in Chinese)
[19] Liu R-Y(刘瑞玉).1992.Ecology and Living Resources of Jiaozhou Bay. Beijing: Science Press. (in Chinese)
[20] Narang F, Mclntosh L, et al.1984. Nucleotide sequence of the ribulose bisphophophotphate carboxylase gene from Rhodospirillum rubrum.Mol Gen Genet,193:220～224
[21] Nei M and Kimura M. 2000. Molecular Evolution and Phylogenetics. New York:Oxford University Press.
[22] Oliveira MC, Bhattacharya D. 2000. Phylogenetic of theBangiophycidae (Rhodophyta) and the secondary endosymbiotic origin of algae plastids. Amer J Bot, 87(4) :482～492
[23] Paul JH, Cesares L, Thurmond J. 1990. Amplification of the rbcL gene from dissolved and particulate DNA from aquatic environments. Appl Environ Microbiol, 56:1963～ 1966
[24] Pichard SL, Campbell L, Carder K, et al. 1997. Analysis of ribulose bisphosphate carboxylase gene expression in natural phytoplankton communities by group-specific gene probing. Mar Ecol Prog Ser, 149:239～253
[25] Pichard SL, Frischer ME, Paul JH. 1993. Ribulose bisphosphate carboxylase gene expression in subtropical marine phytoplankton populations. Mar Ecol Prog Ser, 101:55～65
[26] Pichard SL, Campbell L, Pall JH. 1997. Diversity of the ribulose bisphosphate carboxylase/oxygenase form Ⅰ gene( rbcL)in natural phytoplankton communities. Appl Environ Microbiol, 63: 3600～3606
[27] Polne-Fuller M. 1991. A two-hour method for extraction of DNA from seaweeds. Phycol Newslet, 23: 2
[28] Rowan RS, Whitney M, Fowler A et al. 1996. RubisCo in marine symbiotic dinoflagellates: Form Ⅱ enzyme in eukaryotic oxygenic phototrophs encoded by a nuclear multigene family. Plant Cell, 8:539～553
[29] Shimada A, Kanai S, Maruyama T. 1995. Partial sequence of ribulose-1, 5-bisphosphate carboxylase/oxygenase and the phylogeny of Prochloron and Prochlorococcus sp. (Prochlorales). J Mol Evol,40:661～677
[30] Tabita FR. 1995. The biochemistry and metabolic regulation of carbon metabolism and CO₂ fixation in purple bacteria. In:Blankenship RE, Madigan MT, Bauer CE, eds. Anoxygenic Photosynthetic Bacteria. Dordrecht: Kluwer Acdemic Publishers. 885 ～ 914
[31] Walson GMF, Tabita FR. 1996. Regulation, unique gene organization, and unusual primary structure of carbon fixation genes from a marine phycoerythrin containing cyanobacterium. Plant Mol Biol,
[32] :1103～111632.Watson GMF, Tabita FR. 1977. Microbial ribulose 1, 5-bisphosphate carboxylsse/oxygenase: A molecule for phylogenetic and enzymological investigation. FEMS Microbiol Lett, 146:13 ～ 22
[33] Wyman M, Davies JT, Crawford DW, et al. 2000. Molecular and physiological responses of two classes of marine chromophytic phytoplankton(Diatoms and Prymnesiophytes)during the development of nutrient-stimulated blooms. Appl Environ Microbiol, 66: 2349～ 2357
[34] Xu HH, Tabita FR. 1996. Ribulose-1, 5-bisphosphate carboxylase/oxygenase gene expression and diversity of Lake Erie planktonic microorganism. Appl Environ Microbiol, 62:1913～ 1921
[35] Zhang H-W(张惠文),Zhang Q-R(张倩茹),Zhou Q-X(周启星).2003. Introduction and progress of molecular microbial ecology.Chin JAppl Ecol(应用生态学报),14(2):286～292(in Chinese)
[36] Zingone A, Enevoldsen HO. 2000. The diversity of harmful algal blooms: A challenge for science and management. Ocean Coastal Man, 43:725～748