2012 11, 19(6): 939 945 Journal of Fishery Sciences of China 研究论文 DOI: 10.3724/SP.J.1118.2012.00939 1, 赵永伟 2, 宋文涛 2, 廖小林 2, 牛余泽 2, 庞仁谊 2, 高峰涛 2, 沙珍霞 陈松林 2 1., 266109; 2.,, 266071 2, 摘要 :, (Cynoglossus semilaevis) F 1, 320 92 F 1, 288, 112 (P<0.05) 242, 21, 1 311.9 cm, 4.9 cm, 83.3%; 218, 21, 1 316.2 cm, 5.5 cm, 82% QTL : ; ; ; 中图分类号 : Q959 文献标志码 : A 文章编号 : 1005 8737 (2012)06 0939 07 (Cynoglossus semilaevis),,,,,,,,, [1],, [2],, AFLP [3] [4],,, QTL, Bostein [5],, (Ziphias gladius) (Danio retio) (Cyprinus carpio) (Oncorhynchus mykiss) (Ietalurus Punetaus) (Tilapia) (Oryzias latipes) (Sparus aurata) (Scophthalmus maximus) (Seriola lalandi) (Lates calarifer) (Paralichthys olivaceus) [6 21] 收稿日期 : 2011 12 16; 修订日期 : 2012 02 07. 基金项目 : (200903046);. 作者简介 : (1986 ),,. 通信作者 :,,,. E-mail: chensl@ysfri.ac.cn
940 19, Liao [22] 103 AFLP 33 1 26, 934.6 cm, 8.4 cm;,, PCR PAGE, JoinMap4.0, 288, 1311.9 cm 1 316.2 cm, 4.9 cm 5.5 cm,,, 1 1.1 2008 6 F 1, 2009 10 200, 8,,, 92, 1.2, 1 000 1.3 DNA DNA, 1% (Pharmacia Biotech Ltd.) DNA, DNA 100 ng/μl, 20 PCR PAGE PCR : DNA genomic DNA 1.0 μl; dntps (2.5 mmol/l) 1.0 μl10 PCR buffer ( Mg 2+ ) 1.5 μl; (10 μmol/l) upstream primer 0.5 μl; (10 μmol/l)downstream primer 0.5 μl; Taq DNA polymerase (5 U/μL) 0.1 μl; ddh 2 O 15.4 μl; 20 μl PCR : 95 4 min 1 ; 95 35 s, 57.5 35 s, 72 35 s, 32 ; 72 7 min 1 ; 4 PCR DYCZ-20C : 1 300 V, 150 W, 150 ma 3 h JoinMap4.0 χ 2 1:1,, (P<0.05), CP(cross-pollinating), Locus genot. freq. Exclude selected items, Individual genot. freq., Similarity of individuals (the value>0.95), Similarity of loci (the value=1), Grouping tree Calculate (LOD 3.0) Kosambi (centimorgan, cm),, 2 2.1 SSR 1 000 SSR,, 362, 36.2% 320, 1
6 : 941 1 1 1~46 47~92 F 1, (, ). Fig. 1 The result of amplification of one pair primer in mapping population Lanes 1 46 and 47 92 on the two sides are F 1 individuals respectively; parents are on the middle(left, right ), 1~46 47~92 F 1, (, ),, 1, BC AD, F 1 AB AC BD CD 4 19 ( HK HK), 12 (u<0.05), ; 53, 18 ; 71, 16 ; 157, 80 ; (F-382, F-783) 2.2 320 SSR, 288, 242 214, 21 18.7~111.9 cm, 5~29 ( 1, 2) sex female male number of linkage groups Tab. 1 total number of markers 表 1 半滑舌鳎遗传连锁图谱的特点 The characterization of Cynoglossus semilaevis genetic linkage map number of markers per group /cm average marker interval Gof/cM observed consensus map length Goa/cM estimated consensus map length / % coverage 21 242 11.7 4.9 1092.6 1311.9 83.3 21 214 10.8 5.5 1079.2 1316.2 82.0 3 3.1,, [23 24], SSR, SSR ( 2), 5~29,, 14 41 cm, 4 5 9 17 19,,,,, AFLP SNP, 114, 96, 39.7%, 95, 43.5%, (Ictalurus punctatus R.) [25] (Crassostrea gigas T.) [26], (Cynoglossus semilaevis) [22]
942 第 19 卷 中国水产科学 记的顺序 [28] 分析引起偏分离现象原因, 可能是 生物的遗传连锁图谱 [29 31] 由于半滑舌鳎性成熟 由实验统计误差 取样的随机性以及隐形致死突 需要 2 年时间, 难以建立高世代或回交家系群体, 变的选择等因素所引起的 因此本研究运用 拟测交 策略, 仅用亲代和 F1 构 3.2 作图群体的选择 建其遗传连锁图谱, 必将加快遗传育种改良的 本实验的亲本材料选择中, 作图群体的父本 进程 为渤海近海野生个体, 母本来自于烟台海阳黄海 3.3 图谱分析及评价 水产有限公司人工养殖的半滑舌鳎, 这种亲本组 Liao 等 [22]用 60 个半滑舌鳎 F1 个体建立了半 合, 有利于增加子代群体之间的遗传差异, 同时 滑舌鳎第一代遗传连锁图谱, 其中包含 103 个 选择家系间亲本间遗传差异性较大的家系作为作 AFLP 标记 33 个微卫星标记和 1 个雌性特异的 图家系, 有利于获得更多的多态性标记和遗传信 标记, 该图谱由 26 个连锁群组成, 图谱总长度为 息量, 从而能有效地提高作图效率 934.6 cm, 标记间平均间隔为 8.4 cm, 图谱覆盖 近年来 拟测交 理论 [28] 已广泛用于构建水生 图2 Fig. 2 率为 64.4%; 然而本研究中用了 92 个 F1 个体, 构 半滑舌鳎遗传连锁图谱(待续) Genetic linkage map of Cynoglossus semilaevis(to bo continued)
6 : 943 ( ) 2,. (continued)fig. 2 Genetic linkage map of Cynoglossus semilaevis The adjacent marker spacing is displayed on the left in cm Kosambi and SSR markers are shown on the right of each group.
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