Matas Isabel M, Pérez-Martínez Isabel, Quesada José M, Rodríguez-Herva José J, Penyalver Ramón, Ramos Cayo
Area de Genética, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos s/n, E-29071 Málaga, Spain.
Appl Environ Microbiol. 2009 Feb;75(4):1030-5. doi: 10.1128/AEM.01572-08. Epub 2008 Dec 19.
In this study, Pseudomonas savastanoi pv. savastanoi isolates were demonstrated to contain two iaaL paralogs, which are both chromosomally located in most strains. Comparative analysis of iaaL nucleotide sequences amplified from these two paralogs revealed that one paralog, iaaL(Psn), is 100% identical to iaaL from P. savastanoi pv. nerii, while the other paralog, iaaL(Psv), exhibited 93% identity to iaaL from Pseudomonas syringae pv. tomato (iaaL(Pto)). A 3-nucleotide motif (TAC) comprised of 3 to 15 repeats, which remained stable after propagation of the strains in olive plants, was found in iaaL(Psv). Based on the observed nucleotide sequence variations, a restriction fragment length polymorphism assay was developed that allowed differentiation among iaaL(Psn), iaaL(Psv), and iaaL(Pto)(.) In addition, reverse transcriptase PCR on total RNA from P. savastanoi pv. savastanoi strains demonstrated that both iaaL(Psv) and iaaL(Psn) containing 14 or fewer TAC repeats are transcribed. Capillary electrophoresis analysis of PCR-amplified DNA fragments containing the TAC repeats from iaaL(Psv) allowed the differentiation of P. savastanoi pv. savastanoi isolates.
在本研究中,证明了野油菜黄单胞菌野油菜致病变种(Pseudomonas savastanoi pv. savastanoi)分离株含有两个iaaL旁系同源基因,在大多数菌株中它们均位于染色体上。对从这两个旁系同源基因扩增的iaaL核苷酸序列进行比较分析发现,一个旁系同源基因iaaL(Psn)与野油菜黄单胞菌丁香致病变种(P. savastanoi pv. nerii)的iaaL完全相同,而另一个旁系同源基因iaaL(Psv)与丁香假单胞菌番茄致病变种(Pseudomonas syringae pv. tomato)的iaaL(iaaL(Pto))有93%的同一性。在iaaL(Psv)中发现了一个由3至15个重复序列组成的3核苷酸基序(TAC),该基序在菌株于橄榄植株中传代后保持稳定。基于观察到的核苷酸序列变异,开发了一种限制性片段长度多态性分析方法,可区分iaaL(Psn)、iaaL(Psv)和iaaL(Pto)。此外,对野油菜黄单胞菌野油菜致病变种菌株的总RNA进行逆转录PCR表明,含有14个或更少TAC重复序列的iaaL(Psv)和iaaL(Psn)均被转录。对含有来自iaaL(Psv)的TAC重复序列的PCR扩增DNA片段进行毛细管电泳分析,可区分野油菜黄单胞菌野油菜致病变种分离株。