| 摘要: |
| 利用ISSR-PCR方法对条斑紫菜 (Porphyra yezoensis)5个栽培品系进行遗传多态性分析,优化了PCR反应体系和反应参数。从100个ISSR引物中筛选23个引物扩增 了217个DNA片段,其中201个片段呈现多态性,占总扩增片段的92.6%。依据扩增结果进行遗传相似性和遗传距离分析,构建分子树状图。结果表明, 在条斑紫菜5个品系中,采自南通海区的2个品系首先聚在一起,其遗传距离为0.3897;接着依次和来自青岛的2个野生品系聚类,两个野生品系间的距离为 0.4299;原种来自日本的样品与其他4个样品相距最远,平均遗传距离为0.4469。同时,5个紫菜品系的ISSR分析中产生了一些特有的指纹图谱, 可进一步应用于种质分析的研究。 |
| 关键词: 条斑紫菜(Porphyra yezoensis) ISSR 遗传多样性 |
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| 基金项目:国家自然科学基金资助项目(40206019);江苏省高技术研究项目(BG2002312) |
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| Inter-simple sequence repeat (ISSR) analysis of five cultivars in Porphyra yezoensis |
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| Abstract: |
| 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: 25 μL system containing 20ng/μL template, 1.5 mmol/L MgC l2, 0.2 mmol/L dNTP, 0.3 μmol/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. |
| Key words: Porphyra yezoensis ISSR genetic diversity |
