Kaemmer D, Afza R, Weising K, Kahl G, Novak F J
University of Frankfurt/Main, Department of Biology, Germany.
Biotechnology (N Y). 1992 Sep;10(9):1030-5. doi: 10.1038/nbt0992-1030.
DNA oligonucleotide and amplification fingerprinting have been successfully used to detect genetic polymorphisms in 15 representative species and cultivars of the genus Musa, comprising AA, AAA, AAAA, AAB, ABB, and BB genotypes. In-gel-hybridization of Hinf I-digested genomic banana DNA to the 32P-labeled synthetic oligonucleotides (GATA)4, (GTG)5, and (CA)8 revealed considerable polymorphisms between Musa species and cultivars. The fingerprint patterns proved to be somatically stable and did not show differences between individual plants of 'Grand Nain' (AAA genotype). Dendrograms based on oligonucleotide fingerprint band sharing data proved to be consistent with most of the known features of the history of banana and plantain cultivation and evolution, respectively. DNA samples from the same banana species and cultivars were also amplified by PCR using single or pairwise combinations of short oligonucleotide primers. Amplification products were separated on agarose or polyacrylamide gels and visualized by ethidium bromide or silver staining, respectively. Polymorphic patterns were obtained with some but not all primers. By using the CCCTCTGCGG primer in simplex and/or duplex PCR, the induced mutant 'GN60A' was clearly recognized from its original variety 'Grand Nain'. Both fingerprint techniques allowed the detection of bands characteristic for the A and B genome. This DNA fingerprinting technology has potential application in several areas of Musa improvement.
DNA寡核苷酸和扩增指纹技术已成功用于检测芭蕉属15个代表性物种和品种的遗传多态性，这些物种和品种包括AA、AAA、AAAA、AAB、ABB和BB基因型。用Hinf I酶切的香蕉基因组DNA与32P标记的合成寡核苷酸（GATA）4、（GTG）5和（CA）8进行凝胶内杂交，结果显示芭蕉属物种和品种之间存在相当大的多态性。指纹图谱在体细胞水平上是稳定的，‘贵妃蕉’（AAA基因型）的不同单株之间未表现出差异。基于寡核苷酸指纹条带共享数据构建的聚类图分别与香蕉和大蕉栽培及进化历史的大多数已知特征相符。来自相同香蕉物种和品种的DNA样本也使用短寡核苷酸引物的单引物或双引物组合通过PCR进行扩增。扩增产物分别在琼脂糖凝胶或聚丙烯酰胺凝胶上进行分离，然后分别用溴化乙锭或银染法进行可视化。使用部分而非全部引物可获得多态性图谱。通过在单重和/或双重PCR中使用CCCTCTGCGG引物，可从其原始品种‘贵妃蕉’中清晰识别出诱变突变体‘GN60A’。两种指纹技术都能检测到A和B基因组特有的条带。这种DNA指纹技术在芭蕉属植物改良的多个领域具有潜在应用价值。
