三株赤潮硅藻5.8SrDNA及转录间隔区(ITS)的克隆及序列分析

张宝玉; 王广策; 吕颂辉; 齐雨藻; 邹景忠; 曾呈奎

三株赤潮硅藻5.8SrDNA及转录间隔区(ITS)的克隆及序列分析

1.
中国科学院海洋研究所, 山东青岛 266071;中国科学院海洋研究所, 北京 100089
2.
中国科学院海洋研究所, 山东青岛 266071
3.
暨南大学水生生物研究所, 广东广州 510632

基金项目: 国家自然科学基金资助项目(30170499);中国科学院知识创新重要方向性资助项目(KZCX2-211);"973"项目"我国赤潮高发区赤潮生物多样性及其种群分布特征"资助项目(2001CB409701)

计量
- 文章访问数: 863
- HTML全文浏览量: 27
- PDF下载量: 1137
- 被引次数: 0
出版历程
- 收稿日期: 2004-02-16
- 修回日期: 2004-08-19

The cloning and sequence analysis of the 5.8S rDNA and its region from three strains of marine diatoms

1.
Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;Graduate School, Chinese Academy of Sciences, Beijing 100039, China
2.
Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
3.
Institute of Hydrobiology, Jinan University, Guangzhou 510632, China

摘要

摘要: 对引发赤潮的3株硅藻——1株尖刺拟菱形藻(Pseudo-nitzshia pungens)和2株中肋骨条藻(Skeletonema costatum)的5.8SrDNA和ITS(internal transcribed spacers)序列进行了PCR扩增、克隆和序列测定,并分析了硅藻门10株赤潮藻(7株从GenBank获得)的系统进化关系.研究结果表明,尖刺拟菱形藻的ITS和5.8SrDNA的长度为693bp,SK-1(分离自东海赤潮暴发区)测序得到715bp,除ITS和5.8SrDNA外,还包含部分18SrDNA和28SrDNA;SK-2(分离自青岛养殖场)的ITS和5.8SrDNA的长度为331bp,尖刺拟菱形藻与从GenBank中获得的2株尖刺拟菱形藻相似程度最高,为100%,与该属的多列拟菱形藻相似程度稍低,为82.9%.SK-2的ITS序列与SK-1的相似程度很低,只有51%,但与拟中肋骨条藻的ITS序列相似程度高,为95.5%.SK-1的ITS序列与拟中肋骨条藻的相似程度也低,为56.7%.系统进化树反映的结果与相似性反映的结果一致.研究的该株尖刺拟菱形藻从根据ITS序列研究的结果与形态鉴定的结果看是一致的;SK-2可能属于拟中肋骨条藻;SK-1比较特殊,有待于用其他的方法进一步研究确定其分类地位.
- 硅藻 /
- 尖刺拟菱形藻 /
- ITS /
- 序列分析 /
- 系统进化
Abstract: Pseudo-nitzschia pungens and Skeletonema costatum are two strains of diatoms which cause harmful algal bloom in China.SK-1 and SK-2 are tw o strains of diatoms,assumably species of S.costatum by morphological characteristics.Three strains,SK-1 which was isolated from the East China Sea and SK-2 which was isolated from the Aquatic Farm of Qing dao and one strain of Pseudo-nitzschia pungens which was from the East China Sea,were st udied based on rDNA data.The sequences of 5.8S rDNA and ITS regions of three strains of diatoms were amplified,cloned and sequenced.Meanwhile,seven additional ITS sequences and 5.8S rDNA were included in the data analysis.The results were as follows:(1) The ITS sequence and 5.8S rDNA of P.pungens were 693 bp while the sequence of SK-1 was 715 bp,which contained complete ITS and 5.8S rDNA,partial 18S rDNA and 28S rDNA;The ITS and 5.8S rDNA sequences of SK-2 were 331bp;(2) the sequence of P.pungens had very high level of similarity with that of two strains of P.pungens retrieved from GenBank,and the similarity was 100%;while it showed 82.9% of identity compared with P.multiseries which also belonged to the genus of Pseudo-nitzschia;the ITS sequence similarity of SK-2 and SK-1 was very low,showing 51.5% of identity,but it had very high level of similarity with that of Skeletonema pseudocostatum(95.5%) ;the ITS sequence of SK-1 had lower similarity with S.pseudocostatum;(3) results of evolutionary tree coincided with similarity analysis of all three strains;it was suggested that recognition of P.pungens by morphological characteristics was supported with r DNA and ITS regions;(4) the findings demonst rated that classifications of SK-2 and SK-1 based on morphological criteria maybe were incongruous.SK-2 should be belonged to S.pseudocostatum and more data are needed to classify which groups SK-1 is belonged to.
- diatom /
- Pseudo-nitzschia pungens /
- ITS region /
- sequence analysis /
- phylogeny

HTML全文

参考文献(30)

BATESSS, GARRISON, HORNE R.Bloom dynamics and physiology of domoic-acid-producing Pseu do-nitzschia species [A].ANDERSON D M, et al.Physiological Ecology of Harmful Algal Blooms [C].Heidelberg:Springer-verlag, 1998.267-292.

杜琦.厦门西海域赤潮的研究[J].暨南大学学报(赤潮研究专刊),1989, 57-67.

洪君超, 黄秀清, 袁永进.长江口外及邻近海区赤潮多发区的分析与探讨[J].暨南大学学报(赤潮研究专刊),1989, 40-50.

方绍锦.浙江近海上升流调查区赤潮的初步探讨[J].海洋学报.1984, 6(3):408-414.

齐雨藻, 钱锋, 郑磊.中国东南沿海赤潮的生理生态学研究[A].朱明远, 李瑞香, 王飞.中国赤潮研究[M].青岛:青岛出版社,1996.6-15.

