Characteristics of Isolates of Pseudomonas aeruginosa and Serratia marcescens Associated With Post-harvest Fuzi (Aconitum carmichaelii) Rot and Their Novel Loop-Mediated Isothermal Amplification Detection Methods

Abstract

Fuzi (the lateral root of Aconitum carmichaelii 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 Pseudomonas aeruginosa and Serratia marcescens. 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 (gyrB) gene of P. aeruginosa and the phosphatidylinositol glycan C (pigC) gene of S. marcescens 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 P. aeruginosa and S. marcescens 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 P. aeruginosa and S. marcescens 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.

Strip intercropping with local crops increased Aconitum carmichaeli yield and soil quality

“…Fuzi, which is the lateral root of Aconitum carmichaeli Debx., has been widely used in Asia as an essential herbal drug for 2,000 years (Fu et al, 2021). Researchers have dated the initial record on Fuzi back to the Han dynasty in the earliest Chinese classic medical work "Shennong Bencao Jing" (Singhuber et al, 2009;Zhou et al, 2015a).…”

“…Moreover, Fuzi polysaccharides, which are another pharmacological component, could stimulate murine lymphocyte proliferation and improve the immune function (Zhao et al, 2006). Due to their outstanding clinical effects and the considerable demand for Fuzi and Chuanwu in traditional Chinese medicine, more than eight provinces in China are currently cultivating A. carmichaeli, and the artificial planting regions have rapidly expanded in recent years (Fu et al, 2021).…”

Development and clinical application of a rapid and visual loop-mediated isothermal amplification test for tetM gene in Clostridioides difficile strains cultured from feces

“…LAMP assays have been widely applied to the detection of bacterium (Frisch et al, 2021;Fu et al, 2021), viruses (Li et al, 2021;Ptasinska et al, 2021), parasites (Mesquita et al, 2021;Ruang-Areerate et al, 2021) and fungi ( Karakkat et al, 2018;Choopara et al, 2021), as well as detection of the hypervirulent strain, toxin types, binary toxin, ermB gene of C.difficile (Noren et al, 2011;Boyanton et al, 2012;Lin et al, 2015;Yu et al, 2017), and even in forensics (Arenas et al, 2017). In this study, four sets of primers were designed and LAMP assay for detection of tetM were optimized.…”

Microbial community diversity and function analysis of Aconitum carmichaelii Debeaux in rhizosphere soil of farmlands in Southwest China

“…It plays an important clinical role in treating cardiogenic shock associated with acute myocardial infarction, rheumatism, bruises and injuries, acute or chronic bacterial dysentery, and enteritis ( Zhou et al, 2015 ; Zhang et al, 2020 ). Due to their remarkable curative effects, there is an increasing demand for this medicine ( Fu et al, 2021 ). Currently, A. carmichaeli is cultivated in the southwest region of China as a commercial crop, and most of them are grown in Yunnan province ( Xia et al, 2021 ).…”