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与采后附子()腐烂相关的 和 分离株的特征及其新型环介导等温扩增检测方法。

Characteristics of Isolates of and Associated With Post-harvest Fuzi () Rot and Their Novel Loop-Mediated Isothermal Amplification Detection Methods.

作者信息

Fu Meng, Zhang Xin, Chen Bei, Li Mingzhu, Zhang Guoyan, Cui Langjun

机构信息

The Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest China, The Ministry of Education, College of Life Sciences, Shaanxi Normal University, Xi'an, China.

Chenggu County Qunli Traditional Chinese Medicine Cooperative, Chenggu, China.

出版信息

Front Microbiol. 2021 Aug 20;12:705329. doi: 10.3389/fmicb.2021.705329. eCollection 2021.

Abstract

Fuzi (the lateral root of Debx.) is a traditional Chinese medicine that is cultivated in more than eight provinces in China. However, it can be easily devastated by post-harvest rot, causing huge losses. Therefore, it is extremely important that the primary causal pathogens of post-harvest Fuzi rot are identified and appropriate detection methods for them are developed to prevent and control losses. In this study, two bacterial strains (X1 and X2) were isolated from rotten post-harvest Fuzi. Based on their morphological, physiological, and biochemical characteristics, housekeeping gene homologies, and matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS) results, these isolates were identified as and The pathogenicities of these isolates were confirmed by fulfilling Koch's postulates demonstrating that they were post-harvest Fuzi rot pathogens. Two loop-mediated isothermal amplification (LAMP) methods targeting the gyrase B subunit () gene of and the phosphatidylinositol glycan C () gene of were successfully developed, and it was found that the target genes were highly specific to the two pathogens. These LAMP methods were used to detect and in 46 naturally occurring Fuzi and their associated rhizosphere soil samples of unknown etiology. The two bacterial assays were positive in some healthy and rotten samples and could be accomplished within 1 h at 65°C without the need for complicated, expensive instruments. To our knowledge, this is the first report of and causing post-harvest Fuzi rot. The newly developed methods are expected to have applications in point-of-care testing for the two pathogens under different Fuzi planting procedures and will significantly contribute to the control and prevention of Fuzi rot.

摘要

附子(毛茛科植物乌头的子根)是一种在中国八个以上省份种植的传统中药。然而,它很容易受到采后腐烂的破坏,造成巨大损失。因此,确定采后附子腐烂的主要致病病原体并开发针对它们的合适检测方法以预防和控制损失极为重要。在本研究中,从采后腐烂的附子中分离出两株细菌菌株(X1和X2)。基于它们的形态、生理和生化特征、管家基因同源性以及基质辅助激光解吸电离/飞行时间质谱(MALDI-TOF MS)结果,这些分离株被鉴定为[具体名称未给出]和[具体名称未给出]。通过满足科赫法则证实了这些分离株的致病性,表明它们是采后附子腐烂病原体。成功开发了两种针对[具体名称未给出]的gyrase B亚基(gyrB)基因和[具体名称未给出]的磷脂酰肌醇聚糖C(PIG-C)基因的环介导等温扩增(LAMP)方法,并且发现靶基因对这两种病原体具有高度特异性。这些LAMP方法用于检测46份病因不明的天然附子及其相关根际土壤样品中的[具体名称未给出]和[具体名称未给出]。这两种细菌检测方法在一些健康和腐烂样品中呈阳性,并且可以在65°C下1小时内完成,无需复杂、昂贵的仪器。据我们所知,这是关于[具体名称未给出]和[具体名称未给出]引起采后附子腐烂的首次报道。新开发的方法有望应用于不同附子种植程序下这两种病原体的即时检测,并将对附子腐烂的控制和预防做出重大贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e89/8417746/a0906b94978a/fmicb-12-705329-g001.jpg

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