Ritchie N J, Myrold D D
Department of Crop & Soil Science, Oregon State University, Corvallis, Oregon 97331, USA.
Appl Environ Microbiol. 1999 Apr;65(4):1378-83. doi: 10.1128/AEM.65.4.1378-1383.1999.
Little is known about Ceanothus-infective Frankia strains because no Frankia strains that can reinfect the host plants have been isolated from Ceonothus spp. Therefore, we studied the diversity of the Ceonothus-infective Frankia strains by using molecular techniques. Frankia strains inhabiting root nodules of nine Ceanothus species were characterized. The Ceanothus species used represent the taxonomic diversity and geographic range of the genus; therefore, the breadth of the diversity of Frankia strains that infect Ceanothus spp. was studied. DNA was amplified directly from nodular material by using the PCR. The amplified region included the 3' end of the 16S rRNA gene, the intergenic spacer, and a large portion of the 23S rRNA gene. A series of restriction enzyme digestions of the PCR product allowed us to identify PCR-restriction fragment length polymorphism (RFLP) groups among the Ceanothus-infective Frankia strains tested. Twelve different enzymes were used, which resulted in four different PCR-RFLP groups. The groups did not follow the taxonomic lines of the Ceanothus host species. Instead, the Frankia strains present were related to the sample collection locales.
由于尚未从美洲茶属植物中分离出能够再次感染宿主植物的弗兰克氏菌菌株,因此对感染美洲茶的弗兰克氏菌菌株了解甚少。因此,我们利用分子技术研究了感染美洲茶的弗兰克氏菌菌株的多样性。对栖息于9种美洲茶属植物根瘤中的弗兰克氏菌菌株进行了表征。所使用的美洲茶属植物代表了该属的分类多样性和地理范围;因此,研究了感染美洲茶属植物的弗兰克氏菌菌株的多样性广度。通过PCR直接从根瘤材料中扩增DNA。扩增区域包括16S rRNA基因的3'端、基因间隔区和23S rRNA基因的大部分。对PCR产物进行一系列限制性酶切,使我们能够在所测试的感染美洲茶的弗兰克氏菌菌株中鉴定出PCR-限制性片段长度多态性(RFLP)组。使用了12种不同的酶,产生了4个不同的PCR-RFLP组。这些组并不遵循美洲茶宿主物种的分类界限。相反,所呈现的弗兰克氏菌菌株与样本采集地点有关。