Cigna Jérémy, Dewaegeneire Pauline, Beury Amélie, Gobert Virginie, Faure Denis
Seed Innovation Protection Research Environment, Comité Nord-SIPRE, 62217 Achicourt, France; and Institute for Integrative Biology of the Cell, CNRS CEA Univ. Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette, France.
Seed Innovation Protection Research Environment, Comité Nord-SIPRE, 62217 Achicourt, France.
Plant Dis. 2017 Jul;101(7):1278-1282. doi: 10.1094/PDIS-12-16-1810-RE. Epub 2017 May 3.
Several pectinolytic Pectobacterium and Dickeya species and subspecies are causative agents of blackleg and soft rot diseases on potato plants and tubers. Rapid and accurate identification of these taxa is a crucial issue for the production and international trade of potato seed-tubers. Here, we developed a PCR-sequencing tool to easily characterize the different Pectobacterium and Dickeya taxa. The gapA gene sequences from 53 published genomes were aligned and a phylogeny tree was constructed. A set of 35 signature nucleotides was discovered to distinguish the Pectobacterium and Dickeya genera, species, and subspecies. Then, a PCR-primer couple was designed for amplifying the gapA gene in pectinolytic enterobacteria. The primers were tested on 22 isolates recovered from blackleg symptoms in several potato fields. Amplicons were sequenced and signature-nucleotides were analyzed. A phylogeny that includes gapA sequence specimens confirmed the taxonomical identification of these environmental isolates.
几种能产生果胶酶的果胶杆菌属和迪基氏菌属的菌种及亚种是马铃薯植株和块茎黑胫病和软腐病的病原体。快速准确地鉴定这些分类单元对于马铃薯种薯的生产和国际贸易而言是一个关键问题。在此，我们开发了一种PCR测序工具，以便轻松地对不同的果胶杆菌属和迪基氏菌属分类单元进行特征描述。对来自53个已发表基因组的gapA基因序列进行比对，并构建了系统发育树。发现了一组35个特征性核苷酸，用于区分果胶杆菌属和迪基氏菌属、菌种及亚种。然后，设计了一对PCR引物，用于扩增能产生果胶酶的肠杆菌中的gapA基因。这些引物在从几个马铃薯田的黑胫病症状中分离出的22个菌株上进行了测试。对扩增子进行测序并分析特征性核苷酸。包含gapA序列样本的系统发育树证实了这些环境分离株的分类鉴定。
