[Analysis of genetic diversity in wild populations of Trachidermus fasciatus by RAPD and the transformation of two SCAR markers].
[Analysis of genetic diversity in wild populations of Trachidermus fasciatus by RAPD and the transformation of two SCAR markers].
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DOI：
10.3724/sp.j.1141.2012.02203
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发表时间：
2013-01
期刊：
影响因子：
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通讯作者：
Zhen Zeng;Zhi-zhi Liu;Lian-de Pan;Wenqiao Tang;Qian Wang;Yun Geng
中科院分区：
文献类型：
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作者：
Zhen Zeng;Zhi-zhi Liu;Lian-de Pan;Wenqiao Tang;Qian Wang;Yun Geng
首先，从294个随机引物中筛选出32个具有多态性的10-bp随机引物，对富春江种群(FR)、黄河种群(YR)、滦种群(LR)和鸭绿江种群(YL)4个群体中的扁尾棘皮鱼进行了遗传多样性分析。在每个种群中检测到30只野生个体。结果表明，T.筋膜吸虫相对较丰富。主要研究结果如下：1)共检测到591条谱带，其中多态性谱带515条，占87.14%。多态基因座比例范围(P)为：FR(89.17%)>；YR(87.99%)>；YL(86.63%)>；LR(83.25%).2)群体间的Shannon‘s信息指数(I(T))和Nei’s遗传多样性(H(T))分别为0.3393~0.3566和0.2157~0.2279。与其他三个群体相比，LR群体的值相对较低。总体遗传多样性H(T)为0.2336±0.1643，信息指数I(T)为0.3710±0.2153。3)基因流(N(M))值较高(5.76103~19.84497)，表明四个种群之间存在一定的基因交换。但AMOVA结果在群体间存在显著差异(P<；0.05或P<；0.01)。4)在根据遗传距离构建的UPGMA树中，YL和YR群体先聚类，然后与FR群体聚在一起，最后与LR群体聚在一起。由此可见，YL、YR和FR种群在地理距离上具有较近的亲缘关系，而LR种群与其他3个种群表现出明显的分化。其次，在S(1225)、S(1345)、S(695bp)和S(1345)(825bp)的5条特异扩增带中，S(1255)(605bp)和S(1255)(841bp)分别成功转化了SCAR01(560bp)和SCAR02(443bp)。对两个标记进行大样本检测后发现，YL人群的频率最高(96.67%和93.33%)，FR人群的频率较高(83.33%和90%)，YR人群的频率较高(56.67%和66.67%)，LR人群的频率最低(13.33%和20%)。因此，SCAR01(560个碱基)和SCAR02(443个碱基)可作为群体识别的特殊分子标记。
Firstly, RAPD was conducted to analyze genetic diversity of Trachidermus fasciatus in the Fuchun River population (FR), Yellow River population (YR), Luan River population (LR), and Yalu River population (YL), with 32 polymorphic 10-bp random primers selected from 294 ones. Thirty wild individuals were detected in each population. The results indicated that the genetic diversity of T . fasciatus was relatively rich. The major results were as the following: 1) Altogether, 591 bands were detected and 515 of them were polymorphic, accounted for 87.14%. The range of proportion of polymorphic loci (P) was: FR(89.17%)>YR(87.99%)>YL(86.63%)>LR(83.25%). 2) The Shannon's information index(I(T)) and Nei's genetic diversity(H(T)) among populations were 0.3393-0.3566 and 0.2157-0.2279, respectively. Compare to other three populations, LR population had relative lower values. If took the populations as a whole, the total Nei's genetic diversity(H(T)) and Shannon's information index(I(T)) was 0.2336±0.1643 and 0.3710±0.2153, respectively. 3) The value of gene flow (N(m) ) (5.76103-19.84497) were high, indicating certain gene exchange existed among the four populations. But the AMOVA results exhibited significantly differentiation (P<0.05 or P<0.01) among the populations. 4) In the UPGMA tree constructed according to genetic distance, YL and YR populations clustered firstly, then with FR population, and finally they joined to LR population. Obviously, the YL, YR and FR populations had relatively close relationship according to their geographic distance, whereas LR population showed clear divergence to the other three populations. Secondly, out of the five special RAPD bands (S(1225)(525 bp), S(1225)(605 bp), S(1225)(841 bp), S(1345)(695 bp) and S(1345)(825 bp)), SCAR maker SCAR01(560 bp) and SCAR02(443 bp) were successfully transformed from S(1255)(605 bp) and S(1255)(841 bp), respectively. After large samples examination of the two markers, we found the highest frequency (96.67% and 93.33%) in the YL population, higher frequency (83.33% and 90%) in the FR population, high frequency (56.67% and 66.67%) in the YR population, and the lowest frequency (13.33% and 20 %) in the LR population. Therefore, SCAR01(560 bp) and SCAR02(443 bp) can be used as special molecular markers for the population identification between LR and other three populations.