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Molecular characterization of Xanthomonas species isolated from Araceae and the development of a triplex TaqMan assay for detection of Xanthomonas phaseoli pv. dieffenbachiae
European Journal of Plant Pathology ( IF 1.7 ) Pub Date : 2022-03-22 , DOI: 10.1007/s10658-022-02466-1
Jan van der Wolf 1 , Marjon Krijger 1 , Odette Mendes 1 , Balazs Brankovics 1 , Peter Bonants 1 , Leonie Didden 2 , Ellis Meekes 2
Affiliation  

In total 58 Xanthomonas strains isolated from Araceae worldwide, together with 13 other phylogenetically-related Xanthomonas strains, were characterized using multilocus sequence analysis based on concatenated sequences of seven single copy orthologous genes, extracted from whole genome sequences. The analysis revealed a monophyletic clade of 48 strains, 44 isolated from Anthurium, identified as X. phaseoli pv. dieffenbachiae (Xpd) confirmed by nucleotide identity analysis. The other strains from aroids were identified as Xanthomonas euvesicatoria (2 strains), X citri (5 strains) and Xanthomonas sacchari (3 strains). Two TaqMan assays were designed for specific detection of Xpd, one targeting sequences of a hypothetical protein and one targeting a type I restriction endonuclease subunit S. The two assays showed similar reaction kinetics and were merged with an assay comprising an amplification and extraction control into a triplex assay. The assay was able to detect minimally 100 copies of a target sequence delivered as a gBlock, 100 fg of genomic DNA and 104 cells per mL in an Anthurium leaf extract.



中文翻译:

从天南星科分离的黄单胞菌属物种的分子特征和用于检测黄单胞菌 pv 的三重 TaqMan 测定的开发。万年青

使用基于从全基因组序列中提取的七个单拷贝直系同源基因的串联序列的多位点序列分析,对从全球天南星科分离的总共 58株黄单胞菌菌株以及 13 种其他系统发育相关的黄单胞菌菌株进行了表征。分析揭示了 48 株的单系进化枝,其中 44 株从红掌中分离出来,被鉴定为X.phaseoli pv。通过核苷酸同一性分析证实了万年青(Xpd)其他来自拟南芥的菌株被鉴定为黄单胞菌(2 株)、柑橘黄单胞菌(5 株)和糖黄单胞菌。(3 株)。设计了两种 TaqMan 测定以特异性检测 Xpd,一种靶向假设蛋白质的序列,另一种靶向 I 型限制性内切酶亚基 S。两种测定显示出相似的反应动力学,并与包含扩增和提取对照的测定合并为三重测定。该测定能够在红掌叶提取物中检测至少 100 个作为 gBlock 传递的靶序列拷贝、100 fg 基因组 DNA 和每毫升10 4 个细胞。

更新日期:2022-03-22
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