Gnat Sebastian, Małek Wanda, Oleńska Ewa, Trościańczyk Aleksandra, Wdowiak-Wróbel Sylwia, Kalita Michał, Wójcik Magdalena
Department of Veterinary Microbiology, University of Life Sciences, 12 Akademicka st., 20-950, Lublin, Poland.
Department of Genetics and Microbiology, University of Maria Curie-Skłodowska, 19 Akademicka st., 20-033, Lublin, Poland.
J Appl Genet. 2015 Nov;56(4):551-554. doi: 10.1007/s13353-015-0285-6. Epub 2015 May 1.
We assessed the genomic diversity and genomic relationship of 28 Astragalus glycyphyllos symbionts by three methodologies based on PCR reaction, i.e., RAPD, ERIC-PCR, and AFLP. The AFLP method with one PstI restriction enzyme and selective PstI-GC primer pair had a comparable discriminatory power as ERIC-PCR one and these fingerprinting techniques distinguished among the studied 28 A. glycyphyllos symbionts 18 and 17 genomotypes, respectively. RAPD method was less discriminatory in the genomotyping of rhizobia analyzed and it efficiently resolved nine genomotypes. The cluster analysis of RAPD, ERIC-PCR, and AFLP profiles resulted in a generally similar grouping of the test strains on generated dendrograms supporting a great potential of these DNA fingerprinting techniques for study of genomic polymorphism and evolutionary relationship of A. glycyphyllos nodulators. The RAPD, ERIC-PCR, and AFLP pattern similarity coefficients between A. glycyphyllos symbionts studied was in the ranges 8-100, 18-100, and 23-100%, respectively.
我们通过基于PCR反应的三种方法,即随机扩增多态性DNA(RAPD)、肠杆菌基因间重复共有序列PCR(ERIC-PCR)和扩增片段长度多态性(AFLP),评估了28株甘草黄芪共生菌的基因组多样性和基因组关系。使用一种PstI限制酶和选择性PstI-GC引物对的AFLP方法与ERIC-PCR方法具有相当的鉴别能力,这些指纹技术分别在研究的28株甘草黄芪共生菌中区分出了18种和17种基因型。RAPD方法在分析根瘤菌的基因分型中鉴别能力较弱,它有效地分辨出了9种基因型。对RAPD、ERIC-PCR和AFLP图谱的聚类分析在生成的树形图上得出了测试菌株总体相似的分组,这支持了这些DNA指纹技术在研究甘草黄芪根瘤菌基因组多态性和进化关系方面具有巨大潜力。所研究的甘草黄芪共生菌之间的RAPD、ERIC-PCR和AFLP模式相似系数分别在8 - 100%、18 - 100%和23 - 100%的范围内。