香蕉种质资源的AFLP鉴别与分类中DNA模板的制备

在运用AFLP技术进行香蕉种质资源的鉴别与分类时,成功的关键之一是HMW基因组DNA的成功制备和避免降解,为了获得高纯度的HMW香蕉模板DNA和清晰的AFLP指纹图谱,以粉蕉为试材进行了用不同方法从不同植物体部位提取香蕉DNA的研究。结果表明,以香蕉未展开的幼叶、成熟叶、新根为材料提取的DNA量足以进行AFLP分析,但考虑到取材的方便性,最好选用未展开的幼叶或成熟叶;用改进后的SDS、CTAB法均可以获得HMW基因组DNA和AFLP指纹图谱,SDS法(用氯仿异-戊醇溶液抽提3次)提取的DNA量将近是CTAB法提取的DNA量的2倍,但SDS法提取的DNA纯度不如CTAB法提取的高。两种方法提取的DNA片段大小一致,大约在16kb左右。

Making Template DNA in the Study of Identification and Classification of Banana Germplasm Based on AFLP Fingerprint

One of the key factors for AFLP marker is the successful preparation of HMW template DNA.In order to obtain pure DNA and clear AFLP fingerprint for the further study of identification and classification for banana,extraction of tem-plate DNA from different plant organs of banana was studied.The results showed that the HMW template DNA could be ob-tained from plant organs including white young roots,outspread young leaves and mature leaves.It was more convenient to choose young and mature leaves as plant organs for the excellent quality genome DNA extraction.The results also showed that the HMW tem plate DNA obtained by using SDS and CTAB method were suitable for banana AFLP analy sis.The quantity of DNA extracted by using SDS method was two times as much as that by using CTAB method,but the degree of purity was low-er than using CTAB method.No difference of the molecular weight of DNA extracted by using SDS and CTAB methods was noted(all around16kb).