收稿日期: 2012-04-07
修回日期: 2012-06-05
网络出版日期: 2012-10-24
基金资助
国家“十一五”科技支撑计划项目(2007BAD29B03); 国家自然科学基金(30972253); 广东省自然科学基金(8452408801001272, 10452408801004224); 广东省科技计划项目(2010B020308004)
Study on the AFLP primer selection and genetic diversities of 4 Snappers
Received date: 2012-04-07
Revised date: 2012-06-05
Online published: 2012-10-24
以湛江近海的勒氏笛鲷Lutjanus russellii、紫红笛鲷L. argentimaculatus、红鳍笛鲷L. erythropterus、千年笛鲷L. sebae为研究对象, 采用EcoRⅠ/MselⅠ双酶切组合, 对4种笛鲷进行扩增片段长度多态性(amplified fragment length polymorphism, AFLP)标记的开发和筛选, 并对其进行遗传多样性分析。从50对引物中筛选适合4种笛鲷的AFLP引物, 其中勒氏笛鲷筛选出24对, 获得位点1431个, 多态位点比例为22.85%; 紫红笛鲷筛选出20对, 获得位点1370个, 多态位点比例为17.08%; 红鳍笛鲷筛选出22对, 获得位点1403个, 多态位点比例为17.18%; 千年笛鲷筛选出22对, 获得位点1349个, 多态位点比例为12.90%。Nei氏遗传距离法计算每种笛鲷30个个体之间的平均遗传距离, 勒氏笛鲷为0.1035, 紫红笛鲷为0.0719, 红鳍笛鲷为0.0805, 千年笛鲷为0.0572。选取2对引物对4种笛鲷群体间遗传多样性进行分析, 获得总位点140个, 多态位点104个。4种笛鲷种群间遗传距离分布在0.3773—0.6650, 明显高于各笛鲷种群内遗传距离, 其中紫红笛鲷和千年笛鲷的遗传距离最远, 为0.6650, 勒氏笛鲷和红鳍笛鲷的遗传距离最近, 为0.3773。
刘丽 , 赵洁 , 郭昱嵩 , 刘楚吾 . 4种笛鲷AFLP引物筛选及其遗传多样性分析[J]. 热带海洋学报, 2012 , 31(4) : 112 -116 . DOI: 10.11978/j.issn.1009-5470.2012.04.015
4 species of snappers (Lutjanus russellii, L.erythropterus, L.argentimaculatus and L.sebae) were analyzed in the study using the AFLP technique with restriction endonuclease of EcoRⅠ/MselⅠmix, so as to develop AFLP Markers and analyze genetic diversity. 50 pairs of AFLP (amplified fragment length polymorphism) primers were used, 24 of which were selected for L. russellii, 20 for L. argentimaculatus, and 22 for both L. erythropterus and L. sebae, the polymorphism sites were 1431, 1370, 1403 and 1349 in turn, their proportion separately was 22.85%, 17.08%, 17.18% and 12.90%. The Nei’s genetic diversity was analyzed in 4 snapper species, the average genetic distance was 0.1035, 0.0719, 0.0805 and 0.0572, respectively. AFLP fingerprinting of 4 snappers with 2 primers (A4, C4) revealed a total number of 140 sites, of which 104 ones were polymorphic. Genetic distance among the 4 snappers ranged from 0.3773 to 0.6650, which was significantly higher than the ones within species. The genetic distances and phylogenic trees both show that L.russellii and L.erythropterus are closest and L.argentimaculatus and L.sebae are furthest.
VOS P, HOGERS R, BLEEKER M, et al. AFLP: A new technique for DNA fingerprinting[J]. Nucleic Acids Res, 1995, 23(21): 4407-4414.
WANG Y, YIN G, YANG Q, et al. Identification and isolation of Mu-flanking fragments from maize[J]. J Genet Genomics, 2008, 35 (4): 207-213.
马晋, 李涛, 龙文敏, 等.中华稻蝗不同地理种群遗传多样性的AFLP分析[J]. 遗传, 2010, 32(2): 163-169.
