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利用从中国酸浆中分离的独特的ATP依赖性DNA解旋酶recQ、hrpB1和hrpB2基因对番茄溃疡病菌进行比较基因组分析和快速分子检测

Comparative Genomic Analysis and Rapid Molecular Detection of Xanthomonas euvesicatoria Using Unique ATP-Dependent DNA Helicase recQ, hrpB1, and hrpB2 Genes Isolated from Physalis pubescens in China.

作者信息

Siddique Faisal, Mingxiu Yang, Xiaofeng Xu, Zhe Ni, Younis Haseeb, Lili Peng, Junhua Zhang

机构信息

College of Agriculture, Northeast Agricultural University, Harbin 150030, China.

出版信息

Plant Pathol J. 2023 Apr;39(2):191-206. doi: 10.5423/PPJ.OA.08.2022.0119. Epub 2023 Apr 1.

Abstract

Ground cherry (Physalis pubescens) is the most prominent species in the Solanaceae family due to its nutritional content, and prospective health advantages. It is grown all over the world, but notably in northern China. In 2019 firstly bacterial leaf spot (BLS) disease was identified on P. pubescens in China that caused by both BLS pathogens Xanthomonas euvesicatoria pv. euvesicatoria resulted in substantial monetary losses. Here, we compared whole genome sequences of X. euvesicatoria to other Xanthomonas species that caused BLS diseases for high similarities and dissimilarities in genomic sequences through average nucleotide identity (ANI) and BLAST comparison. Molecular techniques and phylogenetic trees were adopted to detect X. euvesicatoria on P. pubescens using recQ, hrpB1, and hrpB2 genes for efficient and precise identification. For rapid molecular detection of X. euvesicatoria, loop-mediated isothermal amplification, polymerase chain reaction (PCR), and real-time PCR techniques were used. Whole genome comparison results showed that the genome of X. euvesicatoria was more closely relative to X. perforans than X. vesicatoria, and X. gardneri with 98%, 84%, and 86% ANI, respectively. All infected leaves of P. pubescens found positive amplification, and negative controls did not show amplification. The findings of evolutionary history revealed that isolated strains XeC10RQ, XeH9RQ, XeA10RQ, and XeB10RQ that originated from China were closely relative and highly homologous to the X. euvesicatoria. This research provides information to researchers on genomic variation in BLS pathogens, and further molecular evolution and identification of X. euvesicatoria using the unique target recQ gene through advance molecular approaches.

摘要

酸浆(Physalis pubescens)因其营养成分和潜在的健康益处,是茄科中最突出的物种。它在世界各地都有种植,但在中国北方尤为显著。2019年,中国首次在酸浆上发现了细菌性叶斑病(BLS),由BLS病原体野油菜黄单胞菌致病变种(Xanthomonas euvesicatoria pv. euvesicatoria)引起,造成了巨大的经济损失。在此,我们通过平均核苷酸同一性(ANI)和BLAST比较,将野油菜黄单胞菌的全基因组序列与其他引起BLS疾病的黄单胞菌物种进行比较,以发现基因组序列中的高度相似性和差异。采用分子技术和系统发育树,利用recQ、hrpB1和hrpB2基因检测酸浆上的野油菜黄单胞菌,以进行高效精确的鉴定。为了快速分子检测野油菜黄单胞菌,使用了环介导等温扩增、聚合酶链反应(PCR)和实时PCR技术。全基因组比较结果表明,野油菜黄单胞菌的基因组与穿孔黄单胞菌(X. perforans)的亲缘关系比与疮痂黄单胞菌(X. vesicatoria)和加德纳黄单胞菌(X. gardneri)更近,ANI分别为98%、84%和86%。所有酸浆感染叶片均呈阳性扩增,阴性对照未显示扩增。进化历史研究结果表明,源自中国的分离菌株XeC10RQ、XeH9RQ、XeA10RQ和XeB10RQ与野油菜黄单胞菌亲缘关系密切且高度同源。本研究为研究人员提供了有关BLS病原体基因组变异的信息,并通过先进的分子方法,利用独特的靶标recQ基因对野油菜黄单胞菌进行进一步的分子进化和鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78dc/10102564/d899dd219973/ppj-oa-08-2022-0119f1.jpg

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