Inter-simple sequence repeat (ISSR) analysis of five cultivars in Porphyra yezoensis
Inter-simple sequence repeat (ISSR) analysis of five cultivars in Porphyra yezoensis
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发表时间：
2006
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影响因子：
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通讯作者：
X. Pu
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作者：
X. Pu
采用ISSR-PCR技术对条斑紫菜5个品种的基因组DNA变异及亲缘关系进行了分析。研究了影响ISSR的因素，并对实验参数进行了优化。最佳实验条件为：25μL体系，含20 ng/μL模板，1.5μ/L氯化镁，0.2 mmol/L dNTP，0.3 mol/L引发剂。优化的扩增程序为94℃下1个循环5 m in，94℃下45个循环，52℃下45 s，72℃下2 m in，72℃下1个10 m in。共扩增出217条DNA条带，其中201条为多态，占92.6%。聚类分析结果表明，Y-9965与Y-9505的遗传距离为0.3897，与青岛的Y-9308、Y-2035的遗传距离为0.4299，而来自日本的Y-0008与其他4个样品的平均遗传距离为0.4469。同时，基于ISSR标记的DNA图谱揭示了不同品种的潜在指纹。
Inter-simple Sequence Repeat (ISSR)-PCR was used to determine the genomic DNA variations and genetic relationships of 5 cultivars of Porphyra yezoensis. The influence factors of ISSR were studied and the experiment parameters were optimized. The optimal experiment conditions were as follows: 25μL system containing 20ng/μL template, 1.5mmol/L MgCl2, 0.2mmol/L dNTP,0.3μmol/L primer. Optimal amplification program was 1 cycle of 5 min at 94℃; 45 cycles of 45s at 94℃,45s at 52℃,and 2 min at 72℃; 1 cycle of 10 min at 72℃.One hundred ISSR primers were screened, of which 23 polymorphic and informative primers were selected. Total 217 DNA bands were amplified, 201 of which were polymorphic (92.6%). Cluster analysis based on the ISSR results was performed, which indicated that genetic distance between Y-9965 and Y-9505 was 0.3897; and the distance between the two samples from Qingdao (Y-9308,Y-2035) was 0.4299; further more, the cultivar come from Japan (Y-0008) had the average genetic distance 0.4469 compared with other four samples. At the same time, DNA profiles based on the ISSR markers had revealed potential fingerprints for various individual cultivars.