Segregation of RAPD markers in F1 generation of Pinctada fucata family
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摘要:
对RAPD标记在合浦珠母贝F1代全同胞家系共27个样品中的分离方式进行了研究。从21条RAPD随机引物中筛选了8条引物用于扩增分析，结果共扩增出49个位点，其中正常位点47个(95.9%)，异常位点2个(4.1%)，即子代有扩增带而双亲无。47个正常位点中3个位点在F1中不分离(6.4%)，44个位点发生分离(93.6%，即多态位点)。卡方检验表明，44个分离位点中32个符合孟德尔分离规律(72.7%)，12个位点发生偏分离(27.3%)。符合孟德尔分离位点中按1：1分离位点占43.8%，3：1分离位点占56.2%，其中，父本特有标记、母本特有标记和双亲共有标记分别占28.2%，15.6%和56.2%，父本特有标记略多于母本特有标记。根据“双假测交”理论，1：1分离位点是构建F1代家系的作图位点。较高的1：1分离位点表明，利用RAPD标记构建合浦珠母贝遗传连锁图谱是可行的。
Abstract:The mode of segregation of random amplified polymorphic DNA (RAPD) markers in 27 F1 full siblings of Pinctada fucata from a single pair of parents was studied using eight 10-mer primers screened from 21. Of the total 49 loci amplified, 47 loci are normal, accounting for 95.9%, and two loci are unexpectedly amplified in F1 but not in parents. There are 44 loci segregating in the F1 individuals, accounting for 93.6%. Chi square tests indicate that the segregation of 12 loci are deviated significantly from Mendelian fashion, while the segregation of 32 loci are in line with Mendelian fashion, among which 1 : 1 segregating loci account for 43.8% and 3 : 1 segregating loci 56.2%. Among the 32 Mendelian segregation loci, paternal specific, maternal specific and biparental loci account for 28.2%, 15.6%和56.2%, respectively, indicating that the number of male specific markers is slightly larger than that of female specific markers. The high percentage of 1 : 1 segregating loci revealed in this study indicates that RAPD markers are useful in construction of linkage map of P.fucata using F1 family materials based on double pseudo-testcross configuration.
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Keywords:
- full sibling /
- F1 generation /
- Mendelian inheritance /
- genotype /
- polymorphism
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致谢: 华南农业大学水产系2005届毕业生伍珊珊参加了部分实验工作，谨此致谢！
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图 1 OPM14引物在合浦珠母贝亲本及其F1代的扩增与分离情况
0.DNA分子量标准(bp); 1.父本；2.母本；3~29.F1代个体
Figure 1. Amplification and segregation of RAPD markers in parents and F1 generation using primer OPM14
0.DNA ladder(bp); 1.male specific marker; 2.female speific marker; 3~29.F1 progenies
表 1 RAPD位点在F1代的分离与卡方检验
Table 1 Segregation of RAPD loci in F1 generation and Chi square test
编号
noRAPD标记
RAPD marker亲本扩增条带
parentsal bandsF1代期望比值
expected ratio in F1F1代观测值
observed value in F1卡平方值
Chi square values父本
♂母本
♀出现：缺失
presence：absence出现
presence缺失
absence1 OPM2-1359 + + 3：1 23 4 1.00 2 OPM2-1092 + + 3：1 19 8 0.11 3 OPM2-0960 + - 1：1 13 14 0.00 4 OPM2-00836 + - 1：1 22 5 9.48* 5 OPM2-0600 - - 异常分离/abnormal 10 17 - 6 OPM2-0530 + + 3：1 17 10 1.49 7 OPM14-1876 + + 3：1 18 9 0.60 8 OPM14-1705 + + 3：1 11 16 15.12* 9 OPM14-1272 + + 3：1 17 10 1.49 10 OPM14-1180 + + 3：1 22 5 0.31 11 OPM14-1030 + - 1：1 13 14 0.00 12 OPM14-0754 - + 1：1 11 16 0.59 13 OPM14-0668 + + 3：1 21 6 0.01 14 OPM14-0580 + - 1：1 16 11 0.59 15 OPM17-1753 + - 1：1 5 22 9.48* 16 OPM17-1046 + + 3：1 6 21 37.35* 17 OPM17-0836 + + 3：1 23 4 1.00 18 OPM17-0726 + - 1：1 16 11 0.59 19 OPM17-0622 + + 3：1 12 15 11.6* 20 OPM20-1351 + - 1：1 15 14 0.00 21 OPM20-0719 + + 子代不分离/no segregation 22 OPM20-0585 - + 1：1 23 6 8.83* 23 OPM20-0466 + + 子代不分离/no segregation 24 OPM20-0423 + + 3：1 22 7 0.01 25 S4-1542 + - 1：1 5 22 9.48* 26 S4-1327 + + 3：1 5 22 42.98* 27 S4-0940 + - 1：1 11 16 0.59 28 S4-0861 + + 3：1 21 6 0.01 29 S4-0668 + + 3：1 20 7 0.01 30 S4-0583 - + 1：1 18 9 2.37 31 S4-0423 - + 1：1 24 3 14.81* 32 S10-1763 + + 3：1 12 14 10.05* 33 S10-1583 - + 1：1 11 15 0.35 34 S10-1225 + - 1：1 12 14 0.04 35 S10-0828 + + 3：1 23 3 1.85 36 S10-0689 + + 3：1 22 4 0.82 37 S10-0308 + - 1：1 25 1 20.35* 38 S11-1093 + + 3：1 20 7 0.01 39 S11-0889 + - 1：1 13 14 0.00 40 S11-0600 + - 子代不分离/no segregation 41 S11-0485 + + 3：1 14 13 6.53* 42 S11-0376 - + 1：1 13 14 0.00 43 S11-0313 + + 3：1 20 7 0.01 44 S17-1307 + + 3：1 18 9 0.6 45 S17-1085 + + 3：1 22 5 0.31 46 S17-0936 + - 3：1 25 2 3.57 47 S17-0717 - + 1：1 19 8 3.70 48 S17-0500 + + 3：1 25 2 3.57 49 S17-0320 - - 异常分离/abnormal 11 16 - 注：*表示显著偏离孟德尔分离比例Note: * denotes significant deviation from Mendelian segregation ratio. 
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