LI Z, HAINES C J, HAN Y.“Micro-deletions” of the human Y chromosome and their relationship with male infertility[J]. J Genet Genomics, 2008, 35(4): 193-199.
初冠囡, 秦艳杰, 李霞, 等.中间球海胆生长分化相关的AFLP标记[J]. 中国水产科学, 2010, 17(3): 448-454.
代悦, 喻子牛, 赵晓霞, 等.马氏珠母贝4个壳色选育系F1遗传结构的AFLP分析[J]. 基因组学与应用生物学, 2010, 29(2): 252-258.
李飞, 贾智英, 闵霞, 等.鲤鱼cDNA-AFLP技术反应体系的建立[J]. 基因组学与应用生物学, 2011, 30(1): 122-128.
毕承武, 吴初新, 谭婉珑, 等. AFLP分析江西省4个斑点叉尾鮰养殖群体遗传多样性[J]. 基因组学与应用生物学, 2010, 29(5): 871-878.
周遵春, 包振民, 董颖, 等.中间球海胆、光棘球海胆及杂交F-1代(中间球海胆♀×光棘球海胆♂)群体遗传多样性AFLP分析[J].遗传, 2007, 29(4): 443-448.
刘卫东, 鲍相渤, 宋文涛, 等. 虾夷扇贝遗传连锁图谱的初步构建[J].遗传, 2009, 31(6): 629-637.
张俊彬, 黄良民.紫红笛鲷遗传多样性的AFLP分析[J].热带海洋学报, 2004, 23(5): 50-55.
张俊彬, 黄良民, 陈真然.AFLP技术在笛鲷的仔鱼鉴定及其分类学上的研究[J]. 海洋学报: 中文版, 2005, 27(2): 165-171.
DORENBOSCH M, POLLUX B J A, PUSTJENS A Z, et al. Population structure of the Dory snapper, Lutjanus fulviflamma, in the western Indian Ocean revealed by means of AFLP fingerprinting[J]. Hydrobiologia, 2006, 568(1): 43-53.
成庆泰, 郑葆珊. 中国鱼类检索系统: 下册[M].北京. 科学出版社, 1987: 331-1119.
中国科学院南海海洋研究所. 中国海洋鱼类原色图集[M]. 上海: 上海科学出版社, 1992: 105.
卢圣栋.现代分子生物学实验技术[M]. 北京: 中国协和医科大学出版社, 1999: 61.
NEI M, LI W H.Mathematical model for studying genetic variation in terms of restriction endonucleases[J]. Proc Natl Acad Sci, 1979, 76(10): 5269–5273.
JOHNSON M S, HEBBERT D R, MORAN M J. Genetic analysis of population of north- western Australian fish species[J]. Aust J Mar Freshwat Res, 1993, 44(5): 673-685.
GOLD J R, RICHARDSON L R. Mitochondrial DNA variation among ‘red’ fishes from the Gulf of Mexico[J]. Fish Res, 1994 , 20(2-3): 137-150.
CAMPER J D, BARBER R C, RICHARDSON L R, et al. Mitochondrial DNA variation among red snapper (Lutjanus campechanus) from the Gulf of Mexico[J]. Mol Mar Biol Biotechnol, 1993, 2(3): 154-161.
丁少雄, 王军, 全成干, 等.鮸状黄姑鱼养殖群体的遗传多样性[J].科学通报, 1998, 43(21): 2294-2299.
TOHME J, GONZALEZ O D, BEEBE S, et al. AFLP analysis of gene pools of a wild bean core collection[J]. Crop Science, 1996, 36: 1375-1384.
周发林, 江世贵, 苏天凤, 等. 6种笛鲷属鱼类线粒体16S rRNA基因片段的序列比较[J]. 中国水产科学, 2004, 11(2): 99-103.
郭昱嵩, 王中铎, 刘楚吾, 等. 9种常见笛鲷微卫星位点筛选与遗传多样性分析[J]. 热带海洋学报, 2010, 29(3): 82-86.
郭昱嵩, 刘楚吾.南海海域4种笛鲷的微卫星DNA分析[J]. 广东海洋大学学报, 2007, 27(6): 7-12.
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