HASLE G R.Morphology and taxonomy of Skeletonema costatum (Bacillariophyceae) [J].Norw J Bot,1973, 20:109-137.

FUKUYOY.Morphology of Protogonyaulax tamarensis (Labour) Taylor and Protogonyau laxcatenella (Whed on and Kofoid) Tayl or from Japanese coastal waters[J].Bull Mar Sci, 1985, 37:529-537.

陈月琴, 屈良鹄, 曾陇梅, 等.南海赤潮有毒甲藻)-) 塔马亚历山大藻的分子鉴定[J].海洋学报, 1999, 21(3):106-111.

庄丽, 陈月琴, 李钦亮, 等.赤潮叉角藻18S rDNA 和ITS 区序列测定与分析[J].海洋与湖沼, 2001, 32(2):148-153.

ADACHIM, SAKOY, ISHIDA Y.Restriction fragment length polymorphism of ribosomal DNA internal transcribed spacerand 5.8S regions in Japanese Al exandr iumspecies (Dinoph yceae) [J].J Phycol, 1994, 30:857-863.

ADACH I M, SAKOY, ISHIDA Y.Analysis of Alexandrium (Din ophyceae) species using sequences of the 5.8Sribosomal DNA an dinternal transcribed spacer regions [J].J Phycol 1996, 32:424-432.

张宝玉, 王广策, 张炎, 等.东海原甲藻和海洋原甲藻APBM 的5.8SrDNA 及其转录间隔区(ITS) 的克隆和序列分析[J].海洋与湖沼, 2004, 35(3):264-272.

ZH ANG Zhao-duo, GREE NBRT, CAVALIERS.Single genecircles in din of lagel late chloroplast genomes [J].J Nature, 1999, 400:155-159.1 期张宝玉等:三株赤潮硅藻5.8S rDNA 及转录间隔区(ITS) 的克隆及序列分析115

王广策, 孙海宝, 曾呈奎.三角褐指藻磷酸甘油酸变位酶基因可能侧翼序列的筛选、克隆以及序列测定[J].海洋与湖沼, 2002, 33(3):259-264.

萨姆布鲁克J, 弗里奇E F, 曼尼阿蒂斯T.分子克隆实验指南[M],第3版.黄培堂等译.北京:科学出版社, 2002.96-105.

BALDWIN B G.Phylogene sticutility of the internal transcribed spacers of nuclear ribosomal DNA in plants:an exmaple from the vomposite[J].Mol Phylogen Evol, 1992, 1:3-16.

GONZALEZIL, SYLVESTER J E, SMITHTF, et al.Ribosomal RN Agene sequences and hominoid phylogeny [J].Mol Biol Evol ,1990, 7:203-219.

MORALES V M, PELCH ERE, TAYLOR J L.Composition of the 5.8SrDNA and internal transcribed spacer sequences of isolates of Leptosph aeria maculans from different pathogenicity groups [J].CurrGenet,1993, 23:490-495.

陈菊芳, 徐宁, 王朝晖, 等.大亚湾拟菱形藻(Pseud o-nit zschia spp.-种群的季节变化与环境因子的关系[J].环境科学学报, 2002,22(6):743-748.

左冬梅, 韩志国, 武宝酐.铁对尖刺拟菱形藻生长及光合作用的影响[J].暨南大学学报(自然科学与医学版),2002, 23(5):81-87.

虞秋波, 高亚辉.拟菱形藻软骨藻酸的研究进展[J].海洋科学, 2003, 27(8):26-29.

L UNDH OLM N, MOEST RUP O, H ASLE G R, et al.A study of the Pseudo-nitz schia pseud odelicati ssima/cuspidata complex (Bacil lariophy ceae):what is P.pseu dodelicatissima? [J].J Phycol, 2003, 39:797-813.

SCHOLIN A, ROMAN M Ⅲ, MILLER P E.DNA probes and a receptor-binding assay for detection of Pseudo-nitzschia (Bacil larioph yceae) species and domoicacid activity in cultured and naturals amples[J].J Phycol, 1999.35:1 356-1 367.

M IL LER P E, SCH OLIN C A.On det ect ion of Pseu do-nit z schi (Bacill arioph yceae) species using whole cell hybridization:sample fixati on and stability[J].J Phycol, 2000, 36:238-250.

CANGELOSIG A, HAMLIN A M, ROMAB M Ⅲ, et al.Detection of stable Pre-rRNA in toixgentic Pseudo-nitz schia species[J].Applied and Environmental Microbiology, 1997, 63(12):4 859-4 865.

DAUGBJERG N, H ANSEN G, LARSEN J,et al.Phylogeny of some of major genera of din of lagellates based on ultrastructure and partal LSUrDNA sequence data, including the erecti on of three new genera of unarmoured dinoflagellates [J].Phycologia, 2000, 39(4-:302-317.

陈慈美, 周慈由, 郑爱榕, 等.中肋骨条藻增殖的环境制约作用——Fe(Ⅲ) 与N、Mn,光、温交互作用对藻生化组成的效应[J].海洋通报,1996, 15(2):37-42.

陈雄文, 高坤山.CO₂浓度对中肋骨条藻的光合无机碳吸收和胞外碳酸酐酶活性的影响[J].科学通报.2003, 48(21):2 275-2 279.

KORB R E, SAVIKKE P J, JOHNSTON A M, et al.Sources of inorganic carbon for phot osynthesis by three species of marine diatom [J].J Phycol, 1997, 33:433-440.

MEDLIN L K, ELWOOD H J,SH AWN S, et al.Morp hological and genetic variation within the diatom Skelet onema costatum(Bacil lario lariophyta):evidence for a new species, Skeletonema pseudocostatum [J].J Phycol,1991, 27:514-524.

施引文献

资源附件(0)

访问统计